Tropical Fruit

‘Dwarf Cavendish’ is the most common banana cultivar in Pakistan, but plans
are under way to introduce internationally available cultivars such as ‘Grand Naine’
into Pakistan. The use of different substrates for acclimatization of in vitro plantlets
was studied. In vitro plantlets with good shoot-root system were transferred to the
greenhouse onto seven different soil types and their combinations as substrates. These
were river silt, black sand (commonly used in nurseries in Pakistan), clay, desert sand,
hill sand, sugar cane compost, and sugar mill soil residues. Obtained results indicated
that river silt and its combinations with desert sand (1:1 and 2:1, respectively) are
better substrates. The survival percentage on inorganic substrates after one month of
culture was 100%. In addition, acclimatized bananas were growing much better on
the soil mixture of river silt and desert sand at a ratio of 2:1. The recorded vegetative
data of plants grown on these two substrates were: plant length above soil surface 8,
12 cm; number of leaves 4, 4; second leaf length 10, 13 cm; leaf width 5, 7 cm; stem
thickness 0.9, 1.2 cm; number of roots 20, 15; and roots length 18, 29 cm, respectively.
Most likely, the addition of desert sand resulted in better aeration to allow more
vigorous growth of the root system compared to pure silt. Other substrates such as
sugar mill soil residues and sugar cane compost had 0% survival. The addition of sand
did not increase the survival percentage except in the mixture of sugar mill residue
and desert sand at ratio 1:1, where the survival percentage was 80%. Eventually,
using local nutrient-rich soil substrates like river Indus silt could be of additional
benefit on the acclimatization of in vitro cultivated bananas. Acclimatized plants of
‘Grand Naine’ cultivar were successfully transferred to open field and produced an
average of 38 kg/bunch as compared to 12 kg obtained from common ‘Dwarf
Cavendish’ (Basrai). The results of this study will help to reduce costs for in vitro
banana production in Pakistan.

Date palm (Phoenix dactylifera L.) is the third most important fruit crop after citrus and mango in Pakistan. This crop is found in all four provinces of Pakistan on 90,000 ha with a production of around 600,000 mt yr-1. Pakistan’s position is always among the seven largest producers and exporters of dates in the world. Sindh and Balochistan provinces contribute more than 90 % of production and crop area in Pakistan. Date palm has a very strong effect on socioeconomic activities of the area. The top Pakistani cultivars are Aseel, Dhakki, Begum Jangi, Rabai and Muzawati. Pakistani date fruit is being exported as fresh, dry or chohara. This book is published by USAID Pakistan to be used as a manual for date palm growers all over Pakistan describing date palm cultivation in Pakistan, major flood-affected date palms and fruit, protection from monsoon rains, the first commercial trial for date palm micropropagation using inflorescence explants at the Date Palm Research Institute (DPRI) at Khairpur, and major date palm problems generally in Pakistan, including diseases and pests, with emphasis on the Khairpur and Balochistan provinces, and date fruit processing

T
HIS study was carried out during the growing season of 2009 on date palm fruit Hellawi cv. which was pollinated by two pollen parents (Al-Khikri Al-Adi and Al-Ghannami Al-Akhdar) for studying  gene expression  in terms of studying the nature of changes  in protein pattern. The results showed that the process of gene expression had occurred during fruit growth and development as appearance of protein bands which differed in their number, location on the gel and molecular weight during the various stages of fruit development. There was a difference at the appearance timing of the protein bands on the gel of the pollinated fruit by Khikri and Ghannami. Four protein bands were detected in the 13th week of the pollinated fruit by Khikri pollen grains, which preceded fruit entrance into the ripening stage. Whereas, such protein bands appeared in the 15th week of the pollinated fruit by Ghannami pollen grains which coincided with the fruit entrance into the ripening stage (Rutab stage).  The current study showed that the protein pattern could be reliable as a base correlates gene expression of the date palm fruit with their different growth and ripening stages. Therefore, the Metaxenia impact on the fruit quality may be caused by the formation of new proteins that affect the hormone content in the fruit.

Fruit of 85 varieties of Pakistani dates were collected throughout the harvest season.
Appropriate Performa was designed to write the scientific and commonly used properties. These
properties included fruit color at Khalal stage, shape, size (length & diameter), color & height of
perianth, fruit group (soft, semi-dry, dry), edible stage and the micropyle position and split (wide,
narrow, deep). Furthermore, pictures of the whole fruit, longitudinal and transverse sections and
dorsal & ventral sides of seed have been taken. This study aimed to evaluate the quality of different
Pakistani dates. Aseel variety is considered as one of the predominant and commercial varieties in
Pakistan. It is elliptical in shape like most of the Pakistani dates. A few of the evaluated varieties
have good properties. The majority of Pakistani dates fall in the semi-dry group. Most of the
varieties, if not all are sensitive to monsoon rains, which synchronizes with ripening/harvest season.
The harvest season of dates in Khairpur, Pakistan starts from the end of June until the mid of
August. Urgent and rapid demand is being required for the high quality varieties. Whether, this
demand can be achieved by means of micropropagation of offshoots of these evaluated and
selectable varieties by the tissue culture or to import offshoots of best quality dates from abroad for
propagation needs.

