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Annona senegalensis Pers. is one of the wild fruit tree for domestication in southern Africa. An assessment of the genetic diversity in A. senegalensis would assist in planning for future germplasm collection, conservation and fruit domestication programmes. During 2004 to 2006 nine populations were collected from different locations in Malawi and...
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... genetic distances were then clustered using unweighted pair-group method with arithmetic averages (UPGMA). Figure 2 shows typical electrophoregrams obtained with multiple sample loading of the four primers that were test- ed on (A) Bunda population, (B) A. cherimoya from which these primers were adopted, (C) Karonga population and (D) Nkhotakota population. Three loci were examined and the pooled analysis of the primers showed that the green primer was the best followed by the blue and then red. ...
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... These results indicated that the phenotypic diversity of the wild hemp population in the Pakistani Himalayas and Potohar Plateau represents three geographical variation patterns (viz., continuous variation, regional variation, and nominal variation). Such contrasts in the phenotypic diversity were also found in different species that occur in various biogeographical locales with different climates [65][66][67][68][69]. Furthermore, the observed hierarchical inconsistency of trait variations may reflect intraspecific genetic variation [70]. ...
Hemp (Cannabis sativa L.) is an annual, and dioecious herb belonging to the Cannabaceae family. This plant is native to Central and Southeast Asia. The wild races of this species are commonly growing in Khyber Pakhtunkhwa and Punjab provinces, as well as in Islamabad, Pakistan. This study provides crucial insights into how environmental variables influence the wild hemp populations, which can be utilized in for conservation and breeding. The present study was aimed at evaluating the effects of key environmental factors such as altitude, geographical location, precipitation, relative humidity, maximum, minimum, and average temperature on 16 morpho-agronomic traits of a wild population of hemp growing in the Potohar Plateau and Lesser Himalayas. Our findings indicated that high relative humidity (> 64%), low average temperature (< 15 °C), intermediate average temperature (19–22 °C), and high average temperature (> 22 °C) played significant roles in determining the distribution pattern of the wild hemp. Correlation analysis demonstrated that average annual temperature contributed a higher percentage of variation in phenotypic diversity than geographic variables. Additionally, cluster analysis indicated three groups for the selected 35 populations. Clustering and Principal Component Analysis (PCA) of the morpho-agronomic traits indicated that group 1 from the Lesser Himalayas showed high relative humidity (> 64%) and low average temperature (< 15 °C). Conversely, Group 2 populations from the Potohar Plateau demonstrated intermediate average temperature (19–22 °C). There is an existence of Group 3 in the Potohar Plateau with a high average temperature (> 22 °C) compared to Group 1 and Group 2. Our examination highlights the complex interplay between ecological factors, and morphological attributes in native landraces of Cannabis sativa, giving significant insight into knowledge for preservation and breeding initiatives. A study of genetic diversity could complement morpho-agronomic traits in future research to learn more about how genetic variation affects environmental adaptation.
... Molecular markers provide information on DNA polymorphism, to study genetic diversity within/between populations. Simple Sequence Repeats (SSR) [Escribano et al., 2004 (Annona cherimola Mill); Kwapata et al., 2007 (Annona senegalensis pers.); Escribano et al., 2008 (Annona cherimola Mill) and Anuragi et al., 2016 (Annona cherimola Mill, Annona reticulata, Annona muricata L., Annona atemoya and Annona squamosa L.) had been used to characterize Annona genotypes. ...
... Microsatellites are currently the most used marker because they are highly polymorphic even among closely related lines, need little DNA, can be automated, are quickly shared between laboratories, and are highly transferrable between populations. According to Kwapata et al. (2007), SSRs frequently carry large numbers of alleles at extremely low frequencies. The most crucial factor in choosing desired Annona genotypes for domestication in the domain of adaptation is the knowledge of nutritional value. ...
... In this study, amplicon size ranged from 152 bp to 497 bp which is similar to results obtained in A. cherimola using 15 SSRs by Escribano et al., 2004 andEscribano et al., 2008. A similar result is obtained in A. senegalensis (Kwapata et al., 2007) and 152 bp to 497 bp in Annona species (Anuragi et al., 2016). PCR amplification profile of 18 Annona genotypes using SSR markers ASSSR_5 and ASSSR_16 are presented in the plates (Plate 1 and 2). ...
