Genetic diversity comparison between Chinese and Japanese soybeans (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.) revealed by nuclear SSRs

Soybean (Glycine max (L.) Merr.) has a long planting history in both China and Japan. In order to investigate the genetic relationship between Chinese and Japanese soybeans, 205 Chinese soybean accessions, that represent the seven different soybean ecotypes, and 39 Japanese soybean accessions from various regions were analyzed by using 46 SSR loci. In total 745 alleles were detected with an average allele number of 16.2 per locus. The allelic frequencies varied from 0.002 to 0.554 with an average of 0.06. Cluster analysis with UPGMA separated the Chinese accessions from Japanese accessions, suggesting that soybean in these two countries form different gene pools. When comparing the Japanese soybean with that from seven different Chinese soybean ecotypes, 164–200 alleles were specific to the Chinese accessions and 64–112 specific to the Japanese accessions. The comparison of SSRs diversity revealed that accessions from China exhibited more genetic diversity than those from Japan. The data were analyzed to resolve the genetic structure and to interpret the evolutionary relationships between groups. Three distinct groups were identified, corresponding to Japanese soybean, Northern China soybean, Southern China soybean and a mixed group in which most accessions were from central China. The results indicate that accessions from Japan are distinct from Chinese ones, and Japanese accessions had more close relationship with Chinese northeast spring and southern spring ecotypes. We further analyzed five agronomic trait-related SSR loci and found that the preponderant alleles are different in Chinese and Japanese soybean. Our study provides important insights into further utilization of Japanese soybean in Chinese soybean breeding.     

Detection of genetic integrity of conserved maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm in genebanks using SNP markers

Twenty maize landrace accessions regenerated and conserved in five maize genebanks were investigated for genetic integrity using 1,150 Single Nucleotide Polymorphisms (SNPs) and 235 SNP haplotypes. The genetic diversity of three accessions changed significantly in terms of the average number of alleles per locus. Ten out of twenty accessions had significantly different SNP allelic frequencies, either after regeneration or in the same accession held in different genebanks. The proportion of loci with significant changes in SNP allelic frequency was very low (37/1,150). Changes in the major allelic frequency (MAF) for the majority of SNP loci (60.2–75.2%) were less than 0.05. For SNP haplotypes, the genetic diversity of four accessions changed significantly in terms of average number of haplotype alleles and polymorphic information content (PIC) per locus. The proportion of SNP haplotype alleles lost in the later generations ranged between 0 and 22.6%, and at the same time 0–19.9% of the SNP haplotype alleles appeared in later generations, however, these were absent in the earlier generations. Dynamic changes in genetic integrity, in terms of presence and absence of genes (alleles), by both SNP and SNP haplotype analysis were detected during regeneration. A suboptimum number of ears harvested in one generation can be combined with those from another, repeated regeneration to capture the diversity of the previous generation. Use of molecular markers during regeneration of accessions can help in understanding the extent of genetic integrity of the maize accessions in ex situ genebanks and in recommending the best practice for maintaining the original genetic diversity of the genebank accessions.     

Genetic structure and differentiation among grapevines (<Emphasis Type="Italic">Vitis vinifera</Emphasis>) accessions from Maghreb region

