Sources of Resistance to Phytophthora Pod Rot at the International CocoaGenebank, Trinidad

The International Cocoa Genebank, Trinidad (ICG,T) contains about 2000 cacao (Theobroma cacao L.) accessions. The purpose of the collection is to provide a source of genes for the genetic improvement of cacao through breeding. In fulfilling this objective, it is imperative that this collection be evaluated to identify major sources of genes for the genetic improvement of important agronomic–economic traits. In this study, 816 cacao accessions were evaluated for resistance to Phytophthora pod rot (black pod disease), which causes enormous loss of yield in cocoa production throughout the world.The study identifies promising resistant genotypes that could be exploited in cacao breeding programmes. It further compares the levels of resistance to Phytophthora pod rot among two cacao types (wild and cultivated types), three major groups (Forastero, Trinitario and Refractario) and 11 accession groups (B, AM, CL, ICS, IMC, JA, LP, MOQ, NA, PA and TRD) . The distribution of scores for the 816 accessions showed skewness towards the susceptible end of the disease rating scale, indicating that a large proportion (68.9%) of the sample was susceptible (disease rating 6–8) to Phytophthora pod rot. However, 12.9% of the sample population was found to be resistant (disease rating 1–3) and 18.2% moderately resistant (disease rating 4–5). Significant differences were observed between the wild and cultivated accessions and among the Forastero, Refractario and Trinitario groups. Marked differences were also observed among 11 accession groups, each of which was represented by at least 20 genotypes. Higher proportions of resistant (17.7%) and moderately resistant (22.6%) genotypes were observed in the wild accessions than in the cultivated varieties (9.4% resistant and 14.4% moderately resistant). The Forastero group, consisting of many wild accessions, was found to contain more resistant (18.0%) and moderately resistant (23.1%) genotypes than either the Trinitario (4.8% resistant and 13.6% moderately resistant) or Refractario (11.3% resistant and 15.4% moderately resistant). Among 11 accession groups, the largest percentage of resistant (24.2%) and moderately resistant (28.8%) genotypes were found in the PA (Forastero). Different accession groups had varying proportions of resistant and moderately resistant genotypes. This is not unexpected in an out-breeding crop such as cacao with a high level of heterogeneity. This finding reinforces the idea of a pre-breeding programme (germplasm enhancement) to accumulate resistance genes over several populations as a strategy for improving the genetic base of resistance in national cacao breeding programmes. One hundred and five promising resistant genotypes were identified among the 816 accessions evaluated. These accessions are potential sources of resistance genes for the development of high-yielding resistant varieties in cacao.     

Identification of duplicates and fingerprinting of primary and secondary wild annual <Emphasis Type="Italic">Cicer</Emphasis> gene pools using AFLP markers

Wild annual Cicer gene pools contain valuable germplasm for chickpea improvement programs. Previous research showed that duplication might exist in accessions collected from these gene pools, which would hinder chickpea breeding and related research. AFLP (amplified fragment length polymorphism) markers were used to fingerprint the world collections of the primary and secondary gene pools including C. reticulatum Lad., C. bijugum K.H. Rech., C. judaicum Boiss. and C. pinnatifidum Jaub. et Sp. Duplicates were detected in a total of 24 accessions in both the gene pools, highlighting the necessity to fingerprint the germplasm. Genotypic difference was detected as gene pool specific, species specific and accession specific AFLP markers. These were developed into fingerprinting keys for accession identification between and within species and gene pools. Use of AFLP markers to detect duplicates and to identify accessions is a reliable method which will assist in the characterisation and use of wild annual Cicer germplasm in chickpea improvement programs. We recommend the procedure presented in this paper as a standard approach for the precise genetic identification and characterisation of future world collections of wild Cicer, to keep germplasm integrity and to benefit chickpea breeding and related research programs.     