The experiment was conducted during July, 2020–January, 2021 at Agroforestry Research Station, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, Gujarat, India. Annona belongs to the family Annonaceae. The Annonaceae fruits are morphologically uniform but each type has a unique taste, flavour pulp colour and texture. The study revealed three types of leaf shapes (ovate, elliptic and lanceolate), leaf apex (acute, rounded and obtuse), leaf base (acute, rounded and acuminate), fruit shape (cordate, rounded and irregular), medium and late time of harvest maturity, overlapping and smooth segmentation of the surface and only one type for fruit at peduncle end i.e., inflated. The biochemical analysis of Annona species revealed the ranges of TSS (18.39 to 28.32 °B), TA of pulp (0.18–0.80%), reducing sugar (15.02–19.44%), non-reducing sugar (2.77–5.92%), total sugar (17.79–23.70%), ascorbic acid (27.55–43.10 mg 100 g-1) and phenol (0.18–0.36%). The DNA was amplified using fourteen simple sequence repeats primers and twenty-six alleles were found with an average of 1.86 alleles loci-1. The polymorphism information content value of the SSR markers was in the range of 0.09 to 0.38. A dendrogram was developed based on the cluster analysis, which revealed similarity index values varied from 0.58 to 1.00 with an average of 0.83. Based on the dendrogram the genotypes were clustered into two main clusters viz., A and B. Cluster A has sixteen genotypes and cluster B has two genotypes belonging to different Annona species. These results concluded that SSR markers could be efficiently used to study divergence among Annona genotypes.
... We may infer that climate and environmental variables play an essential role in D. innoxia's phenotypic plasticity, which confers a considerable capacity to adjust to ecological challenges [50][51][52]. The detected hierarchical inconsistency of trait variations may reflect intraspecific genetic variation, as has been shown by Ref. [53]. Since the observed variability has a heritable component, quantitative genetic studies can also investigate the variation in morphological and biomass traits. ...
Plant functional traits are consistently linked with certain ecological factors (i.e., abiotic and biotic), determining which components of a plant species pool are assembled into local communities. In this sense, non-native naturalized plants show more plasticity of morphological traits by adopting new habitat (an ecological niche) of the invaded habitats. This study focuses on the biomass allocation pattern and consistent traits-environment linkages of a naturalized Datura innoxia plant population along the elevation gradient in NW, Pakistan. We sampled 120 plots of the downy thorn apple distributed in 12 vegetation stands with 18 morphological and functional biomass traits during the flowering season and were analyzed along the three elevation zones having altitude ranges from 634.85 m to 1405.3 m from sear level designated as Group I to III identified by Ward's agglomerative clustering strategy (WACS). Our results show that many morphological traits and biomass allocation in different parts varied significantly (p < 0.05) in the pair-wise comparisons along the elevation. Likewise, all plant traits decreased from lower (drought stress) to high elevation zones (moist zones), suggesting progressive adaptation of Datura innoxia with the natural vegetation in NW Pakistan. Similarly, the soil variable also corresponds with the trait's variation e.g., significant variations (P < 0.05) of soil organic matter, organic carbon, Nitrogen and Phosphorus was recorded. The trait-environment linkages were exposed by redundancy analysis (RDA) that was co-drive by topographic (elevation, r = −0.4897), edaphic (sand, r = -0.4565 and silt, r = 0.5855) and climatic factors. Nevertheless, the influences of climatic factors were stronger than soil variables that were strongly linked with elevation gradient. The study concludes that D. innoxia has adopted the prevailing environmental and climatic conditions, and further investigation is required to evaluate the effects of these factors on their phytochemical and medicinal value.
... The development of various DNA markers such as Random Amplified Polymorphic DNA, microsatellites, also referred to as simple sequence repeats (SSR), and single nucleotide polymorphism (SNP) has increased the range of characterization of existing genetic resources of diverse crop species more effectively and reliably. Since custard apple is one of the orphan crops of the tropics, there have been very few attempts to develop molecular markers (Yoshida et al., 1998;Kwapata et al., 2007;Escribano et al. 2009;Larranaga and Hormaza et al., 2015) [13,8,4,9] or use of 'omics' tools (Gupta et al., 2015) [5] . Due to their relative lack of commercial value and the paucity of studies on genetic improvement of cultivars, the genetic resources of dry fruit crops have not received the attention they deserve. ...