Three gene pools representative of Vitis vinifera L. subsp. vinifera (=subsp. sativa Beger) growing in the Maghreb regions (North Africa) from Tunisia (44), Algeria (31) and Morocco (18) and 16 wild grape accessions (Vitis vinifera L. subsp. sylvestris (Gmelin) Beger) from Tunisia were analysed for genetic diversity and differentiation at twenty nuclear microsatellites markers distributed throughout the 19 grape chromosomes. 203 alleles with a mean number of 10.15 alleles per locus were observed in a total of 109 accessions. Genetic diversities were high in all populations with values ranging from 0.6775 (Moroccan cultivars) to 0.7254 (Tunisian cultivars). F _st pairwise values between cultivated grapevine populations were low but found to be significantly different from zero. High F _st pairwise values were shown between wild and cultivated compartments. Two parent offspring relationships, two synonyms and two clones of the same cultivar were detected. The rate of gene flow caused by vegetative dissemination of cultivated grapevine plants was not sufficient to genetically homogenise the pools of cultivars grown in different regions. The Neighbour Joining cluster analysis showed a clear separation according to geographical origins for the cultivated grapevines gene pools and revealed a high dissimilarity between cultivated and wild grapevine. However, three cultivars (Plant d’Ouchtata 1, Plant de Tabarka 3 and Plant d’Ouchtata 3) are very close to wild accessions and may result from a hybridisation between cultivated and wild accessions. The high level of differentiation between cultivated and wild accessions indicates that the cultivated accessions do not derive directly from local wild populations but could mostly correspond to imported materials introduced from others regions during historical times or derived from crossing between them.     

Natural introgression from cultivated soybean (<Emphasis Type="Italic">Glycine max</Emphasis>) into wild soybean (<Emphasis Type="Italic">Glycine soja</Emphasis>) with the implications for origin of populations of semi-wild type and for biosafety of wild species in China

Introgression from soybean cultivars to its wild progenitor species is an interesting antidromic recombination in natural ecosystem with many consequences, including the alteration of genetic diversity, the origin of semi-wild soybean, and implication for biosafety of the wild progenitor species with future release of the genetically modified varieties. Although such interspecific introgression is not suspected to be ubiquitous in the sympatric regions of wild and cultivated soybeans, the documentations published based on some molecular experimental analyses on the introgression have been little substantiated by the occurring process morphologically and remain deficient for unquestionable evidence, owing to the lack of actual insight into the population dynamics. Here, we found the phenomenon of gene escape and presented the evidence for occurrence of introgression from soybeans into the wild species and for how originates about the semi-wild type soybean based on morphological investigation of population dynamics. Our results suggest that more attention should be paid to the escape of genetically modified genotypes to safeguard the biosafety of wild soybean gene pool, if GM soybeans are released in China, the place of origin of cultivated soybeans.     

Identification and genetic structure of wild Italian <Emphasis Type="Italic">Humulus lupulus</Emphasis> L. and comparison with European and American hop cultivars using nuclear microsatellite markers

Nine genic SSR loci were used to evaluate the genetic diversity and identify accessions in wild Italian Humulus lupulus L., in comparison with widely cultivated European and U.S. commercial cultivars. A collection of 80 wild hop samples from Italy and 43 hop cultivars from Europe and U.S., were characterized. Allelic frequency analysis revealed 65 distinct Italian genotypes and differentiated all the commercial cultivars; moreover, specific alleles were observed for wild and cultivated hops. The number of alleles identified in the wild population were 104 and 123 within all the accessions. The maximum polymorphic information content was evidenced for locus HlGA23 in the Italian wild population and in the whole set of accessions (0.905 and 0.902 respectively). The dendrogram constructed from Euclidean distance with the UPGMA method showed two main clusters, one including commercial American and European accessions and one mostly composed by wild Italian accessions. Model-based clustering (Bayesian method) placed the accessions into five germplasm groups, one of which was characterized by Italian genotypes only. The study showed for the first time the great biodiversity present in Italy, and the remarkable differences with European and American hops. It was also found that within the population of north-central Italy a large genetic variability is present, suited to be studied and exploited; this genetic wealth could be used in future breeding programs in order to develop new hop varieties carrying characteristics useful for brewers.     

