Sporocarp Survey Protocol For Macrofungi
Sporocarp Survey Protocol For Macrofungi
Sporocarp Survey Protocol For Macrofungi
Version 1.0
December 2008
RUSS HOLMES is the Regional Botanist, USDA Forest Service Region 6, Regional
Office, Portland, Oregon.
Cite as: Van Norman, K., J. Lippert, D. Rivers-Pankratz, R. Holmes, and C. Mayrsohn.
2008. Sporocarp Survey Protocol for Macrofungi, version 1.0. Portland, OR.
Interagency Special Status/Sensitive Species Program. U.S. Department of Interior,
Bureau of Land Management, Oregon/Washington and U.S. Department of Agriculture,
Forest Service, Region 6. 16 pp.
Cover photo: Collecting fungal sporocarps, Roseburg District BLM. Photo by Russ Holmes, 1999.
2
Table of Contents
INTRODUCTION ............................................................................................................. 4
PRELIMINARY FIELD PLANNING................................................................................. 5
FIELD SURVEY METHODS ........................................................................................... 6
SURVEY TYPES .......................................................................................................................................................... 6
Complete Surveys ................................................................................................................................................. 6
Intuitive Controlled Surveys ................................................................................................................................. 7
TIMING OF SURVEYS .................................................................................................................................................. 7
Late Summer/Fall ................................................................................................................................................. 7
Spring ................................................................................................................................................................... 8
EQUIPMENT LIST ....................................................................................................................................................... 8
VOUCHER COLLECTION AND DATA MANAGEMENT ............................................... 9
SURVEYOR SKILLS AND SAFETY............................................................................. 11
GLOSSARY .................................................................................................................. 12
BIBLIOGRAPHY ........................................................................................................... 13
APPENDIX 1 - SPECIAL CONSIDERATIONS FOR HYPOGEOUS FUNGI ................ 15
3
Introduction
The vegetative body (thallus) of a fungus usually grows within a substrate such as wood
or soil with filamentous growth known as hyphae individually and mycelium collectively.
A thallus may be present within a substrate at one location for many years, yet the
mycelium cannot be easily detected or identified to species. Instead, surveyors typically
rely upon finding the fruiting bodies (sporocarps) of macrofungi, those fungal species
whose fruit can be seen by the unaided eye, to locate and identify species.
Because species produce sporocarps at different times throughout the year and in
response to specific weather conditions, multi-season and multiple surveys per season
are recommended to increase the probability of detection and the proportion of fungal
diversity sampled. The greater number of survey visits, the less chance of a missed
detection and the greater chance of sampling maximum species richness (Smith et al.,
2002; Lodge et al., 2004).
This document is intended for use by the Region 6 Forest Service (FS) and
Oregon/Washington Bureau of Land Management (BLM) for strategic surveys of
Special Status fungal species. It is not intended for pre-project surveys. This survey
protocol is written to target more than one fungal species, which maximizes field visits
and efficiency. These methods can also be used for site revisits that are occasionally
needed to monitor continued species presence. The protocol was developed by agency
botanists based upon experience, journal articles, consultation with fungal survey
4
contractors and researchers, and other survey protocols and guidelines for macrofungi
(Castellano et al., 1999; O’Dell, 1999). We do recognize that alternative survey
methods may differ depending on project objectives, but offer these guidelines as a
minimum standard.
There are a combined 62 Sensitive and 139 Strategic fungal taxa on the FS and BLM
Special Status lists (USDA Forest Service, 2008; USDI Bureau of Land Management,
2008). Because the lists are updated annually and there are many species, no specific
habitat information is offered in this protocol. A compilation of FS/BLM Sensitive fungal
species biology, ecology, habitat, threats, and management considerations is available
from the Interagency Special Status/Sensitive Species Program website
(http://www.fs.fed.us/r6/sfpnw/issssp/species-index/flora-fungi.shtml).