... The development of various DNA markers such as Random Amplified Polymorphic DNA, microsatellites, also referred to as simple sequence repeats (SSR), and single nucleotide polymorphism (SNP) has increased the range of characterization of existing genetic resources of diverse crop species more effectively and reliably. Since custard apple is one of the orphan crops of the tropics, there have been very few attempts to develop molecular markers (Yoshida et al., 1998;Kwapata et al., 2007;Escribano et al. 2009;Larranaga and Hormaza et al., 2015) [13,8,4,9] or use of 'omics' tools (Gupta et al., 2015) [5] . Due to their relative lack of commercial value and the paucity of studies on genetic improvement of cultivars, the genetic resources of dry fruit crops have not received the attention they deserve. ...
The current study was conducted at the College of Horticulture in Bagalkot, Karnataka, between 2017 and 2019. A Randomized Complete Block Design was used to set up the experiment, and twelve cultivars were duplicated three times. Horticulturists have traditionally used morphological and physiological descriptors to identify different varieties of fruit crops. Using the allele sizes of nine SSR markers acquired from previously described Annona species, the genetic diversity of the 12 Annona cultivars was evaluated (Larranaga and Hormaza, 2015). Nine microsatellite markers were used to identify a total of 21 alleles from the nine SSR manufacturers that were evaluated across 12 Annona cultivars in the current investigation. In the current study, a total of 11 RAPD markers that were evaluated across 12 Annona cultivars amplified a total of 28 loci. Additionally, to determine how closely related these 12 cultivars were genetically, they were subjected to phylogenetic analysis using the pair-wise genetic matrix generated by RAPD markers. The results were presented in Figure 2 using an unrooted neighbour joining tree and the Mega7.0 software. The findings of the molecular analysis had shown that the genetic diversity of cultivars varied. Twelve Annona cultivars under investigation have been successfully characterized using the SSR and RAPD markers. The findings showed that SSR markers are co-dominant and highly reproducible. The significance of SSR markers in the molecular characterisation of Annona cultivars is indicated by this.
... All the indices presented herein indicated that genetic variabilities within-populations of A. squamosa were generally greater than those between-populations, thereby confirming findings reported previously for this species [44], for A. crassiflora [39] and for A. senegalensis Pers [45]. This finding suggests that efforts to augment the gene pool of A. squamosa in the germplasm bank at Embrapa Meio-Norte should be concentrated on each separate population to prevent genetic redundancy within the collection [46]. ...
The fruits of Annona squamosa L. are highly appreciated for their sweet taste and their nutritional and medicinal properties. Since the species is threatened by habitat loss, germplasm conservation is of the utmost importance. The present study aimed to characterize genetic variability among 19 accessions of A. squamosa maintained in the Active Germplasm Bank (AGB) of Embrapa Meio-Norte, Teresina, Piauí, Brazil, and to determine whether the geographical distribution of the genotypes influenced population structure. The accessions were derived from populations originally located in orchards situated some 200 km apart in three municipalities in the northeast of Brazil, namely Timon, Bom Jesus and Canto do Buriti. Leaf DNA was extracted and analyzed by inter simple sequence repeat (ISSR)-polymerase chain reaction. Nine ISSR primers generated 127 scorable bands of which 101 (79.52%) were polymorphic. The within-population diversity was high (Canto do Buriti), intermediary (Timon) and low (Bom Jesus). Unweighted pair-group method with arithmetic averages and Bayesian analysis of polymorphic bands separated the genotypes into two main clusters according to genetic distance and ancestry (vegetative- and seed-propagated groups), indicating that the geographical location of the populations did not influence the genetic diversity of the accessions. While ISSR marker-based DNA fingerprinting of the 19 studied accessions revealed a degree of genetic diversity between populations, the variability detected within each population was somewhat higher. This information will contribute to a better understanding of genetic variation and population structure of the accessions of A. squamosa in the AGB of Embrapa Meio-Norte, and provide crucial information for future conservation and genetic improvement programs of the species.
... Thus, truly divergent genotypes could be further collected and utilized in crop improvement programmes. Studies on the use of morphological and molecular markers have been reported on members of Annonaceae (Escribano et al., 2007;Kwapata et al., 2007;Ahmad et al., 2010;Pomper et al., 2010 andCota et al., 2011). ...
