云南茶树资源遗传多样性与亲缘关系的ISSR分析

以8个种群134份云南茶树资源为材料,应用ISSR标记方法,进行了遗传多样性与亲缘关系分析。结果表明,18个多态性ISSR引物对全部试验材料进行PCR扩增,共获得475条稳定的谱带,其中多态性谱带470条(占98.9%),遗传多样性丰富;应用Nei-Li相似系数法估算了134个材料间的相似系数为0.445~0.819,平均为0.512,说明茶树资源间的遗传基础较宽;对134份茶树资源的分子系统聚类分析(UPGMA)将资源分为3大组,聚类结果与地理距离没有明显的相关性;主成分分析(PCA)表明主成分分析的结果与系统聚类基本一致,但是主成分分析更加直观清晰地显示各个材料间的亲缘关系;大厂茶等8个种群间的遗传相似系数于0.850~0.987间,平均为0.92,表明不同种群间的遗传差异较小。     

Genetic similarity among European winter triticale elite germplasms assessed with AFLP and comparisons with SSR and pedigree data

Genetic similarities (GS) based on molecular markers are well suited for direct exploration of relationships within a germplasm pool. The objectives of this study were to: (i) assess the genetic diversity in the European winter triticale germplasm by using AFLP markers, and (ii) compare the GS estimates of AFLP markers, simple sequence repeat (SSR) markers and MALÉCOT's coancestry coefficient (f). A representative set of 127 European winter triticale varieties and breeding lines, previously investigated with SSR, was assessed with 10 PstI/TaqI primer combinations (PC). AFLP analysis identified 344 polymorphic fragments with an average polymorphic information content per PC of 0.25 and a marker index of 8.56. GS‐values between genotypes (calculated after DICE) averaged 0.61 for AFLP and 0.43 for SSR. The mean f‐value was 0.06. Dendrograms based on ‘unweighted pair‐group method and arithmetic average’ showed no clear groupings within the triticale germplasm pool, but smaller clusters were consistently found. Both molecular marker systems were superior to the coancestry coefficient for genetic diversity assessment within the elite triticale germplasm.     

Genetic diversity and differentiation of Chinese wild soybean germplasm (G. soja Sieb. & Zucc.) in geographical scale revealed by SSR markers

Wild soybean (Glycine soja), as the progenitor of soybeans (G. max), is widely distributed in China and has been collected as a supplementary germplasm pool of soybeans. In this study, 375 wild soybean accessions from a set of genebank core collection were analysed for genetic diversity by using 42 simple sequence repeat primer pairs. The mean allele number per locus was 19.62. Ten‐percent unique alleles involving 35 or 83.33% loci differentiated among the geographical regions. The mean gene diversity (h) per locus was 0.89. A very low mean coefficient of gene differentiation (G_ST = 0.08) for geographical regions and a high mean within‐region gene diversity (H_S = 0.81) were observed, indicating that most genetic diversity existed within the regions. There was an obvious relationship between genetic distance and geographical distance. The results showed multiple centers of genetic diversity for Chinese wild soybean in North China, the Huanghe River Valley, and Central China as well as the Changjiang River Valley, implicating multiple site origins of soybeans within China.     

北疆早熟陆地棉品种的遗传多样性分析

从SSR水平分析北疆早熟陆地棉品种的遗传多样性,为该棉区种质资源的创新和新品种选育提供依据。利用38个SSR引物对北疆33个早熟陆地棉品种进行遗传多样性分析,通过NTSYS-pc2.11软件计算品种之间的相似系数,并进行聚类和系谱追溯。38个引物在33个品种中共扩增出126个条带,平均每个引物为3.32个。共检测出160个等位基因,等位基因变异的多态信息含量(PIC)在0.1690～0.8503。33个品种间的遗传相似系数在0.4120～0.9560,遗传相似系数在0.57～0.61可将33个棉花品种分为2类。其中,第Ⅰ类包含17个品种,第Ⅱ类包含16个品种。在第Ⅰ类中以贝尔斯诺血统为主,第Ⅱ类中以中棉所17和新陆早16号血统为主。SSR标记分析与系谱分析的结果对北疆棉花品种间的遗传多样性分析结论基本一致。本研究表明北疆棉花品种的遗传基础狭窄。     

