Morphological and genetic diversity analysis of rice accessions (Oryza sativa L.) differing in iron toxicity tolerance

A major emphasis in breeding for iron toxicity tolerance in rice is to identify differences that are associated with resistance and harness them for genetic improvement. In this study, thirty accessions, including IRRI gene bank accessions, two varieties from Brazil, 8 cultivars from West Africa and 10 cultivars from Uganda were analyzed for sensitivity to iron toxicity, and genetic diversity using morphological and SSR markers. Two genotypes, IR61612-313-16-2-2-1 and Suakoko 8 showed significantly high resistance with an average score of ≤ 3.5 on 1–9 scale. The SRR markers were highly informative and showed mean polymorphism information content (pic) of 0.68. The PIC values revealed that RM10793, RM3412, RM333, RM562, RM13628, RM310, RM5749, and RM154 could be the best markers for genetic diversity estimation of these rice cultivars. Diversity at the gene level showed an average of 4.61 alleles ranging from 2 to 12 per locus. Mean gene diversity (H) value for all SSR loci for the 30 genotypes evaluated was 0.69 but was decreased to 0.53 when analysis was performed on Ugandan accessions. The low genetic diversity found among the Ugandan accessions is the evidence of a narrow genetic base, and such a scenario has a potential vulnerability for resistance break down. A low correlation was detected between the observed molecular and morphological datasets. This means that a combination of morphological traits and SSR analysis would be required when assessing genetic variation under iron toxic conditions, and could be a practical strategy for breeders when planning crosses. A distinction between the resistant and susceptible accessions in both phenotyping and SSR datasets suggests the presence of unique alleles that could be harnessed for improvement of rice against iron toxicity.     

Population structure and linkage disequilibrium of ICRISAT foxtail millet (Setaria italica (L.) P. Beauv.) core collection

Use of diverse germplasm is a key factor which allows high level of resolution due to extensive recombination in the history. Therefore, population used in association mapping should posses as many phenotypes as possible. One of the methods to obtain most of the phenotypes is to construct the core collection. The ICRISAT foxtail millet core collection consisting of 155 accessions was genotyped using 72 simple sequence repeat (SSR) markers to investigate the genetic diversity, population structure and linkage disequilibrium (LD). A high degree of molecular diversity among the accessions was found, with an average of 16.69 alleles per locus. STRUCTURE analyses classify the accessions into four subpopulations (SP) based on SSR allelic diversity. The Neighbor joining clustering and the principal coordinate analysis were in accordance with the racial classification. The distribution of molecular genetic variation among and within the four SP and three races showed high degree of variability within each group, and low level of genetic distance (GD) among the groups. LD decay of <40 cM of GD in foxtail millet core collection was observed, which suggests that it could be possible to achieve resolution down to the 40 cM level. From this investigation, it is evident that the foxtail millet core collection developed at ICRISAT is very diverse and could be a valuable resource for trait association mapping, crop breeding and germplasm management.     

黄麻核心种质的遴选

遴选黄麻核心种质可为黄麻种质创新及新品种选育奠定基础。本研究以300份黄麻种质资源为基础,基于SSR分子标记和农艺性状考察,结合地理来源构建核心种质。结果表明,11个农艺性状变异系数变幅在13.06%～84.87%,表现出丰富的遗传多样性。按农艺性状聚类分析可划分为8个类群,按分子标记聚类可划分为10个类群。结合2个聚类分析、地理位置并按比例取样,建立一个由108份品种(系)组成的预选核心种质。采用44对SSR引物对其进行遗传差异分析,在遗传相似系数为0.65时,可把108份品种(系)分成圆果种和长果种两大类。根据遗传差异分析,剔除遗传相似系数大于或等于0.85的遗传冗余,获得84份品种(系)的核心种质,其中圆果种60份和长果种24份。比较84份核心种质与300份种质的农艺性状变异系数及Shannon-Wiener指数发现,两者之间相差不大,表明遴选的84份核心种质可以最大限度代表300份黄麻种质资源的遗传多样性加以利用和保存。     

Assessing the genetic diversity of Portuguese maize germplasm using microsatellite markers