Genetic diversity of winter wheat in Shaanxi province, China, and other common wheat germplasm pools

Cultivated Chinese wheat germplasm has been a valuable genetic resourcein international plant breeding. Landraces endemic to China are a geneticresource that is distinct from other wheat germplasm. Patterns of geneticdiversity among cultivated Chinese accessions and relationship to othergermplasm pools are unknown, despite the proven value and potential novelty. Theobjective of this work was to determine the level of genetic diversity withinimproved Chinese germplasm in the context of several other wheat germplasmpools. We analyzed a set of improved accessions cultivated from the 1940s to the1990s in Shaanxi province, China, using six simple sequence repeat (SSR) primerpairs and 30 restriction fragment length polymorphism - probe enzymecombinations (RFLP-PEC) previously used to characterize 21 geographically basedgermplasm pools. Shaanxi germplasm consists of three groups based on foreignintroductions from Italy, Australia, Denmark, and Russia. There was a decreasein genetic diversity among Shaanxi accessions cultivated in the 1970s and 1980sto the 1990s, and accession classifications based on primary decade ofcultivation were found to be significantly undifferentiated. The analysis of themean genetic distance among 22 geographically based pools of germplasm suggestsseveral regions are significantly undifferentiated. A vast majority of the totalamount of variation was found within pools; therefore, pools appear to belargely differentiated based on small differences in band relative frequency andfew if any unique bands. Previous studies have identified some Chinese landracepools as morphologically and genetically unique. The Shaanxi pool does not havethe same unique morphological or genetic features, nor is it more similar to thelandrace pools than other improved germplasm pools.     

Morphological Diversity of Bambara Groundnut [Vigna subterranea (L.) Verdc.] Landraces in Tanzania

A field experiment was conducted in Tanzania for two seasons to assess the genetic diversity of bambara groundnut landraces based on morphological characters. One hundred accessions collected from a wide range of agricultural zones in the country were evaluated in a 10 × 10 triple lattice block design at Maruku station in Bukoba, Tanzania. For the qualitative characters evaluated, considerable morphological variations were observed for growth habit, pod shape and pod colour. Quantitative morphological characters such as peduncle length, number of leaves per plant, terminal leaflet width, terminal leaflet length, petiole length, plant spread, plant length, pod width, seed length, seed width, number of pods per plant, shelling percentage and days to 50% flowering showed significant variation among accessions for the two test seasons. Multivariate analysis for 15 quantitative morphological traits that showed significant variation indicated that the first four PCs with eigenvalues ≥1 accounted for 63.0 and 65.0% of the total variance among the accessions during the two test seasons, respectively. The most important loadings for PC1 and PC2 for the two seasons were terminal leaflet width, terminal leaflet length, petiole length, plant spread, plant height and pod length, pod width, seed length, seed width, numbers of pods per plant. Cluster analysis grouped bambara groundnut accessions into three major groups with respect to their geographic origins. Based on this present study, bambara groundnut landraces from Tanzania displayed a considerable diversity for morphological and agronomic traits useful for germplasm management and utilization into crop improvement.     

Development of an algorithm identifying maximally diverse core collections

The development of a core collection, one which represents the genetic diversity of a crop with minimal redundancy and increases utility of the collection as a whole, is especially important as the funding for germplasm collections decreases. With limited resources, it is difficult to manage large germplasm collections and disperse genetically diverse germplasm to plant breeders. An algorithm was developed to assist in selection of core collections based on estimates of genetic distance. The criteria for selection of the maximum genetically diverse set were based on rankings of genetic distance between an accession with respect to all other accessions. Depending on the size core which a user wished, a zone around each selected accession was determined and no other accession within these limits was selected. The premise for the algorithm was that the genetic variability represented in the core must be representative of the distribution of genetic distances within the population of interest. In the present study, the algorithm was used with RAPD-marker-based estimates of genetic distance for 270 Theobroma cacao L. accessions and 134 Capsicum accessions that chose a set representing 18.5% of the population and representing the breadth of RAPD-based variation.     

Assessing the Genetic Diversity in the International Cocoa Genebank,Trinidad (ICG,T) using Isozyme Electrophoresis and RAPD

Random amplified polymorphic DNA (RAPD) and isozyme electrophoresis (IE) techniques were used to estimate the level of genetic diversity in a sample of cacao germplasm existing at the International Cocoa Genebank, Trinidad. Twenty-six cocoa populations represented by 459 cocoa genotypes were analysed using IE and 22 populations represented by 353 cocoa genotypes were analysed using RAPD. Despite few differences in the classification of the populations, both techniques revealed three major groups: the indigenous trees, the cultivated Trinitario and the cultivated trees from Ecuador. Two-thirds of the partitioned diversity were found within populations and one-third between the populations, with both techniques.     

