A STABLE ISOTOPE APPROACH TO NEOTROPICAL CLOUD

FOREST PALEOCLIMATOLOGY

by

Kevin John Anchukaitis

A Dissertation Submitted to the Faculty of the

DEPARTMENT OF GEOSCIENCES

In Partial Fulfillment of the Requirements

For the Degree of

DOCTOR OF PHILOSOPHY

In the Graduate College

THE UNIVERSITY OF ARIZONA

2007
 2

THE UNIVERSITY OF ARIZONA

GRADUATE COLLEGE

As members of the Final Examination Committee, we certify that we have read the
dissertation prepared by Kevin John Anchukaitis
entitled A Stable Isotope Approach to Neotropical Cloud Forest Paleoclimatology
and recommend that it be accepted as fulfilling the dissertation requirement for the Degree
of Doctor of Philosophy.

Date: 23 April 2007

Michael N. Evans

Date: 23 April 2007

Julio L. Betancourt

Date: 23 April 2007

Malcolm K. Hughes

Date: 23 April 2007

Julie E. Cole

Date: 23 April 2007

Jonathan T. Overpeck

Final approval and acceptance of this dissertation is contingent upon the candidate’s
submission of the final copies of the dissertation to the Graduate College.
I hereby certify that I have read this dissertation prepared under my direction and
recommend that it be accepted as fulfilling the dissertation requirement.

Date: 23 April 2007

Dissertation Director: Michael N. Evans
 3

STATEMENT BY AUTHOR

This dissertation has been submitted in partial fulfillment of requirements for an advanced
degree at The University of Arizona and is deposited in the University Library to be made
available to borrowers under rules of the Library.

Brief quotations from this dissertation are allowable without special permission,
provided that accurate acknowledgment of source is made. Requests for permission for
extended quotation from or reproduction of this manuscript in whole or in part may be
granted by the head of the major department or the Dean of the Graduate College when
in his or her judgment the proposed use of the material is in the interests of scholarship.
In all other instances, however, permission must be obtained from the author.

SIGNED: Kevin John Anchukaitis

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ACKNOWLEDGEMENTS

There are dozens of people that have helped make this research a reality. First and
foremost, I owe a debt of gratitude to my advisor, Mike Evans, for allowing me to
come to Arizona to work with him on tropical dendroclimatology and paleoclimatology.
Throughout my years at Arizona, Mike has provided me with unlimited opportunities to
expand my education and research experience. He has been unwavering in his optimism
and support, always willing to give me the latitude to explore new ideas but diligent in
his guidance. It has been my extremely good fortune to have him as my mentor. Julio
Betancourt and Malcolm Hughes have been incredibly generous with their wisdom and
their time, and have been unfailingly positive and encouraging. I will miss very much
their enthusiasm and our conversations. Julie Cole and Jonathan Overpeck have always
kept my eye on the larger context of my research.
Laboratory portions of this research were made possible by the diligence, knowledge,
and creative thinking of John Buchanan, MC Foo, Lisa Wade, Greg Eischeid, Heidi
Barnett, Rex Adams, Brooke Rabe, Brianna Muhlenkamp, Chris Jones, Jim Burns, Bruno
Lavettre, David Dettman, Dave Steinke, Ben McElhaney, and Martin Munro. In Costa
Rica, I benefitted tremendously from the assistance of Frank Joyce, Koky Porras, Alan
Pounds, Eladio Cruz, and Arturo Cruz. Access to Monteverde was facilitated through
the Tropical Science Center (Rafael Bolanos, Carlos Hernndez) and the Organization for
Tropical Studies. My collaborators Nat Wheelwright, Dan Schrag, Todd Lange, Dave
Smith, and Steve Leavitt have all contributed important advice, ideas, and data to various
aspects of this work. I also thank Todd Dawson, David Killick, and Henry Diaz for their
continued support of me and of this research.
My housemates, Mary Good, Jerome Guynn, and Tim Shanahan, all of them talented
scholars as well as good friends, have been instrumental in keeping me optimistic, happy,
and sane. I have been very fortunate to be surrounded by a wonderful, supportive group of
friends and colleagues including Toby Ault, Jessica Conroy, Tom Damassa, Matt Devitt,
Nathan English, Anna Felton, Nicole Flowers, Andy Frassetto, Camille Holmgren, Kevin
Jones, Dave Keeler, Lynnette Kleinsasser, Susan Mentzer, Maarten de Moor, Christa
Placzek, Dan Potts, Jessica Rowland, Britt Starkovich, Scott St. George, Aly Thibodeau,
and many others. Indeed, the graduate students, faculty, and staff of the Laboratory of
Tree-Ring Research and Department of Geosciences have given me five wonderful years
in an amazing and generous academic community.
Mary Gagen – my colleague, friend, and partner – has been a constant source of
support, expertise, wisdom, and love, and I would never have made it without her. My
parents, Melanie and Stephen, and my brother, Jeff, have supported me through 25 years
of education with patience, humor, and unquestioning love. This thesis is dedicated to all
of them.
This research was made possible by generous funding from the National Science
Foundation and the Department of Energy. Specific grants and fellowships are acknowl-
edged individually in the attached appendices to this dissertation.
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DEDICATION

This dissertation is dedicated to my parents, Melanie and Stephen, and to my brother,

Jeff, for their patience, love, and unyielding support.
 6

TABLE OF CONTENTS

LIST OF FIGURES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

ABSTRACT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

CHAPTER 1 INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . 14
1.1 Statement of the Problem . . . . . . . . . . . . . . . . . . . . . . . . . . 14
1.2 Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
1.2.1 The Tropics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
1.2.1.1 The Pacific . . . . . . . . . . . . . . . . . . . . . . . . 15
1.2.1.2 The Atlantic . . . . . . . . . . . . . . . . . . . . . . . 17
1.2.2 Tropical climate change: phenomenology and dynamics . . . . . 20
1.2.3 Why tropical dendroclimatology? . . . . . . . . . . . . . . . . . 21
1.2.4 Neotropical montane cloud forests . . . . . . . . . . . . . . . . . 21
1.2.5 The stable isotope approach to tropical dendroclimatology . . . . 22
1.2.6 Monteverde, Costa Rica . . . . . . . . . . . . . . . . . . . . . . 24
1.3 Research Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

CHAPTER 2 PRESENT STUDY . . . . . . . . . . . . . . . . . . . . . . . . . . 26

2.1 Evaluation of a rapid α-cellulose extraction protocol . . . . . . . . . . . 27
2.2 Stable oxygen isotope chronology and climate signal calibration from
Ocotea tenera . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
2.3 Reconstructing dry season climate variability from Sapotaceae . . . . . . 29

REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

APPENDIX A PURITY AND ISOTOPIC RESULTS FROM A RAPID

CELLULOSE EXTRACTION METHOD FOR HIGH-RESOLUTION ISOTOPE
DENDROCLIMATOLOGY . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
A.1 Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
A.2 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
A.3 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
A.3.1 Sample preparation . . . . . . . . . . . . . . . . . . . . . . . . . 39
A.3.2 Stable isotope and infrared analysis . . . . . . . . . . . . . . . . 41
A.4 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
 7

TABLE OF CONTENTS – Continued

A.4.1 Stable isotopes and radiocarbon . . . . . . . . . . . . . . . . . . 42

A.4.2 Infrared analysis of compounds . . . . . . . . . . . . . . . . . . 42
A.5 Analysis and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . 46
A.5.1 Stable isotopes and infrared spectra . . . . . . . . . . . . . . . . 46
A.5.2 Approaches to deacetylation . . . . . . . . . . . . . . . . . . . . 52
A.5.3 Radiocarbon analysis . . . . . . . . . . . . . . . . . . . . . . . . 53
A.5.4 A statistical radiocarbon correction . . . . . . . . . . . . . . . . 55
A.6 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
A.7 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
A.8 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59

APPENDIX B STABLE ISOTOPE CHRONOLOGY AND CLIMATE SIGNAL

CALIBRATION IN NEOTROPICAL MONTANE CLOUD FOREST TREES . 68
B.1 Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
B.2 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
B.3 Hypothesis and Approach . . . . . . . . . . . . . . . . . . . . . . . . . . 72
B.4 Methods and Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
B.4.1 Site description . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
B.4.2 Field and laboratory sampling . . . . . . . . . . . . . . . . . . . 78
B.4.3 Sample preparation . . . . . . . . . . . . . . . . . . . . . . . . . 80
B.4.4 Isotopic analysis . . . . . . . . . . . . . . . . . . . . . . . . . . 81
B.4.5 Forward modeling . . . . . . . . . . . . . . . . . . . . . . . . . 82
B.5 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 86
B.5.1 Plantation trees (NWT, NWH) . . . . . . . . . . . . . . . . . . . 86
B.5.2 Forest canopy trees (MV) . . . . . . . . . . . . . . . . . . . . . 90
B.5.3 Forward δ 18 O modeling . . . . . . . . . . . . . . . . . . . . . . 95
B.6 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
B.6.1 Annual oxygen isotope cycles . . . . . . . . . . . . . . . . . . . 98
B.6.2 Climate analysis . . . . . . . . . . . . . . . . . . . . . . . . . . 100
B.6.3 Forward modeling . . . . . . . . . . . . . . . . . . . . . . . . . 103
B.6.4 Spatial patterns and future work . . . . . . . . . . . . . . . . . . 106
B.7 Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . 108
B.8 Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
B.9 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109

APPENDIX C DRY SEASON CLIMATE VARIABILITY IN A TROPICAL

MONTANE CLOUD FOREST RECONSTRUCTED FROM TREES WITHOUT
RINGS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118
C.1 Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 119
 8

TABLE OF CONTENTS – Continued

C.2 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120

C.3 Methods and Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
C.3.1 Site description . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
C.3.2 Heuristic model . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
C.3.3 Sampling and stable isotope analysis . . . . . . . . . . . . . . . . 125
C.3.4 Radiocarbon . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126
C.4 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
C.4.1 Oxygen isotope chronology . . . . . . . . . . . . . . . . . . . . 127
C.4.2 Climate analysis . . . . . . . . . . . . . . . . . . . . . . . . . . 136
C.5 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 141
C.5.1 Annual and interannual climate patterns . . . . . . . . . . . . . . 141
C.5.2 Climate and ecological change at Monteverde . . . . . . . . . . . 145
C.5.3 Uncertainty and potential biases . . . . . . . . . . . . . . . . . . 147
C.5.4 Multidecadal dynamics . . . . . . . . . . . . . . . . . . . . . . . 150
C.5.5 Future applications . . . . . . . . . . . . . . . . . . . . . . . . . 153
C.6 Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
C.7 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
C.8 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
 9

LIST OF FIGURES

Figure A.1 Boxplots for δ 18 O measurements on replicate Sigma Alpha-

Cellulose (SAC), Arizona/Peruvian Prosopis (APP), and Alaska White
Spruce (AWS) laboratory standards prepared with Leavitt-Danzer and
SBrendel techniques . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Figure A.2 Fraction modern carbon determinations (FMC) from AMS radio-
carbon assays at the University of Arizona’s Accelerator Mass Spectrom-
etry Laboratory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Figure A.3 Area normalized absorbance spectra derived from ATR FTIR anal-
ysis of unprocessed and SBrendel-processed Arizona/Peruvian Prosopis
(APP), compared against Sigma Alpha Cellulose (SAC), and SBrendel-
and MBrendel-processed as well as unprocessed SAC . . . . . . . . . . . 47
Figure A.4 Area normalized absorbance spectra derived from ATR FTIR
analysis of Arizona/Peruvian Prosopis (APP) processed using SBrendel,
Leavitt-Danzer and MBrendel . . . . . . . . . . . . . . . . . . . . . . . 48
Figure A.5 Area normalized absorbance spectra derived from ATR FTIR
analysis of Alaskan White Spruce (AWS) processed using SBrendel,
Leavitt-Danzer and MBrendel. Spectra from untreated Sigma Alpha
Cellulose (SAC) and Alaskan White Spruce (AWS) are shown for
comparison . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Figure A.6 Example of potential error associated with the statistical correction
of the SBrendel radiocarbon bias . . . . . . . . . . . . . . . . . . . . . . 58

Figure B.1 Conceptual model of climatic controls on the annual and interan-
nual patterns of stable oxygen isotope ratios in the α-cellulose of cloud
forest tree radial growth . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Figure B.2 Patterns of mean annual temperature and precipitation for Mon-
teverde from the Campbell weather station and derived relative humidity
(1979-2000,10.2 o N, 85.35 o W, 1540m, Pounds et al. [1999]. . . . . . . 79
Figure B.3 Meteorological data from Monteverde used as input for the
forward model of tree α-cellulose δ 18 O . . . . . . . . . . . . . . . . . . 84
Figure B.4 Isotope ratios as a function of sampling depth for the four
plantation Ocotea tenera from the Trostles and Hoges sites . . . . . . . . 87
Figure B.5 Isotope ratios as a function of time for the four plantation Ocotea
tenera from the Trostles and Hoges sites . . . . . . . . . . . . . . . . . . 89
Figure B.6 Composite isotope timeseries for the Trostles/Hoges calibration set 91
 10

LIST OF FIGURES – Continued

Figure B.7 Isotope timeseries for the canopy tree calibration set and the
observed relationships between elevation, δ 18 O mean, and the amplitude
of the annual δ 18 O cycle . . . . . . . . . . . . . . . . . . . . . . . . . . 94
Figure B.8 Comparison between observed composite mean plantation isotope
timeseries and forward model simulation . . . . . . . . . . . . . . . . . . 97

Figure C.1 Conceptual model of climatic controls on the annual and interan-
nual patterns of stable oxygen isotope ratios in the α-cellulose of cloud
forest tree radial growth. . . . . . . . . . . . . . . . . . . . . . . . . . . 124
Figure C.2 Isotope chronology from MV12 . . . . . . . . . . . . . . . . . . . 128
Figure C.3 Isotope chronology from MV15 . . . . . . . . . . . . . . . . . . . 130
Figure C.4 Probability density functions from radiocarbon assays on wood
and cellulose from MV15C . . . . . . . . . . . . . . . . . . . . . . . . . 133
Figure C.5 Composite oxygen isotope chronology from MV12A and MV15C
based on radiocarbon dates and cross-correlation analysis with the logical
constraints discussed in the text . . . . . . . . . . . . . . . . . . . . . . . 135
Figure C.6 Zonal wind anomalies and annual wet-dry season δ 18 O amplitude
from MV12A . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139
Figure C.7 Field correlations between Monteverde dry season δ 18 O amplitude
and dry season (February to April) sea surface temperature anomalies . . 140
Figure C.8 Field correlations between Monteverde dry season δ 18 O amplitude
and dry season (February to April) zonal wind anomalies . . . . . . . . . 142
Figure C.9 Composite oxygen isotope chronology and climatic analysis of
MV12A and MV15C . . . . . . . . . . . . . . . . . . . . . . . . . . . . 144
 11

LIST OF TABLES

Table A.1 Statistical intercomparison of wood and cellulose prepared with

SBrendel and Leavitt-Danzer techniques . . . . . . . . . . . . . . . . . . 44
Table A.2 Radiocarbon offsets associated with the Brendel technique with no
NaOH step . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56

Table B.1 Site elevation, mean δ 18 O value, and characteristics of annual

oxygen isotope cycles from plantation composite mean and cloud forest
trees from Monteverde . . . . . . . . . . . . . . . . . . . . . . . . . . . 93

Table C.1 Radiocarbon analysis from MV12A and MV15C . . . . . . . . . . 131

 12

ABSTRACT

Many tropical trees do not form reliable annual growth rings, making it a challenge
to develop tree-ring width chronologies for application to paleoclimatology in these
regions. Here, I seek to establish high-resolution proxy climate records from trees

without rings from the Monteverde Cloud Forest in Costa Rica using stable isotope
dendroclimatology. Neotropical cloud forest ecosystems are associated with a relatively
narrow range of geographic and hydroclimatic conditions, and are potentially sensitive to
climate variability and change at time scales from annual to centennial and longer.
My approach takes advantage of seasonal changes in the δ 18 O of water sources used

by trees over a year, a signature that is imparted to the radial growth and provides
the necessary chronological control. A rapid wood extraction technique is evaluated
and found to produce cellulose with δ 18 O values indistinguishable from conventional
approaches, although its application to radiocarbon requires a statistical correction.

Analyses of plantation-grown Ocotea tenera reveal coherent annual δ 18 O cycles up

to 9h. The width of these cycles corresponds to observed basal growth increments.
Interannual variability in δ 18 O at this site is correlated with wet season precipitation
anomalies. At higher elevations within the orographic cloud bank, year-to-year changes

in the amplitude of oxygen isotope cycles show a relationship with dry season climate.
Longer δ 18 O chronologies from mature Pouteria (Sapotacae) reveal that dry season
hydroclimatology is controlled at interannual time scales by variability in the eastern
equatorial Pacific (ENSO) and the Western Hemisphere Warm Pool (WHWP), which

are correlated with trade wind strength and local air temperature. A change in the late
 13

1960s toward enhanced annual δ 18 O amplitude may reflect low frequency changes in the
Atlantic and Pacific ocean-atmosphere system. This study establishes the basis for cloud
forest isotope dendroclimatology and demonstrates that the local climate of neotropical
cloud forests is sensitive to interannual, and perhaps, multidecadal changes in important

large-scale modes of climate variability.

 14

CHAPTER 1

INTRODUCTION

1.1 Statement of the Problem

The tropical ocean-atmosphere system influences climate and weather at a global

scale, and variability in the system across a range of time scales results in anomalous
patterns of both temperature and precipitation around the world. The result can be
the disruption or destruction of habitat, transportation, agriculture, and infrastructure,
and the concomitant economic, political, environmental, and social costs of recovery.

Despite this, considerable uncertainty exists regarding both the potential behavior of and
controls on tropical climate system variability at time scales from years to centuries and
longer. This uncertainty extends to interactions between the Pacific and Atlantic Oceans
as well as the range of potential responses to radiative forcing. Substantial uncertainties

also exist for predictions of spatiotemporal changes in regional hydroclimatology due to

anthropogenic greenhouse gas emissions and the accompanying alterations to the global
energy balance [c.f. Douville, 2006; Douville et al., 2006; Neelin et al., 2006].
Uncertainties concerning tropical climate system state and behavior arise due in

part to the lack of long-term meteorological records from these regions. Even where
continuous climate records exist, they often cover too limited a time span to resolve
ocean-atmosphere variability or feedbacks which may have preferred timescales of
decades or longer. Paleoclimate reconstructions of precipitation and temperature can
help alleviate the limitation of the instrumental, observational record for interpreting

tropical climate dynamics and the potential range of natural variability; however, high-
 15

resolution paleoclimate proxies from the terrestrial tropics are thus far mostly limited to
comparatively few speleothem, ice core, and varved lake sediment records.

1.2 Background

1.2.1 The Tropics

1.2.1.1 The Pacific

The El Niño Southern Oscillation (ENSO), a coupled interaction between the ocean
and atmosphere of the tropical Pacific Ocean, is the leading mode of global interannual
climate variability [Cane, 2005]. During warm ENSO events, anomalously warm waters

move toward the east along the equator into the normally colder waters associated with
deep ocean upwelling and a shallow thermocline. Accompanying the changes in sea
surface temperatures (SST), sea level pressures also shift, with low pressure anomalies
over the eastern tropical Pacific, and anomalously high pressure descending on the

western Pacific, Indonesia, and eastern Australia [Rasmusson and Wallace, 1983; Diaz
and Kiladis, 1992].
These changes result in anomalous zonal atmospheric circulation and wind patterns,
bringing drought to the western Pacific, and heavy rains to the west coast of South
America. Through concurrent changes in atmospheric circulation, the effect of ENSO

are also felt around the globe. During canonical ENSO events, the southwestern and
southeastern United States experiences a wetter winter, while the northwest and northern
Prairies are dry [Diaz et al., 2001]. Canonical teleconnections, however, may conceal
more complicated seasonal to interdecadal spatiotemporal patterns. On the southeastern

coast of the United States, for instance, warm ENSO events are associated not only with a
wetter winter, but also a subsequently drier summer [Diaz et al., 2001]. In the Caribbean
 16

and Central America, warm ENSO events are associated typically with wetter boreal
winter dry seasons and drier summer wet seasons [Diaz et al., 2001], but the sign of
the correlation depends on the modulating influence of the Atlantic Ocean, as well as
whether the Pacific or Caribbean side of the Central American cordillera is considered

[Giannini et al., 2001a].

Decadal-scale variability can be detected within the interannual behavior of the ENSO
system. It is commonly characterized by periods of lesser or greater tropical SST anomaly
(SSTA) amplitudes [Urban et al., 2000; Yeh and Kirtman, 2005], and has a basin-wide
expression [Garreaud and Battisti, 1999; Tomita et al., 2001; Evans et al., 2001; Newman

et al., 2003]. The wavelet-based spectral analysis of Torrence and Webster [1999]
identified periods of high canonical ENSO wavelength variance from 1875 to 1920 and
from 1960 to 1990. A period of low ENSO variance occurred from 1920 to 1960. Periods
of low and high variance in the canonical ENSO band as identified in Torrence and

Webster [1999] agree with those seen in evolutionary spectral analysis of reconstructed
NINO 3.4 SST records by Urban et al. [2000]. The tree-ring based reconstruction of
ENSO by Stahle et al. [1998] also shows decadal-scale fluctuations in the amplitude of
the canonical ENSO frequencies.

There have been a number of attempts to explicitly reconstruct decadal modes of both
tropical and extratropical Pacific climate variability using high resolution proxies. Evans
et al. [2001], Biondi et al. [2001], D’Arrigo et al. [2001] and MacDonald and Case [2005]
have used tree-ring chronologies from western North and South America to develop
reconstructions extending back as far as AD 1000. Stable isotope proxies from corals

have also been successfully used to identify decadal patterns of SST variability [Linsley
et al., 2000; Urban et al., 2000]. The reconstructions are not, however, identical. In fact,
in many cases the independent reconstructions show little coherency prior to the current
 17

century [Gedalof et al., 2002]. Evans et al. [2002] further point out that reconstructed
(coral and tree-ring) and instrumental Pacific sea surface temperatures show different
spectral power at low frequencies. With existing proxy records, largely extratropical tree
rings and marine corals, it is still difficult to estimate what the magnitude and preferred

time scale of interannual to multidecadal variability in the Pacific may have been over the
last several centuries.

1.2.1.2 The Atlantic

The tropical Atlantic also displays variability over a range of time scales. High-frequency
interannual variations in the equatorial Atlantic are associated with the so-called Atlantic
Niño [c.f Chang et al., 2006] and the North American Oscillation [NAO; Seager et al.,
2000]. Since the NAO is related to the strength of the subtropical high pressure cell in

the Atlantic, it also has an influence on trade wind strength and SSTs in the subtropical
and tropical Atlantic and Caribbean [Seager et al., 2000]. Both the Atlantic Nino and
NAO may reflect the influence of remote Indo-Pacific tropical forcing [Hoerling et al.,
2001; Hoerling and Kumar, 2003; Chang et al., 2006]. The cross-equatorial SST gradient
between northern and southern tropical Atlantic influences the position of the ITCZ and

tropical trade wind strength at interannual time scales as well, and therefore both rainfall
and sea-air latent heat feedbacks on both SST anomalies and wind [Wang and Carton,
2003; Xie and Carton, 2004].
At multidecadal timescales, both tropical and extratropical Atlantic variability is

prominently related to the Atlantic Multidecadal Oscillation (AMO), which is centered

on the North Atlantic and characterized by relatively rapid multidecadal shifts between
warmer and colder conditions [Enfield et al., 2001]. From approximately 1930 to 1970,
the Atlantic was in a warm phase. From 1970 to 1995, the Atlantic shifted to a cold phase
 18

[Enfield et al., 2001]. The warm phase of the AMO is related to spatiotemporal SST
patterns similar to those that exist during prolonged negative phases of the NAO [Grosfeld
et al., 2007]. Gray et al. [2004] used tree-ring chronologies from North America, Europe,
and North Africa to reconstruct the index of the AMO over the period 1567 to 1990, which

demonstrated significant mulitdecadal power over nearly the entire period. Evidence for
robust multidecadal variability in the Atlantic coupled with long GCM simulations have
been used to suggest that the AMO is the result of periodic changes in thermohaline
circulation and meridional overturning circulation (MOC) in the North Atlantic [Knight
et al., 2005].