1. Determine the list of target species for the selected survey area based on species
range and habitat requirements. Gather known information about the habitat and
ecology for each targeted species. Known habitat information for FS and BLM
special status fungi has been documented in the Conservation Assessment for fungi
(Appendices 1 and 2) and in the Habitat Summary for Sensitive Fungal Species
spreadsheet (http://www.fs.fed.us/r6/sfpnw/issssp/planning-
documents/assessments.shtml).
2. Determine if suitable habitat exists in the survey area for the target species using
agency GIS records, aerial photos, USGS maps, predictive habitat models, and
individuals knowledgeable with the survey area.
3. Document known sites of target species on all land ownerships within the vicinity of
the survey area. Agency and Natural Heritage Program occurrence databases are
the primary sources for target species locations though other sources, such as
university herbaria, field guides, and knowledgeable individuals may be useful.
4. Determine the ecological preference of fruiting for target species. Some species fruit
in the fall, spring, or both, and others during summer dry periods, but after
precipitation. The conservation assessment for fungi, field guides, and guidebooks
(Castellano et al. 1999, Castellano et al. 2003) are good references for this
information. Also consider elevation, aspect, and recent weather conditions at the
survey location when determining when to survey. Freezing weather can delay or
curtail fruiting. Sporocarps often quickly disintegrate when exposed to extreme
temperatures. (See Timing of Surveys section below)
5
5. Delineate the areas of probable suitable habitat on topographic maps and aerial
photos to be used during field surveys.
This survey protocol is designed to target multiple species. If there are several target
species within a survey area and they fruit at different times of the year such as fall and
spring, then two survey visits in the fall and two in the spring are required.
Local botanists have discretion to determine the appropriate survey timing depending
on local conditions and objectives. Survey visits three weeks apart generally
accommodate the seasonal variation in weather conditions and variations in the
conditions that stimulate fruiting in different species, thereby increasing the chance of
detection. However, if a weather change is imminent such as the arrival of snow or a
hard freeze, then the interval may be shortened to two weeks. If weather conditions are
not appropriate (e.g., too dry), an interval greater than 3 weeks is acceptable.
Survey Types
Complete Surveys
Complete surveys are recommended when the survey area is less than 2.5 acres.
These surveys complete a 100% visual examination of the survey area (distance
between transect lines are close enough to permit an overlapping site distance between
transect lines). For hypogeous fungi this means looking for suitable habitat (see App. 1)
and sampling within those areas.
6
Intuitive Controlled Surveys
Intuitive controlled surveys are recommended when the survey area is greater than 2.5
acres. These surveys are conducted by traversing through and around the survey area
focusing surveys efforts in areas of high probable habitat. Suitable habitat delineated
on topographic maps during preliminary field planning and observed while traversing the
survey area receive complete (100%) visual examination. It is expected that surveyors
will pass through all represented plant associations and significant changes in slope
position, aspect and substrate, especially when there are multiple target species.
Trained and experienced surveyors looking in terrain that is not excessively steep or
brushy should be able to survey on average 40 acres in a day using this methodology,
and even as much as 60 acres per day, depending on the number of target species
found.
Timing of Surveys
Timing of surveys should follow the guidelines provided below unless information is
available to show that conditions that stimulate fruiting of the target fungi within the
survey area are different. Check existing records and with local mycological experts to
obtain this information. Monitor weather conditions and conduct on-the-ground field
reconnaissance. If conditions are not right, do not survey. Continue to monitor and wait
for appropriate survey conditions.
Late Summer/Fall
In general, late summer and fall surveys begin after significant precipitation has
moistened the substrate. Good fall sporocarp production generally occurs when there
has been at least ½ inch of precipitation at least every 3 weeks from late August
through fall until the first frost and when temperatures have been below 90 degrees F.
Consider too that more precipitation may be required to reach the ground thereby
stimulating fruiting in areas where dense canopies or shrub layers occur such as the
Western Cascades. In general, stop conducting fall surveys when there is persistent
snow or a persistent hard freeze that impacts the understory. Depending on the target
species, freezing conditions may end sporocarp production. However, some species
begin to fruit after the first frost.