Genetic diversity and spatial structure in a new distinct <Emphasis Type="Italic">Theobroma cacao</Emphasis> L. population in Bolivia

Cacao (Theobroma cacao L.) is an important economic crop in the Bolivian Amazon. Bolivian farmers both cultivate cacao, and extract fruits from wild stands in the Beni River region and in valleys of the Andes foothills. The germplasm group traditionally used is presently referred to as “Cacao Nacional Boliviano” (CNB). Using DNA fingerprinting technology based on microsatellite markers, we genotyped 164 Bolivian cacao accessions, including both cultivated and wild CNB accessions sampled from the Amazonian regions of La Paz and Beni, and compared their SSR profiles with 78 reference Forastero accessions from Amazonian cacao populations, including germplasm from the Ucayali region of Peru. Results of multivariate ordination and analysis of molecular variance show that CNB cacao has a unique genetic profile that is significantly different from the known cacao germplasm groups in South America. The results also show that cultivated CNB and wild CNB populations in the Beni River share a similar genetic profile, suggesting that the cultivated CNB is of indigenous origin in Bolivia. The level of genetic diversity, measured by allele richness and gene diversity in the Bolivian cacao, is moderately high, but was significantly lower than gene diversity in the other Amazonian cacao populations. Significant spatial genetic structure was detected in the wild CNB population, using analysis of autocorrelation (rc = 0.232; P < 0.001) and Mantel tests (Rxy = 0.276; P < 0.001). This finding is also highly valuable to support in situ conservation and sustainable use of CNB genetic diversity in Bolivia.     

Changes in seed Morphology, Dormancy and Germination from wild to Cultivated Hyacinth bean Germplasm (Lablab purpureus: Papilionoideae)

Morphological and physiological seed characteristics of the hyacinth bean (Lablab purpureus) were investigated in a set of 18 different germplasm accessions, from wild over semi-domesticated forms to landraces and current cultivars. The objective of this study was to improve the understanding of the domestication of this tropical legume crop. Wild accessions were easily distinguished by morphological characters, such as small, greyish-brown, mottled seeds. Cultivated or semi-domesticated forms showed a much wider variation in size, colour and shape. Seed mass of cultivated accessions reached up to almost 10 times that of wild accessions. Most wild accessions showed a greater spread of germination and larger proportions of hard seeds, up to about 70%, than most cultivated and so-called semi-domesticated accessions. Cluster analysis applied separately to morphological and germination data and a subsequent discriminant analysis did not help integrating the morphological variability observed. The presumed occurrence of semi-domesticated accessions within the germplasm tested as well as the crop's origin from Africa alone or both Africa and Asia are debated on the basis of seed germination and hard seed coat dormancy.     

Genetic diversity of potato landraces from northwestern Argentina assessed with simple sequence repeats (SSRs)

Andean potato varieties are cultivated in the northwest of Argentina and constitute the most important staple food for the local farmers. The genetic diversity of 155 accessions conserved at the Genebank of Balcarce (INTA) was tested using four microsatellites. Three commercial potato varieties of Tuberosum group and one accession of Curtilobum group were used as outgroups. The presence of bands was scored for each microsatellite and for each accession and the data were analysed by principal coordinate analysis. The polymorphism information content was obtained for each molecular marker from banding patterns. Analysis of molecular variance was carried out with a variable number of accessions for each landrace, from different departments and sites within departments. More than one genotype was detected in the majority of the potato landraces. Some accessions within each landrace did not differentiate. AMOVA revealed that most of the genetic variation occurred among sites within departments and among local varieties. These findings are discussed considering the agricultural practices carried out in the Andean farming system.     

Variation of cultivated flax (Linum usitatissimum L. subsp.usitatissimum) and its wild progenitor pale flax (subsp.angustifolium (Huds.) Thell.)

The domestication of cultivated flax (Linum usitatissimum L. subsp.usitatissimum) is briefly discussed. Using data documented as a matter of routine in genebank work, 63 accessions of cultivated flax from the flax germplasm collection of the Gatersleben Genebank are compared with 73 accessions of its wild progenitor pale flax (subsp.angustifolium (Huds.) Thell.), which have been observed in systematic field trials. Range of variation, genetically based variation, heritability and correlation of several characters are considered, especially with respect to the influence of domestication. Cultivated flax shows higher variation in the characters of generative plant parts, while pale flax varies more in the vegetative parts of the plant. The character correlations are similar in both subspecies. Of the 29 characters studied in pale flax the parameters describing tillering, height of plant, weight of seeds, width of petals and width of leaves are especially suitable for distinguishing between different accessions. In spite of the high heritabilities of most of the characters, the influence of the environment is significant in most cases.     

