Population structure and phylogenetic relationship of Peach [Prunus persica (L.) Batsch] and Nectarine [Prunus persica var. nucipersica (L.) C.K. Schneid.] based on retrotransposon markers

For effective varietal improvement of horticultural crops peach (Prunus persica) and nectarine (Prunus persica var. nucipersica), information about their population structure and genetic relatedness plays an important role. In this study we used retrotransposon-based markers (iPBS) to estimate the genetic diversity and population structure of 48 peach and nectarine genotypes from various distinct geographical regions of Punjab and Khyber Pakhtunkhwa, Pakistan. A total of 461 alleles were identified from PCR amplicons derived from nine iPBS primer pairs with an average of 8.5 alleles/locus. Among all four groups the genotypes collected from Swat and Hunza had the highest and the lowest expected heterozygosity, unbiased expected heterozygosity and Shannon’s information index, respectively. We constructed a Neighbour-Joining dendrogram and performed principal coordinate analysis based on the distance matrices, and both forms of analysis grouped the 48 genotypes into two distinct clusters. The STRUCTURE software distributed the forty-eight genotypes into two main populations (k?=?2) indicating a low diversity between genotypes collected from Chakwal, Swat, Mansehra and Hunza.

     

Analysis of genetic diversity in the tuber mustard (Brassica juncea var. tumida Tsen et Lee) in the Yangtze river basin of China

In the present study, analyses of SSR molecular markers were performed to investigate the genetic diversity of 133 tuber mustard cultivars. Eighty-one pairs of SSR primers from a total of 600 in Brassica produced stable amplified bands. In addition, 810 bands were detected among the cultivars, and 724 of those were polymorphic (89.38 %). The average number of bands per locus was 10.0 with a range from 5 to 16. Shannon’s information index for each SSR locus varied from 0.52 to 3.72, with an average of 2.74. The coefficients of genetic similarity in the SSR marker patterns among the 133 cultivars ranged from 0.77 to 0.91, with an average of 0.85. The cluster analysis showed that the cultivars could be classified into six clusters when the genetic similarity was 0.83, with 90.23 % of the cultivars included in Clusters 5 and 6. Principal component analysis was carried based on the SSR data. The results showed that the first three principal components could explain the genetic variation with 85.47, 0.67, and 0.61 %, and the 133 cultivars could be divided into six clusters according to the nearest phylogenetic relationship. It was indicated that the similarity was high and the genetic diversity was narrow among the 133 mustard tuber cultivars. 360 individuals from 24 cultivars were analyzed to reveal the genetic structure and genetic diversity within cultivars. A total of 925 alleles were detected in the 24 cultivars. Estimates of the mean number of alleles ‘A’, the effective allelic number ‘A_e’, the observed heterozygosity ‘H_o’, and expected heterozygosity ‘H_e’ were 6.0, 3.6, 0.64, and 0.37, respectively. An obvious genetic deviation from Hardy–Weinberg expectation was observed both among and within cultivars and a considerable genetic variation was revealed within rather than among cultivars. It is necessary to broaden the genetic basis of the breeding germplasm in tuber mustard. Based on their geographical distributions, the tuber mustard cultivars in this study can be divided into up-Yangtze river, mid-Yangtze river, and down-Yangtze river groups. Genetic diversity was highest in mid-Yangtze river group, followed by up-Yangtze river group, and then down-Yangtze river group. It was presumed that the origin center or genetic diversity center of tuber mustard was mid-Yangtze river, and the crop was transmitted along the Yangtze river in both directions.     

Genetic diversity in wild and cultivated black raspberry (Rubus occidentalis L.) evaluated by simple sequence repeat markers

Breeding progress in black raspberry (Rubus occidentalis L.) has been limited by a lack of genetic diversity in elite germplasm. Black raspberry cultivars have been noted for showing very few phenotypic differences and seedlings from crosses between cultivars for a lack of segregation for important traits. Despite these challenges, little molecular work has been done to explore genetic diversity and relationships in wild and cultivated black raspberry germplasm. Microsatellite, or simple sequence repeat (SSR), markers are highly polymorphic codominant markers useful for studying genetic diversity, population genetics, genetic fingerprinting and other applications. We examined genetic diversity in 148 wild and cultivated black raspberry accessions using 21 polymorphic SSR markers. Black raspberry cultivars clustered tightly and showed higher than expected heterozygosity while that of wild accessions was low. Relationships between wild black raspberry accessions were poorly resolved and regional clusters were mostly absent from our analysis. Our results indicated that wild black raspberry germplasm is a relatively untapped resource available for future breeding.     