7
The following table outlines general minimum conditions and recommendations for
timing epigeous and hypogeous fungal surveys by ecoregion in the fall.
Spring
Late winter and spring survey conditions are more variable by locale than autumn. For
both epigeous and hypogeous fungi, late winter and spring surveys should commence
after freezing conditions have ceased. Surveys may continue into early summer, but be
aware that substrate can dry out quickly. Sampling should occur while the substrate is
still moist.
Surveys may be conducted when 50% or more of the suitable habitat within the target
survey area is free of snow and should be scheduled to locate:
• Fungi that fruit just after snow melt
• Fungi that fruit after snow melt, a warming trend and spring rain
Equipment List
• Aluminum foil, wax paper bags (full sandwich size) or brown paper bags and a
roll of aluminum or wax paper for larger specimens (fungi can be placed inside
and the ends twisted to contain the specimen like a tootsie-roll)
• Trowel or large knife to dig up base of sporocarps
• 4-tine garden cultivator; shorten the handle to 1-meter for a built-in measuring
tool
• Sharp knife or razor blade
• Specimen field tags
• Survey data forms and fungal description forms
• GPS unit
• Camera, preferably digital
• Permanent marking pens
8
• Flagging and permanent tags to mark collection sites
• Basket or bucket to carry collected specimens
Further information about handling and vouchering fungi such as drying guidelines and
collecting spore prints can be found in Castellano et al. (1999; 2003).
To voucher a fungal collection, first clean specimens of debris. Photograph the fresh
specimen, preferably digital and in situ. Write the unique collection number legibly on a
piece of paper and photograph along with the specimen. For truffles, show a cut truffle
in the photo (see appendix for cutting recommendations). Identification of some other
genera such as Ramaria is also aided by cutting the specimen. Fill out the entire Field
Tag being sure to include the tentative species identification. Place the specimen and
completed field tag in wax paper bags or heavyweight foil and write the unique
collection number in permanent marker on the outside of the bag. Never collect in
plastic or air-tight containers as sporocarps quickly degrade. Do not mix collections
when collecting and storing. Regularly clean your collecting materials (stray spores can
hinder identification).
At the location where the potential target species was found, mark the site. Marking
sites with labeled metal or hard plastic tags is preferred. Another more ephemeral
method of marking a site is to flag 2-3 nearby trees and hang 3 flags above site. Write
on one of the flags: collection ID, suspected species, date, and surveyor initials. Keep
in mind that flagging has a tendency to break off or be pulled off by animals so write
near the knot or top of the flag. Often surveyors initially mark a site with flagging and
9
then return to the site to establish a permanent site marker once a target voucher
specimen has been verified by an expert.
The fresh characteristics of specimens such as spore color, exterior traits, and bruising
are critical for correct identification. In the office, record characteristics of fresh
specimens on the appropriate Fungal Description Form before drying. Prior to
shipping for expert verification, specimens should be thoroughly dried, usually within 1
day of collection. Once a specimen is thoroughly dried, it can be transferred into a
recloseable “zip” style plastic bag with the field tag and unique collection number written
in permanent marker on the bag (Fig. 1). Fill out the Fungal Collection Tracking
Spreadsheet for the entire fungal shipment being sent to the Interagency Inventory
Coordinator Assistant.
Figure 1. Photos of bagged and labeled fungal collections with filled-out field tags.
Data collected during field surveys must be recorded and can be done on paper data
forms or electronic data recorders. There are three general categories of information
that are collected during BLM/FS species surveys:
• Survey data (information characterizing the survey)
• Detection data (information characterizing site occurrences)
• Supporting data (supporting information including vouchers and accompanying
forms)
All of the data collected during surveys must be entered into agency databases
(GeoBOB and NRIS TESP) following agency guidance issued for each application. If
target species are not encountered during the field survey, list the target species and
document their absence within the survey area (negative surveys). If a target species
is found, mark the species location on a field map (7.5 minute USGS quad or aerial
10
photo), clearly labelling the species found. UTM coordinates should be taken with a
GPS using the North American Datum of 1983 (NAD83) and document the accuracy of
GPS reading.
