Chloroplast DNA non-coding sequences variation in Aegilops tauschii Coss.: evolutionary history of the species

Sequences of four chloroplast DNA non-coding regions, about 3,000?bp in total, were analysed in 112 Aegilops tauschii accessions, 56 of ssp. tauschii and 56 of ssp. strangulata, representing all of the species range. One inversion, 8 insertions/deletions, 18 base pair substitutions and 5 microsatellite loci were found. The data revealed that Ae. tauschii originated in Caucasia. Neither of the two Ae. tauschii subspecies was an ancestor to one another. Aegilops tauschii divided into ssp. tauschii and ssp. strangulata at the very beginning of its existence as a species. Subspecies tauschii was the first to start geographic expansion and relatively rapidly occupied a vast area from Caucasia—eastward up to central Tien Shan and western Himalayas. In contrast to ssp. tauschii, geographic spread of ssp. strangulata was a complicated, multi-stage and slow process. At the beginning of ssp. strangulata evolutionary history its major phylogenetic lineage for a lengthy time span had existed as a small isolated population. Several forms of ssp. strangulata, better adapted to relatively moister and cooler habitats, had originated. Each of these forms has gradually forced out ssp. tauschii from some part of its area in the west, up to central Kopet-Dag.     

Stripe rust resistance in <Emphasis Type="Italic">Aegilops tauschii</Emphasis> and its genetic analysis

Aegilops tauschii Coss., the D-genome progenitor of common wheat (Triticum aestivum L.) includes two subspecies, tauschii and strangulata (Eig) Tzvel. Subspecies tauschii has a wide geographic distribution spreading westwards to Turkey and eastwards to Afghanistan and China, while ssp. strangulata has a narrower distribution occurring only in two disjoined regions, southeastern Caspian Iran and Transcaucasia. A collection of 56 Ae. tauschii accessions was screened at adult stage against a mixture of pathotypes of stripe rust prevalent in the current wheat production in China. The results for three crop seasons indicated that among the 38 ssp. tauschii accessions, 37 were susceptible and only one was resistant, while all the 18 ssp. strangulata accessions were resistant. These results indicated that stripe rust resistance was related to taxonomic origin. Further genetic analysis revealed the resistance of stripe rust in ssp. strangulata accession AS2388 was conferred by a single dominant gene.     

Distribution and diversity of Aegilops tauschii in Iran

The wide morphological variation of Aegilops tauschii has led to the distinction of different subspecies; a typical ssp. tauschii and a second ssp. strangulata. However some researchers pointed out the existance of the intermediate form among morphologically distinguished subspecies. Distribution, diversity and the relationship between different subspecies and the intermediate form were evaluated in the Iranian Ae. tauschii collection. This collection was classified to 15 different populations according to morphological similarities and the collecting origin of accessions. The highest variation was found in tauschii population of Golestan followed by tauschii populations of Gilan and Ardebill, whereas the lowest variation was observed in tauschii populations of central Iran. Two discriminant functions suggested that the length of rachis node and spikelet glume, particularly, the length/width ratios of these traits had the highest impact on identification of different forms. Mahalanobis distances (D ² ) between the two subspecies along with intermediate form on the multidimensional scaling plot showed that the intermediate form is more similar to ssp. tauschii than ssp. strangulata. Although, the diversity within the ssp. strangulata was not very high, it widely affected the diversity of Iranian accessions of Ae. tauschii through continues crossing with the more diversed subspecies, tauschii, during thousands of years. This fact had lead to expansion of its distribution from its origin to Northern Khorasan, Northern Semnan and Eastern Ardebill by producing the intermediate form.     

Geographic patterns of Got1, Got2, Got3 and Est2 enzyme-encoding genes allelic variation in Aegilops tauschii Coss.