Date palms of 10 year old Ajwa, Safawi and Ruthana varieties from Al-Madina, Saudi Arabia were evaluated under the agro-climatic conditions of Khairpur, Sindh, Pakistan throughout the growing seasons from 2009 to 2011. The results obtained indicated better fruit quality similar to those fruits obtained from the original place of origin. The palms of the three varieties were thriving successfully. The edible stage of vars. Ajwa and Safawi is tamer and rutab for var. Ruthana. The fruit was harvested early from 13 to 20th of July before the onset of monsoons. The fruit size of vars. Ajwa, Safawi and Ruthana at their edible stages were 3.16, 4.25 and 3.52 cm long, and 2.31, 2.05 and 2.38 cm in diameter, respectively. The average fruit and seed weight of vars. Ajwa, Safawi and Ruthana were 11.42, 10.49 and 12.42 g, and 1.23, 0.88 and 1.1 g, respectively. The fruit flesh percentage in vars. Ajwa, Safawi and Ruthana reached 89.14%, 90.84% and 90.92%, respectively. It was found that the climatic conditions of Khairpur are suitable for the cultivation of these three exotic varieties. The vegetative, flowering and fruit characteristics of the three varieties were described, and the impact of climatic conditions on fruit quality was discussed in this study.

The date palm (Phoenix dactylifera L.) is considered as one of the most important horticultural crop of the world and
holds a very significant position in the agricultural horizon of district Khairpur, Sindh, Pakistan. The sudden decline
disease of date palm is one of the major problems of the region and has destroyed hundreds of orchards in dispersed
areas of district Khairpur that resulted in huge economic losses. The aim of the current study was to investigate the
incidence of sudden decline disease of date palm and mortality by surveyed seventeen different locations of district
Khairpur and the fungal pathogens associated to the disease were also explored. The sudden decline disease incidence
greatly varied at different locations and ranged from 0.25% to 50.17%. Maximum disease incidence was at Noor pur
followed by Baberloe and Ahmedpur. The minimum disease incidence was recorded at Nizamani followed by Noonari,
Therhi and Shadi shaheed with 1.5%. Six fungi namely, Fusarium solani, Phoma ucladium, Helminthosporium sativum,
Alternaria alternata, Aspergilus niger and Penicillium chrysogenium were isolated from different plant parts collected
from affected date palm trees of various orchards of Khairpur, Pakistan. The study will help further to find out the
control remedies of this devastating disease.

This study aim to use plant growth regulators and PEG to enhance embryo maturation and obtain a vigorous plantlets of Phoenix dactylifera L. cv. Malakaby. Embryonic nodular callus 0.5 g were treated with1 μM ABA or JA with BAP or NAA and PEG at 0 or 0.25%. Addition 1μm ABA into maturation medium was more effective to produce somatic embryo numbers (11.60) compared to 1μm JA (6.11). Addition PEG into maturation medium did not enhance significantly somatic embryo number (10.33 and 7.38 respectively). The embryos length increased by addition ABA into maturation medium compared to JA as well as, addition BA with ABA into maturation medium encouraged embryos length. Osmolarity affected by PEG seemed the main component required for protein synthesis. Furthermore, the plantlets which treated with PEG treatments appeared vigor and healthy compared to the plantlets which untreated with PEG.

Date Palm (Phoenix dactylifera L.) is a salt and drought tolerant fruit crop mainly cultivated in Arabian countries. Shoot
tip explants of offshoots were used traditionally for various micropropagation protocols on research or commercial levels.
However, its main disadvantage was the scarification of the entire plant. Subsequently, this hinders the micropropagation
of male and female recalcitrant individuals with no offshoots or the interesting cultivars with a limited population.
Consequently, current study is investigating the factors affecting commercial production of tissue cultured palms with
cost-effectively and short-production cycle, which may also strongly enhance the transformation protocols. The innovative
way by which the 15 cm long immature inflorescence was excised before emergence between fronds is reported herewith
this study for the first time. Established spikelet explants were able within 2-3 months only without any callus phase
to produce shining globular structures instead of immature florets. These embryogenic structures couldn’t develop
further without maturation process for 1-2 months under full darkness as well. Subsequently, well-matured embryogenic
structures were shifted to the subsequent differentiation medium under illumination conditions. After then, green shoots
and multiple somatic embryos have been subjected to the multiplication stage, then rooting and eventually successfully
transplanted in the greenhouse. All used nutrient media and their sequential usage is reported in this study by which
it became possible from one inflorescence to produce 10000 plantlets in rooting stage right now of Gulistan Pakistani
cultivar without any bad consequences on the mother tree.