A collection of Portuguese maize accessions representing a valuable source of genes for introduction into modern cultivars is stored at the Portuguese Plant Germplasm Bank (Banco Português de Germoplasma Vegetal—BPGV). To assess genetic diversity among inbreds, microsatellite analysis was carried out for 54 inbred lines representing the diversity of Portuguese dent and flint maize germplasm. Fifty American and other European elite inbreds were also analysed for comparison. Fifteen microsatellite loci distributed throughout the maize genome were chosen based on their repeat unit and base composition. A total of 80 alleles were detected with an average allele number of 5.33 per locus. Polymorphism information content (PIC) values and observed genetic distances showed the existence of large variability among inbreds. Cluster analysis indicated that almost all of the inbreds could be distinguished from each other and Portuguese inbreds were present in all clusters formed. These associations were consistent with the known pedigree records of the inbreds, confirming a mixed origin of Portuguese materials. Comparative analysis of microsatellite diversity among groups was established according to important traits for both breeding and line identification. This revealed that, although most of the genetic diversity (>95%) was attributable to differences among inbreds of different groups, the existence of phenotypic differentiation in endosperm colour, kernel type and cob colour could be suggested for grouping. These findings support the joint use of molecular and morphological traits in management of the germplasm collection. In this study, SSR markers proved to be effective to characterise and identify maize inbred lines, and demonstrate associations among them. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular genetic evaluation of sorghum germplasm differing in response to fungal diseases: Rust (Puccinia purpurea) and anthracnose (Collectotrichum graminicola)

To evaluate genetic diversity in relation to rust and anthracnose disease response, ninety-six accessions were randomly selected from the core collection database of the Germplasm Research Information Network (GRIN) and characterized by a set of 40 SSR markers. The mean value of polymorphism information content (PIC) was 0.8228. Two dendrograms were generated from the molecular genetic data and field morphological data, respectively. The genetic dendrogram demonstrates that the accessions can be classified into three main clades and nine subgroups. The branched subgroups correlated very well with the locations where the accessions were collected. Geographical origin of accessions had significant influences on genetic similarity of sorghum germplasm. Out of 96 accessions, only eight accessions were highly resistant to both rust and anthracnose. All the accessions from South Africa and Mali were highly resistant to anthracnose. The information from genetic classification would be useful for choosing parents to make crosses in sorghum breeding programs and classifying sorghum accessions in germplasm management.     

Analysis of population structure and genetic diversity reveals gene flow and geographic patterns in cultivated rice (<Emphasis Type="Italic">O. sativa</Emphasis> and <Emphasis Type="Italic">O. glaberrima</Emphasis>) in West Africa

To fully exploit the diversity in African rice germplasm and to broaden the gene pool reliable information on the population genetic diversity and phenotypic characteristics is a prerequisite. In this paper, the population structure and genetic diversity of 42 cultivated African rice (Oryza spp.) accessions originating from West Africa (Benin, Mali and Nigeria, Liberia etc.) were investigated using 20 simple sequence repeats (SSR) and 77 amplified fragment length polymorphisms (AFLP). Additionally, field trials were set up to gain insight into phenotypic characteristics that differentiate the genetic populations among rice accessions. The analysis revealed considerably high polymorphisms for SSR markers (PIC mean?=?0.78) in the germplasm studied. A significant association was found between AFLP markers and geographic origin of rice accessions (R?=?0.72). Germplasm structure showed that Oryza sativa accessions were not totally isolated from Oryza glaberrima accessions. The results allowed identification of five O. glaberrima accessions which grouped together with O. sativa accessions, sharing common alleles of 18 loci out of the 20 SSR markers analyzed. Population structure analysis revealed existence of a gene flow between O. sativa and O. glaberrima rice accessions which can be used to combine several interesting traits in breeding programs. Further studies are needed to clarify the contributions of this gene flow to valuable traits such as abiotic and biotic stresses including disease resistance.     