... There are limited reports on exploitation of molecular markers for diversity analysis in Annonas. Few of these markers are random amplified polymorphic marker (RAPD) marker (Bharad et al. 2009), amplified fragment length polymorphism (AFLP) markers and simple sequence repeat (SSR) markers (Escribano et al. 2004;Kwapata et al. 2007). ...
... The primers LMCH-1, LMCH-3, LMCH-4, LMCH-5 LMCH-6 and LMCH-9 showed highest percentage of polymorphism i.e. 100 per cent (Table 6). Escribano et al. 2007, Escribano et al. 2009and Kwapata et al. 2007 reported the results for Annona spp. using SSR primers. ...
Background: Annona spp. are one of the underutilized fruit crop having great commercial and medicinal value and large diversity present in Maharashtra which is untapped. The present study conducted to characterize the Annona genotypes by ISSR and SSR markers for future germplasm conservation and crop improvement programme. Methods: Ninety genotypes were collected from various regions of Maharashtra. These ninety genotypes and eleven varieties from All India Coordinated Research Project on Arid Zone Fruits, MPKV, Rahuri were screened for morphological and biochemical characters. After morphological and biochemical characterization twenty-two superior genotypes were characterized using ISSR and SSR markers. Result: Among the 29 ISSR primers used, 20 were found polymorphic which produced a total of 171 reproducible amplicons, in which 109 amplicons (75.43%) were polymorphic. In 20 SSR primers used, 14 primers were found polymorphic which produced a total of 43 reproducible amplicons, in which 36 amplicons (83.72%) were polymorphic. Twenty unique amplicons produced by ISSR primer and eight unique amplicons produced by SSR in studied genotypes is useful in genotype/variety identification in future crop improvement. The dendogram generated based on UPGMA method of cluster analysis using ISSR and SSR marker data revealed little different but similar grouping of genotypes into two major clusters viz., cluster A and cluster B. The UPGMA based cluster analysis using dice similarity coefficient grouped Annona genotypes into two major clusters which differentiate squamosa and atemoya species. The distribution of the genotypes in the dendogram was mostly consistent with the known pedigree information, geographical locations and the morphological attributes. Genotypes collected from Purandar, Sawargaon, Pemgiri and Ajanta districts of Maharashtra formed cluster as per geographic collection. The close relationship across genotypes might be explained by either historical relationship to sharing common ancestor or more likely geographical proximity and large population size which favour genetic interchange. This low genetic difference among genotypes suggests that there was more gene flow through random mating without barrier within agro-ecological zones. ISSR and SSR markers gives direction in characterization of Annona genotypes but more areas need to be survey and explore for tapping genetic diversity to widen the gene pool. The screened material found superior need to be prioritized in terms of in-situ and ex-situ conservation for further evaluation and crop improvement.
... The primer OPP-04 showed a polymorphic band between 600 and 800bp and this particular band is seen in the seedlings 6, 7, 9, 15 and 21. Even the primer OPT-06 showed a polymorphic band around 1000bp, this polymorphic band is seen in 2, 6,7,9,12,16,17,18,19 and 20 seedlings. The seedlings mentioned above could be nucellar seedlings based on the preliminary observations. ...
... They have co-related the wild varieties from island and variety from mainland India by using different molecular markers of RAPD and ISSR. These findings are also in accordance with Shakya et al. (2010) [15] in Syzygium cuminii, and Kingdom et al. (2007) [9] in Annona spp. ...
... They have co-related the wild varieties from island and variety from mainland India by using different molecular markers of RAPD and ISSR. These findings are also in accordance with Shakya et al. (2010) [15] in Syzygium cuminii, and Kingdom et al. (2007) [9] in Annona spp. ...
Jamun (Syzygium cumini Skeels.) is an important minor indigenous fruit of India. It belongs to the Myrtaceae family. Though native to India and Myanmar, it has naturalized throughout the South East Asia and Pacific regions. Jamun is an important fruit crop with a special character called polyembryony. This trait could be effectively utilized for propagation in nursery or hybridization work. The proper identification of zygotic and nucellar seedlings may lead to proper management of the elite germplasms in mass scale propagation. Conventional techniques of 'Off type' rouging and other morphological identification techniques are not full proof. Molecular marker may be used for discrimination of different seedlings. Cheap and cost effective RAPD technique is used in seedlings of a single seed of jamun with 20 non-specific primers. Primer OPP-04, OPT-06 and OPT-07 gave optimum polymorphism as compared to other primers. Total 11 amplicons were identified with these 3 primers that are effective in zygotic and nucellar discrimination. Sequencing of the amplicons could generate nucleotide sequence associated with the trait. Polyembryony character is regulated by several environmental factors, hence specific marker selection for specific Eco-region is crucial. The universal primers identified here are important for detection of polyembryony trait of jamun of this subcontinent.