Geographic patterns of Got1, Got2, Got3 and Est2 enzyme-encoding genes allelic variation were investigated among 322 accessions of Aegilops tauschii Coss., 161 accessions of ssp. tauschii and 161 accessions of ssp. strangulata, representing all the species area. It was found that: (1) in the two ecologically different subspecies, ssp. tauschii and ssp. strangulata, the patterns of allelic variation of the four genes differ greatly; (2) the same allozymes have originated several times independently in different Ae. tauschii local populations; (3) allelic variation of Got1, Got3 and Est2 in ssp. strangulata corresponds to climatic conditions. The data obtained reflected that Ae. tauschii has been inhabiting its area from ancient times; and allelic variation patterns of Got1, Got2, Got3, Est2 loci were mostly formed by natural selection. Further investigations of these loci with molecular genetic methods are prospective for understanding the peculiarities of Ae. tauschii evolution.     

Genetic diversity and gene-pool subdivisions of diploid D-genome <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. (Poaceae) in Iran as revealed by AFLP

The diploid goatgrass Aegilops tauschii is considered the D-genome donor of bread wheat and has probably a centre of diversity in north of Iran. In order to measure the genetic diversity of and the relationships among different populations, varieties and subspecies belonging to Ae. tauschii in Iran, DNA was extracted from 48 accessions of Ae. tauschii collected across the geographic range of the species in the Country and the genetic diversity was assessed using AFLPs based on eight PstI/MseI +3 primer pairs resulted in 277 bands, 198 of which were polymorphic. High level polymorphism was detected, with an average of polymorphism rate of 0.715; relatively low genetic similarity (0.455) between accessions and significant difference between the lowest (0.179) and the highest genetic similarity (0.817). The Iranian Ae. tauschii populations showed high level of genetic diversity. The populations studied were divided into two groups: one group was mainly representing Northern populations collected from Southern Caspian Sea shore and the other group was mainly representing Northeast and Northwest populations. Based on the results of this study, it can be suggested that Ae. tauschii possesses two separate gene-pools in Iran: Northern and Northeastern–Northwestern. Considering the needs for introducing new characteristics and alleles for wheat improvement purposes, Ae. tauschii Iranian gene-pool is assumed to be of high importance for more investigation in the future.     

Evaluation of geographic distribution of high-molecular-weight glutenin subunits (HMW-GS) in wild and cultivated Triticum species

Variation of high-molecular-weight glutenin subunit (HMW-GS) in 632 wild and cultivated Triticum accessions was investigated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A total of 11 alleles of HMW-GS in diploid species, 22 in tetraploid species, and 15 in hexaploid species were detected. Diploid species on Glu-1A locus and tetraploid species Glu-1B locus showed the highest diversity, respectively. Tetraploid species had the highest level of diversity on three Glu-1 loci, followed by hexaploid and diploid, based on Shannon’s information index, Nei’s genetic diversity, and percentage of polymorphic loci. Molecular variance analysis confirmed main variance of HMW-GS within species, regions, and locations, respectively. Variance among species and regions was enhanced gradually with the increase of ploidy. Significant non-random distributions between the phylogenic trees of HMW-GS and the locations of accessions were tested by GenGIS software, indicated that geographic factors played an important role along the different orientations in the spread of Triticum species. We found one original diversified center in diploid what located around Elazig, Malatya, Gaziantep, Urfa, and Kiziltepe in Turkey, and three diversified centers in tetraploid wheat, including Turkey–Armenia–Georgia–Iran, Portugal–Spain, and Ethiopia, respectively, and two diversified adjacent areas between Turkey and Switzerland and around Turkey, Georgia, and Armenia. The original center of diploid species located in southeast Turkey, where the unexpressed 1Ay subunit was mainly distributed in T. urartu, could be one of the candidate regions of polyploidization of Triticum L. The regional distribution of HMW-GS and species also provided geographic evidences for the existence of founder effect on the spread of Triticum species. The present study suggests that integrating genetic diversity with geographic characterization in Triticum could very useful for collection, conservation, and utilization, as well as for research microevolution and domestication.     