AFLP analysis of genetic variability among Stylosanthes guianensis accessions resistant and susceptible to the stylo anthracnose

Stylosanthes guianensis, belonging to the genus Stylosanthes, is one of the most important tropical forage legumes and is native to South and Central America and Africa. Anthracnose, caused by the fungus Colletotrichum gloeosporioides (Penz.) Sacc., is a major constraint to the extensive use of Stylosanthes as tropical forage. Forty‐two accessions of S. guianensis were assessed with amplified fragment length polymorphism (AFLP) for genetic diversity and for resistance to anthracnose. In AFLP analysis, four selective primer combinations screened from 96 primer combinations were used to analyse these accessions, and a total of 225 clear bands were used for genetic similarity (GS) analysis, showing a 95.5% level of polymorphism on average. GS from 31.0% to 95.0% among the accessions was calculated with ntsys ‐pc software. The dendrogram was constructed with unweighted pair group method of averages (UPGMA) based on the AFLP data, and five clusters were defined at 48% GS. Two typical strains of C. gloeosporioides from Stylosanthes in China were used for anthracnose resistance screening. Most of the plant accessions showed variation in the reaction to two strains, and the correlation of resistance had a value of 0.904 (P < 0.01), suggesting common resistance to the two strains. The resistance accessions were randomly distributed in different groups of UPGMA clustering. These results demonstrate that AFLP analysis is an efficient method for evaluating the genetic diversity among S. guianensis accessions.     

Evaluation of genetic diversity in jute (Corchorus species) using STMS, ISSR and RAPD markers

Jute is an important fibre crop that has dominated the packaging sector for over one and a half centuries in India. For sustenance of the trade in the face of tough competition from synthetics, there is an urgent need to redesign the ongoing breeding strategy to improve both the yield and quality of jute fibre. It is therefore, essential to understand the pattern of diversity in this important commercial crop species. In the present study, genetic diversity analysis of 20 exotic germplasm lines and 20 commercial varieties of the two cultivated species (Corchorus olitorius and C. capsularis) and two wild relatives of jute (C. aestuans and C. trilocularis) was carried out using sequence tagged microsatellite site (STMS), inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD) markers. The first set of six STMS markers developed from the genomic sequence of C. olitorius was not fully transferable to the related species C. capsularis. The level of intraspecific polymorphism revealed by these markers was very low. The four ISSR and 22 RAPD primers employed in the study revealed 98.44% and 100% polymorphism, respectively, across all the species, while the level of polymorphism was significantly low within a species. The commercial varieties, particularly those of C. capsularis, had an extremely narrow genetic base that demands immediate effort for diversification. The germplasm accessions in both the cultivated species showed considerably higher levels of diversity and thus should be used in broadening the base of the varieties. All the accessions of C. olitorius together with the wild species C. aestuans clustered separately from those of C. capsularis and C. trilocularis, suggesting a polyphyletic origin of the two cultivated species.     

Vine-1 retrotransposon-based sequence-specific amplified polymorphism for Vitis vinifera L. genotyping

The sequence‐specific amplification polymorphism (S‐SAP) method, derived from the amplified fragment length polymorphism (AFLP) technique, produces amplified fragments containing retrotransposon long terminal repeat ( LTR ) sequence at one end and a host restriction site at the other. The development and application of this procedure to the LTR of the Vine‐1 element from grapevine is reported. Two primers derived from one of the LTR sequences flanking the retrotransposon were used in combination with MseI degenerated primers on 15 grapevine accessions. S‐SAP results were compared with AFLP data. The heterozygosity and gene diversity values were higher for S‐SAP than for the AFLP procedure. Results show that S‐SAP amplification is effective in identifying polymorphisms and defining genetic distances among cultivars, and could be used for fingerprinting and for ‘Traminer’ clone identification. To the contrary Vine‐1 retrotransposon‐based S‐SAP was not able to distinguish ‘Pinot’ clones.     