Genetic structure and history of Swiss maize (<Emphasis Type="Italic">Zea mays</Emphasis> L. ssp. <Emphasis Type="Italic">mays</Emphasis>) landraces

Between 1930 and 2003 with emphasis on the 1940s maize landraces (Zea mays L. ssp. mays) from all over Switzerland were collected for maintenance and further use in a new Swiss breeding program. The genetic relationship and diversity among these accessions stored in the Swiss gene bank is largely unknown. Our hypothesis was that due to the unique geographic, climatic, and cultural diversity in Switzerland a diverse population of maize landraces had developed over the past three centuries. The aims were to characterize the genetic diversity of the Swiss landraces and their genetic relationship with accessions from neighbouring regions as well as reviewing their history, collection, and maintenance. The characterization and grouping was based on analyses with ten microsatellite markers. Geographic, cultural, and climatic conditions explained a division in two distinct groups of accessions. One group consisted of landraces collected in the southern parts of Switzerland. This group was related to the Italian Orange Flints. The other group contained accessions from northern Switzerland which were related to Northern European Flints in particular German Flints. Historic evidence was found for a frequent exchange of landraces within the country resulting in a lack of region-specific or landrace-specific genetic groups. The relatively large separation between the accessions, indicated by high F _ST (0.42), might be explained partly by a bottleneck during the collection and maintenance phase as well as by geographical and cultural separation of north and south of the country. Due to the high genetic diversity, the accessions here are a potential resource for broadening the European flint pool.     

AFLP analysis of genetic diversity in leafy kale (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. convar. <Emphasis Type="Italic">acephala</Emphasis> (DC.) Alef.) landraces,cultivars and wild populations in Europe

AFLP markers were used to characterize diversity and asses the genetic structure among 17 accessions of kale landraces, cultivars and wild populations from Europe. The range of average gene diversity in accessions was 0.11–0.27. Several landraces showed higher levels of diversity than the wild populations and one cultivar had the lowest diversity measures. The landraces that were most genetically diverse were from areas where kales are known to be extensively grown, suggesting in situ conservation in these areas as a supplement to storage of seeds in gene banks. An analysis of molecular variance (AMOVA) showed that 62% of the total variation was found within accessions. For most accessions, genetic distance was not related to geographic distance. Similarities among accessions were probably not caused by recent gene flow since they were widely separated geographically; more likely the relationship among them is due to seed dispersal through human interactions. Our results indicate that a kale population found in a natural habitat in Denmark was probably not truly wild but most likely an escape from a cultivated Danish kale that had subsequently become naturalized.     

Diversity analysis of Central Asia and Caucasian lentil (<Emphasis Type="Italic">Lens</Emphasis><Emphasis Type="Italic">culinaris</Emphasis> Medik.) germplasm using SSR fingerprinting

Diversity analysis was performed among 39 cultivated lentil (Lens culinaris Medik.) accessions of Central Asia and Caucasian origin using five highly polymorphic microsatellite markers. A total of 33 alleles determined ranging from 3 to 8 per locus. Estimated gene diversity value for 33 loci was 0.66. Genetic similarity indices among 39 accessions ranged from 0.24 to 1.0. Cluster analysis using the unweighted pair group method with arithmetic mean method classified accessions into six major groups at 0.5 similarity coefficient. More than half accessions from Tajikistan formed large cluster. On the other hand, a few accessions from each country showed unique genotypes. Overall, most of the accessions, except ones with closely related origin, were distinguished by the present high quality DNA fingerprinting. This molecular diversity information gives important basis for conservation strategy in gene bank and exotic germplasm introduction in breeding programs in Central Asia and Caucasian countries.     

Genetic variation among populations of wild safflower, <Emphasis Type="Italic">Carthamus oxyacanthus</Emphasis> analyzed by agro-morphological traits and ISSR markers

Wild species of safflower, Carthamus oxyacanthus Bieb., is highly crossable with cultivated species, C. tinctorius L. and could be directly exploited in broadening safflower gene pool and improving the crop for biotic and abiotic stress environments. In this study, genetic diversity among accessions of C. oxyacanthus and their relationships with cultivated safflower were evaluated using agro-morphological traits and polymorphic inter-simple sequence repeats (ISSR) markers. Significant variation was observed among accessions particularly for seeds per capitulum, seed yield per plant, harvest index and capitula per plant. Cluster analysis based on agro-morphological traits classified the wild accessions in two groups according to their geographical regions, and separated them from the cultivated genotypes. ISSR marker also revealed a high genetic variation among the accessions, and cluster analysis based on this marker divided genotypes into four groups, with cultivated ones in a separate clade. Genetic variation observed among the wild safflower germplasm at the DNA level was higher than the agro-morphological traits, indicating that ISSR is an effective marker system for detecting diversity among safflower genotypes and their genetic relationships. Accessions of C. oxyacanthus with high genetic relationship to cultivated species could be used for interspecific hybridization in breeding programs of safflower.     