Haplotype variations of gene Ppd-D1 in Aegilops tauschii and their implications on wheat origin

The Ppd-D1 controlling photoperiod response is an important gene for wheat adaptation since it affects heading time. In the present study, three haplotypes, i.e. haplotype I without deletion, haplotype II with a 24?bp deletion, and haplotype III with two deletions of 24 and 15?bp, were identified in the upstream of the coding region in 80 Ae. tauschii accessions. The haplotype distribution was related to subspecies taxon. All typical ssp. tauschii accessions had haplotype I, whereas all ssp. strangulata had haplotype III. The three haplotypes were observed in Ae. tauschii with morphologically intermediate forms between the two typical subspecies. Present results supported that ssp. strangulata or intermediate form was the D-genome donor of common wheat since only haplotype III were found in wheat. Moreover, a 16?bp deletion in exon 8 of gene Ppd-D1 exists in common wheat. However, none of Ae. tauschii accessions analyzed had the 16?bp deletion.     

Evaluation of genetic diversity in a Camelina sativa (L.) Crantz collection using microsatellite markers and biochemical traits

A genomic DNA library enriched with GA/TC repeats from Camelina sativa variety Calena has been analysed. After sequencing of about 200 randomly selected clones, approximately 60 % of them showed to contain simple or compound microsatellites with a high number of repeats. Among all microsatellite markers analysed 15 primer pairs amplified polymorphic fragments. Forty C. sativa accessions of different origin were genotyped with 15 microsatellite markers that generated 134 alleles with an average of 8.93 alleles per locus. The observed heterozygosity (Ho) among the accessions ranged from 0.0 to 0.15 with an average of 0.0370, whereas the average of expected heterozygosity (He) among accessions was 0.2769. The analysis of the average total heterozygosity (H_T = 0.651), the intrapopulation genetic diversity (H_S = 0.260), the interpopulation genetic diversity (D_ST = 0.391) and the coefficient of genetic differentiation among populations (G_ST = 0.574) demonstrated that 57.4 % of the genetic diversity is among the accessions, while 42.6 % resides within them. Phylogenetic tree of the 40 C. sativa accessions was constructed based on Nei’s genetic distance. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram shows, except for CAM108 and CAM170, a clear discrimination among C. sativa accessions grouping them in five subgroups. ANOVA analysis indicates significant differences in some biochemical and agronomic parameters among the C. sativa accessions grouped according to Nei’s genetic distance. The result of the Tukey HSD test demonstrated that the A4 subgroup showed a significant higher TWS and linoleic acid (LA) content, while the subgroup A1 showed a significant higher linolenic and lower LA content compared to the remaining groups.     

Spatially structured genetic diversity of the Amerindian yam (Dioscorea trifida L.) assessed by SSR and ISSR markers in Southern Brazil

Dioscorea trifida L. (Dioscoreaceae) is among the economically most important cultivated Amerindian yam species, whose origin and domestication are still unresolved issues. In order to estimate the genetic diversity maintained by traditional farmers in Brazil, 53 accessions of D. trifida from 11 municipalities in the states of São Paulo, Santa Catarina, Mato Grosso and Amazonas were characterized on the basis of eight Simple Sequence Repeats (SSR) and 16 Inter Simple Sequence Repeats (ISSR) markers. The level of polymorphism among the accessions was high, 95 % for SSR and 75.8 % for ISSR. The SSR marker showed higher discrimination power among accessions compared to ISSR, with D parameter values of 0.79 and 0.44, respectively. Although SSR and ISSR markers led to dendrograms with different topologies, both separated the accessions into three main groups: I—Ubatuba-SP; II—Iguape-SP and Santa Catarina; and III—Mato Grosso. The accessions from Amazonas State were classified in group II with SSR and in a separate group with ISSR. Bayesian and principal coordinate analyzes conducted with both molecular markers corroborated the classification into three main groups. Higher variation was found within groups in the AMOVA analysis for both markers (66.5 and 60.6 % for ISSR and SSR, respectively), and higher Shannon diversity index was found for group II with SSR. Significant but low correlations were found between genetic and geographic distances (r = 0.08; p = 0.0007 for SSR and r = 0.16; p = 0.0002 for ISSR). Therefore, results from both markers showed a slight spatially structured genetic diversity in D. trifida accessions maintained by small traditional farmers in Brazil.     