Interaction between the Pacific and Atlantic Oceans at decadal time scales appears
to play an important role in extratropical climate as well. Cole et al. [2002] found that
some major North American droughts, including that of the 1950s, corresponded to La
Niña (cold) conditions in the Pacific, while other droughts, including the 1930s Dust

Bowl, were only marginally correlated with Pacific SSTs. In their analysis, McCabe
et al. [2004] used instrumental PDSI measurements to hypothesize that patterns of
severe drought were related to the interaction between Pacific and Atlantic SST forcing,
with severe drought occurring when the Atlantic Ocean was in a cold (negative) phase.

Modeling studies have also suggested that Pacific SST anomalies alone are not sufficient
to force droughts with the spatial and temporal characteristics of the Dust Bowl [Schubert
et al., 2004]. The Atlantic could therefore be an important determinant of extratropical
climate at decadal-scales or longer, in addition to potentially mediating the canonical
teleconnections associated with ENSO. However, Mann and Emanuel [2006] have shown

that the residual sea surface temperature signal from the tropical Atlantic after accounting
for global SST patterns does not have a robust multidecadal signal similar to the AMO.
This finding suggests that observed multidecadal trends in the tropical Atlantic are
 19

primarily related to greenhouse gas and aerosol forcing over the 20th century. Whether
endogenous multidecadal tropical variability exists in the Atlantic is difficult to determine
over the period of instrumental and historical observations, since it is inextricable linked
to global scale forcing.

It is an active research question whether variability in the Pacific drives changes in the
Atlantic, or whether multidecadal variability in the Atlantic, perhaps arising from changes
in ocean circulation in the North Atlantic [Knight et al., 2005] can influence the amplitude
of interannual and interdecadal variability in the Pacific. For instance, Dong et al.
[2006] used a coupled ocean-atmosphere GCM with an imposed Atlantic SSTA forcing

to mimic the AMO. In these simulations, easterly surface wind anomalies over the central
and western Pacific forced by latent heat flux in the tropical North Atlantic deepened
the western Pacific themocline, stabilizing the ocean-atmosphere coupled instability in
the tropical Pacific and reducing ENSO variance. The more common view has been

that ENSO influences the Atlantic via atmospheric teleconnections [c.f. Latif , 2001].
Wang and Enfield [2001] have shown that warm ENSO anomalies influence the Western
Hemisphere Warm Pool via the connection of anomalous Walker circulation to changes
in the Atlantic subtropical high. Similarly, Giannini et al. [2001b] hypothesized that

ENSO influenced the tropical Atlantic variability through the stabilization of the tropical
atmosphere via the propagation of the warming signal through the troposphere. These and
numerous other studies point to extant uncertainties regarding the forcing mechanisms
for the observed association between Atlantic and Pacific variability at a variety of time
scales.
 20

1.2.2 Tropical climate change: phenomenology and dynamics

At least three important research questions arise out of observations and modeling of
the Pacific and Atlantic ocean-atmosphere variability discussed above. First, there

are outstanding issues regarding the preferred time scale and stability of decadal scale
variability in the tropical Pacific [Cane and Evans, 2000; Evans et al., 2001], which
might be better resolved by an expanded network of high resolution tropical proxy
records. Second, similar questions exist regarding tropical North Atlantic variability and
its characteristic natural and forced time scales and variability. Removing the global sea

surface temperature signal from the tropical Atlantic reveals no significant multidecadal
variance. However, as Mann and Emanuel [2006] note, this is based on the assumption
that the tropical Atlantic signal does not project significantly onto the global mean.
Whether this assumption is valid over longer time scales than considered by Mann and

Emanuel [2006], or even under current transient forcing, is difficult to resolve using the
instrumental record alone. Finally, understanding the interactions and feedbacks between
the Atlantic and Pacific is critical for interpreting variability in the paleoclimate record
and understanding potential global teleconnections associated with both the tropics and

North Atlantic. For instance, if ocean circulation in the Atlantic forces multidecadal or
longer changes in the Pacific through changes in sea surface temperatures and circulation
anomalies generated in the tropics, this has substantial implications for feedbacks between
thermohaline circulation and the Asian monsoon, and could help explain how regional

signals associated with the MOC can propagate globally. There is clear evidence that
variability across a range of time scales between the Atlantic and Pacific are linked.
However, the mechanisms, stability, and directionality of those linkages remain unclear
and in some cases contradictory. [Schwing et al., 2003] found that North Atlantic and
North Pacific variability at decadal scales was positively correlated until the 1950s, and
 21

negatively correlated after that time. The stability of interbasin interactions may therefore
be a function of other, low frequency components of their variability.

1.2.3 Why tropical dendroclimatology?

Hundreds of tree-ring width and density chronologies provide a spatially extensive data

network for high resolution paleoclimate reconstructions in temperate regions. Because

they have annual or seasonal resolution and are exactly dated, these records can be
empirically calibrated against existing instrumental data in order to develop quantitative
reconstructions of past climatic variables. Relatively few such tree-ring chronologies,
however, have been developed in tropical regions. Despite some notable exceptions [c.f.

Worbes, 2002; Fichtler et al., 2004; D’Arrigo et al., 2006], many trees in the tropics do not
develop reliably annual rings whose variability reflects the influence of climate variability
and can be utilized for paleoclimate reconstructions. Even when they appear to form
annual increment bands, patterns of ring width variability may be incoherent between

individual trees, making both chronology development and climate signal detection
difficult [Worbes and Junk, 1989; February and Stock, 1998; Dünisch et al., 2002; Speer
et al., 2004; Robertson et al., 2004; Bauch et al., 2006]. As a consequence, high-
resolution, long terrestrial proxy climate records from the tropics remain sparse compared

to temperate regions.

1.2.4 Neotropical montane cloud forests

Tropical montane cloud forests cover as much as 50 million hectares worldwide

[Stadtmüller, 1987], about half of which is found in Latin America [Brown and Kappelle,
2001]. Cloud forests are characterized by frequent or persistent ground-level clouds

and fog (horizontal or ‘occult’ precipitation) and have a multicanopy forest laden
 22

with epiphytes [Stadtmüller, 1987; Haber, 2000]. These forests have high rates of
endemism [Brown and Kappelle, 2001] and are important in regional hydrology, since
they intercept cloud moisture, increasing available water and influencing biogeochemistry
within catchments and in areas downstream.

Tropical cloud forests are ecosystems found within a relatively narrow set of both
geographic and meteorological conditions, and as a consequence, they are particularly
sensitive to climate change [Loope and Giambelluca, 1998; Foster, 2001; Bush, 2002].
Computer modeling of climate under doubled CO2 conditions predict higher lifting
condensation levels and reduced cloud contact for tropical montane cloud forests as global

temperatures rise [Still et al., 1999]. Analysis of observational data also suggests that
warmer eastern tropical Pacific sea surface temperatures (SSTs) are related to interannual
decreases in cloud cover at Monteverde [Pounds et al., 1999], but that there also exists
a longer-term trend of reduced moisture potentially related to tropical surface and sea

surface temperature increases. Increases in global temperature may fundamentally alter

the suite of climatic and biophysical conditions that maintain cloud forests environments
[Pounds et al., 1999; Still et al., 1999]. The observed and modeled sensitivity of the
hydroclimatology of tropical cloud forests to broad-scale climate variability and change

indicates that they may be optimal locations at which to develop proxies for paleoclimate
reconstructions.

1.2.5 The stable isotope approach to tropical dendroclimatology

Stable oxygen isotope ratios (δ 18 O) offer an analytical approach to resolving both

chronology and climate signal in tropical trees [Evans and Schrag, 2004]. The advantage

of this approach to tropical dendrochronology is that it does not rely on the formation nor
width variations of annual growth rings, which in tropical trees can be absent or may not
 23

reflect local climate variability. I hypothesize that the δ 18 O value of cellulose in tropical
montane cloud forest trees from Central America preserves a record of plant water use
that reflects regular seasonal changes in hydroclimatology. These cycles can be used for
chronological control in the absence of rings. Annual oxygen isotope cycles have been

previously identified in lowland rainforests [Evans and Schrag, 2004; Poussart et al.,
2004; Verheyden et al., 2004].
The δ 18 O of cellulose in the stem of a tree reflects the original oxygen isotope
composition of the source water (precipitation or cloud water), the isotopic enrichment
via transpiration in the leaves, and the fractionation associated with the synthesis of

α-cellulose from sucrose [Roden et al., 2000; Barbour et al., 2004; McCarroll and
Loader, 2004]. Assuming the latter is constant, cellulose in cloud forest trees should
therefore reflect both the amount of rainfall received and climatic influences on the rate
of evapotranspiration from the leaf. On annual time scales in this conceptual model,

cellulose δ 18 O is controlled by the seasonal change in δ 18 O of source water as determined

by the amount of rainfall and the utilization by trees of available cloud water. On
interannual time scales, departures from the mean annual cycle are likely to result from
anomalous rainfall and changes in temperature, relative humidity, and evapotranspiration.

My selection of tropical cloud forests as a site for tropical paleoclimate reconstruction

parallels, in a somewhat paradoxical manner, the approach of classical dendroclimatology
in seeking out sampling locations where trees are likely to be sensitive to relatively small
changes in annual climate. In temperate regions and for trees with regular annual changes
in morphology or wood density, these are typically dry or cold sites at the limits of a

species range. For tropical isotope dendroclimatology, however, I seek a research site
which is wet enough to allow trees to grow throughout the year, yet is subject to regular
seasonal changes in the stable oxygen isotope composition of available moisture that
 24

provides the means of establishing annual chronology. In cloud forest environments, the
largest interannual changes of interest are likely to be wet season rainfall and dry season
cloudiness. As such, site selection for tropical isotope dendroclimatology is very different
from that of classical field approaches in dendrochronology in terms of climate regimes,

but the guiding principle is the same.

1.2.6 Monteverde, Costa Rica

My study focuses on the Monteverde Cloud Forest in Costa Rica where environmental
and meteorological data available over the last 20 years provides the data necessary to
validate this hypothesis. The existing environmental data from Monteverde also reveal

that its local climatology reflects interannual variability in the Pacific related to ENSO, as
well as apparent long-term trends potentially related to similar trends in tropical air and
sea surface temperatures.
Several recent publications [Still et al., 1999; Pounds et al., 1999; Lawton et al.,

2001] have explored the specific issue of the impact of climate variability and global
environmental chance on these tropical montane cloud forests in Costa Rica. The impetus
for this research was the population crash and eventual extinction of the Monteverde
Golden Toad after the 1987 El Niño event, as well as population declines in other anuran

and lizard populations, and observed changes in the elevational range of several bird
species [Pounds and Crump, 1994; Pounds et al., 1997, 1999; Pounds and Puschendorf ,
2004; Pounds et al., 2006]. Still et al. [1999] used the GENESIS GCM to simulate
relative humidity and lifting condensation levels (LCL) for doubled CO2 conditions.
Using both calculated relative humidity surfaces and warmth index values, they concluded

that anthropogenic global warming could shift the altitude of cloud forests upward by
more than 200 meters. Other studies have found that LCLs at cloud forest sites could
 25

shift upward by as much as 400 to 500 meters [Foster, 2001]. Pounds et al. [1999]
found that dry season mist and cloud frequency at Monteverde was negatively correlated
with Pacific SSTs. A positive trend in dry days (increasing number of dry days/year)
remained even when taking into account the influences of warm ENSO events, apparently

indicated some longer-term trend underlying interannual variability. An earlier study

of the relationship between precipitation at Monteverde and ENSO events [Pounds and
Crump, 1994] had found higher temperatures and lower seasonal rainfall during the 1982-
83 and 1986-87 El Niño events, indicating that warmer SSTs were inducing warmer, drier
conditions. Because they occupy an altitudinal range and topography associated with very

specific climatic conditions, tropical montane cloud forests may also be some of the first
ecosystems to unequivocally show the consequences of anthropogenic global warming,
and may provide early indications of the impacts of global temperature rise.

1.3 Research Approach

My focus on the application of tropical stable isotope dendroclimatology in the cloud

forests of Monteverde in Costa Rica accomplish several complementary goals. First and
foremost, it does not require the identification of trees with reliable annual growth rings,
instead using the isotope hydroclimatology of the cloud forest for both chronology and

paleoclimate signal. Second, I am able to take advantage of the sensitivity of the local
climate of these forests to broad-scale forcing in order to develop the basis for using
cloud forest trees to develop proxy records of past ocean-atmosphere variability. Finally,
a reconstruction of the local, dry season climate conditions at Monteverde allows recent

changes identified from the short instrumental record to be interpreted in the context of
interannual variability and trends over several decades.
 26

CHAPTER 2

PRESENT STUDY

The research presented in this dissertation is focused on the development and application
of a stable isotope approach to extracting annually-resolved climate information from

trees without rings in tropical cloud forests in Central America. Specifically, I first
investigated the purity and isotopic composition of cellulose extracted from whole wood
using a rapid chemical processing procedure. Using this sample preparation method and
analytical protocols for tropical isotope dendroclimatology [Evans and Schrag, 2004],

I then established the basis for developing high resolution δ 18 O records from cloud
forest trees at Monteverde. I used a calibration study to test my hypothesized age
model and identify the existence of a wet season precipitation signal in the isotope time
series. I also conducted an additional pilot survey along an elevation transect to identify
promising candidate species and sites for long climate reconstructions from the upper

cloud forest and transition zone. I identified and selected mature canopy Sapotaceae
from above 1500m and used analyses of these trees to develop a longer δ 18 O chronology
and to investigate the link between local climate at Monteverde and broad-scale ocean
atmosphere forcing. The methods, results and conclusions of this research are presented

in the papers appended to this dissertation and the following is a summary of the methods
used and the most significant results.
 27

2.1 Evaluation of a rapid α-cellulose extraction protocol

I used Fourier Transform Infrared Spectroscopy (FTIR), radiocarbon assays, and stable
oxygen isotope analyses to evaluate a rapid cellulose preparation protocol for wood
samples from tropical trees. Paired comparison of replicate δ 18 O measurements on

sets of laboratory standard wood powders and α-cellulose established that there was no
statistically significant difference between samples processed using traditional methods
[Leavitt and Danzer, 1993] and those prepared using the Brendel method [Brendel et al.,
2000]. Radiocarbon dating, however, revealed a small degree of contamination sufficient

to significantly bias absolute age estimates. My analysis of FTIR spectra revealed that
the contamination was cellulose acetate formed during acid delignification. FTIR and
radiocarbon analysis suggested that contaminant acetyl groups were not removed by
strong alkali saponification, nor by other methods used to prevent acetylation. However, I

proposed a statistical correction to allow wood samples processed to α-cellulose using

the Brendel technique to be corrected for contamination and utilized for chronology
development. My study design enabled me to establish that the Brendel technique
could be used for stable oxygen isotope analysis, to identify the cause and consequences
of sample contamination during processing, and to develop a correction to allow the

technique to be reasonably to radiocarbon dating. My assessment of the Brendel method

for applications in high resolution tropical isotope dendroclimatology is contained in the
article manuscript attached as Appendix A.

2.2 Stable oxygen isotope chronology and climate signal calibration from Ocotea tenera

I used high-resolution stable oxygen isotope analysis on wood from planted and
monitored Ocotea tenera in order to demonstrate that chronological control in trees
without annual rings could be established using δ 18 O cycles imparted to the xylem
 28

of cloud forest trees due to seasonal changes in the moisture regime. Cores that I
collected from these plantation trees were microsampled, extracted to α-cellulose using
the Brendel technique, and their δ 18 O was measured by continuous flow isotope ratio mass
spectrometer. I identified regular δ 18 O cycles in wood of up to 9h, which were associated

as I had hypothesized with the seasonal changes in precipitation and moisture sources.
The calculated annual growth rates I derived from the isotope time series match those
observed from long-term basal growth measurements, confirming my hypothesized age
model. My statistical analysis of the interannual anomalies in the amplitude of the oxygen
isotope cycles in Ocotea tenera revealed that, at this site, they are primarily controlled to

interannual changes in summer precipitation. I also used a forward mechanistic model

of the relationship between monthly climate (precipitation, temperature, and relative
humidity) to independently confirm the detection and identification of the annual δ 18 O
and their relationship to climate.

I sought to identify trees from which I could potentially develop longer δ 18 O

chronologies for paleoclimate analysis using a pilot isotopic survey of nine additional
trees along an elevational transect from the transitional lower forest zone up to the
continental divide. Trees from elevations above the orographic cloud bank showed

larger interannual variability, but less consistently showed coherent annual cycles. The
confirmation of annual chronology and sensitivity to interannual climate anomalies
verified my heuristic, conceptual model of the seasonal, annual, and interannual controls
on cloud forest tree xylem δ 18 O. These data further indicated that trees from higher
elevation sites within the persistent orographic cloud bank at Monteverde could preserve

a record of past variability in dry season climate. My calibration study establishing the
basis for tropical isotope dendroclimatology in the cloud forests of Costa Rica is contained
in the article manuscript attached as Appendix B.
 29

2.3 Reconstructing dry season climate variability from Sapotaceae

Following my validation of the annual chronology and climate signal from the lower
forest plantations, I applied this same analytical approach to mature Sapotaceae trees
from the upper elevation cloud forest. High-resolution isotope measurements along

the growth radius of these trees revealed coherent annual cycles in δ 18 O that I then
used to establish annual chronological control in the absence of growth rings. I also
used radiocarbon dates to verify the age models established using the δ 18 O time series,
including the identification of a growth hiatus not evident from the δ 18 O data alone.

The annual maxima and amplitude of the oxygen isotope cycles reflected interannual
variability in dry season orographic cloud cover, moisture advection, and temperature.
The minima of the annual δ 18 O reflected summer precipitation anomalies. Positive dry
season δ 18 O anomalies are associated with El Niño events and weaker northeasterly trade

winds. I interpreted lower frequency (multidecadal) variability in the amplitude of the

δ 18 O time series be potentially related to climate trends in the tropical Atlantic and
changes in the frequency of ENSO events in the tropical Pacific. According to my oxygen
isotope chronology from cloud forest Sapotaceae, the extinction of the Monteverde
Golden Toad after 1987 occurred during one of the driest periods in the last 50 years,

and potentially in the last century. The development of longer isotope chronologies from
Sapotaceae and my analysis of these records in the context of local and global patterns of
climate variability are contained in the article manuscript attached as Appendix C.
 30

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 36

APPENDIX A

PURITY AND ISOTOPIC RESULTS FROM A RAPID CELLULOSE EXTRACTION

METHOD FOR HIGH-RESOLUTION ISOTOPE DENDROCLIMATOLOGY

Manuscript for submission to Analytical Chemistry

 37

Purity and isotopic results from a rapid cellulose extraction method

for high-resolution isotope dendroclimatology

Kevin J. Anchukaitis1,2, Michael N. Evans1,2, Todd Lange3 , David R. Smith4, Daniel P.

Schrag5 , and Steve W. Leavitt1

1. Laboratory of Tree-Ring Research, The University of Arizona, Tucson, AZ

2. Department of Geosciences, The University of Arizona, Tucson, AZ

3. Accelerator Mass Spectrometry Laboratory, The University of Arizona, Tucson AZ

4. Department of Chemistry, The University of Arizona, Tucson AZ

5. Department of Earth and Planetary Sciences, Harvard University, Cambridge, MA

A.1 Abstract

Light stable isotope analyses are increasingly becoming part of multiproxy, high-
resolution approaches to the reconstruction of past climates from trees. The extraction of
α-cellulose from whole wood is now the primary time-limiting factor in the development
of long and well-replicated tree-ring isotope chronologies. Here, we use infrared,

radiocarbon, and stable isotope analyses to investigate a rapid processing technique

for extracting cellulose from whole wood for isotope dendroclimatology. Replicate
laboratory standards processed using the standard Brendel method are not significantly
different with respect to δ 18 O from those prepared using traditional techniques, although

the process does result in a slight acetylation of the wood samples. Radiocarbon
comparisons, however, show significant differences. We conclude that the Brendel
method is appropriate for developing stable isotope time series for high-resolution
isotope dendroclimatology, but must be used with caution for precision radiocarbon
measurements. We propose a simple statistical correction for those instances where

Brendel must be used for radiocarbon analyses.

 38

A.2 Introduction

Stable isotope analysis of tree-rings for the study of paleoenvironmental and plant
physiological processes has typically focused on the α-cellulose component of wood,
necessitating the removal of resins, lignins and non-celluloic polysaccharides, or

hemicellulose, prior to measurement. α-cellulose is by definition the components

of a cellulosic material which are insoluble in a 17.5% solution of NaOH at 20o C,
which includes primarily cellulose but also small amounts of lignin, hemicellulose, and
extractives which cannot be removed even under these conditions [Green, 1963; Leavitt

and Danzer, 1993; Gaudinski et al., 2005]. The goal of extracting cellulose for stable
isotope analysis is not, however, to arrive at α-cellulose simply by processual definition,
but rather to remove as much of the unstable, translocated, or exchangeable noncellulosic
components as possible in order to consistently isolate the most stable component of the

wood.
The use of specific wood components is often preferred because the noncellulosic
compounds can be deposited after the time the ring itself is formed, may be radially
translocated, and have different isotopic signatures [Gray and Thompson, 1977; de Water
et al., 1994]. While recent studies suggest that for both stable carbon and oxygen

isotopes the climate signal is preserved in whole wood [Borella et al., 1999; Saurer et al.,
2000; Barbour et al., 2001; Saurer et al., 2002; Loader et al., 2003; Verheyden et al.,
2005], analysis of specific compounds is still desirable or necessary in some applications,
including radiocarbon analysis [Hoper et al., 1998], analyses of fossil wood that may

have experienced heterogeneous decomposition [Schleser et al., 1999; Poole et al., 2004],
and those instances where mechanistic models of α-cellulose synthesis will be used to
interpret the climatic influence on stable isotope ratios [Roden et al., 2000; Evans, 2007].
Analyses using α-cellulose may also be preferable to avoid potential biases in isotope
 39

time series as a function of spatiotemporal changes in the relative contribution in whole

wood from lignins, hemicellulose, and α-cellulose.
Here, we investigate a procedure for rapidly producing α-cellulose for use in high-
resolution stable isotope dendrochronology. The Brendel method [Brendel et al., 2000;

Evans and Schrag, 2004] allows on average 100 samples per day to be extracted to
α-cellulose and prepared for analysis the following day. In its modified form [Evans
and Schrag, 2004], it also permits the use of very small initial wood samples (∼400
µg). Furthermore, the Brendel method also uses smaller quantities of less toxic reagents,
reducing both chemical waste (less than 1.5 mL of waste per sample) and the potential

hazards to laboratory personnel. Unlike traditional extraction procedures, however, it

uses a hot nitric-acetic acid step to simultaneously remove both lignins and hemicellulose
from wood samples. It is important to understand how cellulose extracted using the
Brendel procedure compares to samples prepared using traditional techniques [Green,

1963; Leavitt and Danzer, 1993; Loader et al., 1997] and to determine the efficacy
of the procedure in removing extractives, lignins, and hemicellulose. Potential biases
introduced by alternative chemical preprocessing protocols could include a difference in
means or a reduction in variance [Gaudinski et al., 2005]. The former could complicate

intercomparison with other studies, while the latter might lead to underestimation of
environmental variability as reflected in isotope time series.

A.3 Methods

A.3.1 Sample preparation

We used three laboratory wood and alpha cellulose standards in this study. We prepared
our Arizona/Peruvian Prosopis (APP) laboratory wood standard by repeatedly grinding
a radial cross section collected in 2001 from a living Prosopis sp. near Piura, Peru,
 40

until the wood powder could pass through a 125µm sieve. The Alaska White Spruce
(AWS) standard was previously prepared by grinding to 250µm a cross-section (UA-
BCLV-132) of Picea glauca with a total of 172 annual rings collected in the early 1990s.
We used Sigma-Aldrich (Batch 024K0329) Alpha Cellulose (SAC) for our α-cellulose

control standard.
Samples (8 mg) of the whole wood standards (APP and AWS) were homogenized and
separated using a microsplitter (ACS Scientific, Jamestown, RI) and sealed in commercial
digestion pouches (ANKOM Technology, Boston, MA). These were processed to
holocellulose in a soxhlet apparatus [Sheu and Chiu, 1995; Hoper et al., 1998] using a

modified Jayme-Wise procedure [Green, 1963; Leavitt and Danzer, 1993]. Samples were
progressively extracted with toluene/ethanol, ethanol, and boiling deionized water. They
were then delignified in an sodium chlorite-acetic acid solution at 70o C and thoroughly
rinsed in deionized water. The resulting holocellulose samples were then converted to

α-cellulose by treatment in a 17% NaOH solution [Green, 1963; Borella et al., 1998] and
dried in a warm oven.
Replicate samples (1 – 1.5 mg) of the whole wood and industrial α-cellulose (APP,
AWS, and SAC) were processed following the standard Brendel method [Brendel et al.,

2000] as modified for small sample processing [Evans and Schrag, 2004, , hereafter
‘SBrendel’]. Samples were heated at 120o C in a 10:1 mixture of 80% acetic acid and
70% nitric for 30 minutes. Samples were then washed with 100% ethanol and de-ionized
water and subsequently dehydrated with consecutive additional washes of pure ethanol
and acetone. Samples were dried for 30 minutes in a warm oven, and then overnight in a

vacuum desiccator [Evans and Schrag, 2004].