The genetic characterization of Turkish watermelon (<Emphasis Type="Italic">Citrullus lanatus</Emphasis>) accessions using RAPD markers

Genetic diversity of the Turkish watermelon genetic resources was evaluated using different Citrullus species, wild relatives, foreign landraces, open pollinated (OP) and commercial hybrid cultivars by RAPD markers. The germplasm was consisted of 303 accessions collected from various geographical regions. Twenty-two of 35 RAPD primers generated a total of 241 reproducible bands, 146 (60.6%) of which were polymorphic. Based on the RAPD data the genetic similarity coefficients were calculated and the dendrogram was constructed using UPGMA (Unweighted pair-group method with arithmetic average). Cluster analysis of the 303 accessions employing RAPD data resulted in a multi-branched dendrogram indicating that most of the Turkish accessions belonging to var. lanatus of Citrullus lanatus (Thunb.) Matsum et Nakai were grouped together. Accessions of different Citrullus species and Praecitrullus fistulosus (Stocks) Pangalo formed distant clusters from C. lanatus var. lanatus. Among 303 accessions, a subset of 56 accessions was selected representing different groups and a second dendrogram was constructed. The genetic similarity coefficients (GS) within the Turkish accessions were ranged from 0.76 to 1.00 with 0.94 average indicating that they are closely related. Taken together, our results indicated that low genetic variability exist among the watermelon genetic resources collected from Turkey contrary to their remarkable phenotypic diversity.     

Assessment of genetic diversity in Egyptian barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) genotypes using SSR and SNP markers

A barley core collection can be studied extensively and the derived information can be used to identify loci/genes for the genetic improvement of quantitative and qualitative traits. To assess genetic diversity, allelic variation and population structure of Egyptian barley, 134 barley genotypes collected from a different region along with 19 cultivated genotypes obtained from of the Egyptian Agricultural Research Center. All genotypes were analyzed with 261 polymorphic SSR and SNP alleles. The mean number of alleles per locus was 4 and PIC was 0.49, while the level of genetic diversity was 0.55 ranging from 0.03 to 0.82. The genotypes were assigned to three subpopulations that were consistent with their origins. The genetic variation within population was higher (51%) than among population at the molecular levels (F_ST =?0.491 when P?<?0.10). The level of polymorphic variation was highest in subpopulations-II, due to collected from different regions with different ear-types thus, expected to contain more diversity than local genotypes in subpopulations-I and subpopulations-III. The structured study found that the 153 barley genotypes are in harmony with clustering approaches using the SSR and SNP genotypic data in a neighbor-joining tree and principal components analysis, which identified three subpopulations. These results demonstrated genetic diversity among the Egyptian barley genotypes can be applied to suggest approaches, such as association analysis, classical mapping population development, and parental line selection in breeding programs. Therefore, it is necessary to use the exotic genotypes as the genetic resources for developing new barley cultivars in Egypt.     

The influence of European and American wild germplasm in hop (<Emphasis Type="Italic">Humulus lupulus</Emphasis> L.) cultivars

Microsatellite variation at the nuclear and chloroplast genomes was evaluated for wild European and wild American hops, in order to assess the genetic diversity and origin of cultivated hops. Seven nuclear loci and 32 chloroplast loci were used in the analysis of 182 hop accessions including wild European (68), wild American (48), and cultivars (66). A total of 116 alleles were identified using 7 nuclear microsatellites showing different averages of polymorphism and distribution in the wild American and European accessions and cultivars. Two main groups were established as revealed by several statistical analyses; one including European wild accessions and cultivars and a second group consisting of American wild accessions. Three polymorphic chloroplast microsatellite loci were detected, six alleles were scored which defined a total of five haplotypes that were exclusive or presented different distribution between American and European wild accessions. A major influence of the wild European haplotypes was detected among hop cultivars. To the best of our knowledge, this is the first work reporting the use of chloroplast microsatellites in hops.     