PCR Marker-based Analysis of Wild and Cultivated Yams (<Emphasis Type="Italic">Dioscorea</Emphasis> spp.) in Nigeria: Genetic Relationships and Implications for <Emphasis Type="Italic">ex situ</Emphasis> Conservation

Reliable characterization of the variation among wild and cultivated yams in Nigeria is essential for improved management and efficient utilization of yam genetic resources. RAPD and double stringency PCR (DS-PCR) analyses were used to investigate genetic relationships and the extent of redundancy among 30 accessions of two cultivated, and 35 accessions of four wild yam species collected from Nigeria. Twenty-five selected random decamer and two microsatellite primers were used individually and in combination to generate DNA profiles for each accession of the six Dioscorea species. The number of amplified fragments varied from 7 to 18 fragments per primer/primer combination. Different levels of intraspecific genetic diversity were found, with Dioscorea rotundata Poir. being the most variable. Based on identical profiles for the RAPD and DS-PCR primers, 12 duplication groups consisting of a total number of 37 accessions were observed in the present study. An UPGMA analysis grouped the majority of plants according to the species. Cultivated yams belonging to the D. cayenensis–rotundata species complex, which were classified into seven morphotypes/varietal groups, could be clearly separated into two major groups corresponding to D. rotundata Poir. and D. cayenensis Lam. D. cayenensis cultivars exhibited a low level of intraspecific variation and were genetically close to the wild species Dioscorea burkilliana J. Miège. D. rotundata cultivars classified into six varietal groups showed a high degree of DNA polymorphism and were separated into two major groups that appeared most closely related to Dioscorea praehensilis Benth. and Dioscorea liebrechtsiana de Wild. We propose, based on these results, that cultivars classified into D. cayenensis should be considered as a taxon separate from D. rotundata. The implications of intraspecific variability for the ex situ conservation of wild and cultivated yam germplasm in Nigeria are discussed.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.     

Wild melon diversity in India (Punjab State)

We present here the first comprehensive genetic characterization of wild melon accessions from northern India. The genetic diversity among 43 wild melon accessions collected from the six agro-ecological regions of the Punjab State of India was assessed by measuring variation at 16 Simple Sequence Repeat (SSR) loci, morphological traits of plant habit and fruit morphological traits, two yield-associated traits, root nematode resistance and biochemical composition (ascorbic acid, carotenoids, titrable acidity). Variation among accessions was observed in plant habit and fruit traits and wild melon germplasm with high acidity and elevated carotenoid content and possessing resistance to Meloidogyne incognita was identified in the collection. A high level of genetic variability in wild melon germplasm was suggested by SSR analysis. Comparative analysis using SSRs of the genetic variability between wild melons from the north and other melons from the south and east regions of India and also reference accessions of cultivated melon from Spain, Japan, Korea, Maldives, Iraq and Israel, showed regional differentiation among Indian melon accessions and that Indian germplasm was not closely related to melon accessions from other parts of the world. A highly drought tolerant accession belonging to var. agrestis Naud. was also identified.     

Distinguishing among Pisum accessions using a hypervariable intron within Mendel’s green/yellow cotyledon gene

Distinguishing among commercial varieties or accessions in a germplasm repository presents considerable challenges when dealing with common crops or large collections. In the genus Pisum L. the third intron of the gene STAYGREEN (SGR), responsible for the green/yellow cotyledon variation, contains a complex of repeat sequences that display considerable indel and single nucleotide (SNP) polymorphism. In a sample of 137 Pisum L. accessions obtained from various germplasm collections 76 alleles were identified, ranging from about 600 to 1500 nucleotides in length. In most cases alleles differed by insertion/deletion (indel) polymorphisms in addition to single base changes. The amplicon was particularly useful for distinguishing wild accessions (those with dehiscent pods and dormant seeds). Among the 51 P. sativum L. accessions examined with a definite wild phenotype there were 44 SGR alleles. When this allelic variation was combined with location data for the wild accessions, each allele could be assigned a unique geographical location. Similarly, of the 10 P. fulvum Sibth. & Sm. accessions analyzed, only a group of three had the same allele, two of which are known to be replicates. We were able to confirm the identities of several P. sativum accessions from different germplasm collections, as well as identifying two cases where supposedly synonymous accessions gave different intron sequences. In addition, a group of wild pea accessions from Greece, Sicily, and Portugal exhibited a unique repeat sequence not found in any other wild accession, suggesting that these accessions constitute a distinct lineage. The rate of nucleotide site variation in this fragment of the SGR locus was found to be about 2.22 × 10^–8 substitutions per site per year. This value is similar to those found for many other sequences in herbaceous papilionoid tribes, although it is at the faster end of the distribution of herbaceous legume substitution rates. We propose that both a fast substitution rate as well as the formation and rearrangement of indels have contributed to the high level of variation we detected at SGR. This PCR product should prove valuable in the identification of Pisum accessions, for quality control operations in germplasm repositories, and for exploring the evolutionary relationships within the genus.