We also processed a set of 1 to 1.5mg wood and cellulose samples (SAC, APP,
and AWS) using the ‘Modified Brendel’ method [Gaudinski et al., 2005, , hereafter,
 41

‘MBrendel’], which adds a 10 minute 17% NaOH extraction and several additional water
rinses to the published Brendel procedure [Brendel et al., 2000; Evans and Schrag, 2004].

A.3.2 Stable isotope and infrared analysis

For stable oxygen isotope analysis, 100 to 150 µg cellulose samples were loaded in

silver capsules and converted to CO online by pyrolysis in a ThermoFinnigan Thermal

Conversion/Elemental Analyzer (TC/EA) at 1450o C. The stable isotopic composition of
the CO was then analysed on a ThermoFinnigan Delta Plus XL continuous flow isotope
ratio mass spectrometer. Data are reported relative to Vienna Standard Mean Ocean
Water (VSMOW). External precision for oxygen isotopic measurements during these

experiments based on a Baker α-cellulose standard was 0.45h (VSMOW).

For radiocarbon analysis, carbon was extracted from the samples in the form of CO2
using conventional combustion methods. The volume of the evolved and purified gas was
measured and the CO2 was reduced to an iron carbide powder over hot zinc. Radiocarbon

measurement was performed on a National Electrostatics Accelerator Mass Spectrometer

at the University of Arizona-NSF AMS Facility [Donahue et al., 1990].
Fourier Transform Infrared Spectroscopy (FTIR) [Putzig et al., 1994; Moore and
Owen, 2001] was performed using a Thermoelectron (Nicolet) Avatar spectrometer at

a resolution of 4 cm−1 . The spectra were all obtained using a single-bounce diamond
Attenuated Total Reflectance (ATR) accessory. At least two duplicate spectra were
obtained in every case to verify the results.
 42

A.4 Results

A.4.1 Stable isotopes and radiocarbon

Stable oxygen isotope measurements on the paired groups of replicate laboratory wood

and cellulose standards shows no statistically significant difference (two-tailed t-test, α

= 0.05) between those processed using the SBrendel method and those extracted using
the Leavitt-Danzer procedure (Figure A.1, Table A.1), although the intercomparison
for the industrial Sigma Alpha Cellulose is statistically different at a lower significance

threshold (α = 0.10). Significant differences are observed (Figure A.2), however, in the
fraction of modern radiocarbon (FMC) measured in replicate samples processed using
SBrendel and MBrendel, Leavitt-Danzer, and a standard acid-base-acid (hydrochloric
and sodium hydroxide) [DeVries and Barendsen, 1954] procedure. Wood and cellulose

samples processed using Leavitt-Danzer and acid-base-acid (ABA) pretreatments have

consistently higher fractions of modern carbon (FMC), whereas samples processed using
SBrendel and MBrendel have reduced FMC, indicating contamination by radiocarbon-
depleted sources. For both wood and cellulose standards, the offset between Leavitt-
Danzer and SBrendel is consistently 0.07 to 0.08. Wood samples processed with

MBrendel show FMC values inconsistently intermediate between SBrendel and Leavitt-
Danzer and ABA (0.05 to 0.14). The Sigma Alpha-Cellulose processed with MBrendel
actually shows further decreases in FMC relative to Leavitt-Danzer/ABA.

A.4.2 Infrared analysis of compounds

Figures A.3, A.4, and A.5 show the FTIR spectra for in-house wood and α-cellulose
samples. Consistent with previous studies [Brendel et al., 2002], SBrendel-processed
whole wood shows removal of resins (Figure 3, Peak 1; ∼1600 cm−1 ), lignin (Figure 3,
 43

34
Brendel
Leavitt-Danzer
32

30
δ18O (SMOW)

28

SAC APP AWS

26

24

22

Figure A.1: Boxplots for δ 18 O measurements on replicate Sigma Alpha-Cellulose (SAC),

Arizona/Peruvian Prosopis (APP), and Alaska White Spruce (AWS) laboratory standards
prepared with Leavitt-Danzer and SBrendel techniques. Following Tukey [Tukey, 1977],
boxes show the interquartile range, with the notch-line indicating the median. Whiskers
demarcate values within 1.5 times the interquartile range. In all three cases, there is
no significant difference (two-tailed t-test, 95% significance level) between the samples
processed by the two methods.
 44

Standard Material x̄ B (h) x̄ L D (h) d.f. t-statistic p-value

SAC 30.99 31.51 37 -1.99 0.054
APP 31.26 31.16 28 0.25 0.808
AWS 23.03 23.26 37 -0.88 0.387

Table A.1: Statistical intercomparison of wood and cellulose prepared with SBrendel and
Leavitt-Danzer techniques. x̄ B is the mean of the samples processed using the SBrendel
technique, x̄ L D is the mean of those processed using Leavitt-Danzer, d.f. is the number
degrees of freedom, which is based on the number of samples measured that passed data
quality control for minimum voltage. Also shown are the result of the Student’s t-test
and the significance level ( p > 0.05 indicates no statistically significant difference in the
means).
 45

1.2

1.18
Fraction Modern Carbon

1.16

1.14

1.12

1.1

1.08
Whole Wood ABA
1.06 Brendel
Modified Brendel
Leavitt Danzer
1.04
APP AWS SAC
Standard Type
Figure A.2: Fraction modern carbon determinations (FMC) from AMS radiocarbon
assays at the University of Arizona’s Accelerator Mass Spectrometry Laboratory.
SBrendel-processed samples are consistently offset from Raw, ABA, and Leavitt-Danzer
processed samples suggesting a 6% contribution of 14 C dead carbon from a contaminant,
most like acetyl groups inherited for acetic acid). MBrendel-processed methods show an
inconsistent offset.
 46

Peak 2; 1550 to 1450 cm−1 ) and noncelluloic polysaccharides (Figure 3, Peak 3; 1230-
1180 cm−1 ) (Figure A.3a) However, samples (both wood and α-cellulose) processed
with the SBrendel method are characterized by infrared spectra with 3 peaks that are
not present in wood or cellulose processed using Leavitt-Danzer at ∼1720, 1245, and

1110 cm−1 (Figure A.3, Peaks 4, 5, 7, and 8). Additionally, the peak near ∼900 cm−1
(Figure 3, Peak 9) seen in all samples is less distinct in those processed using SBrendel.
The peak near 1720 cm−1 is reduced and shifted toward a maximum near 1710 cm−1 ,
but is not fully removed by the MBrendel NaOH extraction (Figure A.3b, A.4, A.5).
This peak reduction and shift is consistently detected despite being close to instrument

wavelength resolution. The NaOH wash does appear to sharpen the peak near 900 cm−1 ,
but interestingly also removes the peak near 1430 cm−1 , which is present in both the
unprocessed and SBrendel-processed SAC samples, and adds an additional peak near
3430 cm−1 , something which is also seen in the Leavitt-Danzer processed samples (not

shown). For the samples that were put through the additional 17% NaOH wash, the peaks
at 1245 and 1110 cm−1 also disappear (Figure A.3b).

A.5 Analysis and Discussion

A.5.1 Stable isotopes and infrared spectra

Statistical analysis of the stable oxygen isotope measurements on laboratory standard

material reveals that no significant bias ( p < 0.05) is introduced by our small-sample
SBrendel method for δ 18 O analysis. This finding is consistent with previously reported

results [Evans and Schrag, 2004] for a small set of δ 18 O samples, as well as for δ 13 C
as reported by Brendel et al. [2000], but differs from the conclusions of Gaudinski et al.
(2005) [Gaudinski et al., 2005]. This discrepancy may be the result of comparing different
techniques across tissue types, (leaves, wood, roots), instead of considering xylem wood
 47

−3 −3
x 10 x 10
2.5 2.5
a). APP Brendel SAC Brendel b).
APP Whole Wood SAC MBrendel
SAC Raw SAC Raw
2 2

8
Area Normalized Absorbance

1.5 1.5
4

1 1
7
3 9
1 5
2 6
0.5 0.5

0 0

1800 1600 1400 1200 1000 1800 1600 1400 1200 1000
wavelength (cm −1) wavelength (cm −1)

Figure A.3: Area normalized absorbance spectra derived from ATR FTIR analysis of
(a) unprocessed and SBrendel-processed Arizona/Peruvian Prosopis (APP), compared
against Sigma Alpha Cellulose (SAC), and (b) SBrendel- and MBrendel-processed as
well as unprocessed SAC. Number labeled spectral peaks or bands show characteristic
wavelengths [Putzig et al., 1994; Stewart et al., 1995; Krasovskii et al., 1996; Vazquez
et al., 1997; Pandey, 1999; Brendel et al., 2000; Adebajo and Frost, 2004b; Sun and
Tomkinson, 2004; Silverstein et al., 2005; Rinne et al., 2005] for (1) resin, (2) lignins, (3)
hemicellulose and cellulose acetate/acetic acid, (4,8-9) soluble cellulose acetate/acetic
acid, and (5) insoluble cellulose acetate. Peak (6) is removed by the Leavitt-Danzer
method, even though it is still present in SBrendel-processed and raw α-cellulose(See
Figure 4 and 5). Slight differences in the normalized area under the curve reflect
between-sample differences in cellulose adsorbed water, whose bands (not shown) are
disproportionately strong and vary with relative humidity.
 48

SAC
Area Normalized Absorbance

APP Whole Wood

APP Brendel

APP MBrendel

APP Leavitt-Danzer

2000 1800 1600 1400 1200 1000 800 600

wavelength (cm -1)

Figure A.4: Area normalized absorbance spectra derived from ATR FTIR analysis
of Arizona/Peruvian Prosopis (APP) processed using SBrendel, Leavitt-Danzer and
MBrendel. Spectra from untreated Sigma Alpha Cellulose (SAC) and Arizona-Peruvian
Prosopis (APP) are shown for comparison. Shaded vertical bars show wavelengths
discussed in the text associated with the acetylization of α-cellulose.
 49

SAC
Area Normalized Absorbance

AWS Whole Wood

AWS Brendel

AWS MBrendel

AWS Leavitt-Danzer

2000 1800 1600 1400 1200 1000 800 600

wavelength (cm -1)

Figure A.5: Area normalized absorbance spectra derived from ATR FTIR analysis of
Alaskan White Spruce (AWS) processed using SBrendel, Leavitt-Danzer and MBrendel.
Spectra from untreated Sigma Alpha Cellulose (SAC) and Alaskan White Spruce (AWS)
are shown for comparison. Shaded vertical bars show wavelengths discussed in the text
associated with the acetylization of α-cellulose.
 50

separately. Leaves and roots, in particular, may pose a particular challenge for cellulose
extraction. In this respect, however, concerns raised by Gaudinski et al. [2005] apply
equally for any methods used to extract α-cellulose from these tissues.
Peaks in the FTIR absorbance spectra for SBrendel samples at ∼1720, 1245, and

1110 cm−1 are associated with carbonyl group C=O and C-O bond stretching, and are
most likely related to the presence of cellulose acetate or triacetate esters [Cherian
et al., 1994; Krasovskii et al., 1996; Matsumura and Saka, 1992; Adebajo and Frost,
2004a; Sun and Tomkinson, 2004; Silverstein et al., 2005]. The appearance of FTIR
peaks in the regions associated with cellulose acetate were originally interpreted by

Brendel et al. [2000], using DRIFT spectroscopy, as arising from modified non-cellulosic
polysaccharides (glucimannans) present in the wood of Pinus sylvestris. Our infrared
analysis, however, indicates that it is a slight acetylation of cellulose that is causing the
differences detected in the radiocarbon results and NMR spectroscopy in previous studies

[Gaudinski et al., 2005]. If it were instead acetic acid residue, as has been alternatively
suggested [Gaudinski et al., 2005], we would expect the diagnostic peaks in the carbonyl
band (C=O) near 1700 cm−1 and the C-O band near 1250 cm−1 [Sun and Tomkinson,
2004; Silverstein et al., 2005].

Furthermore, it is highly unlikely that acetic acid residue would persist following
treatments in strong NaOH and subsequent repeated solvent washes. Differences in
SAC processed with SBrendel and MBrendel (Figure 3b) might be partially related to
acetic acid residue or perhaps to removal of acetylized acid-insoluble hemicelluloses,
but the diagnostic peak near ∼1720 cm−1 is almost certainly indicative of cellulose

acetate formation. The shift of this carbonyl peak toward a maximum at ∼1710 might
indicate partial hydrolysis of the acetylized α-cellulose [Sun et al., 2005b], but it is clear
that saponification is incomplete and a portion of the cellulose acetate or triacetate ester
 51

remains insoluble [Matsumura and Saka, 1992]. We believe that the acetylation found
here is similar in nature to that reported previously for stable isotope analyses of pollen
grains [Amundson et al., 1997; Loader and Hemming, 2000], where it occurs occurs
during standard acetolysis procedures for pollen extraction and preparation [Charman,

1992], and in industrial chlorine-free pulping applications [Sun and Tomkinson, 2004].
The SBrendel method does successfully delignify and remove most hemicelluloses
from our whole wood samples. The method, however, slightly acetylates some of the
non-extracted cellulose fraction. A similar conclusion about the efficacy of the cellulose
extraction and the concomitant acetylation has been reached for industrial applications

of the nitric/acetic acid delignification approach [Sun and Tomkinson, 2004; Sun et al.,
2004a, b, c, 2005a, b; Xu et al., 2005]. One hypothesis concerning the shape of the
peak near 900 cm−1 and the presence of additional bands near 1240 and 1110 cm−1
could be that they are related to incomplete removal of some residual (alkali-soluble)

hemicellulose; however, since the SAC processed with the SBrendel method also has
these features (Figure A.3b), it must instead be predominantly related to the creation
and presence of acetylized cellulose. The additional of an NaOH step [Brendel et al.,
2002], or use of the MBrendel technique [Gaudinski et al., 2005] may indeed remove

some remaining acid-insoluble hemicelluloses and any partially acetylized hemicelloses

[Liu and Hsieh, 2002; Son et al., 2004], but still incompletely removes the acetylized
α-cellulose. That the diagnostic peak near 1720 cm−1 largely remains even after the
alkaline hydrolysis suggests perhaps that either a portion of the cellulose acetate cannot
be deacetylized by saponification in NaOH, possibly because the C=O bonds cannot be

readily hydrolyzed, or that partial hydrolysis of the cellulose acetate does occur, but
an additional solvent treatment is necessary to subsequently dissolve and remove the
remaining acetate.
 52

Gaudinski et al. [2005] conclude that the SBrendel potentially leaves behind
hemicelluloses, lipid and/or waxes, as well as acetic acid contamination which can
potentially bias stable isotope measurements. As we have indicated here, however, the
primary contaminating residue is most likely a small amount of insoluble cellulose acetate

which persists even after the application of a strong alkali treatment and additional water
washes in the MBrendel [Gaudinski et al., 2005] protocols. The NMR spectra which
Gaudinski et al. [2005] identified as diagnostic of lipids and waxes is also diagnostic
for the presence of cellulose acetate [Torii et al., 1994; Ohkoshi et al., 1997; Kono
et al., 2002; Czimczik et al., 2002; Bootten et al., 2004], which would also explain the

apparent inconsistency between the higher δ 18 O values they observed for Brendel samples
in contrast to the expected lighter values that would result from incomplete removal
of these compounds. Because the dominant contamination is from acetyl groups, with
one oxygen atom and two carbon, there is no biochemical reason to conclude that 18 O

analyses utilizing wood processed with SBrendel techniques is any more problematic
than for 13 C isotope ratios. That there is no difference in δ 13 C rations between SBrendel
and traditional α-cellulose preparation methods despite this contamination suggests that
the carbon isotope signature of the contaminating acetyl groups is similar to that of the

cellulose of C3 plants. This could be tested by using a C4 standard like maize or sugar
cane, as opposed to wood from C3 trees as has been previously done [Brendel et al., 2000;
Gaudinski et al., 2005].

A.5.2 Approaches to deacetylation

Our experiments with a variety of pre- and post-treatments have failed to completely

remove the small amount of acetylized cellulose as indicated by the 1710-1720 cm−1
peak. Our experimental goal was to either prevent acetylation during the delignification
 53

step, or to remove it post hoc following acid digestion. Approaches to solubilizing

the refractory cellulose acetate using extended, hot alkali saponification with NaOH
(0.05 to 4N) in various combination of water and in ethanol [Liu and Hsieh, 2002;
Son et al., 2004] failed to further reduce or shift the diagnostic FTIR peak near 1710

cm−1 . There was no detectable difference as a function of temperature, time, solvent,

or alkali normality. Adding extended vortex mixing between NaOH washes likewise has
no discernible effect. We also attempted to prevent acetylation using sulfolane as an
additional solvent during delignification [Zaleski et al., 2002], but FTIR spectra indicated
that acetyl group substitution still occurred. The addition of other solvents during the

acid delignification step could in theory help reduce the eventual acetylation, but it may
be impossible to prevent the conversion of some number of the hydroxyl groups to acetyl
groups [Ramsden and Blake, 1997; Saka et al., 1998]. The overall inefficacy of these
experimental methods may be a consequence of the low-order, heterogeneous or random

nature of the minor acetylization of the cellulose structure, which could render the acetate
fraction largely insoluble [Sassi and Chanzy, 1995; Klemm et al., 1998].

A.5.3 Radiocarbon analysis

Radiocarbon assays demonstrate significant differences between samples processed using

SBrendel and those prepared with conventional pretreatments. Furthermore, these

differences are not resolved using the MBrendel procedure [Gaudinski et al., 2005],
indicating that both SBrendel and MBrendel protocols introduce contamination from
radiocarbon-dead sources, and should be used with considerable caution. It remains
unclear whether the MBrendel [Gaudinski et al., 2005] technique should be used for either

radiocarbon or stable isotope analysis, though, since it appears to inconsistently remove

acetylized cellulose components (Figure A.2), at least across the wood and cellulose types
 54

examined in our study. The use of a weak NaOH wash by Brendel et al. [2002], without
additional solvent washes, has a similar effect on the extracted cellulose as the MBrendel
technique, but also leaves a considerable alkali residue which can be readily detected in
the FTIR spectra (results not shown).

Using our radiocarbon measurements (Figure A.2, Table A.2), a simple mass balance
equation comparing samples processed using Leavitt-Danzer and SBrendel suggests that
approximately 6% of the carbon (and therefore, approximately 3% of the oxygen) in
the SBrendel samples comes from the contaminating acetyl groups. This assumes that
petroleum-derived acetic acid possesses no detectable 14 C. This percentage is also

consistent with preliminary tests of the Brendel method conducted by Poussart [2004].
Ultimately, however, the choice of α-cellulose extraction protocols for environmental
and paleoenvironmental studies will depend rather specifically on the particular
application. The Brendel technique, even without the additional of an NaOH wash, is

acceptable for stable oxygen isotope analysis, but not for many radiocarbon applications.
Other, non-dendroclimatological uses, especially those employing plant tissues besides
wood, need to be closely examined [Gaudinski et al., 2005], although this issue is
generalizable to all cellulose extraction procedures, and not limited to the Brendel

procedure. Wood types with considerable resin content, particularly pines and other
conifers, often require additional treatments even using standard Leavitt-Danzer type
protocols [Miller et al., 2006]. We continue to investigate changes to the SBrendel
procedure which preserve its advantages in terms of speed, sample size, and safety for
removing the insoluble fraction of the acetylized cellulose, to allow this technique to be

used in the future for radiocarbon analyses.

 55

A.5.4 A statistical radiocarbon correction

Radiocarbon measurements on a range of wood types and ages consistently shows that
the contamination with 14 C dead carbon due to the SBrendel method is approximately

6±0.8% (Table A.2). The similarly between the mass balance calculations of the
percent of radiocarbon-dead contamination suggests that, for the delignification duration,
temperature, and acid molarity used in the Brendel protocols described by Evans and
Schrag [2004], the degree of acetyl group substitution is similar between samples. This
finding leads us to believe that, in the temporary absence of a procedural chemical solution

to the acetylation bias, a statistical correction can be applied to develop a corrected

radiocarbon date.
An example of this correction is shown in Figure A.6. Accounting for the observed
6±0.8% contribution from a carbon source with FMC = 0,

F MCcorr ected = F MCmeasur ed /(1 − 0.06) (A.1)

results in calibrated calendar ages [Bronk Ramsey, 1995] with a difference of less

than 1 year during the post-1955 atmospheric 14 C peak that resulted from aboveground
atomic weapons testing [Hua et al., 1999; Reimer et al., 2004a; Hua and Barbetti,
2004], compared to radiocarbon dates on wood processed using the traditional Leavitt-
Danzer technique (Figure A.6). Errors could be somewhat larger for portions of the

radiocarbon curve between 1700 and 1950, during which time large oscillations in
atmospheric 14 C production make precise calibration difficult [Reimer et al., 2004b];
however, the calibration errors associated with the wide range of probability density
functions for single radiocarbon determinations during that time period are substantially

larger than those arising from the uncertainty in the SBrendel correction. Whenever
possible, replicate radiocarbon assays on samples processed using both SBrendel and
 Sample Identification F MCm F MCa % contamination reference
AA64893 1.114 (0.0088) 1.1908 (0.0058) 5.9±0.6% this study
AA64894 1.0693 (0.0047) 1.1558 (0.0056) 7.4±0.4% this study
AA64895 1.0889 (0.0054) 1.1573 (0.0062) 6.5±0.5% this study
Brendel-A02 1.5247 (0.0061) 1.6453 (0.0108) 5.0±1.0% Poussart, 2004; Poussart et al., 2004
PG1-1 1.0941 (0.0043) 1.174 (0.0045) 6.8±0.4% Poussart, 2004; Poussart et al., 2004
AA74380 1.0143 (0.0083) 1.0842 (0.0088) 6.4±1.0% Anchukaitis et al., 2007
Mean (σ ) 6.0 ±0.8%

Table A.2: Radiocarbon offsets associated with the Brendel technique with no NaOH step. F MCm is the measured fraction
modern carbon, F MCa is the ‘actual’ fraction modern carbon measured on traditionally extracted α-cellulose [Leavitt and
Danzer, 1993; Westbrook et al., 2006]. Percent contamination is the calculation of the contribution from a radiocarbon dead
source (FMC = 0).

56
 57

Leavitt-Danzer or ABA can help verify the correction factor proposed here. Alterations
to the SBrendel procedure, such as a change in the time or temperature of the acid
delignification step, or a change in the strength of the acetic acid, would be expected
to change the degree of acetylation and therefore the magnitude of the contamination and

the necessary correction factor.

A.6 Conclusions

Wood and cellulose samples prepared using the standard Brendel methodology [Evans
and Schrag, 2004] have δ 18 O values statistically indistinguishable than those processed

using traditional Leavitt-Danzer type protocols. More importantly, there is no indication

that the Brendel method biases the range of δ 18 O variability between samples. α-cellulose
prepared using this technique can therefore be securely used for paleoclimate and other
time series applications, and absolute mean values can be compared between samples
processed with either method. The Brendel method does result in a slight acetylation of

the α-cellulose that is sufficient to bias radiocarbon measurements by approximately 6%,

but which can be corrected statistically if necessary. MBrendel does appear to remove
some of the acetyl group contamination, but results in an inconsistent shift in the offset
between measured radiocarbon ages on samples extracted with different methods. The

SBrendel technique accomplishes the goal of rapidly extracting very small whole wood
samples to stable α-cellulose, which is important for the application of high-resolution
isotope dendroclimatology in the tropics [Evans and Schrag, 2004].

A.7 Acknowledgements

We thank Greg Eischeid, Jonathan Buchanan, Chris Jones, Brianna Muhlenkamp, Jim
Burns, Li Cheng, and Sonya Issaeva for valuable laboratory support. We also benefitted
 58

1958
1957
Calibrated Date (cal AD)
OxCal v4.0.1 Bronk Ramsey (2006); Post-bomb atmospheric NH2 curve (Hua and Barbetti 2004)

1956
1955
Corrected Brendel (+1б)

Corrected Brendel (-1б)

alpha-cellulose

1954

Figure A.6: Example of potential error associated with the statistical correction of the
SBrendel radiocarbon bias. Within the post-1950 bomb radiocarbon curve, the influence
of the uncertainty in the correction factor applied to the calibrated calendar age is < 1
year.
 59

from helpful discussions with and advice from with Pascale Poussart, Iain Robertson,
Mary Gagen, Julia Gaudinski, and Todd Dawson. KJA was supported by a graduate
training fellowship from the NSF IGERT Program (DGE-0221594) and a Graduate
Research Environmental Fellowship from the US Department of Energy. The research

was supported by a grant from NSF-ESH (NSF/ATM0349356).