Sampling strategies and screening of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) germplasm for salt tolerance

Six hundred accessions of chickpea (Cicer arietinum L.) landraces and its wild relatives from 28 different countries, available at Australian Temperate and Field Crops Collection (ATFCC) were screened for tolerance to salt under greenhouse conditions using three sampling strategies; (1) random sampling of 200 accessions from different countries, (2) restricted random sampling of 200 accessions from geographical regions with salinity problems and high diversity (Middle East and West & South Asia) and (3) as for strategy 1 but with a reduced representation of accessions from the geographical regions used in strategy 2. Degree of salt tolerance was based on necrosis scores and shoot biomass reduction relative to unstressed controls at harvest after subjecting stressed plants to salt treatment from 21 to 42 days after sowing. There was a wide variation in salinity tolerance determined by both measures. For sampling strategies 1, 2 and 3 respectively; 24, 28 and 14% of accessions were salt tolerant. Accessions from the middle east and south Asian (regions with salinity problem, a long history of chickpea cultivation and high diversity) gave a higher probability (P < 0.01) of getting salt tolerant accessions.     

Origin of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) domestication genes

A number of genes that contribute to the domestication traits of cultivated rice have been identified. These include Sh4, Rc, PROG1 and LABA1, which are associated with non-shattering rachis, white pericarp, erect growth and barbless awns, respectively. The mutations giving rise to the “domestication alleles” of these genes are either invariable in cultivated rice, or have variability that is strictly associated with the phenotypic trait. This observation forms the basis to those current rice domestication models that envisage a single origin for the domesticated phenotype. Such models assume that the domestication alleles are absent or rare in wild rice, emerged under cultivation and spread across all rice groups by introgressive hybridization. We examined whole-genome sequencing datasets for wild and cultivated rice to test the former two assumptions. We found that the rc and laba1 alleles occur in wild rice with broad geographical distribution, and reach frequencies as high as 13 and 15%, respectively. These results are in agreement with previous observations of the prog1 and sh4 domestication alleles in wild populations. We also show that the diversity of the genomic regions surrounding the rc, laba1, prog1 and sh4 alleles in wild accessions is greater than that in cultivated rice, suggesting that these alleles emerged prior to domestication. Our findings indicate that the possibility that independent rice groups obtained identical domestication alleles directly from the wild population needs to be considered.     

Genetic differentiation and diversity of phenotypic characters in Chinese wild soybean (<Emphasis Type="Italic">Glycine soja</Emphasis> Sieb. et Zucc.) revealed by nuclear SSR markers and the implication for intraspecies phylogenic relationship of characters

Genetic diversity is reduced from wild soybean to cultivars and from landraces to modern varieties. However, intraspecies genetic diversity loss between characters or phenotypes also existed in wild soybean. We revealed the phylogenic relationship of character types in Chinese wild soybean using 42 SSR markers. We conjectured that white flower, no-seed bloom, grey pubescence, and four seed coat colours were evolutionarily acquired phenotypes. There were a small decrease (∆H = 0.1–6.46%) of gene diversity and a moderate reduction (∆Na = 10.81–53.54%) of number of alleles but a violent loss (80.74–98.59%) of unique alleles in the acquired phenotypes. Our results seemed to suggest that ovoid and elliptic leaves were differentiated at the earliest and subsequently lanceolate leaf appeared before the domestication of soybeans within wild soybean, and that G. gracilis type was another earliest type, maybe emerged since the appearance of soybeans or it was concomitant with the domestication of soybean.     