     

Variability and selection on different <Emphasis Type="Italic">Argyrolobium roseum</Emphasis> accessions for morphological traits and yield

Variability studies were carried out among different accessions of Argyrolobium roseum for 12 metric traits under natural and cultivated conditions. Almost all characters showed higher values in cultivated than natural population. Leaf breadth recorded highest CV (30.59%). Highest percentage of vitexin and D-pinitol (0.208 and 0.773% dwb) was observed in RAR-7 and RAR-6, respectively, under cultivated conditions. Phenotypic coefficient was higher than genotypic coefficient of variation. The association analysis revealed that root yield had positive significant correlation with 100 seed weight (r=0.839) biomass yield (r=0.601), where biomass yield had a positive significant correlation with leaf length, single pod weight. High heritability (97.4–99.8%) coupled with moderate genetic advance ranged between (49–67%) as a percent of mean was observed for leaf breadth, pod length, total foliage biomass yield and total root yield/plant. This suggests that direct selection for these traits is suitable for the improvement of this crop.     

Analysis of Genetic Diversity Among Saccharum spontaneum L. From Four Geographical Regions of India, Using Molecular Markers

Saccharum spontaneum L. a wild relative of sugarcane is the most variable and diverse among the Saccharum species. This species had been successfully exploited in sugarcane improvement programmes since 1915 and most of the present day commercial varieties are derivatives of interspecific hybrids involving S. spontaneum. The S. spontaneum germplasm available today in the World collections is diverse and represent different geographical groups. In the present investigation, an attempt was made to characterize 40 S. spontaneum clones collected from 4 different geographical areas in India using 20 random, 2 ISSR and 2 telomere primers. Of the 491 bands generated by these primers 83.9% were polymorphic. The genetic diversity estimated based on these markers was found to be moderate (48.9%). The pair-wise genetic distance between the accessions ranged from 29.8 to 60.0. The accessions from Arunachal Pradesh were found to be the most diverse, while Tamil Nadu accessions showed relatively less diversity. Diversity between Tamil Nadu and Kerala collections was found to be low, while the diversity between the Orissa group and the rest was found to be high. The collections from Mayurbanj and Cuttack regions of Orissa were found to be distinct. Arunachal Pradesh accessions, being more diverse, are a potential source for exploitation in sugarcane breeding programmes.     

Genetic Diversity of the Vigna Germplasm from Thailand and Neighboring Regions Revealed by AFLP Analysis

Thailand is a center of diversity for section Angulares of the Asian Vigna (genus Vigna subgenus Ceratotropis) and 4 Vigna species are cultivated in Thailand. Using newly collected wild and cultivated germplasm of Vigna from Thailand and outgroup accessions AFLP analysis was conducted to clarify genetic diversity and relationships. The results suggest that cultivated V. umbellata and V. mungo evolved from wild relatives in a single domestication event. Vigna umbellata is poorly differentiated from its wild and weedy relatives compared to V. mungo. Results suggest northern Thailand and the neighboring Shan state, Myanmar, is the probable center of domestication for V. umbellata as wild accessions from this area and cultivated rice bean from a wide area in Asia are not greatly diverged. Vigna minima, V. tenuicaulis and V. exilis accessions in Thailand are well differentiated with considerable intra-specific variation. Vigna hirtella consists of two well differentiated subgroups, suggesting taxonomic revision may be necessary. Close genetic relationships between V. radiata and V. grandiflora, and between V. mungo and V. trinervia are confirmed. Naturally growing V. mungo populations in northern Thailand appear to be true wild species as they are well differentiated from Indian wild and Thai cultivated populations. The origin of naturally growing cowpea in Thailand needs to be further studied using a more comprehensive set of materials. This study clarifies inter and intra-specific genetic diversity and inter species relationships of Thai Vigna species.     