DNA profiling and genetic diversity of Korean soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merrill) landraces by SSR markers

Approximately 7,000 accessions of Korean soybean (Glycine max (L.) Merrill) landraces, largely composed of three collections, the Korea Atomic Energy Research Institute’s soybean (KAS), the Korean Crop Experiment Station’s soybean (KLS) and the Korean Agricultural Development and Technology Center’s soybean (KADTC) collections, have been conserved at the Rural Development Administration (RDA) genebank in Korea. The accessions within collections were classified based on their traditional uses such as sauce soybean (SA), sprouted soybean (SP), soybean for cooking with rice (SCR), and OTHERS. A total of 2,758 accessions of Korean soybean landraces were used to profile and to evaluate genetic structure using six SSR loci. A total of 110 alleles were revealed by at the six SSR loci. The number of alleles per SSR locus ranged from 9 to 39 in Satt187 and Satt_074, respectively. The number of alleles ranged from 87 in the KADTC collection to 96 in the KLS collection, and from 63 in the SCR group to 95 in the SP group. Nei’s average genetic diversity ranged from 0.68 to 0.70 across three collections, and 0.64 to 0.69 across the usage groups. The average between-group differentiation (G _st) was 0.9 among collections, and 4.1 among the usage groups. The similar average diversity among three collections implies that the genetic background of the three collections was quite similar or that there were a large number of duplicate accessions in three collections. The selection from the four groups classified based upon usage may be a useful way to select accessions for developing a Korean soybean landrace core collection at the RDA genebank. DNA profile information of accessions will provide indications of redundancies or omissions and aid in managing the soybean collection held at the RDA genebank. The information on diversity analysis could help to enlarge the genetic diversity of materials in breeding programs and could be used to develop a core collection.     

Establishment of Chinese soybean Glycine max core collections with agronomic traits and SSR markers

It is very important to efficiently study and use genetic diversity resources in crop breeding and sustainable agriculture. In this study, different sampling methods and sample sizes were compared in order to optimize the strategies for building a rationally sized core collection of Chinese soybean (Glycine max). The diversity in the core collection captured more than 70% of that in the pre-core collection, no matter what sampling methods were used, at a sampling proportion of 1%. Core collections established with both simple sequence repeat (SSR) marker data and agronomic traits were more representative than those chosen on an independent basis. An optimal sampling method for a soybean core collection was determined, in which strategy ‘S’ (allocating accessions to clusters according to the proportion of square root of the original sample size within each ecotype) was used based on SSR and agronomic data. Curve estimation was used to estimate the allelic richness of the entire Chinese soybean germplasm and a minimum sample size for a core collection, on which a sampling proportion of about 2% was determined to be optimal for a core collection. Further analysis on the core collection with fourteen agronomic traits and allelic constitution at 60 SSR loci suggested that it highly represented the entire collections both on genetic structure and diversity distribution. This core collection would provide an effective platform in proper exploitation of soybean germplasm resources for the study of complex traits and discovering important novel traits for crop genetic development.     

Microsatellite (SSR) variation in a collection of Malus (apple) species and hybrids

A collection of 142 accessions of 23 Malus species, derived hybrids and cultivar accessions from the USDA-ARS Plant Genetic Resources Unit's core collection, which represents an extensive range of Malus species, was screened with a set of previously described SSR (simple sequence repeat) markers. The markers were used to determine genetic identities, estimate genetic diversity, identify genetic relationships among the accessions, and determine the utility of SSR primers developed from Malus ×domestica for making genetic assessments across the whole Malus genus. All eight primer pairs amplified multiple fragments when used in polymerase chain reactions with DNA from these accessions. High levels of variation were detected with a mean of 26.4 alleles per locus and a mean direct count heterozygosity across all eight loci equal to 0.623. The eight primer pairs used in this study unambiguously differentiated all but five pairs of accessions in this collection of 142 accessions of 23 Malus species, derived hybrids and cultivars. These SSR data were not useful in identifying genetic relationships among this diverse collection of accessions, with the majority of the accessions not clustering in ways concordant with taxonomic information and/or geographic origin. The resulting phenogram resolved only two meaningful clusters, for the taxonomically isolated Section Chloromeles and for M. fusca accessions, reflecting genetic relationships arising from geographic origin. The detection of identical accessions in the collection, which were previously considered to be unique, highlights the critical need to further bolster collections of certain Malus species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Changes in allelic frequency over time in European bread wheat (Triticum aestivum L.) varieties revealed using DArT and SSR markers