Inferring geographic origin of barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. subsp. <Emphasis Type="Italic">vulgare</Emphasis>) accessions using molecular markers

The USDA Agricultural Research Service (ARS) National Small Grains Collection (NSGC) has 207 landrace barleys obtained from a nursery grown in the Ukraine in 1930 by N.I. Vavilov, many of which have multiple resistance (MR) to disease similar to accessions from Ethiopia. The original collection locations of the accessions from the Vavilov nursery are unknown. The purpose of this study was to determine, using molecular markers, if the MR accessions of unknown origin from the Vavilov nursery are genetically related to MR accessions from Ethiopia. The genetic relatedness among susceptible accessions of unknown origin and among a selection of landrace accessions from the NSGC barley core subset was also assessed. Simple sequence repeat (SSR) and Diversity Array Technology PL (DArT) marker data were used to generate similarity matrices with Dice’s similarity coefficients. Cluster analysis from these results showed that the unknown-origin MR accessions grouped with accessions from Ethiopia. Susceptible accessions of unknown origin were genetically similar to accessions from western Asia, especially near the Caucasus, and from both southern and northern Europe. Based on similarity of marker profiles, some accessions from the core collection are likely duplicates. Future work will seek to identify the probable origin of the remainder of the unknown origin accessions and to more thoroughly characterize the genetic diversity within NSGC barley core subset.     

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.     

Biodiversity of <Emphasis Type="Italic">Secale strictum</Emphasis> in Iran measured using microsatellites

The mountain rye Secale strictum is native to the Middle East and is the progenitor of the cultivated rye. Regarding lack of information about the genetic diversity of this species in Iran, this study was aimed to evaluate its genetic variation and to examine the patterns of diversity related to the varieties and geography. Fifteen wheat and rye derived microsatellite markers were used to achieve this aim. High levels of diversity, with an average number of 6.1 alleles per locus (ranging up to 11) and high level polymorphism with polymorphism rate averaging 0.624 (between populations) and 0.357 (within populations) were observed among 125 individuals from 19 populations collected from various regions of Iran. The Northwestern populations showed the highest and the Northern populations showed the lowest polymorphism and diversity. One population of the Northwest of the country was notably closer in its allele range to the S. cereale accessions used as outgroup. No taxon or geographic specific marker was detected, suggesting high gene flow between the populations, however some groupings which can be related to the geographic regions and varieties, were evident. The analysis of molecular variance attributed same portions of genetic diversity to the within and between populations with no significant variation among different geographic regions. The results of this study indicated that the Iranian genepool of Secale strictum is valuable to search for new useful alleles for crop improvement.     

Genetic diversity and relationships between wild and cultivated olives (<Emphasis Type="Italic">Olea europaea</Emphasis> L.) in Sardinia as assessed by SSR markers

The genetic relationships within and between wild and cultivated olives were examined and clarified in an isolated and restricted area, such as the Mediterranean island of Sardinia. Wild (21 individuals) and cultivated olive trees (22 local cultivars from a germplasm collection and 35 ancient trees) were genotyped by means of 13 SSR loci. Five cases of synonymy were observed and nine distinct genotypes were identified in the collection. Five novel genotypes were also detected among the ancient trees. Differences on the allelic composition and heterozygosity levels were found between wild and cultivated trees. Model-based clustering method classified the olive trees into two major gene pools: (a) wild genotypes and (b) local cultivars from the collection and from heritage olives. Regarding the cultivated plant material, we observed that: (a) most of the Sardinian cultivars shared the same allelic profiles with the ancient cultivated trees and (b) the majority of these cultivars and all the novel genotypes were not related to any other cultivars included in this study. These findings as well as the detection of unique alleles and a certain wild genetic background at some cultivars revealed by the Bayesian analysis may indicate their autochthonous origin. The synonymy cases found between local cultivars and Italian mainland cultivars indicate interchange of genetic material among these growing areas, suggesting thus a possible allochthonous origin. The information obtained can assist in the management of an olive collection and sheds some light on the survival of true oleasters and the origin of Sardinian cultivars.