... Besides its uses, phytocompounds, and biological activities, other interesting aspects of A. senegalensis, including distribution and ecology (Pinto et al. 2005) and molecular genetic diversity (Kwapata et al. 2007) have been documented. However, few studies, to our knowledge, have explored the morphological variation within this shrub species (Folorunso & Olorode 2006) despite the importance of a quantitative evaluation of the plant's morphological variability in planning conservation and domestication policies (Pauku et al. 2010;Gouwakinnou et al. 2011;Ewédjè et al. 2012;Padonou et al. 2017). ...
... This study examined the morphological traits of shrub, leaves, fruits, and seeds of the wild soursop, and assessed their variations across contrasting environmental conditions in Benin. Considering the high genetic diversity observed in natural populations of A. senegalensis across a latitudinal gradient in Malawi (Kwapata et al. 2007), we expect a high variability in all morphological traits. In addition, we expect a significant differentiation between A. senegalensis populations across climatic zones as an expression of phenotypic plasticity or local adaptation of plants to variation in environmental conditions (Pigliucci 2006). ...
... 2018; Wang et al. 2020). The observed multilevel variability may nevertheless also express intraspecific genetic diversity as observed for the species in Malawi (Kwapata et al. 2007). Therefore, quantitative genetic investigations about the studied populations are necessary to examine the heritable part of the observed variability. ...
Background and aims-Geographic patterns of phenotypic variability can inform understanding of the resilience potential of plant species to environmental hazards such as climate change. Such understanding provides support for conservation and domestication efforts. Here, we investigated natural morphological variation of the individuals, fruits, seeds, and leaves of the tropical shrub Annona senegalensis Pers. along a climatic gradient. Methods-Morphological data were collected on shrubs, fruits, seeds, and leaves of 150 shrubs from five populations in the three climatic zones of Benin. Linear mixed effects models were used to test the variability of the morphological traits of the species and also to estimate the variance components in order to tease apart the importance of each source of variation. The most important morphological descriptors discriminating climatic zones were identified using a stepwise discriminant analysis. Redundancy analysis was then used to determine the relationships between discriminant morphological traits and bioclimatic variables. Key results-Morphological traits of A. senegalensis varied greatly both within and among climatic zones. A substantial part (42%) of the among-climatic zones phenotypic variability in the species was attributable to climate, mainly rainfall and temperature. Morphological traits such as big shrubs, big fruits, and high number of seeds per fruit were associated with high mean annual rainfall and low mean temperature of the warmest quarter. Conclusions-The findings suggest an important zonal adaptation of the species to climate variability. The phenotypic diversity pattern that we highlighted can be useful when designing conservation policies for the species. However, quantitative genetics through common garden or reciprocal transplantation experiments related to the species' populations would enable to explore the heritable part of the observed variability to support effective conservation and domestication efforts.
... There is ample information available in literature for use of various molecular markers to diversify various germplasms. There have been some work carried out using RAPD markers (Ronning et al., 1995;Thompson et al., 1997;Bharad et al., 2009;Chang et al., 2011, SSR (Jarret andBowen, 1994;Buteler et al., 1999;Escribano et al., 2004;Zhang et al., 2001;Gichuru et al., 2006;Kwapata et al., 2007), ISSR markers (Njuguna, 2005;Huang and Sun, 2000;Hu et al., 2003) and AFLP (Rahman et al., 1998;Zhang et al., 2000Zhang et al., , 2004Fajardo et al., 2002) to study the genetic diversity of Dioscorea alata. However, very little information is available on use of molecular markers in India for diversity analysis in greater yam. ...