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APPENDIX B

STABLE ISOTOPE CHRONOLOGY AND CLIMATE SIGNAL CALIBRATION IN

NEOTROPICAL MONTANE CLOUD FOREST TREES

Manuscript for submission to Journal of Geophysical Research – Biogeosciences

 69

Stable isotope chronology and climate signal calibration in neotropical

montane cloud forest trees

Kevin J. Anchukaitis1,2, Michael N. Evans1,2, Nathaniel T. Wheelwright3 , and Daniel P.

Schrag4

1. Laboratory of Tree-Ring Research, The University of Arizona, Tucson, AZ

2. Department of Geosciences, The University of Arizona, Tucson, AZ

3. Department of Biology, Bowdoin College, Brunswick, ME

4. Department of Earth and Planetary Sciences, Harvard University, Cambridge, MA

B.1 Abstract

Tropical montane cloud forests are ecosystems intrinsically linked to a narrow range of
geographic and meteorological conditions, making them potentially sensitive to small

changes in precipitation or temperature. We investigate the potential application of stable

isotope analysis to cloud forest dendroclimatology at Monteverde in Costa Rica, in order
to be able to extract both chronological and paleoclimate information from trees without
annual growth rings. High-resolution δ 18 O measurements are used to identify regular

cycles in wood of up to 9h, which are associated with seasonal changes in precipitation
and moisture sources. The calculated annual growth rates derived from the isotope time
series match those observed from long-term basal growth measurements. Interannual
variability in the oxygen isotope ratio of lower forest trees is primarily related to
interannual changes in summer precipitation. Forward modeling independently supports

our detection of both annual chronology and a climate signal. Trees from elevations
 70

above the orographic cloud bank show larger interannual variability, but less consistently
show coherent annual cycles. The confirmation of annual chronology and sensitivity to
interannual climate anomalies suggests that tropical cloud forest dendroclimatology can
be used to investigate local and regional hydroclimatic variability and change.

B.2 Introduction

In the absence of long instrumental records, scientists investigating the causes and
consequences of climate variability and change depend on proxy records, which can be
used to reconstruct past ocean-atmosphere conditions. In temperate latitudes, extensive

networks of tree-ring width and density time series (‘chronologies’) provide a high-
resolution proxy record of past climate state and variability. Relatively few such
chronologies, however, have been developed in tropical regions. Despite some notable
exceptions [c.f. Worbes, 2002; Fichtler et al., 2004; Brienen and Zuidema, 2005;
D’Arrigo et al., 2006; Therrell et al., 2006], tropical trees seldom develop reliable annual

rings whose variability reflects the influence of climate variability and can be used
to reconstruct past climate. Even when they appear to form annual increment bands,
patterns of ring width variability may be incoherent between individual trees, making
both chronology development and climate signal detection difficult [Worbes and Junk,

1989; February and Stock, 1998; Dünisch et al., 2002; Speer et al., 2004; Robertson
et al., 2004; Bauch et al., 2006]. As a consequence, high-resolution, long terrestrial proxy
climate records from the tropics remain sparse compared to temperate regions.
Tropical montane cloud forests cover as much as 50 million hectares worldwide

[Stadtmüller, 1987; Hamilton et al., 1995], about half of which is found in Latin America
[Brown and Kappelle, 2001]. These forests have high rates of endemism and are
important in regional hydrology, because they intercept and capture cloud moisture and
 71

nutrients, increasing available water and influencing biogeochemistry within catchments

and in areas downstream [Bruijnzeel, 1991; van Dijk and Bruijnzeel, 2001; Bruijnzeel,
2004]. Tropical cloud forests are ecosystems found within a relatively narrow set of both
geographic and meteorological conditions, and as a consequence, they are particularly

sensitive to climate change [Loope and Giambelluca, 1998; Foster, 2001; Bush, 2002].
Rising tropical surface and sea surface temperature associated with anthropogenic
global climate change may be fundamentally altering the suite of environmental
conditions that create and maintain unique cloud forest ecosystems. At Monteverde,
in the mountains of Costa Rica, the extinction of the endemic Golden Toad in 1987

and subsequent species disappearances have been linked to apparent decreases in cloud
cover and moisture availability and related to rising temperatures [Pounds et al., 1999].
Indeed, higher temperatures in montane regions throughout the tropics have been linked to
widespread species extinction and alterations to cloud forest biogeography [Pounds et al.,

2006]. A rise in tropical air temperatures above 1000 meters has been observed since
1970 [Diaz and Graham, 1996]. Climate change appears to be exposing plant and animal
communities to increased environmental stress, which may exacerbate other proximal
threats, including disease and habitat destruction [Root et al., 2003; Pounds et al., 2006].

Limited long-term instrumental climate records from the tropics, and in particular
tropical montane regions [Bradley et al., 2006], impede efforts to better understand
both global climate variability and its influence on local hydroclimatology and ecology.
Without a historical baseline and the context provided by long-term climate records, it is
difficult to attribute the observed changes in climate and ecological systems at Monteverde

and other tropical mountain forests to (1) anthropogenic climate forcing [Still et al., 1999;
Pounds et al., 2006], (2) mesoscale influences including land clearance [Lawton et al.,
2001; Nair et al., 2003], or (3) natural variability in tropical and extratropical climate.
 72

Limited observational data and the lack of high resolution proxies from these ecosystems
also restrict any opportunity to validate GCM predictions for future change [Foster,
2001]. Both observed and potential future changes, and their hypothesized ecological
consequences, need to be placed in the context of low-frequency (decadal to centennial)

climate variability and trends over the recent past.

We hypothesize that the unique hydroclimatic conditions associated with cloud forests
creates an annual climate signal to the δ 18 O in tree stems that can be used to develop
a proxy record of climate variability from tropical montane regions. High-resolution
oxygen isotope measurements can be used for both chronological control and climatic

interpretation and reconstruction. Our study focuses on the Monteverde Cloud Forest in
Costa Rica where environmental and meteorological data available over the last 20 years
provides the data necessary to validate this hypothesis. We seek to develop the basis for
proxy reconstructions of interannual climate variability using stable oxygen isotope series

from cloud forest trees without annual rings.

B.3 Hypothesis and Approach

Tropical montane cloud forests are ecosystems characterized by frequent or persistent

orographic cloud immersion [Bruijnzeel and Veneklaas, 1998]. During the dry season,

water stripped from the cloud bank by vegetation provides sufficient soil moisture such
that trees may experience no effective water deficit [Bruijnzeel and Proctor, 1995; Kapos
and Tanner, 1985; Holder, 2004]. Precipitation, which provides the bulk of available
water during the rainy season, and cloud water, which is the primary moisture source

during the dry season, are isotopically distinct [Ingraham, 1998]. Phase transitions
between liquid and vapor result in fractionation effects, which create isotopic differences
between these two water sources. Condensation of clouds by orographic uplift results in
 73

liquid water which has relatively heavy δ 18 O values as compared to rainfall, which has
a lower δ 18 O value due to progressive Rayleigh distillation [Ingraham, 1998]. Similarly,
the limited amount of rainfall received during the dry season has a relatively heavy δ 18 O
signature. This is related primarily to the ‘amount effect’, where the δ 18 O of precipitation

is negatively correlated to the quantity, and is one of the primary causes of intra-annual
patterns in the isotopic composition of rainfall throughout the tropics [Gat, 1996]. The
amount effect is evident in the stable isotopic composition of Costa Rican surface waters
[Lachniet and Patterson, 2002].
Data from both Monteverde [Feild and Dawson, 1998] and other fog-dependent

environments [Ingraham and Matthews, 1990, 1995; Dawson, 1998] indicate that
differences of 4h to 6h δ 18 O exist between cloud water and rainfall. Rhodes et al.
[2006] also report differences between dry (winter) and wet (summer) season volume-
weighted precipitation δ 18 O as great as 8.2 h. The waters available to plants in cloud

forest ecosystems are therefore distinctly different between seasons. Feild and Dawson
[1998] confirmed that the xylem water of canopy trees at Monteverde had the isotopic
signature of rainfall during the summer wet season. Cloud water should be the primary
source water available to trees during the winter dry season [c.f. Dawson, 1998].

The seasonal changes in the isotopic composition of the primary water sources for
trees in cloud forests should produce a seasonal cycle in the oxygen isotope ratio of
cellulose in the secondary radial growth of these trees [Evans and Schrag, 2004]. Fine-
scale analysis of isotope ratios of cellulose along the growth radius of a tree’s main trunk
will yield a time series of water use by the trees. The seasonal cycle can be used to identify

the growth increment from an individual year (with 1 cycle = 1 calendar year), even in the
absence of visible annual banding. Significant departures from the regular isotope cycle
should result from local hydroclimatic alterations, including precipitation anomalies and
 74

changes in cloudiness, moisture advection, relative humidity, and temperature (Figure

B.1).
The relationship between the local hydroclimatology of tropical montane cloud forests
and the resulting δ 18 O value of cellulose in trees can be understood using the mechanistic

model developed by Roden et al. [2000]:

δ 18 Ocellulose = f o ·(δ 18 Owx + ǫo ) + (1 − f o )·(δ 18 Owl + ǫo ) (B.1)

Where δ 18 Ocellulose is the oxygen isotope ratio of tree-ring cellulose, δ 18 Owx is that of
the portion of xylem water that does not experience evaporative enrichment in the leaves,
δ 18 Owl is the oxygen isotope ratio of leaf water at the sites of photosynthesis, fo is the
fraction of oxygen in stem water that does not exchange with the enriched sucrose that
results from transpiration in the leaf, and ǫo is the fractionation that results from the

synthesis of cellulose. The model shows that α-cellulose δ 18 O is influenced primarily

by the δ 18 O of the source water taken up by the plant, and by the amount of isotopic
enrichment of that water that subsequently occurs in the leaf, and whose signature is
inherited by the sucrose used in the eventual biosynthesis of cellulose. As detailed above,

source water differences in cloud forests will be a function of the seasonal regimes of
rainfall and cloud water. Lower relative humidity levels during the peak of the dry season
should result in higher evapotranspiration rates and enrichment of the δ 18 O value of leaf
water and the resultant cellulose.

Barbour et al. [2004] modified the model of Roden et al. [2000] to include the
potential influence of the Peclet effect, a diffusion of isotopically-enriched water away
from the sites of transpiration in the leaves back into the unenriched source water, which
results in lower δ 18 O value for bulk leaf water than would be predicted from a simple

calculation based on evapotranspiration in the leaf [Barbour et al., 2000]:

 75

Warmer Temperatures
Reduced Cloud/Precipitation
Reduced Moisture Advection
OXYGEN ISOTOPE (δ18O) RATIO

DRY SEASON
Fog, Reduced WET SEASON
Precipitation Precipitation
Reduced Relative Higher Relative
Humidity Humidity

Higher Rainfall
Higher %RH
Cooler Tempertures

ANNUAL ISOTOPE CYCLE IN WOOD

Figure B.1: Conceptual model of climatic controls on the annual and interannual patterns
of stable oxygen isotope ratios in the α-cellulose of cloud forest tree radial growth. The
annual cycle is generated primarily by the seasonal change in the δ 18 O of rainfall, the use
of cloud water by trees in the dry season, and the isotopic enrichment of source water
by evapotranspiration during the dry season. Interannual variability in annual maximum
values is expected to be related to temperature, relative humidity, cloudiness, and moisture
advection during the dry season. Anomalies in the annual minima are likely related
primarily to wet season precipitation amounts.
 76

1c = 1l (1 − px pex ) (B.2)

For our purposes, 1c is the enrichment of the cellulose δ 18 O above that of the original

source water, 1l reflects the enrichment of the source water signal in the leaf at the sites
of photosynthesis, as a function of the vapor pressure difference within the leaf, the δ 18 O
of atmospheric water vapor, the fractionation from the liquid-vapor phase change, and
the Peclet Effect. pex is the proportion of oxygen atoms exchanged during the formation

of cellulose from sucrose, equivalent to f 0 from Equation (1). px is the proportion of

unenriched water at the site of cellulose formation. For large trees and for cellulose
formation in the xylem, it is believed that px approaches 1, as the site of cellulose
synthesis is sufficiently distal from the diffusion of enriched leaf water into unaltered
source water that it is primarily those waters that provide the exchangeable oxygen during

cellulose biosynthesis [Barbour et al., 2002]. In this case, px pex is equivalent to f 0 from
Equation (1) and represents the mixing between enriched δ 18 O signatures from the leaf
and δ 18 O from the original source water.
We hypothesize (Figure B.1) that the isotopic difference between wet season

precipitation and dry season rainfall and cloud water is sufficient to generate annual
isotope cycles in the xylem of cloud forest trees. We rely on the seasonal shift in primary
water source to drive concomitant changes in cellulose, enabling identification of annual
cycles and chronology development. Interannual variability should be manifested as

enhancement or suppression of the amplitude of the mean annual isotope cycle through
changes in precipitation, temperature, and relative humidity. Year-to-year differences
in annual maximum values are expected to be related to temperature, relative humidity,
cloudiness, and moisture advection during the dry season. Anomalies in the annual

minima are likely related primarily to wet season precipitation amounts.

 77

B.4 Methods and Materials

We test our hypotheses using samples and data collected from the Monteverde Cloud
Forest Reserve in Costa Rica. Long-term vegetation monitoring and daily climate records
from the Reserve can be used to calibrate, model, and interpret the δ 18 O content of

tree cellulose, making the site an ideal location in which to establish if the necessary
chronological control and climate signal exist. Establishing both of these is a necessary
prerequisite for the development of robust climate reconstructions.

B.4.1 Site description

The Monteverde Cloud Forest (10.2o N, 85.35o W, 1500m) is draped over the Cordillera
de Tilarán in northwestern Costa Rica. The cloud forest extends from ∼1500 m on the
Pacific slope upward to the continental divide, and down the windward Atlantic slope as
low as 1300 m [Haber, 2000]. Below the cloud forest and downslope to 700m elevation on
the leeward Pacific slope the vegetation is predominantly evergreen tropical premontane

wet forests with a distinct dry season. Between 1400 and 1500m, the premontane forest
transitions to lower montane wet forests with greater orographic cloud influence [Haber,
2000]. The cloud forest above 1500m receives an average of 2500mm of precipitation in
a year and has a mean annual temperature of 18.5o C [Figure B.2; Pounds et al., 1999;

Clark et al., 2000]. These forests are characterized by the presence of tradewind-driven
orographic clouds in the transitional (November through January) and dry (February
through April) seasons. During the summer (May through October), the precipitation
regime is dominated by the position of the Intertropical Convergence Zone (ITCZ) and

characterized by strong convective rainfall and thunderstorms [Clark et al., 2000; Rhodes
et al., 2006]. A midsummer drought, common to the Pacific Coast of tropical Central
America [Magaña et al., 1999], is present in July and August (Figure B.2). The cloud
 78

forest receives the majority of its rainfall during the summer wet season, but the annual
moisture regime has substantial and seasonally important inputs from orographic cloud
water between November and April. Whereas the Atlantic slope is perpetually wet,
the leeward slope depends on the northeasterly trade wind-driven moisture over the

continental divide as a critical dry season water source. Clark et al. [1998] found that at
Monteverde these moisture inputs accounted for perhaps 22% of the annual hydrological
budget. However, estimates of the contribution from cloud water at different tropical
montane cloud forest sites vary from 6 up to 154% of precipitation [Bruijnzeel and
Proctor, 1995; Bruijnzeel, 2001]. Cloud water inputs are consistently more important

during the dry season, accounting for up to 75% of total moisture inputs in some forests
[Bruijnzeel, 2001].
Samples for our calibration study of cloud forest isotope dendroclimatology come
from two locations. We first take advantage of two stands of planted and monitored

Ocotea tenera (Lauraceae) from forest plots in the transitional zone between lower
montane and cloud forest [Wheelwright and Logan, 2004] in order to calibrate and
confirm our hypothesized age model and precisely evaluate potential climate signals. We
also analyze a set of pilot samples from mature cloud forest canopy trees above 1500m

to allow us to identify potential candidate species and sites for future long-term climate
reconstructions. Together, the combination of different sites and different species allows
us to investigate spatiotemporal variability in cloud forest hydroclimatology as reflected
in the δ 18 O of cloud forest trees.

B.4.2 Field and laboratory sampling

Increment cores from O. tenera were collected in February 2004 from the Trostles (NWT)
and Hoges (NWH) plantation sites [Wheelwright and Logan, 2004] (1410 and 1430m).
 79

Monteverde, 10.3N, 84.8W, 1540m

600 90 19.5

500
19

400 85

Relative Humidity (%)

Precipitation (mm)

18.5

Temperature (C)
300

18

200 80

17.5
100

0 75 17
J F M A M J J A S O N D
MONTH

Figure B.2: Patterns of mean annual temperature and precipitation for Monteverde from
the Campbell weather station and derived relative humidity (1979-2000,10.2 o N, 85.35
o W, 1540m, Pounds et al. [1999].
 80

Three cores were obtained from the Trostles plantation, with two collected from a single
tree (NWT02A/B), and one from a separate individual (NWT01A). A fourth core was
collected from the Hoges plantation (NWH03A). This sampling approach was designed to
evaluate the fidelity and consistency of the chronological and climatic signal both within

and between trees, and amongst sites.

Seven additional cores or entire cross-sectional discs were also opportunistically
collected in and around the Monteverde Cloud Forest Reserve (1500-1660m), largely
from mature canopy trees killed or damaged in a December 2003 windstorm. This
group included individuals from the genus Quercus (tropical oaks) as well as from the

cosmopolitan Sapotaceae and Lauraceae families (Table B.1). The forest trees were
sampled along a rudimentary transect up the Pacific slope toward the continental divide.
This strategy was intended to accomplish two complementary goals. First, to permit
discovery of sites or species that have maximum sensitivity to both annual and interannual

changes in climate. Second, the comparison of δ 18 O values from both below and within
the cloud bank should allow us to estimate the relative importance across the landscape
of changes in precipitation, temperature, and relative humidity to the α-cellulose δ 18 O of
cloud forest trees.

B.4.3 Sample preparation

The cores were subsampled in the laboratory on a rotary microtome at 20 µm increments.

Ten slices were then aggregated into a single sample for a sampling depth resolution of
200 µm. Radial subsections approximating the diameter of the cores (5mm) were cut
from the large full stem discs using a bandsaw prior to microtoming. Our subsampling

interval resulted in approximately 700 to 1500 µg of wood per sample, depending on

wood density, structure, and moisture content.
 81

Raw wood was then extracted to α-cellulose using the standard Brendel technique
[Brendel et al., 2000; Gaudinski et al., 2005; Anchukaitis et al., 2007b] as modified
for small samples [Evans and Schrag, 2004]. The Brendel procedure uses a hot
10:1 acetic:nitric acid delignification extraction that also removes most hemicelluloses

and mobile resins, followed by progressive solvent washes and sample drying in
distilled water, ethanol, and acetone [Evans and Schrag, 2004; Anchukaitis et al.,
2007b]. Following α-cellulose extraction, samples were dried in a warm oven, and
then overnight under vacuum. Use of the Brendel technique allows for single-day
chemical preprocessing of samples and, most critically, permits the use of very small

( < 1 mg) samples with a sufficient final sample yield [Evans and Schrag, 2004] for
replicate isotopic measurements. The Brendel technique results in α-cellulose which
is not significantly biased in its isotopic composition compared to traditional methods
[Evans and Schrag, 2004; Anchukaitis et al., 2007b].

B.4.4 Isotopic analysis

100 to 150 µg of α-cellulose of each sample from the plantation O. tenera core
were wrapped in silver capsules and converted to CO in a ThermoFinnigan Thermal
Conversion/Elemental Analyzer (TC/EA) at 1450 o C. The oxygen isotope composition

of the CO gas was analyzed on a ThermoFinnigan Delta XL continuous flow mass

spectrometer at Harvard University. Measurement precision for several hundred Baker
α-cellulose standards was 0.45h Long-term precision on the instrument is better than
0.1h on internal reference gas standards.
For the analysis of pilot samples from canopy trees in the Monteverde Cloud Forest

Preserve, 300 to 350 µg of α-cellulose were loaded in silver capsules and converted
online to CO in a Costech High Temperature Generator/Elemental Combustion System
 82

(HTG/ECS) system with a quartz outer reactor and molybdenum crucible packed with
graphite [Evans et al., 2007]. Our HTG pyrolysis system at The University of Arizona
is a 1MHz radio frequency induction heater which quickly brings the molybdenum
susceptor inside the reactor assembly to >1500o C, at which time the sample is

introduced to the crucible and pyrolysed. Use of the HTG peripheral reduces laboratory
consumables, simplifies reactor maintenance and replacement, and results in an efficient
high-temperature conversion of the sample to CO [Evans et al., 2007]. This study is
the first application of this technology to paleoenvironmental research using oxygen
isotopes. The isotopic ratio of the CO gas was measured on a ThermoFinnigan Delta+XP.

Precision on our Sigma Alpha Cellulose (SAC) solid standard material was 0.34h, and
internal precision consistently 0.08h or better. All δ 18 O results from both instruments are
reported with respect to the international Vienna Standard Mean Ocean Water (VSMOW).

B.4.5 Forward modeling

We use the Barbour et al. [2004] model of the environmental controls on the stable
isotope composition of wood, as modified and adapted for time series prediction in
tropical environments by Evans [2007], to simulate a theoretical stable oxygen isotope
time series based on local meteorological data. Synthetic isotope time series are then

compared with our actual measured δ 18 O chronology from O. tenera to independently

test the age model and climate signal detection. The model takes as input monthly
temperature, precipitation, and relative humidity data. There are 6 parameters in the
model, many of which are only weakly constrained by observations, particularly for
tropical species and environments. Following Evans [2007], we parameterized leaf

temperature as a function of air temperature [Linacre, 1964] and atmospheric water vapor
as a function of condensation temperature and the fractionation related to the vapor-
 83

liquid phase change. Evans [2007] derived stomatal conductance as a function of vapor
pressure deficit calculated from leaf and air temperature and relative humidity. This
allows stomatal conductance, and therefore transpiration, to vary temporally in response
to environmental conditions. Two additional parameters provide the coefficients for the

model relating precipitation to δ 18 O of meteoric waters (see below).

Precipitation and temperature data are available from the Campbell (1540m) weather
station [Pounds et al., 1999] for the period 1977 to 2005, and from the Monteverde
Institute [Rhodes et al., 2006] from 2004 through 2006 (1420m). We calculated monthly
source water δ 18 O as a function of the observed relationship between rainfall amount

and the δ 18 O composition of rainfall (the amount effect) from Rhodes et al. [2006].
We regressed the δ 18 O on rainfall amounts for those data with aggregate collection
periods of two weeks to two months. Single-storm events and shorter collection periods
showed a higher variability which likely reflected the time of sampling and the trajectory

and history of individual weather systems, while longer periods excessively smoothed
monthly differences related to the timing and onset of precipitation seasonality. Three
samples which were observed by Rhodes et al. [2006] to have algae growing on the
collection container were excluded from the regression model. Source water δ 18 O values

were then calculated based on monthly total precipitation values from the Campbell data
set:

δ 18 Osw h = −0.0155 × P(mm/month) − 1.2614 (B.3)

The regression model accounts for 56% of the variance in the observed dataset and is
significant at p < 0.01 with 14 effective degrees of freedom.
The Rhodes et al. [2006] data also includes relative humidity for July 2005 through the

end of 2006. In order to develop modeled synthetic chronologies which spanned the full
 84

21

20

Temperature (C)
19

18

17

1000 16
Precipitation (mm)

800

600

400

200

0 100
NCEP Campbell

% Relative Humidity
90

80

70
1980 1985 1990 1995 2000 2005
Year

Figure B.3: Meteorological data from Monteverde used as input for the forward model
of tree α-cellulose δ 18 O. (a) Temperature (o C) and (b) Precipitation from the Campbell
meteorological station [Pounds et al., 1999], and (c) relative humidity derived from the
Campbell data and from the NCEP Reanalysis 2 [Kanamitsu et al., 2002]. The NCEP
relative humidity has been scaled to give it the same mean as that observed in Rhodes
et al. [2006], but allowed to retain its variance. The horizonal lines of the same color
show the original overall mean value. Sensitivity to the choice of relative humidity data
is 0.20h per month per percent mean relative humidity.
 85

timescale of our oxygen isotope chronologies from the O. tenera plantations, we derived
a relative humidity time series to use with the model by regressing the daily relative
humidity measurement from Rhodes et al. [2006] on the maximum daily temperature and
daily precipitation values for the same station. This model was then applied to create a

daily relative humidity time series spanning the full length of the Campbell [Pounds et al.,
1999] daily temperature and precipitation record:

R H % = (−1.0191 × T max(C)) + (0.2459 × P(mm)) − 105.1180 (B.4)

The regression model accounted for 43% of the variance, and was significant at p <
0.01. Relative humidity values were then adjusted so that the long-term seasonal mean

and variance matched those from the observations by Rhodes et al. [2006]. Daily values
from all three variables were then combined to form monthly means and total precipitation
for model input. We also extracted a time series of monthly relative humidity from the
NCEP Reanalysis II gridded data [Kanamitsu et al., 2002] for the latitude, longitude, and

atmospheric pressure level (850mb) which most closely corresponds to the Monteverde
Cloud Forest, in order to test the sensitivity of our results to the choice of available relative
humidity data.
Evans [2007] used the forward model to reconstruct the patterns of interannual

variability in δ 18 O cycles in a tropical trees from La Selva Biological Station in Costa

Rica. In those simulations, the mean and variance of the simulation were adjusted
to that of the observed δ 18 O time series. Here, we use a simple single-component
soil mixing model in place of the variance adjustment to capture the influence of the
temporal smoothing of individual monthly water δ 18 O as a result of soil water residence

times. Following Evans [2007], we use a Monte Carlo procedure (1000 simulations) and
 86

randomized adjustments (up to 20%) of the 6 model and 2 source water coefficients to
estimate the sensitivity of the interannual patterns of variability to the selection of the
parameter values.