A collection of 189 bread wheat landraces and cultivars, primarily of European origin, released between 1886 and 2009, was analyzed using two DNA marker systems. A set of 76 SSR markers and ~7,000 DArT markers distributed across the wheat genome were employed in these analyses. All of the SSR markers were found to be polymorphic, whereas only 2,532 of the ~7,000 DArT markers were polymorphic. A Mantel test between the genetic distances calculated based on the SSR and DArT data showed a strong positive correlation between the two marker types, with a Pearson’s value (r) of 0.66. We assessed the genetic diversity and allelic frequencies among the accessions based on spring- versus winter-wheat type as well as between landraces and cultivars. We also analyzed the changes in genetic diversity and allelic frequencies in these samples over time. We observed separation based on both vernalization type and release date. Interestingly, we detected a decrease in genetic diversity in wheat accessions released over the period from 1960 to 1980. However, our results also showed that modern plant breeding have succeeded in maintaining genetic diversity in modern wheat cultivars. Studying allelic frequencies using SSR and DArT markers over time revealed changes in allelic frequencies for a number of markers that are known to be linked to important traits, which should be useful for genomic screening efforts. Monitoring changes in the frequency of molecular DNA markers over time in wheat cultivars may yield insight into alleles linked to important traits that have been the subject of positive or negative selection in the past and that may be useful for marker-assisted breeding programs in the future.     

Identification and characterization of EST-SSRs in finger millet (<Emphasis Type="Italic">Eleusine coracana</Emphasis> (L.) Gaertn.)

In recent years, microsatellites have become the most used markers for studying population genetic diversity. The increased availability of the DNA sequences has given the possibility to develop EST-derived SSR markers. A total of 1,927 ESTs of Eleusine coracana available in the NCBI database were mined for SSRs. Di-nucleotides are the most occurring motifs accounting for more than 50% of the repeats, of which GA was the most abundant motif and tetra-nucleotides are the least occurring motifs. Of the 132 markers identified, 30 primer pairs based were synthesized. SSR markers were used for variety discrimination and genetic assessment in 15 finger millet accessions; 20 primers showed polymorphism and 13 primers were identified as having a PIC value above 0.5. On the basis of the distribution of these polymorphic alleles, the 15 accessions were classified into two groups. This study has demonstrated the potential of EST-derived SSR primer pairs in finger millet. These primers will serve as valuable source for further breeding programs.     

Mining Elite Sea-Island Cotton Germplasm Based on Phenotyping and SSR Markers

[Objective] An elite germplasm resource of sea-island cotton with outstanding traits was mined in order to accelerate the breeding process of new varieties. [Method] The core collections of sea-island cotton germplasm consisted of 178 accessions were used as experimental materials in this study. Analyses of variability and diversity were performed through detecting phenotypic data of six main breeding-targeted traits, including boll weight, boll number per plant, lint percentage, fiber length, fiber strength, and micronaire. The elite germplasm of sea-island cotton was selected according to 10% optimal sampling strategy based on the phenotypic value of each trait. The 120 pairs of polymorphic simple sequence repeat (SSR) primers were used to analyze the polymorphism of 178 accessions of the core collections. Then, we conducted the population structure and clustering analysis based on the genotyping results. According to the results of cluster analysis, the primary elite germplasm was further selected, and the final elite germplasm of sea-island cotton was identified. [Result] The results showed that there was a high variability and abundant genetic diversity in the 6 studied traits. In 178 accessions of sea-island cotton, 262 alleles were detected by 120 pairs of SSR primers, with an average of 2.18 loci. The average polymorphism information content (PIC) was 0.067 8-0.630 0, with an average of 0.296 0, showing moderate polymorphism. The cluster analysis showed that the core collection of sea-island cotton was divided into six groups. twenty-three elite germplasm resources of sea-island cotton were identified based on phenotypic value and cluster analysis of SSR markers. [Conclusion] The germplasm of sea-island cotton can be analyzed and evaluated based on the phenotyping and SSR markers, and then the elite germplasm of sea-island cotton can be identified. These results provided the material basis for the genetic breeding of sea-island cotton, as well as the important reference and basis for the mining and identification of crop elite germplasm.     