The objective of this study was to determine the appropriate growth stage of inflorescence (spathe) explants
among different cultivars of the date palm (Phoenix dactylifera L.) and to define the optimum period of
incubation on high-auxin medium. Inflorescence explants were excised from seven Pakistani cvs. at different
growth stages and cultured on a basal MS medium containing 100 mg l-1 2,4-D for 3, 6 and 24 weeks (Batches
I, II and III) before being transferred to 10 mg l-1 2,4-D medium. The cultures were incubated at 25 ± 2°C in the
dark and transferred onto fresh culture medium every 3 weeks. Results revealed that spikelet explants isolated
from different cultivars with the same length were not necessarily at the same stage of development. Length
varied according to cultivar, time of excision, location of the inflorescence in the crown and location of the
spikelets in the inflorescence. Accordingly, the response differed among the cultivars tested. After 24 weeks
from initial culture, some Batch II explants produced only unfriable callus and embryos. On the other hand,
explants comprising Batches I and III failed to induce organs even after 42 weeks (14 subcultures); instead they
turned blackish brown. Multiple somatic embryos were subjected to proliferation at the multiplication stage.
Individual shoots were rooted and successfully transplanted in the greenhouse with about 90% survival.

The present study was conducted with an aim to analyze the genetic diversity and phylogenetic relationship among twenty five date palm (Phoenix dactylifera L.) cultivars from Pakistan. The leaf samples for DNA isolation were collected from four provinces of Pakistan i.e., Sindh, Punjab, Khyber Pakhtoonkhwa and Balochistan. DNA was isolated by modified CTAB method. The study was carried out by using six universal RAPD primers. The RAPD primers showed polymorphism among all date palm cultivars. The bands obtained were successfully used to differentiate the genotypes. Based on the pair-wise comparison of amplification products, the genetic relationship was estimated. All date palm cultivars showed variation at the DNA level. The average of genetic diversity among the date palm cultivars ranged 79.4%. A dendrogram was constructed using NTSYSpc programme, on the basis of this analysis, the populations were clustered into two main clusters and sub-clusters. Therefore, the high level of polymorphism was detected and its reproducibility suggests that RAPD markers are powerful and reliable tool for genetic diversity analysis of date palm cultivars.

مو�شم جمع الثمار يبداأ من �شهر يونيو ويوليو شبتمبر � في منطقة خيربور ويمتد لأغ�شط�ض في وليتي البنجاب وبلوت�ش�شتان. حيث ترتفع درج��ات الح��رارة ب�شدة في و�شط باك�شتان في  50منطقة «�شكر» و «خيربور» لحدود تزيد عن درجة مئوية في منت�شف ف�شل ال�شيف. �شقوط الأمطار المو�شمية في ف�شل ال�شيف خا�شة في شبب خ�شائر � أا�شهر يونيو، يوليو، اأغ�شط�ض جمة لمزارعي النخيل في ه��ذه المناطق بلغت % من المح�شول خا�شة 100في بع�ض ال�شنوات انه ياأتي في فترة ن�شج الثمار علي الأ�شجار. يتفاوت ال�شرر من عام لعام آاخر تبعا لموعد ومكان �شقوط الأمطار وعدد �شاعات �شقوط المطر (كمية المطر). في منطقة خيربور محل الدرا�شة عند �شقوط المطر في بداية �شهر يونيو ل�شاعة اأو �شاعتين فقط فاإنه ل يوؤثر اإل علي  ”Gajarالأ�شناف مبكرة الن�شج مثل �شنف « وه��ي من الأ�شناف ”Kasho-wari“و�شنف الن�شف جافة التي ل يمكن أاكلها في مرحلة الخال طازجة لرتفاع المادة التانينية بالثمار. وتتمثل الأع��را���ض ال�شلبية في ت�شقق الق�شرة الخارجية للثمرة كما حدث بالمو�شم الحالي حيث ت�شققت  ”أاوطاقن“ لثمار �شنف 2010 الق�شرة الخ��ارج��ي��ة للثمار وه��ي في مرحلة