B.5 Results

B.5.1 Plantation trees (NWT, NWH)

δ 18 O from all four plantation cores show regular isotope cycles as large as 9h (Figure
B.4). Based on our conceptual model (Figure B.1, we assigned the local maxima of each

peak to the month of April of each year. Age models were developed individually for
each tree (Figure B.5), and confirmed based on the sampling date and incremental growth
measurements over the last two decades [Wheelwright and Logan, 2004]. We were able to
detect missing years primarily using these growth observations, and secondarily through
identification of apparently truncated cycles in the raw δ 18 O record, a rudimentary

crossdating [Fritts, 1976] between the four cores, and the results of the forward model
simulations. These missing years are not obviously associated with any climatic cause,
however, nor are they common amongst the four cores from the plantations.
Annual radial growth increments calculated from the age modeled δ 18 O chronologies

match those from the basal diameter measurements [Wheelwright and Logan, 2004].
The mean of the radial growth rate from sample NWT01A is 4.5mm year−1 , while the
estimate from basal diameter measurements is 5.4mm year−1 . The overall mean growth
rate from tree NWT02 is 4.3mm year−1 from the age modeled isotope time series, and

approximately 4.4mm year−1 from the repeated diameter measurements. Tree NWH03A
from the Hoges plantation has a δ 18 O derived mean growth rate of 4.6mm year−1 and
4.4mm year−1 from observations. The mean interseries correlation [c.f. Fritts, 1976]
between the monthly δ 18 O series for the three trees is 0.75, and 0.68 when the amplitudes
 87

35

δ18O %o VSMOW
NWT01A
30

25

35 20
δ18O %o VSMOW

NWT02A
30

25

20 35

δ18O %o VSMOW
NWT02B
30

25

35 20
δ18O %o VSMOW

NWH03A
30

25

20
80 70 60 50 40 30 20 10 0
Depth from Cambium (mm)

Figure B.4: Isotope ratios as a function of sampling depth for the four plantation Ocotea
tenera from the Trostles and Hoges sites. Individual samples are indicated by dots. In
NWT02B and NWH03A, samples across or close to the pith can be seen in the δ 18 O
plateau near the center of the tree.
 88

of the annual cycles are compared. The samples from the Trostles plantation are better
correlated with one another than with the single core from the Hoges plantation, almost
entirely because NWH03A does not show the reduction in growth rate and δ 18 O amplitude
after 1999 observed in the older trees from the Trostles plantation (see below).

There are some notable features in the δ 18 O series as a function of depth. Cores
NWT02A and NWT02B, although from the same trees, show different mean growth rates,
although in combination their average growth rate is similar to observations. At least
two cores show evidence of a reduced cycle amplitude near the center of the tree, with
NWH03A showing a clear plateau in δ 18 O and a subsequent resumption of regular cycles.

Both the variance reduction and plateau are likely related to either passing through the pith
of the tree, or as a result of sampling tangential to the growth radius near the center of the
tree. At least three out of the four cores (NWT01, NWT02A, and NWT02B) also show
growth reductions in the outermost 10mm of growth. This is also consistent with the basal

growth observations from Wheelwright and Logan [2004], which show a clearly reduced
growth rate (as small as 2.2mm year−1 in NWT01A) after 1997. A suppression of the
amplitude of the annual δ 18 O cycle is also observed in the most recent xylem growth.
The mean annual δ 18 O cycle from each of the individual trees has the same amplitude

within the instrumental precision of measurements (Figure B.6a). The seasonal maximum
is set by the age model to April, while the annual mean minimum is in September for
NWT01A and NWH03A, and in October for NWT02. These correspond to the peak
of the climatological dry and wet seasons, respectively. The composite mean time series
formed from the overlapping portions (1990 to 2001) of the time series of each core shows

an average annual cycle of 5.72h. The δ 18 O chronology shows patterns of interannual

variability as anomalous amplitude in the annual cycles, ranging from 2.40h to 7.55h
and a standard deviation of = 1.45h (Figure B.6b).
 89

35

δ18O %o VSMOW
30

25

35 20
δ18O %o VSMOW

NWT01A
30

25

20 35

δ18O %o VSMOW
NWT02A NWT02B
30

25

35 20
δ18O %o VSMOW

NWH03A
30

25

20
1988 1990 1992 1994 1996 1998 2000 2002 2004
Year

Figure B.5: Isotope ratios as a function of time for the four plantation Ocotea tenera
from the Trostles and Hoges sites. ‘Missing’ years in the age model chronologies reflect
both known growth hiatuses from the annual diameter measurements [Wheelwright and
Logan, 2004] and their confirmation with rudimentary crossdating between cores, and is
supported by the results of forward modeling
 90

The interannual variability in annual cycle amplitude is driven primarily by a

combination of year-to-year changes in the annual minimum value (σ = 0.95h) and
a declining trend in the value of the annual cycle maximum, most clearly influenced by
a decrease after 1996 (Figure B.6c), which accounts for the majority of the variance in

the annual maxima. Interannual variability in δ 18 O is most clearly linked to precipitation

(Figure B.6d), with wet season precipitation anomalies showing a clear association with
detrended amplitude anomalies and a significant relationship that explains more than 50%
of the variance, despite the small degrees of freedom (r = 0.74, R 2 = 0.56, p < 0.01).
The declining trend in maximum annual δ 18 O values mirrors somewhat the declining

trends in Monteverde maximum temperature [Pounds et al., 2006] and dry season relative
humidity over the same period. The decline in maximum δ 18 O after 1996 (Figure B.6c)
in the composite plantation time series is also seen in the relative humidity corresponding
to the location of Monteverde from the NCEP II reanalysis. However, the individual δ 18 O

series maxima at the calibration site do not in turn mirror the large summer anomalies
seen in both the temperature and relative humidity data in, for example, 1992 and 1998
(Figure B.3).

B.5.2 Forest canopy trees (MV)

Whereas all four plantation samples showed distinct, large magnitude changes in δ 18 O
associated with annual changes in precipitation, pilot samples from forest trees showed
a range of temporal stable oxygen isotope patterns, most without consistent annual
isotope cycles (Figure B.7a-f, Table B.1). Of the seven pilot forest trees examined here,
only three (MV03, MV05, and MV14B) show periodic indications of annual cyclicity

in δ 18 O than could be used for chronology. Because of this limited sample set, we
include here in subsequent analyses data from two additional trees (MV12A and MV15C)
 91

31
NWT01A Annual Max Annual Mean Annual Min
30 NWT02A+B
32
δ18O %o VSMOW

29 NWH03A

δ18O %o VSMOW
30
28
28
27

26 26

25 24
a) c)
24 22
J F M A M J J A S O N D 1992 1994 1996 1998 2000
Month Year

34 9 1000
Mean Chronology

Precipitation Anomaly (mm)

Isotope Cycle Amplitude

32 8
500
δ18O %o VSMOW

30
7
28
6 0
26
5
24
-500
4 δ18O
22
Precipitation
b) d)
20 3 -1000
1992 1994 1996 1998 2000 1992 1994 1996 1998 2000
Year Year

Figure B.6: Composite isotope timeseries for the Trostles/Hoges calibration set. (a)
Mean δ 18 O annual cycle (‘climatology’) for the three trees and the mean of all trees.
The amplitude and seasonal patterns are indistinguishable within measurement precision,
with temporal age model differences of approximately 1 month. (b) Composite timeseries
mean of the three trees (four cores), showing variability (1σ ) around the overall mean. (c)
Annual maxima, minima, and mean values from the composite mean site chronology.
Most of the interannual variability (is related to the minima, while the trend in the mean
is a result of patterns observed in the annual maxima. (d) The amplitude of the mean
annual δ 18 O cycle, which shows a clear relationship to interannual patterns in the wet
season rainfall (R 2 = 0.56, p < 0.01, d. f. = 10)
 92

with confirmed coherent annual cycles over their full length, which we have analyzed
elsewhere [Anchukaitis et al., 2007a].
Quercus could be an excellent candidate genus for tropical isotope dendroclimatology,
because it is relatively common, is straightforward to identify in the field, and is a long-

lived mature canopy tree. However, neither of our two samples from Quercus, one at
1540m and the other higher in the cloud forest at 1660 meters shows consistent cycles
in δ 18 O that could be used for chronological control. The lower forest sample (MV05)
shows periods of identifiable cyclicity, but much longer periods with no coherent temporal
pattern for chronology. The upper cloud forest oak (MV11) shows no detectable δ 18 O

cycles at all. Similarly, our pilot sample (MV14B) from Podocarpus shows short periods
of coherent large magnitude changes comparable to those from the plantation calibration
set, but sustained periods with no detectable cycles in δ 18 O. Samples from Sideroxylon
(MV23A, Sapotaceae) from ∼1500m and Pouteria (MV15A, Sapotaceae) at 1580m

likewise showed no apparent coherent δ 18 O cycles in pilot isotopic measurements. The

most promising evidence of cycles outside of the plantation trees came from a Lauraceae
(MV03) in which they could be detected for approximately 12 years. However, the tree
appears to have experienced extremely suppressed growth in the most recent period (0 to

18mm depth), making it impossible to tie the chronology to the meteorological record and
develop an absolute chronology. Moreover, an individual of the same species growing at
the same location (MV02) showed no obvious signs of annual δ 18 O cycles.
There is an increasing but statistically insignificant trend in total mean δ 18 O value
with increasing altitude in the Monteverde Cloud Forest (Figure B.7g). The two oak

trees both show lower average δ 18 O with respect to elevation, and overall, than any
of the plantation or forest trees. There is a distinct and statistically significant pattern
(R 2 = 0.99, p < 0.01, n = 5) associated with elevation in the mean amplitude of the
 Annual Cycle (h)
Sample Taxon Elevation(m) Mean (h) Amplitude (n) σ Amplitude σ Max/Min Reference

NWT/NWH Ocotea tenera 1410 27.27 5.72 (11) 1.45 0.89/0.95 This study
MV02 Lauraceae 1530 28.06 - - - This study
MV03 Lauraceae 1530 27.71 3.23 (13) 1.34 1.00/1.05 This study
MV05 Quercus 1540 25.16 1.72 (13) 0.84 0.81/0.6 This study
MV11 Quercus 1660 26.23 - - - This study
MV12 Sapotaceae (Pouteria) 1560 27.73 2.59 (55) 1.08 0.99/0.64 Anchukaitis et al. [2007a]
MV14B Podocarpus 1580 28.88 2.19 1.42 (8) 0.71/0.97 This study
MV15A Sapotaceae (Pouteria) 1580 27.28 - - - This study
MV15C Sapotaceae (Pouteria) 1580 26.97 1.79 (70) 1.08 0.83/0.72 Anchukaitis et al. [2007a]
MV23A Sapotaceae (Sideroxylon) 1480 26.95 - - - This study

Table B.1: Site elevation, mean δ 18 O value, and characteristics of annual oxygen isotope cycles from plantation composite
mean and cloud forest trees from Monteverde. Elevations are estimated from altimeter readings and topographic maps. Overall
mean δ 18 O is based on all measured values, irrespective of the presence or absence of annual cycles. The characteristics of the
annual cycle are calculated from the portion of the age modeled series where they could be detected, with n equal to the number
of identified annual cycles.

93
 94

30

Estimated Elevation (m)

25 a). 1700
10 5 0
1600
30
1500
25 b).
40 20 0
h).
1400
28 0 2 4 6
18
26 Amplitude δ O %o VSMOW
24
c).

Estimated Elevation (m)

22 1700
100 50 0
δ18O %o VSMOW

28
1600
26
d). 1500
24
10 5 0
i).
1400
30 24 26 28 30
18
Mean δ O %o VSMOW
e).

Estimated Elevation (m)

25
20 10 0 j). 1700

28 1600

26 f ). 1500
20 10 0
1400
28 0 0.5 1
26 Ratio, Interannual:Annual
g).
24
20 10 0
Depth from bark (mm)

Figure B.7: Isotope timeseries for the canopy tree calibration set and the observed
relationships between elevation, δ 18 O mean, and the amplitude of the annual δ 18 O cycle.
δ 18 O by depth from (a) MV02, (b)MV03, (c) MV05, (d) MV11, (e) MV14B, (f) MV15A,
and (g) MV23A (see Table B.1 for details). (h) Amplitude of the annual cycle (including
data from (b,c,e), see text for details), and (i) overall mean δ 18 O versus elevation. (j) The
ratio of the interannual amplitude variance to the mean isotope climatology increases with
elevation, showing greater year-to-year climate variability above the orographic cloud
bank (indicated by the gray line in (h-j).
 95

annual cycle in trees (MV05, MV14B, MV03, MV12A, and MV15C) in those instances
where it could be detected (Figure B.7h). Annual mean δ 18 O amplitude ranges from
5.72h for the plantation species at approximately 1410m to 1.79h for sample MV15C
at approximately 1580 meters. The lower forest oak (MV05) appears to have cycles of, on

average, 1.72h. Both of the oak samples (MV05 and MV11) show lower overall mean
δ 18 O values and MV05 has a suppressed annual cycle amplitude compared to those at
similar elevations. The ratio of the mean annual δ 18 O cycle to the standard deviation of the
annual amplitude decreases with elevation, indicating that as the annual cycle is reduced
with increasing elevation, the magnitude of the year-to-year variability (anomalies) with

respect to the annual cycle increases and a greater proportion of the total variability is in
the interannual or longer time scales.

B.5.3 Forward δ 18 O modeling

Simulations using our forward model produce annual cycles similar to those seen in

the measured δ 18 O from the plantation trees (Figure B.8a-b). The best match between
overall variance in the simulated and actual isotope chronologies for the plantation
trees is achieved using a soil water mixing ratio of approximately 30:70 (30% new
precipitation mixing with 70% of the previous month’s soil water). Using the Campbell

meteorological data, including the derived relative humidity series, the observed and
modeled chronologies are significantly correlated (r = 0.73, R 2 = 0.52, p <
0.01), taking into account the high degree of serial autocorrelation (ar (1, 2)obs =
[0.83, 0.45], ar (1, 2)model = [0.77, 0.46]) in both series [Ebisuzaki, 1997]. Simulations
using the NCEP Reanalysis II relative humidity (Figure B.8) for the grid cell associated

with Monteverde are similarly correlated (r = 0.74, R 2 = 0.55, p < 0.01), but are not
significantly different. This is largely because the correlation between the simulations
 96

and the observed δ 18 O time series reflects the large proportion of the total variance in the
annual cycle, which is predominantly controlled by the seasonal amount effect reflected
in all plantation trees. Overall, there is no statistically significant (two-tailed t- and F-test,
α = 0.05) difference in the means or variances of the residuals (obser ved − modeled)

using either NCEP or Campbell-derived relative humidity.

The observed and modeled annual amplitude show similar patterns, but are weakly
correlated at this scale (NCEP %RH: r[model,obs] = 0.55, p = 0.08; Campbell %RH:
r[model,obs] = 0.51, p = 0.11), largely influenced by the mismatch between the measured
and simulated values for 1991 (Figure B.8c). The modeled amplitude is significantly

correlated over the same period with the wet season precipitation anomalies (NCEP %RH:
r[model, pr eci p] = 0.71, p < 0.01; Campbell %RH: r[model, pr eci p] = 0.91, p < 0.01), with
a substantially stronger correlation between precipitation anomalies and the simulation
using the lower amplitude Campbell derived RH record. A downward trend in the

annual maximum δ 18 O values is seen for model simulations, irrespective of the relative
humidity input used, although the NCEP data results in a slightly better match to the
actual, observed isotope time series, particularly the annual maxima δ 18 O values (Figure
B.8d). The simulated chronology that results from using this dataset, however, has larger

positive excursions in maximum annual δ 18 O that are not seen in the actual chronology
(1992,1994), and results in a overall mean δ 18 O value approximately 1.35h enriched
above that of the simulation using the Campbell derived relative humidity when the
simulations are not adjusted to the observed mean. These differences indicate a model
sensitivity of approximately 0.20h for every percent relative humidity.

When compared over the limited period for which both climate data and volume-
weighted seasonal δ 18 O of meteoric waters at Monteverde are available, simulations using
calculated and observed source water isotope ratios show similar seasonal patterns and
 97

35

30
δ18O %o VSMOW

25

20 model error
modeled
observed a).
15
1988 1990 1992 1994 1996 1998 2000
Year
3
observed
model
2
Normalized Anomalies

Wet Season Precipitation Anomalies

-1

b).
-2
1988 1990 1992 1994 1996 1998 2000
Year
31 31
Actual Source Water
Campbell %RH Plantation Composite
30 NCEP %RH 30 Model
δ18O %o VSMOW

δ18O %o VSMOW

29 29

28 28

27 27

26 26

25 25
c). d).
24 24
2004.4 2004.6 2004.8 2005 2005.2 2005.4 J J A S O N D J F M A M
Decimal Year Month

Figure B.8: Comparison between observed composite mean plantation isotope timeseries
and forward model simulation [Barbour et al., 2004; Evans, 2007] (see text for details).
Input to the model was observed precipitation and temperature from Monteverde [Pounds
et al., 1999], and the mean monthly relative humidity from the NCEP/NCAR Reanalysis
[Kanamitsu et al., 2002]. (a) The simulated isotope time series is significantly correlated
with the observed δ 18 O from the plantation trees (r = 0.74, R 2 = 0.55, p < 0.01).
(b) Observed and modeled chronologies are correlated with wet season precipitation
anomalies (robs = 0.75, p < 0.01; rmodel = 0.71, p < 0.01). (c) The source water model
and actual source water produce annual cycle of similar magnitude, and the magnitude
of the modeled annual cycle is similar irrespective of the relative humidity data set.
(d) The seasonal patterns and isotope climatology of the mean adjusted oxygen isotope
climatology are also similar, with an estimated age model error of 1 month.
 98

amplitudes (Figure B.8c). The amount effect model (Equation 3) reproduces the seasonal
pattern of observed meteoric water δ 18 O on which it was based, with a slight loss of
variance at the annual maxima and minima (Figure B.8c). The modeled α-cellulose δ 18 O
cycle amplitude is also similar whether observed or modeled source water is used, and

irrespective of the relative humidity dataset. However, uncertainties in model parameters

result in a confidence interval up to 1.5h wide, particularly at the local maxima and
minima of the simulated annual δ 18 O cycles.

B.6 Discussion

B.6.1 Annual oxygen isotope cycles

Annual δ 18 O cycles are clearly present in our set of plantation trees, and are sufficiently
large and well-defined that they are easily distinguished from the occasional smaller
positive excursions at the time of the annual minima, which are probably the result

of the existence and magnitude of the Central American mid-summer drought, which
can be seen in some years (i.e. 1993) in both the observed and modeled α-cellulose
δ 18 O time series. The similarity between the measured radial growth rates from long-
term monitoring and those derived from the age modeled isotope time series, and

supported by the forward model simulations, demonstrates conclusively that the proxy
δ 18 O chronology can be securely established using these cycles. Given the potential
uncertainties associated with both individual repeated basal growth measurements [Sheil,
2003; McLaughlin et al., 2003] and the increment estimates from age modeled δ 18 O,

the consistency between the two datasets indicates the age model (1 cycle = 1 year) is
particularly robust. There is no indication from the isotope time series that the Ocotea
tenera have a significant growth hiatus during the year, which is supported by comparison
to our forward modeling results.
 99

Unlike traditional dendroclimatological approaches using tree-ring widths or density,

where massive sample replication and robust crossdating result in an overall composite
age model error which is effectively negligible [Fritts, 1976], the smaller sample depth
and limited opportunity for crossdating in our plantation site δ 18 O chronology leaves

open the possibility of error in assigning the isotope time series to calendar years.
This is exacerbated here by the short length of the chronologies developed from our
calibration set. Even in our O. tenera calibration samples, development of the age model
is complicated by years where individual trees did not have appreciable basal growth, the
reduced growth rates since 1997, and gaps in the annual observational data. This latter

factor is particularly important between 1997 and 2002, because sometime between 2001
and 2004 all basal growth in several trees in Trostles plantation had ceased or slowed
to imperceptible increments. For our O. tenera chronology, we estimate an age model
error no larger than ± 2 years, based on the basal growth measurements and comparison

with the forward model simulations, for our plantation calibration tree set. In non-
plantation trees and particularly in trees growing at average rates less than 2mm year−1 ,
age model error can be determined by the range of possible realistic age models, and may
be complemented by high-resolution radiocarbon assays on the period since A.D. 1955

[Worbes and Junk, 1989; Anchukaitis et al., 2007a]. Unfortunately, while the plantation
O. tenera allow us to test and calibrate our proposed age model and detect the imprint
of climate variability on the δ 18 O cycles, this early successional species is unlikely to
provide the material for long isotope chronologies.
There are several factors which could potentially complicate identification of annual

isotope cycles. Long residence times for meteoric waters in the soil would result
in temporal mixing of different seasonal water sources, which would both smooth
and dampen the amplitude of the annual signal we seek for chronological control.
 100

Likewise, trees which access primarily deeper sources of ground water would be relatively
insensitive to the intra-annual change in the δ 18 O of available moisture [c.f. Evans and
Schrag, 2004]. However, shallow rooting depths for cloud forest trees [Matelson et al.,
1995] in response to soil nutrient availability should mean that shallow soil water is

their primary moisture source and they do not have access, or need to access, deeper
groundwaters. One hypothesis for the lack of annual cycles in some of the forest trees
analyzed here is that they are able to access deeper groundwater or well-mixed stream
water, and are relatively insensitive to the seasonal δ 18 O cycle of meteoric waters.
At our current sampling resolution and given requirements for α-cellulose yield,

growth rates must be sufficiently rapid, ∼2mm per year, to allow for enough samples
per year in order to sufficiently resolve the annual cycle and its amplitude, but not so
rapid as to limit the ultimate length of the reconstruction. Slower growth rates would
currently make it difficult to resolve the full amplitude of the annual cycle given current

analytical constraints. This can be partially addressed by using larger-diameter increment

borers, whole cross-sections, or improved analytical procedures that require less cellulose
for a precise measurement.