In India, greater yam (Dioscorea alata L.) is one of the major cultivated yam species and it provides a very good source of dietary carbohydrates in regions of tropic and subtropics. This research work attempts molecular characterization of 27 genotypes collected from different districts of Gujarat and India for understanding of the distribution and extent of genetic variation present within the germplasm. The genetic diversity on molecular basis was determined using 16 RAPD and 15 ISSR markers. All the RAPD primers and ISSR primers studied showed very high polymorphism. The RAPD primers on an average produced 8 polymorphic alleles with polymorphism and values of polymorphism information content (PIC) 93.04% and 0.563 respectively. The ISSR primers on an average produced 6.33 polymorphic alleles. The polymorphism and PIC values obtained were 72.96% and 0.516 respectively. Highest number of bands was obtained for the ISSR primer UBC 875 and lowest in UBC 878, While in RAPD OPA-1 and OPM-4 respectively. Clusters were made using the UPGMA method and their genetic relatedness was studied using pair wise matrix of genetic distances.
... Considering cherimoya microsatellites as reference database, the genetic diversity of A. senegalensis Pers. populations was studied by SSRs and found that there is large-scale geographical and ecotypic differentiation between the species collected from Malawi (Kwapata et al., 2007). Integrating the molecular marker data into Geographic Information Systems allows visualization and understanding spatial patterns of genetic diversity, to further augment conservation (Van Zonneveld et al., 2012). ...
... eru.Van Zonneveld et al., 2012Fifty-two single locus polymorphic microsatellites were developed in cherimoya cv. Fino de Jete.Escribano et al., 2008 In total, 23 alleles were detected in nine populations of Annona senegalensis Pers. collected from different locations in Malawi and genetic diversity parameters revealed high levels of heterozygosity.Kwapata et al., 2007 Ten microsatellite primer pairs developed in Annona crassiflora Mart.Genetic differentiation among populations of A. squamosa collected from Rajasthan (India) analyzed using 15 ISSR primers. ...
Fruit trees are important as additional sources of revenue and are important diet supplements. Besides yielding edible fruits, there are several other useful products available from these fruit trees. However, despite their significance, these plants have not been researched thoroughly for their propagation, cultivation and molecular characterization. Unlike the annual agricultural crops, fruit trees present certain unique problems in terms of their improvement. The long pre-bearing age and highly heterozygous cross pollination nature of these crops constitute a serious problem for their rapid improvement and assessment of available variability. With changing socio-economic and environmental values, there is now a worldwide burgeoning of interest and support for research on the maintenance, genetic improvement and efficient use of fruit tree resources and their products. However, some of the useful fruit trees have not been investigated in detail. In this review, biotechnological intervention in the area of tissue culture, molecular markers and genetic transformation on three under-researched tropical fruit tree crops viz. Jackfruit, Guava, and Custard apple are described.
Abbreviations
°C:degree Celsius; 2, 4-D- 2, 4:Dichlorophenoxyacetic acid; ABA:Abscissic acid; AC: Activated charcoal; ACO: Aconitase; ADH: Alcohol dehydrogenase; AFLP: Amplified Fragment Length Polymorphism; BA: Benzyladenine; BAP: 6-Benzyl aminopurine; CaMV: Cauliflower mosaic virus; cpDNA: Chloroplast DNA; CTAB: Cetyl Trimethyl Ammonium Bromide; cv.: Cultivars, GA3: Gibberellic acid; GC-MS: Gas chromatography mass spectrometry; GOT: Glutamate oxaloacetate transaminase; GUS: β-glucuronidase gene; HCL: Hydrochloric acid; H2SO4: Sulphuric acid; IAA: Indole-3-acetic acid; IBA: Indole-3-butyric acid; IDH: Isocitrate dehydrogenase; ISSR: Inter-simple sequence repeat; KIN: Kinetin; MDH: Malate dehydrogenase; Me: Malic enzyme; mg/L- Milligrams per liter; MS: Murashige and Skoog medium; NAA: α-Naphthalene acetic acid; PGI: Phosphoglucoisomerase; PGM: Phosphoglucomutase; ppm: Parts per million; PVP: Polyvinylpurrolidone; PVPP: Polyvinylpolypyrrolidone; QSAR: Quantitative structure activity relationship; RAPD: Random Amplified Polymorphic DNA; RFLP: Restriction Fragment Length polymorphism; SSR: Simple sequence repeat; TDZ: Thidiazuron (N-phenyl-N’-1,2,3-thiadiazol-5ylurea); TPI: Triosephosphate isomerase; WPM: Woody plant medium; ZEA: Zeatin[6-(trans-4-hydroxy-3-methyl-2-butenylamino) purine]