Purification and Characterization of the Glutenin Subunits of Triticum tauschii,Progenitor of the D Genome in Hexaploid Bread Wheat

N-terminal amino acid sequences and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) molecular weights have been determined for high-performance liquid chromatography (HPLC)-purified high molecular weight (HMW) and low molecular weight (LMW) glutenin subunits (GS) of Triticum tauschii ssp. strangulata, contributor of the D genome to hexaploid bread wheat. The use of three different extraction procedures resulted in similar glutenin preparations. On the basis of N-terminal sequences, the same types of glutenin subunits that have been reported in bread and durum wheats (HMW-GS of both the x and y types and LMW-GS of the LMW-s, LMW-m, α-, and γ-types) were found in T. tauschii. However, the HMW-GS in T. tauschii were in greater proportion relative to LMW-GS when compared to reported values for a bread and durum wheat. Our results support the likelihood that differences in the proportions of the various subunits contributed by the A, B, and D genomes, rather than qualitative differences in the types of subunits, are responsible for the major differences in quality characteristics between bread wheat and durum wheat.     

Microsatellite analysis of genetic diversity and population genetic structure of <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss. in Northern Iran

Genetic diversity and population genetic structure of Aegilops tauschii in Northern Iran were studied based on nine microsatellite loci. A high level of genetic diversity was observed from the accessions collected from six regions (provinces). These accessions include 79 samples of the two subspecies (tauschii and strangulata), the intermediate form (among morphologically distinguished subspecies) and ten accessions of Triticum aestivum. The nine microsatellites revealed a total of 141 alleles, with an average of 15.7 alleles per locus. A comparison of the parameters showing genetic diversity, including the observed heterozygosity (Ho), gene diversity (He) and Shannon’s information index (I) of Ae. tauschii accessions from different provinces in Northern Iran, indicated that subsp. tauschii possesses the highest genetic diversity, followed by intermediate form. Genetic distance between subsp. strangulata and subsp. tauschii was low, confirming high gene flow between these two subspecies. However, intermediate form was more distinct from both of them. It was also found that the genetic diversity of T. aestivum is obviously lower than that of Ae. tauschii accessions. Moreover, the level of genetic diversity for Gilan, Golestan and Mazanderan provinces was higher than for Ardebil, Ghazvin and Semnan provinces, suggesting that these regions may provide a readily available source of potentially useful variation for wheat improvement.     

Genetic diversity among Polish landraces of common oat (Avena sativa L.)

All over the world about 220,000 accessions of the genus Avena is preserved in gene banks. Polish oats collection, in the National Centre for Plant Genetic Resources, consists of about 2,500 accessions. More than 80 % of them belong to the species Avena sativa. Only 136 accessions have the landrace or traditional cultivar status and 91 of them were collected in Poland. The main objective of this study was to estimate genetic diversity of a set of 67 Polish landraces using inter simple sequence repeat (ISSR) markers. We also tried to determine whether genetic diversity depended on the region and altitude of collection site and if there was a relationship between DNA polymorphism and some morphological traits. The accessions were collected between years 1973 and 1999 from areas of Northern, Southern and Eastern Poland. The obtained results indicated a relatively low diversity (0.09–0.37) of the common oat landraces. Analysis of molecular variation (AMOVA) showed that there were no significant differences within designated geographic regions. It indicated that at the same time the altitude of collection site had significant effect on the genetic differentiation of the accessions. AMOVA also showed the presence of variance between groups formed on the basis of the colour of lemma. White lemma accessions were also characterized by highest genetic distinctiveness. Considering the evident impact of local climatic conditions on the genetic distinctiveness of Polish landraces utility of them in the Polish breeding programs should be considered.     