B.6.2 Climate analysis

Interannual anomalies in the amplitude of the plantation O. tenera δ 18 O cycles are

dominated by variability in the annual minima value, which in agreement with our
conceptual model (Figure B.1) is primarily controlled by the amount of rainfall received
during the wet season (Figure B.6d). There is little year-to-year variance in the annual
maxima in the composite mean δ 18 O series from the plantation sites, with only a slight

downward trend related primarily to a slight step change between 1996 and 1997.
This downward trend might be related to decreases in relative humidity or maximum
 101

temperatures [Pounds et al., 2006] over the common period, but missing from the δ 18 O
maxima are distinct large positive anomalies which could be related to interannual
temperature or relative humidity anomalies associated with El Niño events.
The most likely reason for the lack of a clear dry season δ 18 O anomaly signal in the

O. tenera series is the elevational position of our pilot calibration site at ∼1410m in the
transitional region between premontane wet and cloud forest. At this lower elevation,
the persistent cloud immersion that characterizes forests along the continental divide
above 1500m is considerably reduced, although mist can still be an important moisture
source for vegetation. Trees in the premontane wet forest and below the mean lifting

condensation level would be relatively insensitive to changes in orographic clouds and

relative humidity since the magnitude of the local hydroclimatic alterations to fluctuations
would therefore be relatively small. At higher elevations above the mean lifting
condensation level, where forests are on average consistently within or at the margin of the

prevailing orographic cloud bank, fluctuations between cloud and cloud-free conditions
would be accompanied by rather large local changes in temperature, relative humidity,
water vapor, and solar irradiance. Support for this interpretation comes from analysis
of the canopy trees in which annual cycles were detected. In 3 out of the 5 forest trees

with annual cycles, variance in annual maxima is greater than the annual minima (Table
B.1). Additional data supporting the importance of elevation in determining controls on
interannual δ 18 O variability in cloud forest trees has been developed by Anchukaitis et al.
[2007a], who identify cycles in two Sapotaceae (MV12 and MV15C) where, contrary to
our premontane wet forest calibration set, the interannual δ 18 O amplitude anomalies are

most strongly controlled by the annual maxima and associated with dry season climate
variability.
Two robust features characterize the annual cycles in the forest trees considered here.
 102

The amplitude of the annual cycle decreases with elevation, as does the ratio of the mean
annual cycle amplitude to the variability in the interannual amplitude. The reduction in
the amplitude of the annual cycle is likely related to longer soil water residence times and
increased mixing of seasonal water sources, which in turn is probably a result of lower

temperatures, increased cloud cover, and reduced solar irradiance at higher elevations
within the orographic cloud bank. The decrease in the ratio of the mean and anomalous
annual cycle amplitude indicates that the higher elevation trees may be more sensitive to
year-to-year variability than trees from the lower premontane forest. There are indications
that this is related in some trees to larger interannual variations in the annual cycle

maxima.
There is an upward trend in the mean oxygen isotope ratio with elevation for all the
trees, when Quercus is excluded. While the trend is not statistically significant, it suggests
that higher elevation trees may on average use more 18 O-enriched cloud water, which

would be consistent with their position progressively closer to the continental divide and
within the region of persistent cloud. Quercus appears to be a special case, with overall
mean δ 18 O values substantially lower than other species, and a smaller average amplitude
in MV05. Collectively, these data suggest that our Quercus samples preferentially sample

soil water with a more negative δ 18 O. This could arise if the trees did not add basal
growth during part of the transition or dry season, when δ 18 O values are higher. If
Quercus ceased growth during part of the dry season, both the mean isotopic value and the
annual cycle, where present, would preferentially reflect the more negative δ 18 O during
the rainy season. Alternatively, this species may have access to some deeper soil water

sources, which would also explain the suppressed amplitude of the mean annual cycle.
Finally, if transpiration were more restricted in Quercus, potentially through particular
characteristics or structure of the leaves, enrichment of the source water in the leaves
 103

would be limited and the resultant cellulose would have a more negative δ 18 O value. In
general, though, there are several reasons some of the cloud forest trees sampled do not
display annual rings, with the most likely cause, however, being that our sampling interval
of 200 µm failed to adequately resolve the annual cycle in very slow growing trees.

B.6.3 Forward modeling

The six environmental parameters from the model of Barbour et al. [2004] are assumed
to be temporally stable [Evans, 2007], and therefore changes in the parameter set are
predominantly reflected in changes in the overall mean of the series, and only secondarily
in the annual maximum and minimum value and in the overall amplitude of the seasonal

cycle. The results from Figure B.8c demonstrate that the interannual difference in the
annual amplitude of the δ 18 O cycle imparted by uncertainties in the model parameter set
may be as large as 1.5h. In agreement with the findings of Evans [2007], however,
we find that the overall structure and mean amplitude of the annual cycle is driven

predominantly by the seasonal change in the δ 18 O related to changes in water sources

as driven primarily by the amount effect and, in this case, the contribution of cloud water
during the dry season. It is clear, however, from interannual differences observed between
model simulations using the two difference relative humidity data sets, that difference in

the input data can have an important influence on interannual patterns of variability. In
general, neither relative humidity dataset produces a substantially improved overall match
to the observed plantation δ 18 O time series, although the simulation using the higher
variance NCEP data has a slightly improved correlation between observed and simulated
interannual amplitude patterns.

More interesting is the disparity between the correlations with each simulation and wet
season anomalies. Modeled δ 18 O values using the NCEP relative humidity dataset show
 104

a weaker correlation with precipitation anomalies than the δ 18 O time series simulated
from the calculated Campbell relative humidity, although the coefficient is more similar
to the relationship between the observed plantation δ 18 O and summer precipitation. This
indicates that increased variability in relative humidity can have a substantial influence

on the controls on the interannual patterns of variability. This, and the observations
of increasing interannual variability relative to the mean annual δ 18 O with increases in
elevation, suggest that samples within or at the boundary of the orographic cloud bank
are likely to show great sensitivity to, and control by, changes in relative humidity and
moisture advection than those below it. This finding can be used to guide future sampling

strategies for the development of long chronologies.

In Evans [2007], the Barbour et al. [2004] model was used to construct the interannual
variability in the amplitude of the seasonal cycle, while the mean and variance were
scaled to that of the observed δ 18 O time series. Here, using the default parameterization

from Evans [2007], even without mean adjustment, and using the relative humidity
derived from the Campbell meteorological data [Pounds et al., 1999], the model actually
reproduces the mean of the series within the precision of the instruments (x̄model =
26.52h, x̄obs = 26.27h), although this may simply be fortuitous, since large shifts in

the overall mean can be the result of changing the model and source water parameters
(see below). The use of the soil water model in the place of variance adjustment also
produced a simulated isotope series with similar mean amplitude and variance as the δ 18 O
from O. tenera, and generally reproduced the visually coherent cyclicity of the actual
time series. Most encouragingly, applying the soil water model reproduces the leading

autocorrelation structure of the observed δ 18 O chronologies, as well as the pattern of

interannual variability and the relationship between the annual amplitude anomalies and
wet season precipitation amount (Figure B.8c).
 105

The monthly mixing ratio of 30:70 (precipitation to prior soil water) implies a mean
water residence time somewhat longer than would be calculated from the turnover rate
reported from lowland tropical rainforest soils derived from tritium measurements by
McGinnis et al. [1969] in Panama. However, McGinnis et al. [1969] estimated soil

water flux for only the top 30cm. Somewhat deeper soil water available to plants would
presumably have a longer residence time. Additionally, reduced evapotranspiration due
to persistent cloud cover, cooler temperatures, and higher relative humidity would likely
increase water residence times in soils in tropical montane cloud forests.
The most obvious discrepancy between the O. tenera composite mean time series, the

simulated model, and local climate variability is the wet season minimum of 1991. There
are several possible reasons for this difference. One hypothesis is that the age model
is incorrect, and that the cycle currently dated to 1991 corresponds to an anomalously
wet summer such as occurred in 1988. However, the 1991 negative anomaly is apparent

in the δ 18 O from the most securely dated cores, those from tree NWT02, and there is
no evidence from basal growth analysis to suggest an age adjustment of several years is
warranted or realistic in this case. An additional, albeit entirely speculative, possibility is
that the eruption of Mount Pinatubo in the summer of 1991 influenced cloud formation

at that time. However, there is no obvious signal of the consequences of the Pinatubo
eruption in the Campbell temperature record for either 1991 or 1992. Zonal winds across
the Costa Rican cordillera were strongly anomalous, however, during the summer wet
season in 1991 [Kanamitsu et al., 2002], but it is not clear whether the local effect at our
lower moist forest calibration site would be to increase cellulose δ 18 O (by increasing the

availability of isotopically-heavier source waters) or decrease it (through increased cloud

cover and reduced evapotranspiration).
 106

B.6.4 Spatial patterns and future work

There are clear patterns in both annual mean δ 18 O cycle amplitudes and their interannual
variability related to elevation. Annual variability, as measured by the standard deviation

of the annual amplitudes, is lower with increasing elevation. However, particularly

when expressed as a ratio with respect to the mean annual cycle amplitude, there are
indications that higher elevation trees show a greater sensitivity to year-to-year variability,
particularly to dry season climate likely related to the influence of orographic clouds.
This suggests that optimal sampling locations for reconstructing both cloudiness and the

regional-scale forcing associated with changes in cloud coverage and moisture advection
will be at those elevations where the annual cycle is coherent and of a sufficient magnitude
to be differentiated from short term fluctuations, but where high-frequency variance
suggests sensitivity to these changes [Anchukaitis et al., 2007a]. Additional complicating

factors may arise from the heterogeneous nature of cloud cover [Clark et al., 2000; Haber,
2000]. Gaps in the cordillera can allow orographic clouds to pass to lower elevations on
the leeward side, often along stream courses. Likewise, topography may dictate the extent
to which individual trees are exposed to direct tradewind moisture advection, and soil type

and depth may influence the amount of source water buffering and soil water mixing that
determines the baseline amplitude for the formation of annual δ 18 O cycles.
Our selection of tropical cloud forests as a site for tropical paleoclimate reconstruction
parallels, in a somewhat paradoxical manner, the approach of classical dendroclimatology

in seeking out sampling locations where trees are likely to be sensitive to relatively
small changes in annual climate. In temperate regions and for trees with regular annual
changes in morphology or wood density, these are typically dry or cold sites at the
limits of a species range. For tropical isotope dendroclimatology, however, we seek sites
which are wet enough to allow trees to grow throughout the year, yet are subjected to
 107

seasonal changes in the stable oxygen isotope composition of available moisture which
provides the means of establishing annual chronology. In cloud forest environments, the
largest interannual changes of interest are likely to be wet season rainfall and dry season
cloudiness. As such, site selection for tropical isotope dendroclimatology is very different

from that of classical field approaches in dendrochronology in terms of climate regimes,

but the guiding principle is the same.
Our approach to tropical isotope dendroclimatology [Evans and Schrag, 2004] has a
closer methodological and procedural affinity to paleoclimate analysis using speleothems
or corals than to classical dendrochronology. Despite rapid advances [Kornexl et al.,

1999; Brendel et al., 2000], the analytical requirements for sample preparation and mass
spectrometry still limit the number of samples and measurement replication that can be
realistically achieved. The result is relatively large uncertainties (several years) in the age
modeled chronologies. However, as demonstrated by McCarroll and Pawellek [1998] and

Gagen et al. [2004], stable isotope ratios in tree-ring chronologies often have a higher
signal-to-noise ratio that rings width data. As a consequence, fewer chronologies are
required to achieve a robust common signal. For the Monteverde O. tenera chronology, 4
cores over the common period of overlap are sufficient to exceed an Expressed Population

Signal threshold [EPS; Wigley T. M. L., 1984] of 0.85. Furthermore, the protocols allow
for the development of high-resolution terrestrial proxy records that bypasses some of the
extant challenges to developing ring width chronologies in tropics, and can be applied
even when appropriate species for traditional dendrochronology cannot be identified.
Continued application of these techniques will necessarily be guided by the specific

research question and the ability of complementary proxies to provide the information
necessary to address them. Cloud forests, given their biogeographical and hydrological
importance at the regional scale, and their sensitivity to broad-scale mode of climate at
 108

the global-scale [Loope and Giambelluca, 1998; Still et al., 1999; Pounds et al., 1999;
Foster, 2001; Bush, 2002; Pounds et al., 2006] represent a particular ecosystem where the
application of tropical isotope dendroclimatology can support an improved understanding
of critical environmental processes across a range of spatiotemporal scales.

B.7 Summary and Conclusions

The results from our plantation study clearly identify an annual isotope cycle in trees
growing at lower cloud forest elevations that can be used to develop an annual chronology.
Interannual variability in the amplitude of the annual cycle is most closely related to wet

season precipitation anomalies at our premontane wet forest calibration site. Our forward
model simulations successfully reproduce the annual pattern of δ 18 O observed in our
plantation trees, and using the soil water mixing model demonstrate a similar leading
autocorrelation structure in both synthetic and actual isotope time series. The model also
reproduces quite well the dependence of the interannual patterns of δ 18 O amplitude on the

amount of summer wet season rainfall, although there is at least one notable discrepancy
observed in 1991. Using two different relative humidity datasets does result in a change in
the mean and interannual variance, indicating that the forward model is partially sensitive
to the selection of input data in reproducing year-to-year variability.

Analysis of canopy trees presents a more complicated picture, however. Five out of
nine mature canopy trees considered here from sites ranging from 1500 to 1660m show
an annual isotope cycle, although only two of these have δ 18 O cycles that can be detected
over their entire length [Anchukaitis et al., 2007a]. Four trees showed no annual cycles at

all. These pilot results demonstrate the potential importance of species, site, and sampling
resolution, but can help guide the future development of long climate reconstructions from
older cloud forest trees. Collectively, the results of our calibration study at Monteverde
 109

demonstrate that annual stable oxygen isotope cycles in tropical cloud forest trees can be
used for both chronology development and the detection of climate variability that can
then be applied to the development of climate reconstructions and the interpretation of
recent trends in tropical montane forest hydroclimatology.

B.8 Acknowledgments

We are grateful for comments and suggestions from Julio Betancourt, Malcolm Hughes,
Mary Gagen, Alan Pounds, Julie Cole, Jonathan Overpeck, and Tim Shanahan. Greg
Eischeid, John Buchanan, Mau-Chuang Foo, Brianna Muhlenkamp, Frank Joyce, Rex

Adams, Arturo Cruz, Eladio Cruz, Koky Porras, and Lisa Wade provided excellent
laboratory and field assistance and support. We appreciate technical instrumentation
support and advice from Bruno Lavettre. Our thanks to the Organization for Tropical
Studies (OTS) for assistance with permits and to the Tropical Science Center (CCT) for
access to the Monteverde Cloud Forest Reserve (Rafael Bolanos and Carlos Hernandez).

The Trostles and Hoges families generously allowed us access to the Ocotea plantations
on their property at Monteverde. This research was supported by a graduate training
fellowship from the NSF IGERT Program (DGE-0221594) (to KJA), a Graduate Research
Environmental Fellowship (to KJA) from the US Department of Energy, grants from the

National Science Foundation: NSF/ATM-0349356 (CAREER to MNE) and NSF/ATM-

0321348 (MRI), and a pilot grant from the Biogeography Specialty Group of the
Association of American Geographers (AAG) to KJA.

B.9 References

Anchukaitis, K. J., M. N. Evans, and T. Lange (2007a), Dry season climate variability in
a tropical montane cloud forest reconstructed from trees without rings, in preparation.
 110

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 118

APPENDIX C

DRY SEASON CLIMATE VARIABILITY IN A TROPICAL MONTANE CLOUD

FOREST RECONSTRUCTED FROM TREES WITHOUT RINGS

Manuscript for submission to Global Change Biology

 119

Dry season climate variability in a tropical montane cloud forest

reconstructed from trees without rings

Kevin J. Anchukaitis1,2, Michael N. Evans1,2 , and Todd Lange3

1. Laboratory of Tree-Ring Research, The University of Arizona, Tucson, AZ

2. Department of Geosciences, The University of Arizona, Tucson, AZ

3. Accelerator Mass Spectrometry Laboratory, The University of Arizona, Tucson,

AZ

C.1 Abstract

We use stable oxygen isotopes to investigate the evidence for trends in anomalously
dry winters at the Monteverde Cloud Forest as preserved in the wood of tropical
trees. Reconstructions of past climate are necessary to the interpretation of these recent

climatic changes in the context of interannual and multidecadal variability in the ocean-
atmosphere system. High-resolution isotope measurements along the growth radius reveal
coherent annual cycles in δ 18 O that are used to establish annual chronological control in
the absence of growth rings. The annual maxima and amplitude of these cycles reflects

interannual variability in dry season orographic cloud cover, moisture advection, and
temperature. The minima reflects summer precipitation anomalies. Positive dry season
δ 18 O anomalies are associated with El Niño events and weaker northeasterly trade winds.
Lower frequency variability may be related to multidecadal climate trends in the tropical

Atlantic. According to our oxygen isotope chronology, the extinction of the Monteverde
Golden Toad after 1987 occurred during one of the driest periods in the last 50 years, and
potentially in the last century.
 120

C.2 Introduction

In Central America, northeasterly moisture-laden tropical trade winds blow across the
isthmus, encountering and ascending mountain ranges and resulting in the formation of
orographic cloud banks. Higher relative humidity, increased moisture, persistent cloud

cover, and reduced temperatures support montane forests rich in endemic organisms and
important for local hydrology [Brown and Kappelle, 2001; Bruijnzeel, 2001]. Tropical
montane cloud forests are ecosystems intrinsically linked to a relatively narrow range
of geographic and meteorological conditions, and are therefore particularly sensitive to

relatively small changes in precipitation or temperature [Loope and Giambelluca, 1998;

Foster, 2001].
The population crash and apparent extinction of the Monteverde Golden Toad (Bufo
periglenes) in Costa Rica in 1987 as well as other observed changes in neotropical

cloud forest ecology are believed to have been a consequence of global warming
[Pounds et al., 1999, 2006]. Indeed, general circulation model (GCM) simulations
of climate under doubled CO2 conditions predict higher lifting condensation levels
and reduced cloud contact for tropical montane cloud forests as a result of increasing
tropospheric temperatures [Still et al., 1999]. Increases in global surface and sea

surface temperatures (SST) may therefore already be fundamentally altering the suite of
climatic and biophysical conditions that maintain cloud forest environments in Central
America [Pounds et al., 2006]. Analysis of the limited available observational data
since the late 1970s suggests that a trend toward decreasing cloud cover at Monteverde

reflects the influence of increasing tropical air and sea surface temperatures [Pounds
et al., 1999, 2006]. However, imprinted on the long-term instrumental drying trend at
Monteverde, El Niño events also cause local increases in temperature and reductions in
cloud cover [Pounds and Crump, 1994]. The strong warm El Niño-Southern Oscillation
 121

(ENSO) event in 1986-1987 immediately preceded the rapid disappearance of the

Monteverde Golden Toad [Crump, 2000], suggesting that temperature anomalies may
have contributed to the species’ demise.
Limited long-term instrumental climate records from the tropics, and particularly

montane regions, impede an improved understanding of past, present, and future climate
variability and trends and their relationship with montane forest hydroclimatology and
biogeography [Bradley et al., 2006]. Without the context provided by long-term climate
records, it is difficult to confidently conclude whether the observed changes in neotropical
cloud forest ecology are the result of anthropogenic climate forcing [Pounds et al., 2006],

land-surface feedbacks [Lawton et al., 2001; Nair et al., 2003], or the interaction of an
introduced pathogen and natural variability in tropical climate [Pounds and Crump, 1994;
Crump, 2000]. The observed changes in climate in tropical montane cloud forests and
their hypothesized biogeographic consequences need to be placed in the context of low-

frequency climate variability.

Here, we seek to develop annually-resolved proxy records of interannual variability
and trends in dry season climate from the tropical montane cloud forest at Montverde
in Costa Rica using stable isotope dendroclimatology [Evans and Schrag, 2004]. The

advantage of our approach to tropical dendrochronology is that it does not rely on the
formation nor width variations of annual growth rings, which in tropical trees can be
absent or may not reflect local climate variability. Annual oxygen isotope cycles have
been previously identified in lowland rainforests [Evans and Schrag, 2004; Poussart
et al., 2004; Verheyden et al., 2004]. At Monteverde, Anchukaitis et al. [2007b]

have demonstrated that the seasonal cycle between the summer wet season and cloud-
dominated winter dry season is sufficient to induce an annual δ 18 O cycle along the radial
xylem growth of cloud forest trees, and that the record from higher elevation cloud forest
 122

trees is sensitive to interannual changes in dry season climate.

The goals of our current study are necessarily two-fold. First, we seek to establish the
basis for interpreting the δ 18 O ratio of the radial growth of long-lived tropical canopy trees
as a high-resolution proxy of cloud forest hydroclimatology. The presence of annual δ 18 O

cycles has been established in plantations [Anchukaitis et al., 2007b], but not in mature
canopy trees, nor in this species. We can then apply this record toward an understanding
of the broad-scale ocean-atmosphere phenomena associated with these local climatic
conditions at Monteverde.

C.3 Methods and Materials

C.3.1 Site description

The Monteverde Cloud Forest (10.2 o N, 85.35 o W, 1500m) sits astride the continental
divide in the Cordillera de Tilaran in northwestern Costa Rica. Above 1500m

elevation, vegetation is classified montane wet cloud forest [Haber, 2000] and its unique
hydroclimatology is characterized by persistent immersion in the orographic cloud bank
formed as the northeasterly trade winds moving across the warm waters of the Caribbean
are forced to rise over the cordillera. During boreal summer (May to October), cloud

forests receive most of their rainfall as the northward movement of the Intertropical
Convergence Zone (ITCZ) brings strong convective storms to the region. During the
dry season (February through April), moisture advection and cloud water deposition are
an important component of cloud forest hydrology [Clark et al., 2000], particularly on

the leeward Pacific slope which would otherwise experience the effects of a pronounced
dry season.
 123

C.3.2 Heuristic model

The origin of annual cycles in the stable oxygen isotope composition of the wood of
tropical cloud forest trees is the differential δ 18 O composition of dry and wet season

moisture sources (Figure C.1) [Evans and Schrag, 2004; Anchukaitis et al., 2007b]. This
hypothesis is not specific to any particular species. Annual and interannual oxygen
isotope ratios in tropical meteoric waters are primarily controlled by the ‘amount effect’,
the inverse relationship between the amount of precipitation and its δ 18 O value [Gat,
1996]. The amount effect is evident in surface waters from Costa Rica and Panama

[Lachniet and Patterson, 2002] and in seasonal precipitation from Monteverde [Rhodes
et al., 2006; Anchukaitis et al., 2007b]. In montane forests, moisture inputs from clouds
are an additional source of water for trees, particularly in the dry season. Cloud water has
an enriched isotopic signature similar to tropical dry season rainfall [Feild and Dawson,

1998; Ingraham and Matthews, 1990; Ingraham, 1998; Rhodes et al., 2006]. Dry season
sources of moisture provide trees with sufficient water to avoid water stress and potentially
the need for a seasonal growth hiatus, but have a distinct isotopic signature.
The δ 18 O of cellulose reflects the original oxygen isotope composition of the source

water (precipitation or cloud water), the isotopic enrichment via transpiration in the
leaves, and the fractionation associated with the synthesis of α-cellulose from sucrose
[Roden et al., 2000; Barbour et al., 2004; McCarroll and Loader, 2004]. Assuming the
latter is constant, cellulose in cloud forest trees should therefore reflect both the amount

of rainfall received and climatic influences on the rate of evapotranspiration from the
leaf. On annual time scales in our model (Figure C.1), cellulose δ 18 O is controlled by the
seasonal change in δ 18 O of source water as determined by the amount effect and the trees’
use of 18 O-enriched cloud water. On interannual time scales, departures from the mean
annual cycle are likely to result from anomalous rainfall and changes in temperature,
 124

Warmer Temperatures
Reduced Cloud/Precipitation
Reduced Moisture Advection
OXYGEN ISOTOPE (δ18O) RATIO

DRY SEASON
Fog, Reduced WET SEASON
Precipitation Precipitation
Reduced Relative Higher Relative
Humidity Humidity

Higher Rainfall
Higher %RH
Cooler Tempertures

ANNUAL ISOTOPE CYCLE IN WOOD

Figure C.1: Conceptual model of climatic controls on the annual and interannual patterns
of stable oxygen isotope ratios in the α-cellulose of cloud forest tree radial growth. The
annual cycle is generated primarily by the seasonal change in the δ 18 O of rainfall, the
use of cloud water by trees in the dry season, and the isotopic enrichment of source
water during the dry season. Interannual variability in annual maximum values is related
to temperature, relative humidity, cloudiness, and moisture advection. At Monteverde,
temperature is strongly related to Pacific sea surface temperatures and tracks tropical
surface air temperatures [Pounds and Crump, 1994; Pounds et al., 2006]. Anomalies
in the annual minima are likely related to wet season rainfall amounts and temperature.
 125

relative humidity, and evapotranspiration. We expect that these will be related to changes
in the intensity of moisture advection over the continental divide into the Pacific slope
forests and to the amount of cloud cover. Interannual changes in the annual cycles can
therefore be interpreted as changes in overall cloudiness and moisture inputs during the

winter, and at Monteverde specifically during the February to April dry season.
In tropical cloud forests, cloud water inputs can vary from 6 to 154% of precipitation
[Bruijnzeel, 2001]. Clark et al. [1998] estimated that moisture inputs from cloud and
mist accounted for perhaps 22% of the annual hydrological budget of Monteverde. More
recently, Schmid [2004] however, estimated that fog deposition was only 4% - 12% of

rainfall during a dry period in the winter and spring of 2003, although for individual days
cloud water could still account for 100% of short-term measured precipitation throughfall.
Thus, while orographic clouds are critical determinants of tropical montane cloud forest
ecosystems, large magnitude changes in cloud immersion can occur rapidly as well as for

extended periods of time, even during the dry season when they are the critical element
of cloud forest hydroclimatology.