For surveys and occurrences of Special Status fungal species located on BLM
administered lands, complete the GeoBOB Flora Survey and the GeoBOB
Observation Forms. For surveys and occurrences located on national forest system
lands, complete the NRIS TES Plants Forms. Both the BLM and Forest Service data
field forms are located at:
http://www.fs.fed.us/r6/sfpnw/issssp/inventories/monitoring.shtml
Figure 2. Checklist of items to send to the ISSSSP Inventory Coordinator Assistant with
vouchered fungal specimens.
11
Follow all agency safety rules and polices for field work. Mushroom hunters often get
lost because they are not paying attention to their location as they wander through the
forest. Documenting the survey route and paying attention to the survey area map and
aerial photos will help surveyors remain aware of their location. Do not eat any
mushrooms unless you are absolutely certain of the identification and their edibility.
Glossary
(from Castellano et al., 1999)
Macrofungi: fungi with sporocarps large enough to be seen without a hand lens
Sequestrate: sporocarps that normally retain their spores within until they decay in
place or are eaten.
12
Bibliography
Brown, M. 2002. Sampling intensity and statistical power in a survey of epigeous
ectomycorrhizal fungi. Unpublished report. On file with Forest Service Regional
Office, P.O. Box 3623, Portland, Oregon 97208.
Castellano, M.A., J.E. Smith, T. O’Dell, E. Cazares, and S. Nugent. 1999. Handbook to
strategy 1 fungal species in the Northwest Forest Plan. Gen. Tech. Report PNW-
GTR-476. USDA Forest Service, Pacific Northwest Research Station, Portland,
Oregon. 195 pp. Accessed online at: http://www.fs.fed.us/pnw/pubs/gtr476.pdf
Castellano, M.A., J.M. Trappe, and D.L. Luoma. 2004. Chapter 10. Sequestrate fungi.
Pp. 197-213. In: Mueller, B.M., Bills, G.F., and Foster, M.S., eds. Biodiversity of
fungi: Inventory and monitoring methods. Elsevier Academic Press, London.
777 pp.
Lodge, D.J., J.F. Ammirati, T.E. O’Dell and G.M.Mueller. 2004. Collecting and
describing macrofungi. In Biodiversity of Fungi, Inventory and Monitoring,
G.M.Mueller, G.F Bills and M.S. Foster, editors. Elsevier Academic Press,
London.
Molina, R. 2008. Protecting rare, little known, old-growth forest-associated fungi in the
Pacific Northwest USA: a case study in fungal conservation. Mycological
Research 112: 613-638.
North, M., J.Trappe and J. Franklin. 1997. Standing crop and animal
consumption of fungal sporocarps in Pacific Northwest forests. Ecology
78(5): 1543-1554.
O’Dell, T.E. 1999. Survey Protocol for Bondarzewia mesenterica (=B. montana),
Otidea leoporina, O. onotica, O. smithii, Polyozellus multiplex, Sarcosoma
mexicana, and Sowerbyella (= Aleuria) rhenana Version 1.3. U.S. Department of
Interior, Bureau of Land Management, Oregon/Washington and U.S. Department
of Agriculture, Forest Service, Region 6. pp 8. Accessed online at:
http://www.blm.gov/or/plans/surveyandmanage/SP/Fungi/PBFungi/m2000-
018.htm
13
Smith, J.E., R. Molina, M.M.P. Huso. D.L. Luoma, D. McKay, M.A. Castellano, T. Lebel,
and Y. Valachovic. 2002. Species richness, abundance, and composition of
hypogeous and epigeous ectomycorrhizal fungal sporocarps in young, rotation-
age, and old-growth stands of Douglas-fir (Pseudotsuga menziesii) in the
Cascade Range of Oregon, U.S.A. Can. J. Bot., 80:186–204.