Agro-morphological and molecular variability in <Emphasis Type="Italic">Triticum boeoticum</Emphasis> accessions from Zagros Mountains,Iran

Genetic erosion in cultivated wheat provides a good reason for studying genetic diversity in crop wild relatives. In the present study, genetic diversity of 32 accessions belonging to T. boeoticum species collected from different parts of Iran were evaluated using 13 morphological traits as well as ten inter-simple sequence repeat primers. Statistical analysis for morphological traits showed significant differences among accessions (except number of fertile tillers and total tillers per plant). In principal component analysis, the first three PCA showed 82.65 % of the total morphological variation. Based on the morphological traits, accessions were separated into two main groups by cluster analysis. In molecular analysis, polymerase chain reactions amplified 105 DNA fragments, out of which, 95 (90.47 %) were polymorphic. From geographic perspective, the accessions sampled from western and southwestern of Iran showed the highest and lowest polymorphism, respectively. However, the maximum values of effective number of alleles (Ne), Nei’s gene diversity (He) and Shannon’s information index (I) was related to accessions collected from NW regions. Also, according to cluster analysis and PCoA plot genetic diversity was not related to geographical distribution. Overall, our results revealed a remarkable level of genetic diversity among studied Iranian T. boeoticum accessions; especially accessions collected from Kermanshah and Lorestan provinces, which can be of interest for future breeding programs. So, conservation of germplasm of these areas is recommended.     

Genetic diversity and relationships of wild and cultivated <Emphasis Type="Italic">Zanthoxylum</Emphasis> germplasms based on sequence-related amplified polymorphism (SRAP) markers

The genus Zanthoxylum, belonging to Rutaceae, has a long history of cultivation both for economic and chemical values in China. To effectively conserve and sustainably utilize this genus resource, a study on genetic diversity and relationships of Zanthoxylum germplasms was carried out by employing SRAP markers. We used 16 primer combinations to assess genetic variations and relationships among 175 accessions from eight cultivated provenances, including Shandong, Henan, Shanxi, Shaanxi, Gansu, Sichuan, Guizhou and Yunnan. A total of 145 clear repetitive and intense bands were yielded, and the percentage of polymorphic bands was 100 % for per primer combination, indicating a relatively high diversity among Zanthoxylum germplasms. From a geographic perspective, the highest genetic diversity level was observed within Guizhou provenance (N _a  = 1.97, Ne = 1.52, H = 0.31, I = 0.46) while Henan provenance had the lowest genetic diversity (N _a  = 1.68, Ne = 1.45, H = 0.25, I = 0.37). Based on AMOVA results, the abundant genetic variation was mainly caused by variation of intra-provenances (84.96 %), rather than among provenances (15.038 %). The results indicated low genetic differentiation (G _st  = 0.133) and high gene flow (N _m  = 3.2605) among provenances. The neighbor-joining tree revealed that the 175 accessions could be divided into four groups, and groupings indicated a divergence between the cultivated accessions of Zanthoxylum bungeanum Maxim. and Z. armatum DC. Moreover, three accessions of Z. piperitum DC. var. inerme without prickles introduced from Japan gathered one cluster. Cluster IV is composed of accessions of different geographical origin, including 11 wild species and 10 cultivated accessions of Z. bungeanum. The cluster analysis also reflected a relatively close relationship between the geographical origins and the classification of accessions in cluster I. Structure analysis indicated that collected Zanthoxylum accessions could be divided into two major groups. The information obtained from our research would benefit to make use of Zanthoxylum germplasms and assist the management of a Zanthoxylum germplasms collection.     