C.3.3 Sampling and stable isotope analysis

Increment cores (5mm diameter) from two individual mature Sapotaceae (Pouteria) were

collected in 2004 and 2006 at 1560m (MV12A) and 1580 (MV15C). MV12A was struck
by lightning and subsequently felled in 2002. MV15C was living when cored, with a full
canopy of leaves and abundant latex sap. The cores were subsampled in the laboratory at
200 µm increments using a rotary microtome. The raw wood samples were chemically
processed to α-cellulose using the Brendel Method [Brendel et al., 2000; Anchukaitis

et al., 2007a] as modified for high-resolution sampling [Evans and Schrag, 2004]. 300
to 350 µg of α-cellulose were wrapped in silver capsules and converted online to CO
 126

in a Costech High Temperature Generator/Elemental Combustion System (HTG/ECS).

The HTG uses a radio frequency induction heater with a thin molybdenum crucible
susceptor packed with graphite for pyrolysis at >1500o C. Measurement precision on
a total of 320 Sigma Alpha Cellulose (SAC) solid standards was 0.28h. Data was

corrected based on the mean value of the solid standard material and monitoring gas
values before and after each sample measurement. Data quality assurance was assessed
based on sample peak voltages, peak shape, background stability, and repeated monitoring
CO gas measurements between and during sample analysis.

C.3.4 Radiocarbon

In order to provide independent chronological control for our stable oxygen isotope series,
three samples from each core were selected for radiocarbon analysis to support age model
development (Table C.1). Samples were selected in order to provide high-resolution dates
using the 14 C signature of atmospheric atomic weapons testing [Hua et al., 1999]. A

total of four samples were whole wood and were preprocessed using the acid-base-acid
(ABA) method developed for tropical trees by Westbrook et al. [2006]. Two samples
had been previously processed to α-cellulose using the Brendel Method [Brendel et al.,
2000]. A replicate sample of one of our ABA treated samples was intentionally selected

and prepared using the Brendel technique, in order to provide a confirmation of the
radiocarbon-bias correction for 114 C measurements on α-cellulose extracted using the
Brendel Method [Anchukaitis et al., 2007a]. Extracted wood and cellulose samples were
combusted and the purified carbon dioxide reduced to an iron carbide powder over hot
zinc for 114 C measurement by tandem accelerator mass spectrometer.
 127

C.4 Results

C.4.1 Oxygen isotope chronology

δ 18 O time series from both cores show coherent annual oxygen isotope cycles that can be

used to construct the annual chronology. MV12A (Figure C.2) has distinct oxygen isotope
cycles with a mean amplitude of 2.7h. MV15C (Figure C.3) has cycles with a mean
amplitude of 2.1h. We developed an age model for both series by assigning the maxima
of each discrete cycle to the month of April of the corresponding year and interpolating

the δ 18 O time series between these tie points to monthly resolution. Mean growth rates
derived from these age models are 5.l6 mm year−1 for MV12A and 2.9 mm year−1 for
MV15C, although growth rates vary considerably between the inner and outer portion of
the latter core. These are within the range of median incremental basal diameter growth

observed for tropical trees from La Selva Biological Station in Costa Rica [Lieberman
et al., 1985, 0.34 to 13.14 mm year−1 ], and consistent with radial growth rates from long-
term monitoring and isotopic analysis of of Ocotea tenera in the premontane wet forest
near the Monteverde Cloud Forest reserve (0 to 10 mm/year; Wheelwright and Logan
[2004]; Anchukaitis et al. [2007b]).

The δ 18 O time series from MV15C shows an abrupt change in growth rates at
approximately 121mm depth in the core. Accompanying this change is a substantial
increase in the amplitude of the oxygen isotope cycles. The rapid growth rates which
correspond with the inner portion of the core result in an increase in the number of

individual samples per annual cycle. The higher sampling rate may translate into larger
amplitude δ 18 O cycles simply by virtue of more precisely resolving the monthly and
seasonal changes in water use in the tree. Because this could potentially bias our climatic
interpretation of the oxygen isotope time series, for the purpose of climate analysis we
 128

MV12A (Sapotaceae)
32

δ O %o VSMOW
a).
30
28
26
18

24
300 200 100 0
Depth (mm)

32
δ O %o VSMOW

b).

DRIER WETTER
30
28
26
18

24
1950 1960 1970 1980 1990 2000
Year
Isotope Climatology
32
δ O %o VSMOW

c). ENSO
Non ENSO
30 La Nina

28
18

26
2 4 6 8 10 12
Month

Figure C.2: Isotope chronology from MV12. (a) δ 18 O as a function of depth from the
bark showing the annual isotope cycle. (b) Age modeled δ 18 O series. (c) Composite mean
annual δ 18 O isotope cycle for warm ENSO (El Niño; winter/dry season 1958; 1969; 1977;
1983; 1987; 1992; 1993; 1998), non-ENSO, and cold ENSO (La Niña) years. Warm
ENSO events show a higher amplitude than neutral or La Niña years, slightly beyond the
1σ confidence range.
 129

interpolated the sampling resolution to 6-10 samples per cycle to simulate that of the outer
portion of the core (red line, Figure C.3a-b). Despite this correction, however, the inner
part of MV15C has larger amplitude cycles than much of the outer core, and therefore we
interpret these years cautiously, as they may still contain a growth rate sampling bias.

Radiocarbon measurements (Table C.1) on samples from MV12A confirmed our age
modeled δ 18 O time series, which spans the period 1949 to 2002. We estimate that
age model error for this core is ± 2 years, based on the range of realistic age models
as constrained by the 2 σ resolution of the post-1955 radiocarbon dates [Hua et al.,
1999; Reimer et al., 2004a; Hua and Barbetti, 2004]. However, despite our assessment

during field sampling, the radiocarbon measurements on three samples from MV15C
revealed that the outer portion of the core does not correspond to its sampling date in
2006, suggesting that the tree experienced strongly suppressed basal growth in the recent
decades, or the tree had ceased to add secondary xylem around part of its circumference.

Because they occur during a period of large fluctuations in atmospheric radiocarbon, the
114 C measurements for MV15C calibrate to a large range of potential calendar dates
between A. D. 1700 and A.D. 1950 [Reimer et al., 2004b]. The magnitude of this
uncertainty is also substantially larger than uncertainties associated with the correction

applied to the Brendel-processed samples, and also exists for the sample from MV15C
prepared with ABA.
The actual calendar dates that could be realistically associated with the radiocarbon
assays on MV15C can be better constrained using Bayesian probability estimates
and ‘wigglematching’ between the radiocarbon calibration curve and the sample

measurements [Bronk Ramsey and van der Plicht, 2001; Galimberti et al., 2004;
Robertson et al., 2004]. The likely dates are restricted foremost by the growth orientation
of the tree – samples from near the bark must be more recently formed than those near
 130

MV15C (Sapotaceae)
32

δ18O %o VSMOW
a).
30

28

26

24
200 150 100 50 0
Depth (mm)

32
δ18O %o VSMOW

b).
30

28

26

24
1900 1920 1940 1960 1980
Year
Isotope Climatology
29
δ18O %o VSMOW

c).
28

27

26
2 4 6 8 10 12
Month

Figure C.3: Isotope chronology from MV15. (a) δ 18 O as a function of depth from the
bark showing the annual isotope cycle. (b) Age modeled δ 18 O series. In these two panels,
the red line shows the isotope chronology that results from statistically resampled the
earlier period of rapid growth to better reflect a measurement resolution similar to the
outer portion of the core . (c) Composite mean annual δ 18 O isotope climatology.
 Sample AA74375 AA74376 AA74377 AA74381 AA74379 AA74378

Core MV12A MV15C

Depth (mm) 167 230 242 57 69 121
Initial Data
Fraction modern carbon (F14C) 1.5111 1.0842 0.992 0.9859 0.9882 0.9833
2 σ calibrated date (A.D.) 1969-1972 1955-1957 1951-1956 1526 ( 2.7) 1556 1646 (21.0) 1710 1669 (41.1) 1780
1632 (77.1) 1894 1716 (57.5) 1890 1798 (39.0) 1893
1905 (15.6) 1954 1909 (16.9) 1954 1906 (15.2) 1944

Bayesian Calibration

Posteriori 2 σ range A.D. (% probability) 1724 (67.5) 1814 1716 (67.5) 1807 1694 (67.5) 1784
1834 (9.3) 1888 1828 (9.3) 1881 1804 (9.3) 1858
1926 (18.7) 1954 1918 (18.7) 1948 1896 (18.7) 1924

% Agreement 119.8 113.8 94.4

Sequence Model Agreement (threshold) 115.6 (40.8)

Table C.1: Radiocarbon analysis from MV12A and MV15C. Calibrated dates are shown with the percentage of the probability
density function associated with each range of years. Post-bomb calibrated dates include any additional uncertainty associated
with using from different atmospheric 114 C curves [Reimer et al., 2004a; Hua and Barbetti, 2004].Bayesian analysis for
development of a posteriori probability functions was performed using OxCal, applying the known temporal (stratigraphic)
order of the dates and the estimated annual increment based on the δ 18 O chronology [Bronk Ramsey, 1995; Bronk Ramsey and
van der Plicht, 2001]. Agreement scores are uniformly above 60% for the individual dates and exceeds the lower minimum
threshold (∼ 40%) for the model as a whole [Bronk Ramsey, 1995]. F14C values have been corrected for those samples prepared

131
with the Brendel method as described by Anchukaitis et al. [2007a] and discussed in the text.
 132

the center of tree – and secondly using the number of identified annual cycles between
each 114 C measurement in order to determine the most probable interval spanned by the
core. Within these temporal, self-consistent constraints, the radiocarbon measurements
and their individual probability density functions are then matched against the calibration

curve and a posteriori probability functions calculated. Applying Bayesian analysis and
these additional restrictions reveals that the core from MV15C most likely grew either
during the middle of the 18th century, or during the early 20th century (Figure C.4). If we
further assume that it is highly unlikely that the tree ceased growth for several hundred
years along part of its intact and apparently healthy circumference, we can cautiously

conclude that the MV15C sequence is anchored within the period 1886 to 1990, with a
rather large residual uncertainty of at least ±30 years associated with the range of possible
radiocarbon dates and age model uncertainty from annual δ 18 O cycle identification.
In addition to this logical constraint, we can attempt to cross-correlate the floating

MV15C sequence with the well-dated MV12A chronology. We used the computer
program COFECHA [Holmes, 1983; Grissino-Mayer, 2001] and the constraints from the
2 σ wiggle-matched 114 C calibration to statistically compare potential periods of overlap
between the two cores. Two likely matches for MV15C were found using both annual

maximum and annual amplitude δ 18 O time series, which results in a tentative age model
for the periods 1905–1972 and 1896–1962. The former of these age ranges has a larger
cross-correlation for monthly (r = 0.44, p < 0.01), minimum (r = 0.65, p < 0.01),
and maximum (r = 0.25, p = 0.18) δ 18 O and is supported by the visual crossdating of
the amplitude of the overlapping monthly and annual δ 18 O segments (Figure C.5), but the

uncertainties associated with the radiocarbon dates, the validity of our logical constraints,
and the pattern matching are sufficiently large that the composite chronology should be
considered tentative and its interpretation placed in the context of the substantial age
 133

1400
1500

MVR-4 (121mm)
MVR-7 (57mm)

MVR-5 (69mm)
1600
Calendar Date (AD)
1700
1800
1900
2000

Figure C.4: Probability density functions from radiocarbon assays on wood and
cellulose from MV15C. Markov Chain Monte Carlo wiggle-matching applied to the three
dates, constrained constrained by stratigraphic sequence and spacing between samples.
Assuming the logical constraints discussed in the text are correct, we estimate the absolute
age model error at ± 6 years, but the full chronology is only weakly anchored to calendar
years.
 134

model uncertainties (Figure C.9, Table C.1).

In traditional dendrochronology, the proxy measurement of interest is a characteristic
associated with the annual ring (width, maximum latewood density) and corresponding to
a discrete year. Our isotopic approach to tropical dendroclimatology, however, requires

that we first develop a continuous series of measurements in order to adequately resolve

the annual cycle. From these cycles, we then need to extract information that can be
statistically and mechanistically related to annual climate anomalies (Figure C.1). We
do so by defining 5 characteristics of each annual cycle. The first 3 are the simple
maximum, minimum, and mean value for each annual cycle, which correspond to the

dry season, wet season, and average annual climate conditions, respectively (see Figure
C.1). Our continuous δ 18 O time series have a high degree of low order autocorrelation and
therefore closely sampled prior values may influence the isotope ratio of the successive
samples which follow it. For example, an anomalously dry wet season would result in a

heavier δ 18 O value, with the consequence that even if the dry season that followed was
statistically average in terms of rainfall and temperature, we would expect the maximum
value for that year to be elevated relative to others simply as a result of the autocorrelation
between measurements and the already enriched δ 18 O value which occurred in the prior

increment. This persistence arises from the mixing of discrete precipitation events in the
soil and as a result of short-term carbohydrate storage in the tree. The former phenomenon
has been reproduced by forward modeling of monthly δ 18 O values in tropical trees from
Monteverde [Anchukaitis et al., 2007b]. They show that the high degree of temporal
autocorrelation observed in the actual isotope time series is best reproduced by including

a simple soil water model that results in the mixing of precipitation events over several
consecutive months. We apply two simple normalizations in order to compensate for
the degree of autocorrelation in the continuous δ 18 O series. We calculate a ‘dry season
 135

32
a).
30
δ18O %o VSMOW

28

26

24

22
1900 1910 1920 1930 1940 1950 1960 1970 1980 1990 2000
Year

28 32
b). c).
27.5
31
δ O %o VSMOW

δ18O %o VSMOW
27

26.5 30

26 29
18

25.5
28
25

24.5 27
1900 1920 1940 1960 1980 2000 1900 1920 1940 1960 1980 2000
Year Year

Figure C.5: Composite oxygen isotope chronology from MV12A (black line) and
MV15C (blue line; red line is variance-adjusted), based on radiocarbon dates and cross-
correlation analysis with the logical constraints discussed in the text. (a) Overlap the
potential tie points for the two age modeled monthly chronologies. Triangles show
radiocarbon dates and 2 sigma ranges for MV15C (bottom) and MV12A (top). The
horizontal bar below the period 1962 to 1972 shows the potential range of overlap
from cross-correlation analysis with COFECHA. (b) Overlap and pattern matching for
composite δ 18 O annual chronology and (c) annual maximum chronologies. Correlation
for period of overlap is rmonthl y = 0.44 ( p < 0.01) for monthly δ 18 O, rminimum = 0.65
( p < 0.01), and rmaximum = 0.25 ( p < 0.18). See text for details.
 136

amplitude’, which normalizes the maxima value of a given year with the value of the
preceding wet season, and an ‘annual amplitude’, which is the difference between the
maximum and minimum value of a given annual cycle and has the effect of normalizing
the wet season δ 18 O value. In nearly all cases, however, climate signal detection is not

sensitive to the chosen annual metric (see below).

C.4.2 Climate analysis

Both MV12A and MV15A have interannual difference between wet-to-dry season δ 18 O
variability (σ ) of 1.08h, which is approximately 50% of their mean annual cycle
amplitude. Most of the interannual variability in the chronologies come from variance

in the annual maxima (MV12A = 0.99h, MV15C = 0.83h), suggesting that dry season
variations largely determines the interannual anomalies recorded in these trees. MV12A
shows a distinct change in the amplitude of the annual cycle after 1970, which is related
to a change in both the wet season minimum and the dry season amplitude, suggesting

a reduction in summer rainfall and an increase in dry season moisture prior to the early
1970s, if the cause was climatic.
Wet season δ 18 O anomalies in MV12A are negatively associated with anomalous
summer precipitation (δ 18 O annual mean: r = 0.46, R 2 = 0.22, p < 0.01, d. f = 22;

1977 to 2000; δ 18 O annual amplitude: r = 0.536, R 2 = 0.29, p < 0.01, d. f = 22),

consistent with our heuristic model (Figure C.1) and with earlier findings [Anchukaitis
et al., 2007b]. Dry season amplitude anomalies in MV12A show a clear relationship
to interannual climate variability in sea surface temperature anomalies (SSTA). ENSO
years in the δ 18 O chronology show a dry season isotopic enrichment (mean amplitude

= 3.8h) above the mean of both neutral and La Niña years (1σ ; Figure C.2c). The
local meteorological expression at Monteverde of strong ENSO events is a prolonged
 137

period of elevated temperatures, particularly in 1982-1983, 1986-1988, and 1997-1998.

These years correspond to discernable positive anomalies in the monthly δ 18 O chronology
(Figure C.2a), further consistent with our heuristic model (Figure C.1). In addition to dry
season anomalies associated with strong El Niño events in 1983, 1987, and 1998, positive

δ 18 O anomalies are seen in ∼1960, 1969, 1993, and 1995. These anomalous peaks in
dry season δ 18 O correspond within age model error to years of significant positive ENSO
and Western Hemisphere Warm Pool (WHWP; Wang and Enfield [2001]) temperature
anomalies and spatial extent [Wang and Enfield, 2003]. 1993 and 1995 are also winters
associated with a high index value of the North Atlantic Oscillation (NAO) [Wang, 2002].

In addition to evidence of the influence of local temperature and regional SSTA,

NCEP/NCAR Reanalysis II data from the grid cell and geopotential height corresponding
to the Monteverde Cloud Forest [Kanamitsu et al., 2002] reveal that a weakening of the
northeast trade winds consistently accompanies the δ 18 O anomalies. Years with increased

annual oxygen isotope cycle amplitude and anomalous maximum δ 18 O values are related
to dry season (February to April, 1979 to 2002) westerly zonal wind anomalies (Figure
C.6). Because the slackening of the northeasterly tradewinds at Monteverde occur during
both ENSO and WHWP warm events, the relationship between the dry season amplitude

and zonal wind anomalies accounts for the largest portion of the total variance in δ 18 O
amplitude of any single local climate variable (δ 18 O dry season amplitude/maximum:
r = 0.47, R 2 = 0.23, p = 0.01, d. f. = 22; δ 18 O annual mean r = 0.53, R 2 = 0.29,
p = 0.01, d. f. = 18) (Figure C.6b). Winter (December through February) winds have
a slightly stronger relationship with the δ 18 O peak anomalies for El Niño events in 1983,

1987, and 1998, while winter-spring dry season (February through April) wind anomalies
better account for peaks in 1993 and 1995. The regression statistics with δ 18 O dry season
amplitude improve somewhat (r = 0.53, R 2 = 0.29, p = 0.01, d. f. = 18) for boreal
 138

winter (DJF) winds, but the increased skill is artificial and is leveraged by the anomalies
associated with the three strong ENSO events of the period from 1979 to 2002.
Annual δ 18 O cycle amplitude variability is related to sea surface temperature
anomalies (SSTA) in both the eastern Pacific and the western Atlantic (Figure C.7).

From the early 1970s to the end of the chronology in 2002, the amplitude of the δ 18 O
cycle is positively correlated (r = 0.55, R 2 = 0.30, p < 0.01, d. f. = 28) with an
index of SSTA in the region of the WHWP (50 to 110o W, 7 to 27o N; Wang and Enfield
2001; Kaplan et al. 1998), which includes portions of the eastern equatorial Pacific,
Caribbean, and tropical Atlantic (Figure C.7). Prior to that period (1949-1970), and

concurrent with a period of reduced amplitude in the δ 18 O annual isotope cycles, there
is a weaker relationship with equatorial Pacific SSTA, and an enhanced association with
tropical Atlantic SSTA (Figure C.7). A lagged cross-correlation analysis failed to find
an improved match between either an index of the WHWP or NINO3.4 [Kaplan et al.,

1998], however, even when the MV12A chronology is incrementally adjusted up to 10

years backward or forward, a shift which is more than twice the age model error. The
annual wet-dry amplitude time series from MV12 shows statistically significant spectral
peaks [Mann and Lees, 1996] in the high frequency ENSO band (∼ 3 years) as well

as a trend component (>40 years). The MTM spectrum from MV15C alone shows no
significant peaks.
Correlation between the annual δ 18 O chronology from Monteverde and the dry season
850mb (∼1500m) zonal wind field from NCEP Reanalysis [Kanamitsu et al., 2002]
shows coherent regions of positive correlation (increases in δ 18 O are therefore associated

with weaker tradewinds) over the WHWP and the central and eastern Pacific (Figure C.8).
Correlations of the opposite sign exist through the tropical equatorial Atlantic stretching
from Brazil to west Africa. Unlike the SST field, the pattern of correlations between
 139

3 7
a).
6

Dry Season Amplitude

2

Zonal Wind Anomaly

5
1 4

0 3

2
-1
1

-2 0
1980 1985 1990 1995 2000
Year

6
b).
5
Wet-Dry δ18O %o

0
-2 -1 0 1 2 3
Zonal Wind Anomaly

Figure C.6: Zonal wind anomalies and annual wet-dry season δ 18 O amplitude from
MV12A. (a) Temporal covariability of NCEP Reanalysis 850mb zonal wind anomaly
[Kanamitsu et al., 2002] corresponding to the Monteverde Cloud Forest and the annual
oxygen isotope anomaly. Positive wind anomalies (weaker tradewinds) are associated
with enriched δ 18 O values and drier conditions. Several peaks (1983, 1987, and
1998) correspond to ENSO events, whereas other (1993, 1995) are not. El Niño
tradewind anomalies occur primarily during winter (December-February) while non-
ENSO anomalies occur in spring (February-May). The 1993 wind anomaly occurs in the
dry season to wet season transition in May, and is therefore obscured by the winter dry
season mean shown here. (b) Spring wind anomalies account for 23% of the variance in
the δ 18 O series (p = 0.03). Because they strongly covary, largely with the occurrence
of warm ENSO events, temperature and relative humidity display similar correlation
patterns.
 140

Figure C.7: Field correlations between Monteverde dry season δ 18 O amplitude and dry
season (February to April) sea surface temperature anomalies from Kaplan et al. [1998].
Correlations with mean and maximum δ 18 O show similar patterns. (a) Correlation field
for 1970 to 2002, a period of high amplitude the δ 18 O chronology from MV12A, related
to eastern equatorial Pacific and tropical Atlantic SSTA. (b) The structure of the early
period (1949 to 1969) δ 18 O-SSTA correlations are associated with the Atlantic SST-zonal
wind tripole pattern [Seager et al., 2000]. (c) Detrended full chronology from MV12A
shows the combined influence of Pacific and Atlantic SSTA. Correlations between the
MV12A δ 18 O chronology and the SST field without detrending have a stronger expression
in the Indian Ocean, similar to (d), the correlation of SSTA with the full composite
(MV12A+MV15C) dry season δ 18 O chronology. Significance levels for 1970 to 2002
are 0.34 ( p < 0.05) and 0.27 ( p < 0.05) for 1949 to 2002.
 141

zonal winds and δ 18 O appears to be stable over the period covered by the Reanalysis and
Reanalysis II datasets and our isotope chronology (1979 to 2002; 1948 to 2002; Kalnay
et al. [1996]; Kanamitsu et al. [2002]).

C.5 Discussion

C.5.1 Annual and interannual climate patterns

Annual δ 18 O cycles can be successfully detected in both MV12 and MV15, although the
age model for MV15 is less secure due to the reduced amplitude of the annual cycle, the

slower growth rate of the tree, and the wide range of the calibrated radiocarbon dates on
the cores. Annual maximum δ 18 O and the annual oxygen isotope cycle amplitude show an
upward trend over the period 1949 to 2002 in MV12A, which is caused almost entirely by
the change in the variance of the annual cycle that occurs in the late 1960s. Local summer
precipitation is a consistent influence on the annual amplitude and is similarly associated

with the annual mean δ 18 O value. Interannual variability in dry season amplitude is
associated with positive SSTA in the WHWP and ENSO regions, and directly related
to the strength of the trade winds. Trade wind strength in turn is related to moisture
advection over the continental divide.