Trappe, M., F. Evans, and J. Trappe. 2007. Field guide to North American truffles:
hunting, identifying, and enjoying the word’s most prized fungi. Ten Speed
Press, Berkeley, California. 136 pp.
USDA Forest Service. 2008. Transmittal memorandum dated January 31, 2008 to
Forest Supervisors. Update of the Regional Forester’s Sensitive Species Lists
and Transmittal of Strategic Species List. On file with Forest Service Regional
Office, P.O. Box 3623, Portland, Oregon 97208. Accessed online at
http://www.fs.fed.us/r6/sfpnw/issssp/agency-policy
14
Appendix 1 - Special Considerations for Hypogeous Fungi
The key to locating fungal sporocarps is knowing in general what the target species
sporocarps looks like (i.e., size, color), what type of habitat it tends to occur in, and in
particular what type of substrate it grows on. Epigeous fungi (mushrooms) fruit
aboveground and can be located through visual examination of suitable habitat.
Species grow on different substrate such as wood, soil, litter, cones, and dung. In
general more people are familiar with looking for and finding epigeous fungi.
Hypogeous fungi (truffles), on the other hand, fruit belowground. They can potentially
be found almost anywhere there are trees. Truffles fruit throughout the fall, winter,
spring, and summer depending on species and locality. They usually occur at the
interface between the organic litter and the mineral soil, about one to six inches deep,
but can emerge to the surface or be more than a foot deep. Evidence that small
animals have been digging in an area recently is often a good indication that truffles
may be about. For more information see Castellano et al. (1999) and Trappe et al.
(2007).
Sample by raking roughly ½ meter around potential truffle locations using a four tine
rake (mark ½ meter on the rake handle for accurate measurement). Rakes should be
used to gently peel back the litter layer (remember to replace the litter when you are
done and try to leave the area as you found it). Dig into the soil and work through the
tailings (removed soil). Truffles appear like small potatoes, often beige, yellowish, or
reddish brown (Fig. 3; for more photos see http://www.natruffling.org/photo.htm).
Truffles may appear as small clumps with roots extending out or balls of yarn with loose
strands (Castellano, pers. comm.). If you find a truffle, continue to rake in that area. If
not, move to the next probable area.
While sampling, use your sense of smell and touch. Truffles are fragrant. Arcangeliella
smells garlicky or like maple syrup. Leucogaster smells fruity-sweet. Destuntzia rubra
smells like fish, and Nivatogastrium smells like bubble gum. DO NOT EAT any truffle
sample! Use your hands to touch and feel anything that may appear like a truffle.
15
Some species are very small, less than the size of a dime and appear like small clumps
of dirt. Press these with your fingers. If it is a truffle, it will not break apart. Many
truffles are fairly solid, but some are hollow in the center so be gentle.
Other considerations:
• 1-2 weeks after a flush of large, fleshy epigeous fungi (Boletes, Amanitas, and
late Russulas) is a good time to go truffling. If the weather is warm (approx. 70
degree F or warmer), wait 1 week; if cooler (approx. 60 degree F or cooler), wait
2 weeks.
• Mycelia build up, primordia forms and then fruit when conditions are right.
Rotting mushrooms may indicate the end of the fruiting period. Watch weather
patterns.
• Rake across mycelial fans when they are located.
• Rake small humps as you walk - only takes a few seconds.
• The more sites sampled, the higher likelihood of finding hypogeous fungi.
When cutting open a hypogeous specimen, try to cut it from the base to apex (Fig. 3).
This helps to show diagnostic characteristics such a sterile base or columella that may
easily be missed if cut in the wrong plane. Cutting fresh specimens is important
because it will show features such as lactation, bruising of the gleba, or enhancement of
any odor present that could otherwise go undocumented. A photo of both the peridium
and gleba is ideal such as a cut specimen with one half facing ‘gleba-up’ and the other
half ‘gleba-down’.
Figure 3. Truffles can be quite small, obscure, and difficult to differentiate from dirt clods
except by feel and brushing off the dirt. Cut open truffles from the base to the apex and
photograph both the peridium and gleba.
16