Spatial patterns of adenylate kinase,catalase, endopeptidase and fructose-1,6-diphosphatase encoding genes allelic variation in <Emphasis Type="Italic">Aegilops tauschii</Emphasis> Coss.

Investigation of spatial patterns of adenylate kinase, catalase, endopeptidase and fructose-1,6-diphosphatase encoding genes (Ak, Cat1, Cat2, Ep, Fdp) allelic variation in Aegilops tauschii was carried out. About 300 accessions, representing all the species range were taken for the study. Cat2 and Fdp loci are completely monomorphic in ssp. strangulata and in the western part of ssp. tauschii range, as well. Both Cat2 and Fdp are highly polymorphic in the eastern part of ssp. tauschii range, with the patterns of this polymorphism being discordant in these two loci. Ak ¹⁰⁸, a rare allele with sporadical spatial occurrence, was found in ssp. tauschii only. On the contrary, Ak ⁹² is absent in ssp. tauschii: it is the most common Ak allele in ssp. strangulata in Precaspian Iran, the most moist part of the area, and is very rare in other parts of ssp. strangulata area. Ep is a highly polymorphic locus with the highest level of variation in the west of Ae. tauschii area, where this species had originated. Ep allele variation patterns are rather similar in ssp. tauschii and ssp. strangulata. The data reveal the adaptive nature of Ak, Cat2, and Fdp allele variation, while Ep polymorphism seems to be mostly neutral.     

Differentiation between fiber and drug types of hemp (Cannabis sativa L.) from a collection of wild and domesticated accessions

Accessions of wild and domesticated hemp (Cannabis sativa L.) originating from Colombia, Mexico, California, Bolivia, Thailand, Afghanistan, Serbia, Hungary, south Africa and different regions of China, were studied by means of DNA polymorphisms in order to discriminate between drug and fiber types. Analysis of molecular variance (AMOVA) was used to partition the total genetic variance within and among populations. The significance of the variance components was tested by calculating their probabilities based on 999 random permutations. AMOVA revealed 74 % variation among accessions and 26 % within accessions, all AMOVA variation was highly significant (P < 0.001). The cluster analysis of molecular data, grouped accessions into eight clusters and gave a matrix correlation value of r = 0.943, indicating a very good fit between the similarity values implied by the phenogram and those of the original similarity matrix. In this study, DNA polymorphisms could discriminate the fiber and drug types, and accessions were grouped in accordance to their classification and uses. In addition, seed size variation and micromorphological characters of seeds were studied by means of a scanning electron microscope (SEM). Seeds varied significantly in size, and were bigger in the fiber types. SEM analysis exhibited variation of micromorphological characters of seeds that could be important for discriminating the fiber or drug types.     

Water yam (Dioscorea alata L.) diversity pattern in Brazil: an analysis with SSR and morphological markers

Substantial progress was made in the last decade in understanding the diversity in Dioscorea species. However, most of the studies so far concentrated in the ‘yam belt in Africa’. We present a genetic diversity and structure analysis among commercial and local varieties of water yam (Dioscorea alata L.) in Brazil using microsatellite and morphological markers. Twelve microsatellite primers were used to generate DNA profiles of 72 local varieties and 17 commercial accessions of water yam collected in four different regions in Brazil. Also, four morphological traits were evaluated on individual plants under field conditions. The morphological characterization showed considerable diversity. High polymorphism was found with 100 % polymorphism observed for 11 primers and a discriminating power value of 0.92, on average. We did not observe a strong population structure among sampling regions, while most of the genetic diversity was concentrated within regions (95.9 %). Analysis of the relationship between accessions did not clearly separate the local and commercial genotypes. However, the molecular analyses of D. alata showed high intraspecific diversity in local accessions from different regions in Brazil, with the highest Shannon index value (H′ = 0.41) for the Southeast region. These results confirmed an admixture of accessions in all sampling regions, consistent with the lack of a significant correlation between geographic and genetic distances, suggesting that farmers exchanged water yam materials extensively. The genetic diversity can be explained by the result of a continuous exchange of accessions by farmers throughout Brazil.     