Genetic variability of Coffea arabica L. accessions from Ethiopia evaluated with RAPDs

The genetic diversity of 50 wild and semi-wild accessions of the Coffea arabica L. germplasm collection, gathered by the FAO and ORSTOM missions to Ethiopia, and maintained in Colombia by CENICAFE, was evaluated with RAPD markers. The evaluation was carried out in two phases: In phase one, the polymorphism of 8 Ethiopian accessions of different geographic origin, plus the cultivated variety 'Caturra' was assessed with the RAPD technique with forty-two 10-mer oligonucleotides. In phase two, 51 accessions were assessed with a set of 5 polymorphic primers that reproduced, with a correlation of 95%, the groups generated by the 24 polymorphic primers found in phase one. Principal Coordinate Analysis of molecular data revealed that a closely related group consisting of 86% of the Ethiopian C. arabica accessions evaluated are significantly different from the Caturra variety and could be used in a genetic breeding initiative to increase the variability of cultivated varieties. The results also indicate that a larger polymorphism is present in the Colombian replica of FAO Ethiopian coffee germplasm collection than previously reported.     

Analysis of genetic diversity in Indian taro [Colocasia esculenta (L.) Schott] using random amplified polymorphic DNA (RAPD) markers

Taro [Colocasia esculenta (L.) Schott] germplasm accessions collected from different parts of India were subjected to RAPD (Random Amplified Polymorphic DNA) analysis to assess the genetic diversity prevalent and also to test the genetic basis of morphotypic classification. Thirteen random decamer primers out of the 22 tested were used to analyse 32 taro accessions belonging to 28 morphotypes. Three out of these thirteen primers analysed showed 100 per cent polymorphism. Per cent polymorphism varied from 60 to 100 among the polymorphic primers. High genetic diversity was revealed as the similarity coefficient values ranged from 0.50 to 0.98. No two accessions analysed in the present study showed a similarity coefficient value of one thereby indicating their distinctness and presence of high genetic diversity in Indian taro germplasm. Dendrogram obtained from UPGMA analysis grouped 32 accessions in four clusters and three accessions were placed as outliers. Clustering pattern did not show any strict relationship with geographical distribution, morphotype classification and genotypic diversity. Further, accessions classified, as belonging to the same morphotypic group did not always cluster together. Presence of a very close genepool of the wild, weedy and cultivated forms with extreme levels of phenotypic and genotypic variation is suggested as the reason for high genetic diversity reported. Usefulness of DNA markers such as RAPD in characterising and assessing the genetic diversity in Indian taro germplasm is hereby demonstrated.     

Genetic diversity and parentage in farmer varieties of cacao (Theobroma cacao L.) from Honduras and Nicaragua as revealed by single nucleotide polymorphism (SNP) markers

Cacao (Theobroma cacao L.) is the main source for chocolate with an annual production of four million tons worldwide. This Neotropical tree crop was domesticated in Mesoamerica as far back as 3,000 years ago. Knowledge of genetic diversity and population structure in farmer varieties of cacao in the center of domestication is essential for sustainable production of fine-flavored cacao beans and contributes to in situ/on-farm conservation of farmer varieties. Based on 70 single nucleotide polymorphism markers, we analyzed 84 fine-flavored farmer varieties collected from traditional cacao farms in Honduras and Nicaragua. The study also included 31 clones from the international cacao collections to serve as references. The SNP based multilocus matching identified six synonymous groups, including 14 Criollo and two Amelonado varieties. A moderately high level of genetic diversity was observed in these farmer varieties, indicating the possibility to further explore intra-population variation and breed for fine-flavored cocoa. Multivariate analysis showed clustering of the 84 farmer accessions in five genetic groups: ancient Criollo, Amelonado, Trinitario (including Nicaragua Trinitario and Honduras Trinitario) and Upper Amazon Forastero (only one accession). The Honduras Trinitario differed from the Nicaragua Trinitario group. The clustering results largely supported the perceived classification of cacao by local farmers and researchers, which was mainly based on morphological traits. However, the well known traditional variety “Indio” in this region was identified as synonymous with Amelonado. Parentage analysis showed that the variety “Indio” (or Amelonado) contributed more to the Trinitario type farmer varieties, whereas ancient Criollo had less influence. The present study demonstrates the efficacy of using a small set of SNP makers for cacao germplasm characterization, and further depicts the diverse origins and parentage in farmer varieties from Mesoamerica. This information thus will be highly useful for conservation and utilization of cacao germplasm from this region.