ICRISAT花生微核心种质青枯病抗性鉴定

对ICRISAT花生微核心种质155份材料进行抗青枯病鉴定,获得ICG9249和ICG12625两份抗青枯病材料,平均抗性达到了91.7%。与中国核心高抗5份种质共同作为材料,以SSR技术对其亲缘关系和遗传多样性进行了分析。在选用的58对SSR引物中,24对引物能扩增出稳定的多态性条带,这些引物能在抗青枯病花生种质基因组DNA中扩增出2~7个DNA片段。结果表明,7份抗青枯病种质的遗传距离为0.17~0.71,平均为0.49。其中ICG12625和撮豆、富川大花生的遗传距离最大（0.71）,嵊县小红毛和富川大花生的遗传距离最小（0.17）。经过统计,两两品种之间遗传距离达0.50以上的有12个组合,说明了这些抗青枯病种质资源的遗传多样性。根据DNA扩增结果,绘制了抗青枯病种质的指纹图谱,明确了其SSR分子特性。通过聚类分析结果和植物学性状调查结果显示,ICG12625与其它种质距离较远,且具有优良的植物学性状,可以作为疏枝亚种的亲本材料加以研究利用。     

Comparison of molecular markers and coefficients of parentage for the analysis of genetic diversity among spring bread wheat accessions

The comparison of different methods of estimating genetic diversity could define their usefulness in plant breeding and genetic improvement programs. This study evaluates and compares the genetic diversity of 70 spring wheat accessions representing a broad genetic pool based on molecular markers and parentage relationships. The sample was composed of 32 accessions from the International Maize and Wheat Improvement Center (CIMMYT) and 38 from other breeding programs worldwide. Eight AFLP-primer combinations and 37 pairs of SSR primers were used to characterize the accessions and the Coefficients of Parentage (COP) were calculated from registered pedigrees. The average genealogical (COP) similarity (0.09 with a range of 0.0–1.0) was low in comparison to similarity calculated using SSR markers (0.41 with a range of 0.15–0.88) and AFLP markers (0.70 with a range of 0.33–0.98). Correlation between the genealogical similarity matrix (excluding accessions with COPs = 0) and the matrices of genetic similarity based on molecular markers was 0.34≤r≤0.46 (p <0.05). It is concluded that AFLP and SSR markers are generally in agreement with estimates of diversity measured using COPs, especially when complete pedigree data are available. However, markers may provide a more correct estimate due to some unrealistic assumptions made when calculating COPs, such as absence of selection. Furthermore, both COP and marker distances indicate that CIMMYT accessions are different from the worldwide group of accessions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and association mapping of forage quality in diverse bermudagrass accessions

Bermudagrass is a warm season grass widely cultivated for turf and fodder. Nonetheless, the grass has poor forage quality because animals that consume it fail to assimilate its organic matter efficiently. Thus, identification of the marker-trait association between simple sequence repeat (SSR) markers and forage-quality-related traits in diverse bermudagrass accessions would enable efficient selection of high forage quality bermudagrass cultivars. Association mapping of 8 forage-related-quality traits with 1474 markers was conducted in 60 diverse bermudagrass accessions from five geographical regions in China. Significant variations in eight phenotypic and physiological traits were observed among the 60 accessions. A total of 1474 alleles were amplified by 104 SSR primers. The average gene diversity and polymorphic information content for the study sample were 0.2097 and 0.1748 respectively. The clustering analysis suggested that geographic origin influenced genetic distances between accessions. A total of 76 markers significantly associated with traits at P < 0.01; 73 with a single trait and 3 with two traits each. Nevertheless, only 41 significant marker-trait associations (MTAs) were observed after Bonferroni test was separately conducted for each trait. Forty-one microsatellites had significant associations with 8 forage-quality-related traits. These markers provide a feasible means of genetically improving forage quality in bermudagrass after further authentication.     