Presence of phylogeographic structure among wild diploid alfalfa accessions (Medicago sativa L. subsp. microcarpa Urb.) with evidence of the center of origin

The genetic structure of wild germplasm on a macrogeographic scale has implications for collection and conservation of wild genetic resources of economically important plants. Cultivated alfalfa is derived from a taxonomic group called the Medicago sativa–falcata complex, which includes a number of morphologically and genetically differentiated diploid and tetraploid subspecies representing the primary gene pool for alfalfa improvement. Although the origin of the taxa included in M. sativa–falcata complex has been described broadly, the center of diversity for each taxon is not clear. In this study, 60 individual genotypes from 16 accessions of the diploid taxon M. sativa subsp. microcarpa Urb. [M. sativa subsp. caerulea (Less. ex Ledeb.) Schmalh.] were obtained from the two proposed centers of diversity, Caucasia and Central Asia, and were genotyped with 89 simple sequence repeat markers. Phylogenetic reconstructions together with population genetics statistics (heterozygosity, allelic diversity, and F_ST) were used to describe the pattern of present genetic diversity, to deduce biogeographic history, and to infer the center of diversity. Our results revealed that the Central Asian accessions were distinct from the Caucasian accessions, and that overall accessions, genetic distance was positively correlated with geographical distance, indicating a spatial genetic structure. The accessions from the Caucasian region had higher mean F_ST values, higher allele diversity and higher heterozygosity, suggesting that the Caucasian region is the likely center of diversity for M. sativa subsp. microcarpa [M. sativa subsp. caerulea] and the accessions recolonized Central Asia from Caucasia after the glacial retreat.     

Genetic diversity of cultivated and wild Ussurian Pear (<Emphasis Type="Italic">Pyrus ussuriensis</Emphasis> Maxim.) in China evaluated with M13-tailed SSR markers

Eight genomic SSR markers with a M13 tail attached were used to assess the genetic diversity of 72 Ussurian Pear accessions (Pyrus ussuriensis Maxim.) in China. The M13-tailed method was effective in discriminating all the 32 wild accessions. All the 40 Ussurian Pear cultivars could be successfully discriminated with the exception of 4 sets of synonymies or spots. A total of 108 alleles were obtained with an average of 13.5 per locus. The expected heterozygosity, observed heterozygosity, and power of discrimination were 0.78, 0.63, and 0.86 respectively. Three triploid cultivars (‘Anli’, ‘Ruan’er’, and ‘Pitaiguo’), and one wild accession, P. ussuriensis ‘Xilin-3’, showed three alleles at some SSRs. The number of alleles and observed heterozygosity per locus for 40 Ussurian Pear cultivars were 9.1 and 0.62, respectively, lower than the values of 32 wild accessions which were 11.3 and 0.65, respectively. A dendrogram based on the SSR genotypes was obtained, showing two major groups corresponding to cultivated group and wild group. All the cultivars fell into the cultivated group. Some subgroups (Nanguoli subgroup, Zhibazi subgroup, Xiangshuili subgroup, Balixiang subgroup, Anli subgroup) could be found in the cultivated group. A very close relationship between ‘Huagaili’ and ‘Miansuan’, and a close relationship between ‘Anli’ and a wild accession, P. ussuriensis ‘Huangshanli’ could be found in Anli subgroup. ‘Nanguoli’ and ‘Xiaowuxiang’ showed a close relationship with at least one identical allele at each locus with the exception of NH015a.     

Level and Transmission of Genetic Heterozygosity in Apricot (Prunus armeniaca L.) Explored Using Simple Sequence Repeat Markers

In this study, 17 peach simple sequence repeat (SSR) sequences were used in the exploration of the genetic heterozygosity level of several apricot cultivars from Spain, France, Greece, and the USA, and 23 descendants. The genotypes can be classified in three groups as a function of their genetic heterozygosity (1) local cultivars from Murcia (Spain) (‘Gitanos’ and ‘Pepito del Rubio’) and several descendants from crosses among these cultivars, with very low genetic heterozygosities (less than 0.30); (2) cultivars from France and Spain (‘Moniquí’, ‘Currot’ and ‘Bergeron’) and several descendants, with intermediate levels of genetic heterozygosity (around 0.45); and (3) cultivars ‘Orange Red’ and ‘Goldrich’ from North America and ‘Lito’ from Greece, with the remaining descendants, having genetic heterozygosities higher than 0.50. The results showed the high increase of genetic heterozygosity in the case of descendants from complementary crosses. The use of cultivars from North America could increase greatly the genetic heterozygosity in the Spanish apricot breeding programs, enlarging the genetic variability of the local cultivars. On the other hand, in the case of transgressive crosses among local Spanish cultivars, the increase of genetic heterozygosity was much lower.     

Genetic characterization of guava (Psidium guajava L.) germplasm in the United States using microsatellite markers