The trend in the number of days without rainfall (in runs > 5 days) identified by
Pounds et al. [1999] and Pounds et al. [2006] as being indicative of a reduction in cloud
cover and an elevated orographic cloud base at Monteverde appears to be a result of (1)
the rapid increase in the number of annual dry days which began with the strong 1983 El

Niño and (2) the occurrence of long periods of drought during the 1998 dry season, also
associated with ENSO. Obvious peaks in the number of consecutive dry days also occur
during other El Niños, particularly in 1987 and concurrent with the subsequent extinction
of the Monteverde Golden Toad. Over the period covered by both the instrumental
 142

Figure C.8: Field correlations between Monteverde dry season δ 18 O amplitude and dry
season (February to April) zonal wind anomalies from Kalnay et al. [NCEP1: 1996]
and Kanamitsu et al. [NCEP2: 2002]. The correlations are nearly identical if mean or
maximum δ 18 O is used. Patterns are broadly similar irrespective of the time period and
the generation of NCEP data. (a) The period 1979 to 2002 covered by the chronology
from MV12A and Reanalysis 2 [Kanamitsu et al., 2002]. (b) The full period (1948 to
2002) over which MV12A and NCEP Reanalysis [Kalnay et al., 1996] overlap. (c) The
earlier period of the MV12A δ 18 O chronology, during which time Atlantic SSTA are
the most important remote forcing of dry season hydroclimate at Monteverde. (d) The
composite δ 18 O chronology with the overlapping NCEP Reanalysis data (1948 to 2002).
Significance levels for 1979 to 2002 are 0.39 ( p < 0.05) and 0.27 ( p < 0.05) for 1949 to
2002.).
 143

weather observations from Monteverde (1977 to 2000) and our δ 18 O chronology, there
is no discernable long-term upward trend in annual oxygen isotope values, nor the overall
annual maxima (Figure C.5a,c). Instead, the isotope chronology is dominated by the
positive δ 18 O anomalies associated with ENSO years. Large amplitude δ 18 O cycles in

1983, 1987, and 1998 are associated with an increased number of consecutive dry days
and positive local temperature anomalies and correspond to El Niño events. In these years,
periods of dry season drought (consecutive dry days) increase and there are temperature
anomalies of a degree Celsius or more [Pounds et al., 1999]. On the other hand, two dry
season peaks in the isotope chronology in 1993 and 1995 are not obviously anomalous

in either the local temperature observations nor dry-day calculations. They do however
correspond to late spring weaker zonal winds, and are years of warmer than average
temperatures in the Atlantic warm pool [Wang and Enfield, 2003] and years of high
index values for the NAO [Wang, 2002]. This follows from our finding that the climate

phenomenon which is most consistently related with dry winters at Monteverde is the
slackening of the northeasterly trade winds and a reduction in moisture inputs across the
continental divide.
The largest positive δ 18 O anomalies are also those which correspond to both warm

ENSO and large Western Hemisphere Warm Pool SSTAs in 1983, 1986-1987 and 1998
[Wang and Enfield, 2001, 2003], as well as other notable events within age model error
at ∼1958, 1969, 1993, and 1995. Indeed the spatial pattern of δ 18 O-SSTA correlation
over the period from at least the late 1960s to the present suggests the dominant SST
forcing on local conditions at Monteverde is related to same-sign anomalies in the WHWP

spanning the Central American Isthmus and into the eastern equatorial Pacific cold
tongue. However, the weakening in the correlation between WHWP temperatures and
dry season conditions prior to that period, which does not appear to be the result of age
 144

MV12A

δ18O %o VSMOW
6
MV15C
4

2
a).
6 0

δ18O %o VSMOW
4

b).
0 0

δ18O %o VSMOW
2

c).
6
SST Anomaly (C)

0.5 d).

-0.5

e). 16 Period (yr)

12
8
4
1900 1920 1940 1960 1980 2000
Year

0.5 2.5 4.5 6.5 8.5

Figure C.9: Composite oxygen isotope chronology from MV12A and MV15C. (a)
Dry season amplitude overlap based on radiocarbon dates and crosscorrelation pattern
matching between the two chronologies (see also Figure C.5). (b) Composite detrended
dry season δ 18 O amplitude (normalized dry season). Smoothed red line is the leading
reconstructed component from Singular Spectrum Analysis (SSA; M = 21) associated
with the low-frequency variability and trend (c) Detrended composite annual δ 18 O
amplitude (normalized wet season) series and the leading reconstructed component from
SSA, which has a periodicity of >40 years. (d) Atlantic Multidecadal Oscillation Index
derived from Kaplan et al. [1998]. A warm period in the Atlantic between 1940 and
1970 occurs during a cold period in the Pacific, and with a period of decreased amplitude
in the composite and MV12 δ 18 O chronologies from Monteverde. (e) Wavelet analysis
[Torrence and Compo, 1998] of tropical Pacific SSTA (5o S- 5o N, 110o W- 170o W)
[Kaplan et al., 1998]. Time-frequency plot is developed using a Morlet wavelet (ω0 = 6).
Diagonal black lines show the time-frequency region where zero-padding influences the
power spectrum.
 145

model error, and the change in the spatial structure of the correlation pattern, may suggest
a change in the relationship between the ocean-atmosphere conditions forcing dry season
climate at Monteverde. Even more interestingly, the correlation between the δ 18 O time
series and the concurrent zonal wind field retains its spatial pattern and magnitude (Figure

C.8), suggesting again that the most stable and proximal control on dry season moisture
advection and cloudiness at Monteverde is the strength of the northeasterly trade winds.
The change in the late 1960s and early 1970s in the amplitude of the annual δ 18 O
cycles cycles in MV12A and the resulting upward trend is caused by an increase in the
mean annual minimum values of the annual cycles as well as a reduction in the magnitude

of the dry season δ 18 O enrichment. The pairing of the increase in the annual minimum
with a smaller difference between wet season δ 18 O and the subsequent dry season annual
minimum in midcentury leads to a rising trend in the annual amplitude (Figure C.9a) in
the time series from MV12A and in the composite chronology over the period from the

1930s and 1940s to the present that is caused primarily by the relatively sharp change
between 1960 and 1970. According to our heuristic model (Figure C.1), increases in the
minimum annual δ 18 O and a reduction in the annual amplitude is related to an overall
drier wet season and a wetter (or cloudier) dry season.

C.5.2 Climate and ecological change at Monteverde

Most of the long-term trend over the last several decades as detected in our isotopic record
from Pouteria is due to a change in the amplitude of the annual δ 18 O cycles that occurs
around 1970. Variability at the interannual scale dominates the dry season signal during
the period of increased ENSO variance since the mid 1970s, with 1983, 1987, and 1998

as the three driest winters in the last three decades. Spectral analysis confirms statistically
significant canonical ENSO band power. The period from 1986 to 1988, associated with
 146

the strong El Niño event, appears to be the driest in at least the last 50 years, with reduced
cloudiness and moisture inputs during the two dry seasons bracketing an abnormally dry
wet season. If the absolute age model of MV15C is correct, this period is among the
driest in the last century. This observation from the isotope record is confirmed by the

recollections of a field biologist working in Monteverde at the time [Crump, 2000]. This
suggests that the proximal cause of the well-documented extinction of the Monteverde
Golden Toad [Pounds and Crump, 1994; Pounds et al., 1999] was the combination of
the abnormally strong ENSO-forced dryness and the lethality of the introduced fungus
Batrachochytrium dendrobatidis which causes chytridiomycosis, and eventually death, in

amphibians [Crump, 2000; Lips et al., 2003, 2006]. It is possible that the fungus may
have already been present in Monteverde prior to 1986 [c.f Puschendorf et al., 2006] and
that physical stresses or behavioral changes associated with the anomalous dry conditions
during the 1987 ENSO event created the conditions for a nonlinear interactions between

environment, disease vectors, and interannual climate [Pounds and Crump, 1994; Bosch
et al., 2007].
Nair et al. [2003] and Lawton et al. [2001] used a regional atmospheric model to
demonstrate that deforestation in the tropical lowland upwind from the Monteverde Cloud

Forest could potentially result in increased cloud base height. Pounds et al. [2006] have
subsequently argued that the major period of deforestation in the San Carlos Plains in
lowland Atlantic Costa Rica occurred too early and would not in any case have forced
sufficient changes in cloud formation to account for the change in dry season moisture
inferred from the daily meteorological records during the 1980s and 1990s. Our δ 18 O

chronology is not sufficiently anchored in time to detect potential changes in dry season
climate associated with earlier periods of deforestation prior to the ∼1950s, and the
clearest evidence of climate variability is at interannual time scales associated with
 147

ENSO. However, we interpret the isotope evidence from MV12A to indicate that the most
important shift in climate at Monteverde in the last several decades was primarily related
to the change in annual amplitude that occurred in the late 1960s and early 1970s. The
timing of this change suggests two possible causes. As discussed further below, the rapid

shift in δ 18 O amplitude occurs at approximately the same time as a shift from warm to
cold conditions in the tropical Atlantic and just slightly before a change from cold to warm
conditions in the Pacific. Alternatively, from the 1960s through the 1970s, deforestation
rates in Costa Rica peaked [Rosero-Bixby and Palloni, 1998], and could have contributed
to mesoscale alterations of boundary layer interactions sufficient to influence downwind

montane cloud formation and base heights, as suggested by the simulation modeling by
Nair et al. [2003].
The relative suddenness of the change is therefore suggestive of either a large-scale,
substantial shift in climate or an ecological change (either human-mediated or natural) in

the local or regional environment. With our current δ 18 O chronology, we cannot rule out
either cause, nor can we eliminate the possibility that land use or ecological changes in the
immediate vicinity of the tree that could have changed its local hydrological conditions.
Isotope chronology replication and temporal extension will be necessary to better resolve

questions of natural and anthropogenic climate change and variability at Monteverde.

C.5.3 Uncertainty and potential biases

While high resolution oxygen isotope measurements allow us to resolve the annual cycle
of water use by cloud forest trees, and therefore to establish chronological control even
in the absence of tree-rings, there are a number of limitations and uncertainties in the

record that could bias the climatic interpretation from the δ 18 O time series. Some of the
largest uncertainties could be associated with potential errors in the age model. In the
 148

case of MV12A, the use of high-precision, post-bomb radiocarbon measurements helps

to constrain the chronology and confirmed the age model established using the annual
δ 18 O cycles. For MV15C, the radiocarbon dates alerted us to the apparent growth hiatus
and helped us delimit the most probable time span covered by the isotope time series

from the core. However, the limitation of 114 C calibration over the recent historical
period prevents us from achieving a high-precision chronology from the radiocarbon
measurements alone, despite the application of Bayesian statistics.
Our analysis of both local climate variability and its association with broad-scale
forcing at Monteverde is necessarily tentative. Our understanding of the physical

(climate, hydrology) and biological controls, across a range of spatial and temporal
scales, on the eventual δ 18 O of wood in cloud forest trees is still rudimentary. Moreover,
analytical requirements thus far limit the degree of replication and development of
multiple overlapping proxy chronologies that is de rigueur in traditional approaches

to dendrochronology, although tree-ring isotope series typically have a higher signal-

to-noise ratio than ring width series [McCarroll and Pawellek, 1998; Gagen et al.,
2004]. As a consequence, we must assume that age model error exists and could be
substantial enough to degrade our high-frequency climatic analysis. There are also

potential individual tree-specific responses that are present in our isotope chronology
that may not represent either local or global climate and which potentially bias our
interpretation of temporal trends in our record. For example, because of the short period
of the hypothesized overlap between MV12A and MV15C, it is not possible to definitively
establish the extent to which the lower amplitude annual δ 18 O cycles in the outer portion

of MV15 also represents a reduced climate seasonality at that time, or whether it is the
result of the relative topographic position, hydrology, or microsite conditions. Similar
uncertainties apply to the interpretation of low frequency variability in δ 18 O amplitude
 149

within MV12A. Analysis of additional samples which span this period can aid in more
distinguishing climatological from ecological or biological causality.
Interestingly, our two Pouteria cores show quite different behavior approaching the
center of each tree. MV12A has a reduction in the amplitude of the annual cycle, which

is not apparently related to a reduction in growth rate nor a sampling bias. MV15C shows
the opposite behavior, with an increase in annual δ 18 O amplitude and a much higher
growth rate. In the application of stable carbon isotopes to dendroclimatology, a ‘juvenile
effect’ is often observed, which is characterized by more depleted δ 13 C values caused
by young subcanopy trees using already depleted CO2 resulting from organic respiration

[McCarroll and Loader, 2004]. Treydte et al. [2006] identified what they believed to be
a juvenile trend in δ 18 O in montane forests in Pakistan characterized by enriched oxygen
isotope ratios. At Monteverde, and in tropical trees in general, we hypothesize that the
juvenile trend might be manifest as an increased annual δ 18 O amplitude, as younger

trees would not have the fully developed root system of mature canopy trees, and would
therefore rely on shallow soil water with short residence times and minimal mixing of
distinct meteoric waters. This could be the cause of the large amplitude cycles in the
inner most portion of MV15C, and might be reflected in the magnitude of the annual cycle

in the Ocotea tenera analyzed by Anchukaitis et al. [2007b]. The reduced cycles in the
deepest part of the core from MV12A, however, appear to be related to an environmental
or climatic cause, and are not consistent with a juvenile effect.
The change from low to high amplitude annual cycles which occurs in the 1960s
might have several causes, and we cannot rule out a biological or ecological influences

unrelated to climate or analytical or sampling bias. However, the change in amplitude

is separately observed in both cores, before the composite mean chronology is created.
Furthermore, in MBV12A, the reduction in amplitude in the early part of the chronology
 150

is not consistent with a hypothetical juvenile effect. These lines of evidence, in addition to
the observed relationships at interannual time scales between climate and δ 18 O, suggest a
potential climatic cause.

C.5.4 Multidecadal dynamics

The timing of the change from low to high amplitude δ 18 O cycles in the late 1960s
coincides with the change from a warm to cold phase of the the Atlantic Multidecadal
Oscillation [AMO Enfield et al., 2001; Dong et al., 2006], a decadal-scale oscillation
toward an increase in interannual band (ENSO) variance in the tropical Pacific [Gu
and Philander, 1995; Wang and Wang, 1996; Torrence and Compo, 1998; Torrence and

Webster, 1999; Dong et al., 2006], and a change in the seasonality of ENSO [Mitchell and
Wallace, 1996] (Figure C.9d,e). When the full composite δ 18 O chronology is considered,
a second inflection point marking the change from higher to lower amplitude annual
isotope cycles in the earliest part of the 20th century is detectable around ∼1930, which

is coincident within age model errors with prior mode shifts in the Pacific and Atlantic.
These observations are not inconsistent with an interpretation of the midcentury changes
in δ 18 O amplitude as reflecting the influence of multidecadal changes in the broad-scale
forcing in the Atlantic and Pacific affecting the local climate conditions at Monteverde.

Are these temporal patterns in the δ 18 O chronology consistent with the climatic
consequences of a change from a warm (cold) Atlantic (Pacific) to a cold (warm)
Atlantic (Pacific)? First, we expect that dry season moisture in the period from 1949
to 1970 in MV12A (or, extended to ∼1930 to 1970 if the composite chronology is
considered) should be increased relative to the more recent period, since the dry season

δ 18 O amplitude is reduced. An examination of the differences in February to April 850mb

winds (not shown) reveals stronger northeasterly trade winds over Costa Rica and the
 151

central and eastern Pacific in the earlier period compared to the later, consistent with our
heuristic model and our analysis of the interannual controls on dry season δ 18 O. This
might also be consistent with that observation that at the interannual scale, a cold Pacific
was associated with greater moisture advection via the northeasterly trade winds. When

forced by warmer tropical Atlantic SSTs, coarse-resolution fast GCM (FOAM) simulates
an increase in precipitation over southern Central America and northern South America
at the peak of the dry season (February through April) [Wu et al., 2007], which would
also be expected to suppress maximum annual δ 18 O values in trees at Monteverde.
However, evidence for a climatic cause for the increased δ 18 O minimum wet season

values during the middle part of the century is equivocal. The two climate stations from
the Pacific side of northwestern Costa Rica (GHCN: Monteverde and Puntarenas) that
span the period of the transition do in fact show an increase in precipitation between 1960
and 1970 [Fleming, 1986; Peterson and Vose, 1997]. The Monteverde climate station

(which is different than the Campbell station, at lower elevation and not in the cloud
forest), shows an increase in mean summer precipitation of approximately 200 mm over
the period 1965, which would translate into a change in δ 18 O of up to ∼1.2h, which
is slightly larger than is observed in MV12A and in the composite chronology. Gridded

precipitation data [Mitchell and Jones, 2005], however, show a slight increase in late
summer precipitation, the opposite of what the isotopes and limited station data suggest.
While a decrease in late wet season (September-November) precipitation on the Pacific
side of Central America is simulated for warm periods in the tropical North Atlantic
using the FOAM climate model [Wu et al., 2007], other general circulation model (GCM)

simulations predict a strong increase in summer precipitation over all of Central America
and Caribbean in response to a warming of the Atlantic associated with the positive phase
of the AMO [Sutton and Hodson, 2005]. Different models and different data lead to
 152

different interpretations of midcentury climate in western Costa Rica.

The correlation between the δ 18 O chronology from MV12A and the SSTA field
(Figure C.7) for the period 1949 to 1969 resembles the tripole zonal banding observed in
SST and wind fields that are part of the coupled spatial expression of the North Atlantic

Oscillation (NAO) [Seager et al., 2000] with SSTA. The warm phase of the AMO is
related to patterns similar to the negative phase of the NAO [Grosfeld et al., 2007]. The
correlation structure between Monteverde winter dry season climate and Atlantic sea
surface temperatures therefore suggests that during the midcentury period coinciding with
an observed warm phase of the AMO and the reduction in the interannual SST variability

in the eastern tropical Pacific, it was Atlantic Ocean variability that determined the year-
to-year changes in winter cloudiness. We hypothesize then that positive (negative) NAO
anomalies would result in the weakening (strengthening) of the trade wind flow across
Central America and a decrease (increase) of moisture advection to the leeward side

of the Monteverde Cloud Forest and a positive (negative) δ 18 O anomaly in the isotope
chronology. Warm subtropical waters in the Atlantic lead to weaker trade winds, which
would be reflected in enriched δ 18 O for that year in the Monteverde isotope record.
Following the shift from warm to cold AMO in the 1960s and the increase in equatorial

Pacific SSTA variance, interannual variability at Monteverde predominantly reflects

ENSO and the tropical Pacific. Mechanistically, at least, this interpretation would not be
inconsistent with our model (Figure C.1), our interpretation of the interannual controls on
cloud forest tree δ 18 O, and observed and modeled climate dynamics. However, potential
patterns in the chronology not related to climatic causes prevent us from proving these

linkages with any great certainty.

Whether variability in the Pacific drives concomitant changes in the Atlantic, or vice
versa, remains an open question. The common view has been that ENSO influences the
 153

Atlantic via atmospheric teleconnections [c.f. Latif , 2001]. Wang and Enfield [2001]
have shown that warm ENSO anomalies influence the Western Hemisphere Warm Pool
via the connection of the anomalous Walker circulation to changes in the strength of
the Atlantic subtropical high. Similarly, Giannini et al. [2001] hypothesized that ENSO

influenced tropical Atlantic variability through the stabilization of the tropical atmosphere
via the propagation of the warming signal through the troposphere. More recent model
results focusing on the role of the Atlantic Ocean suggest that changes in the Meridional
Overturning Circulation (MOC) in the North Atlantic [Knight et al., 2005] can influence
the amplitude of interannual and interdecadal variability in the Pacific. Dong et al. [2006]

used a coupled ocean-atmosphere GCM with an imposed Atlantic SSTA forcing to mimic
the AMO. In these simulations, easterly surface wind anomalies over the central and
western Pacific forced by latent heat flux in the tropical North Atlantic deepened the
western Pacific themocline, stabilizing the ocean-atmosphere coupled instability in the

tropical Pacific and reducing ENSO variance. The relative directional influence of one
ocean basin on the other may in fact be time scale dependent. These and numerous other
studies point to extant uncertainty regarding the forcing mechanisms for the observed
association between Atlantic and Pacific variability at a variety of time scales.

C.5.5 Future applications

Costa Rica, and Central America in general, sit at the very crossroads of the complex
and still unresolved interaction between the Atlantic and Pacific Ocean. Our results
here suggest that neotropical cloud forests might be sensitive to changes in the ocean-
atmosphere interactions across the isthmus and could therefore be optimal locations to

develop high-resolution paleoclimate proxies. Climate reconstructions from this region

could provide the necessary long-term context for improving our understanding of the
 154

feedbacks and coupling between the two oceans at interannual, decadal, and perhaps
multidecadal time scales. The distribution of montane cloud forests along the American
cordillera from Mexico through northern Argentina provides climate-sensitive locations
at which a network of paleoclimate proxy reconstruction could be developed, in order

to understand local and interannual changes in cloud forest hydroclimatology, as well as

potential interactions between the tropical Atlantic and the Pacific on multidecadal time
scales.

C.6 Conclusions

We have used a high-resolution, annually-resolved δ 18 O record from tropical trees

without rings from Monteverde to reconstruct temporal change in local climate during
the last century and identity the relationships between cloud forest hydroclimatology and
large-scale modes of ocean-atmophere variability. One sample (MV12A) is securely
dated using annual cycles in δ 18 O and high-precision radiocarbon measurements. A

second floating chronology (MV15), relatively dated using Bayesian analysis of multiple
radiocarbon dates, likely overlaps with the latter half of our securely dated δ 18 O
chronology, and can be used to tentatively extend our record of climate variability at
Monteverde through the 20th century. The results of our analysis suggest the following

conclusions:

• Annual δ 18 O cycles in the radial growth of Pouteria (Sapotaceae) at Monteverde

can be used to develop chronological control in trees without identifiable annual
growth bands.

• Local meteorological data suggest that interannual variability in dry season δ 18 O

variability is positive related to temperature and negatively related to moisture

 155

advection via trade wind strength. As in our previous study [Anchukaitis et al.,
2007b], wet season δ 18 O is related to summer rainfall anomalies.

• El Niño years are associated with anomalously enriched δ 18 O peaks in the stable
isotope chronology. The years of the largest anomalies – 1969, 1983, 1987, 1998,

and possibly ∼ 1958 – are years in which there were both warm ENSO events and
a subsequent positive SSTA anomaly in the Western Hemisphere Warm Pool. The
years associated with the 1986-1987 ENSO event are the driest period in our isotope
chronology since at least 1949, and potentially one of the driest in the last century,

although with considerable uncertainty at the longer timescale. The extinction of

the Monteverde Golden Toad therefore appears to have coincided with one of the
most severe, extended droughts in the last several decades or more.

• A change in the amplitude of the annual δ 18 O cycle occurs between 1960 and 1970.
There are multiple potential biological, ecological, analytical, and climatic factors

that might have contributed to this change. The timing of the change is consistent
with the timing of a switch from warm to cool SSTs in the Atlantic and with a
change toward increased interannual (ENSO) variance in the Pacific. If the cause of
the shift is indeed climatic, this could suggest a relationship between interannual to

multidecadal variability in the δ 18 O chronology of cloud forest trees at Monteverde

and patterns of Atlantic and Pacific ocean-atmosphere variability.

• Neotropical cloud forests, situated as they are at the boundary of two oceans and
influenced by broad-scale modes of climate variability in each, may be promising

sites from which to develop records of past climate in support of research to

better understand Pacific interannual variability and perhaps interbasin climate
interactions and coupling across a range of time scales. The development of high
 156

resolution stable isotope proxies from the mountains of Central America could help
address outstanding uncertainties and provide a validation target for coupled GCM
experiments.

C.7 Acknowledgements

We are grateful for comments and suggestions from Julio Betancourt, Malcolm Hughes,
Alan Pounds, Mary Gagen, Nat Wheelwright, Julie Cole, Jonathan Overpeck and Tim
Shanahan. We benefitted tremendously from excellent laboratory and field assistance
from John Buchanan, Mau-Chuang Foo, Lisa Wade, Frank Joyce, Rex Adams, Jim Burns,

Arturo Cruz, Eladio Cruz, and Koky Porras. Technical instrumentation support and advice
from Bruno Lavettre, David Steinke, and Ben McElhaney. We thank the Organization for
Tropical Studies for help with permits and the Tropical Science Center (CCT) for access
to the Monteverde Cloud Forest Reserve (Rafael Bolanos and Carlos Hernandez). This
research was supported by a graduate training fellowship from the NSF IGERT Program

(DGE-0221594) (to KJA), a Graduate Research Environmental Fellowship (to KJA) from
the US Department of Energy, grants from the National Science Foundation: NSF/ATM-
0349356 (CAREER to MNE) and NSF/ATM-0321348 (MRI), and a pilot grant from the
Biogeography Specialty Group of the Association of American Geographers (AAG) to

KJA.

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