绿豆SSR标记的开发及遗传多样性分析

SSR标记以其数量丰富、多态性好、共显性遗传等优点在基础研究和育种工作中发挥了重要作用,但目前绿豆基因组中的SSR标记依然较少。本研究将磁珠富集法和测序技术相结合高通量检测绿豆基因组SSR位点,鉴定出3,275,355个SSR位点,开发了2742个SSR标记。选取其中157个SSR进行PCR验证,发现有90个(57.33%)标记在10份材料中表现出多态性。挑选40个条带清晰、多态性高、染色体上均匀分布的标记对90份绿豆资源进行遗传多样性分析,单个位点检测到的等位变异数为2~8个,平均为3.0个,有效等位基因数为1.31~4.21个,平均为2.16。Nei’s基因多样性指数在0.23~0.76之间,平均为0.51。多态性信息含量为0.22~0.72,平均为0.43。聚类分析将90份材料分为2个类群,包含4个组。第I组主要由北方资源组成,第Ⅱ组种质来源较为分散,第Ⅲ组主要由山东的资源构成,第Ⅳ组包含多数河北的种质资源。本研究开发的多态性SSR标记不仅可以用于绿豆种质资源的遗传多样性分析,也将在高密度遗传图谱构建、基因定位和分子标记辅助育种中发挥重要作用。     

半野生棉棉仁含油量与SSR标记的关联分析

为探究半野生棉群体材料的遗传变异和群体结构,挖掘与含油量相关的标记位点,利用215对SSR引物,对89份半野生棉自然群体进行遗传多样性、群体结构和亲缘关系分析,并进行棉仁含油量性状与标记的关联分析。结果表明,位点多态信息含量(PIC)为0.0222~0.7390,平均为0.3452;基因型多样性(H′)介于0.0616~1.4412,平均为0.5614。群体结构分析将89份材料分为2个亚群,群体结构简单,遗传变异比较丰富,可以用于半野生棉目标性状的关联分析。应用关联分析,在3个年份环境下重复检测出12个与棉仁含油量显著相关的位点(P0.05),这12个位点可能与含油量性状存在稳定的关联,可为棉仁含油量性状的分子标记辅助选择提供依据,为开展半野生棉其它性状的关联分析提供参考。阔叶棉141平均棉仁含油量为41.90%,为高油份含量品种的遗传改良提供理想材料。     

Molecular diversity and population structure of the Ethiopian lentil (<Emphasis Type="Italic">Lens Culinaris</Emphasis> Medikus) genotype assessment using SSR markers

Knowledge of genetic diversity in germplasm is essential for formulating effective germplasm collection, conservation, utilization strategies in and crop improvement programs. It also provides an opportunity to take corrective steps infusing new genes to avoid risks associated with a narrow genetic bases. Genetic diversity analysis of 119 lentil genotypes of including 83 germplasm and 36 exotic genotypes from International Center for Agricultural Research in the Dry Areas was studied using 27 primers of simple sequence repeat (SSR) marker. Molecular analysis of variance showed variations of 82% within and 18% of the among population variance was explained. Degree of polymorphism observed among the populations was 100%. A total 122 alleles were detected, with 2 to 7 alleles per locus, with a mean of 4.52 alleles per locus. The estimated gene diversity value for 27 loci was 0.64. The average Shannon’s information index value of 1.19 was obtained showed the existence of high genetic variation within the genotypes. The genetic similarity indices ranged from 0.21 to 1.00. The SSR markers showed an average polymorphic information content (PIC) value of 0.58. Cluster analysis grouped the genotypes into five major clusters as distinct genetic populations. Diversity analyses revealed the existence of a high level of genetic variation among genotypes. This molecular diversity information provides a basis for future germplasm collection, utilization, and conservation strategies in gene banks and introducing exotic germplasm to widen the genetic base of the current lentil breeding population.     

Association mapping of yield-related traits and SSR markers in wild soybean (Glycine soja Sieb. and Zucc.)

Wild soybean, the progenitor of cultivated soybean, is an important gene pool for ongoing soybean breeding efforts. To identify yield-enhancing quantitative trait locus (QTL) or gene from wild soybean, 113 wild soybeans accessions were phenotyped for five yield-related traits and genotyped with 85 simple sequence repeat (SSR) markers to conduct association mapping. A total of 892 alleles were detected for the 85 SSR markers, with an average 10.49 alleles; the corresponding PIC values ranged from 0.07 to 0.92, with an average 0.73. The genetic diversity of each SSR marker ranged from 0.07 to 0.93, with an average 0.75. A total of 18 SSR markers were identified for the five traits. Two SSR markers, sct_010 and satt316, which are associated with the yield per plant were stably expressed over two years at two experimental locations. Our results suggested that association mapping can be an effective approach for identifying QTL from wild soybean.