The improved in vitro protocol of monoembryonic mango would facilitate the commercial micropropagation of cultivars and rootstocks. The nucellus tissue of mono-embryonic seeds of world-known mango cultivars 'Saroli', 'Langra' and 'Chaunsa' were used to induce the direct shoots. The simple procedure used for surface sterilization of the immature fruit resulted in 95% free-contaminants explants. The initial fruit size was found to be crucial for getting the DS within a short period of time (2 months). Full half of seeds from immature fruit without zygotic embryo were used as an initial explant. Browning of initial explants was controlled by anti-oxidants solution, activated charcoal containing-media and incubation in dark till maturation. The developed medium formula to induce cluster of shoots (0.5 cm long) was micro-salts of MS and macro of B5 supplemented with 4.52 μM 2,4-D and 9.83 μM 2iP within 3 months under dark. The appropriate fruit size used to induce the DS (caullogenesis) was 5-6 cm long. The smaller fruit size 1 cm long could not develop and died while the fruit size 2-4 cm long developed clusters of embryogenic callus under dark and 27±2°C. The induced clusters of shoots were cultured on the basal medium supplemented with 2.3 μM Kin and 0.49 μM 2iP in order to grow into small healthy plantlets under light and 27±2°C. Caullogenesis was detected only in 'Langra' and 'Chaunsa' whereas 'Saroli' explants failed throughout the two seasons' experiment. The step-wise protocol and morphology of the induced DS were described in the current study.

The present work aimed at improving pineapple (Ananas comosus (L.) Merrill) cv. Smooth Cayenne production throughout the tissue culture technique. Some factors affect this process were studied. The method of surface sterilization, the adventitious and axillary bud formation versus N6-Benzyladenine (BA) concentration, type of medium (liquid or solidified), rooting and acclimatization were evaluated. Explants have been surface sterilized with sodium hypochlorite (NaOCl) at 0.5 %, then with mercuric chloride (Hg Cl2) at 0.1 % both for 5 minutes, with survival percentage of 95%. All explants were cultured onto MS basal medium supplemented with different concentrations of BA and 1.0 mg l-1 Kinetin (Kin). All cultures were maintained in a growth room at a 16 h photoperiod (40 mmol.m-2 s-1), 27 ± 2°C. After eight weeks, number of initiated shoots was counted and the cultures were transferred for multiplication on the best treatment, 2.0 mg l-1 BA and 1.0 mg l-1 Kin. Therefore, using the liquid medium during multiplication stage speed up the growth of proliferated shoots compared to solidified medium. Utilizing 1.0 – 3.0 mg l-1            α-Naphthaleneacetic acid (NAA) or Indole-3-butyric acid (IBA) enhanced the root formation of individual shoots. Finally, the well rooted plantlets, around 12 cm in length, were transplanted onto the Peat moss as a soil bed under the greenhouse. After 4 weeks, the survival percentage was 100 %. By this improved protocol, more than 10000 pineapple plants were produced from one initial plant in less than 9 months.  Introduction

This experiment was carried out in plastic green house at the farm of Horticulture Research Institute during 2009 and 2010 seasons to evaluate the response of Pineapple (Ananas comosus) (stickm.) Merril to bio-fertilizers. The plants were about two years old selected as to be healthy & nearly uniform in vigour produced from sprouts were planted during March and April of seasons (2008 and 2009). The experiment included ten treatments of three bio-fertilizers (control, three levels of each of Microbien and Phosphorene at(40, 80 and 120g/pot) and three levels of EM(1.2,2.4 and 3.6cm/pot) ). All treatments were supplied with a mixture of Amonium sulphate (20.5%N), superphosphate(16.5% P2 O2 )  and Potassium sulphate (48% K2O) in ratio to 10:5:10. Only 16 grams of this mixture was applied to each plant every two months during the first ten months
The obtained results showed that, the three treatments biofertilizers improved plant growth i.e., plant height, stem thickness, number of leaves, leaf area and dry weight as compared to control (without  treated by bio-fertilizers).
In the two seasons, data indicated that, the highest values of height, number of leaves and lesf area were obtained by plants treated with EM at 3.6 cm/pot then 2.4 cm/pot, while the high level of Microbien treatment (120g/pot) gave the thickness plants. Moreover Phosphorene at 120 and 80 g/pot caused highest dry weight of plants. Concerning the effect of biofertilizers on flowering% and fruit parameters , it is noticed that, the three substances differ in this effect while Phosphorene at low level (40g/pot) gave the highest percentage of flowering in both seasons, it is noticed that, Microbein at first concentration increased the length and diameter of fruit, while the high level gave highest TSS and total sugar%, EM treatments at low level 1.2 cm/pot increased the fruit weight and TSS of fruit whereas, the high level (3.6 cm/pot) gave highest acidity of fruit, this is true for the two seasons of study.
Concerning the chemical analysis of leaves it is revealed that, EM treatment at high and medium level 3.6 and 2.4 cm/pot increased the N% while the highest P% was obtained by treated plants by Phosphorene at high level 120g/pot.