Analysis of genetic diversity and population structure of 135 dill (Anethum graveolens L.) accessions using RAPD markers

Dill (Anethum graveolens L.) leaf, seed and their essential oil are rich source of antioxidants. The plant is native in Southwest Asia and is cultivated in Europe, India and the United States. This study evaluated the genetic diversity structure of 135 accessions of A. graveolens from different continents, based on random amplified polymorphic DNA (RAPD) markers. The selected 10 RAPD primers generated a total of 142 highly reproducible bands, among which 89 were polymorphic. Percentage of polymorphism varied from 41.17 % (OPB20) to 92.85 % (OPB15) with an average of 77.74 %. A relatively high genetic diversity was detected among all the accessions with the Nei’s genetic diversity (H) values ranged from 0.346 (OPB07) to 0.444 (OPB18) with a mean of 0.401. When estimated for Shannon’s information index (I), it has ranged from 0.530 (OPB12) to 0.652 (OPB18), the mean was observed as 0.581. The respective values of H and I were found to be the highest value for primer OPB18. Cluster analysis of RAPD data using UPGMA algorithm based on Nei’s genetic similarity matrix placed the 135 accessions into two main clusters. Although a number of groups can be identified, the clusters show little to no association with the geographic origin of the material. The implication of the results of this study in developing a strategy for the conservation and breeding of dill germplasm are discussed.     

Aegilops tauschii: Genetic Variation in Iran

All the 79 Aegilops tauschii Coss. accessions of Iranian origin from Prof. Kihara’s collection were analyzed electrophoretically. Of 23 enzyme-encoding loci studied, 11 were polymorphic. In Iran Ae. tauschii is presented by ssp. tauschii and ssp. strangulata which distinctly differ genetically, morphologically and ecologically. Variation patterns of low polymorphic locus Aco2 and highly polymorphic Ep are similar in both subspecies. In contrast, variation of Acph1, Ak, Est2, Est5, Got1, Got2, Got3 and Lap is a set of diverse patterns which markedly differ between subspecies and natural regions also, implying that natural selection is involved.     

Target region amplification polymorphism (TRAP) for assessing genetic diversity and marker-trait associations in chickpea (Cicer arietinum L.) germplasm

The effective utilization of crop diversity held in genebanks depends on our knowledge of useful traits and available markers associated with the target traits. Target region amplification polymorphism (TRAP) was used to evaluate the genetic diversity and underlying relationships among 263 accessions of chickpea landraces maintained by the USDA-ARS Western Regional Plant Introduction Station in Pullman, WA, USA. Two-hundred sixty-two TRAP markers were amplified by eight primer combinations. Altogether, 110 (42 %) markers were polymorphic, the other 152 (58 %) displayed no variation. These polymorphic markers revealed important differences among the accessions, with an estimated, mean pair-wise genetic distance of 25.82 %, ranging from 2.8 to 50.0 %. Genetic distance analysis divided the accessions into two major groups, with 113 and 150 accessions each, and substantial association between molecular diversity and geographic origin was evident. Bayesian analysis of population structure revealed two groups (K = 2) with evidence for six sub-groups. Additionally, the population structure of a subset of 110 lines was determined (K = 3) for testing marker-trait associations (MTAs). Phenotypic traits included the concentrations of protein and nine mineral elements in the seeds. Two MTAs were significant (p < 0.01) for concentrations of Ca and K, and three MTAs were significant for Cu and Ni concentrations. The results indicate that this population is useful for genome-wide association studies on other economic traits given the level of genetic diversity uncovered and the marker-trait associations in seed minerals discovered.