Genetic diversity of 35 Psidium guajava L. accessions and three related species (P. guineense Sw., P. sartorianum (O. Berg) Nied. and P. friedrichsthalianum (O. Berg) Nied.) maintained at the U.S. Department of Agriculture (USDA), National Plants Germplasm System, Hilo, HI, was characterized using 20 simple sequence repeat (SSR) markers. Diversity analysis detected a total of 178 alleles ranging from 4 to 16 alleles per locus. The observed mean heterozygosity (0.2) and inbreeding coefficient (0.8) indicated a high level of inbreeding among the accessions tested. Multi-locus DNA fingerprints based on the 20 SSR loci unambiguously differentiated all accessions and revealed the absence of duplicated samples. Ordination and cluster analyses suggested that the genetic relationships between majorities of the accessions could be explained by geographic origin, mainly including tropical America, Southeast Asia and Hawaii. A Bayesian cluster analysis partitioned the accessions into two groups and the partition was largely compatible with the result of ordination analyses. Distance-based cluster analyses further indicated that accessions from same geographical region or breeding programs grouped together in spite of the inter-regional exchange of germplasm. Accessions from Southeast Asia were dominantly white fleshed, whereas accessions from tropical America and Hawaii exhibited diverse flesh colors. The results also indicated that accessions from the same region were likely derived from a small number of common ancestors or progenitors. All 20 SSRs were transferable to P. guineense, P. sartorianum and P. friedrichsthalianum, indicating a close relationship between the cultigens and wild relatives. Application of SSR markers in the USDA/Agricultural Research Service germplasm collection provides new insight into the diversity of the guava germplasm, which will be valuable in future breeding endeavors and the conservation of guava genetic resources.     

Identification of unknown apple (<Emphasis Type="Italic">Malus</Emphasis> × <Emphasis Type="Italic">domestica</Emphasis>) cultivars demonstrates the impact of local breeding program on cultivar diversity

Apple (Malus?×?domestica Borkh.) trees, either abandoned or cared for, are common on the North American landscape. These trees can live for decades, and therefore represent a record of large- and small-scale agricultural practices through time. Here, we assessed the genetic diversity and identity of 330 unknown apple trees in northern Minnesota with 9 simple sequence repeat (SSR) markers. The unknown (not identified by cultivar name) trees were compared to >?1000 named cultivars in the U.S. Department of Agriculture Plant Genetic Resources Unit Malus collection and also to each other to identify repeated genotypes. Overall, the 330 unknown trees had high diversity (average H_e?=?0.75), and consisted of 264 unique genotypes. A total of 76 of the unknown trees were matched to 20 different named cultivars, and these cultivars were mainly derived from either the local breeding program at the University of Minnesota, or were Russian cultivars imported for horticulture in the northern US. This study demonstrates the importance of local breeding programs, and also the challenges associated with identifying clones in a genetically diverse crop like apple.     

A study of genetic diversity among Indian bread wheat (Triticum aestivum L.) cultivars released during last 100?years

Genetic diversity was examined in a collection of 263 Indian bread wheat (Triticum aestivum L.) cultivars using 90 SSR markers. These cultivars were classified into Group I (pre-green revolution cultivars) and Group II cultivars (post-green revolution cultivars). SSR markers were also classified into Set-I SSRs (42 random genomic SSRs) and Set-II SSRs (48 SSRs associated with QTLs for grain weight). The SSRs belonging to Set-II exhibited relatively low level of polymorphism, suggesting that the SSRs associated with QTL for grain weight in wheat were probably under selection pressure during wheat breeding. AMOVA indicated that proportion of the variation within each of the two groups of cultivars accounted for most (87.59%) of the molecular variance, while the variation between the two groups of cultivars accounted for only 12.41% of the variance. The estimates of the average number of alleles/locus and gene diversity due to each of the two sets of SSRs suggested increase in overall genetic diversity after green revolution in Indian bread wheat cultivars. Differences were also observed in genetic diversity among cultivars from each of the six wheat growing agro-climatic zones of India. However, decade-wise analysis of genetic diversity among the post-green revolution cultivars indicated a progressive decline in genetic diversity, thus suggesting a need for involving diverse exotic, synthetic and winter wheat germplasm in Indian wheat breeding programs.     

Investigation of the genetic structure of some common bean (Phaseolus vulgaris L.) commercial varieties and genotypes used as a genitor with SSR and SNP markers

Common bean is a species belonging to the Phaseolus genus of the Leguminosae family. It has economic importance due to being rich in protein, vitamin A and C, and minerals. Being one of the most cultivated species of legumes, the determination of genetic diversity in bean genotypes or populations has an important role in terms of our genetic resources. The objective of this study was to evaluate the genetic structure of 94 genotypes which were cultivated in different parts of the world and our country with SSR and SNP markers. 10 SSR loci and 73 SNP primers were used for the determination of genetic structure in commercial cultivars and breeding lines. All of the SSR and SNP loci used in the study were found to be polymorphic. A total of 89 alleles were identified for 10 SSR loci. Mean number of alleles per locus (Na?=?8.9), effective allele number (Ne?=?3.731), Shannon information index (I?=?1.468), observed heterozygosity (Ho?=?0.023), and expected heterozygosity (He?=?0.654) were calculated based on SSR analysis. According to the results of Bayesian-based STRUCTURE analysis using SSR and SNP data, 94 bean genotypes were genetically divided into three main clusters. According to genetic distance based UPGMA dendrogram obtained from SNP analysis, 94 bean genotypes were divided into 2 main clusters corresponding Mesoamerican and Andean gene pools. The obtained results provide important information about the genetic structures of the studied bean cultivars and breeding lines. With the obtained results, it will be possible to develop breeding programs to develop new cultivars by using our gene resources.