Molecular diversity and genome-wide linkage disequilibrium patterns in a worldwide collection of <Emphasis Type="Italic">Oryza sativa</Emphasis> and its wild relatives

Genetic diversity analysis within a species is vital for understanding evolutionary processes at the population and genomic levels. We report a detailed study of molecular diversity, polymorphism and linkage disequilibrium in three groups of rice (Oryza) germplasm accessions based on 176 SSR markers. The first group included 65 rice (O. sativa L.) accessions introduced from seven countries, including five regions of China. The second group included 58 US rice varieties released in the past 25 years. The third group consisted of 54 accessions of rice wild relatives represented by ten different species. The number of alleles per SSR marker ranged from 4 to 32 with a mean of 16 alleles and the polymorphism information content values ranged from 0.43 to 0.91 with a mean of 0.70. The variation in SSR alleles was a significant contribution to the genetic discrimination of the 177 accessions within the three Oryza groups. Analysis of molecular variance identified deviation from Hardy–Weinberg equilibrium. Principal coordinates analysis clearly separated the accessions into their respective three groups. Neighbor-joining phylogenetic cluster reflects the ordination of each accession. Linkage disequilibrium (D′) averaged 0.75 in wild Oryza spp., and about 0.5 in both US and international O. sativa accessions. Our results showed that LD among adjacent loci in both O. sativa and Oryza spp. accessions is strong enough to be detecting marker-trait association via genome-wide scans.     

利用SSR分析四倍体棉种多态性

利用SSR标记对60份四倍体棉种材料进行遗传多样性分析,其中包括42份陆地棉野生种系。结果显示, 从1 050对SSR引物筛选出的95对SSR引物均能在60份材料间扩增出稳定明显的多态性条带,共检测出660个片段,其中多态性片段584个,占88.5%。每个位点的等位基因为2~12个,平均每对引物6.1个。UPGMA聚类分析显示, 42份材料的相似性系数(GS)在0.306~1.000之间,平均成对相似系数为0.493。95对引物的多态信息含量(PIC)的变幅为0.278~0.905,基因多样性(H¢)的变幅为0.451~2.451, 有效等位基因数(Ne)在1.385~10.490之间变动。SSR标记在陆地棉野生种系及四倍体棉种间均可反映丰富的遗传多样性信息,其中陆地棉与陆地棉野生种系中阔叶棉的亲缘关系最近,海岛棉和达尔文棉的亲缘关系非常相近,黄褐棉和毛棉相对较近,陆地棉与其他4个棉种的亲缘关系最远。     

Genomic variation and genetic relationships in Ipomoea spp.

Random amplified polymorphic DNA (RAPD) markers were used to develop genetic fingerprints and analyse genetic relationships among 29 Ipomoea accessions from different geographical locations around the world, including unique wild species, and reproducible profiles were obtained for all accessions using random decamer primers. The primers generated 46 polymorphic markers, one primer alone having 10 products, enabling the discrimination of all 29 accessions. A high level of genetic variability in sweet potato collections was suggested by the degree of polymorphism. Half of the Japanese land races were closely related while accessions from Papua New Guinea and The Philippines were distinct and exhibited the greatest genetic diversity. The wild species Ipomoea gracilis and Ipomoea tiliacea formed a group distinct from the cultivated sweet potato. The wild tetraploid accession K233 and the species Ipomoea trifida were progressively more related genetically to the cultivated sweet potato and are the probable progenitors of Ipomoea batatas, and may be suitable as germplasm for genetic enhancement. RAPDs proved to be useful for sweet potato systematics and should be valuable for germplasm management, gene tagging and efficient choice of parents in breeding programmes.     

Genetic analysis of forage grasses based on heterologous RFLP markers detected by rice cDNAs

Genetic polymorphism within and between three species of forage grasses, perennial ryegrass (Lolium perenne L), meadow fescue (Festuca pratensis Huds.) and tall fescue (Festuca arundinacea Schreb.), was analyzed using restriction fragment length polymorphism (RFLP) markers detected by rice cDNA probes developed at the Rice Genome Research Programme of Japan (RGP). One hundred and ninety‐seven rice cDNA clones were used for hybridization to genomic DNA of forage grasses. Many of the rice cDNA clones produced no visible band or only a smear with no discrete bands. Twenty‐three clones showed high efficiency cross‐hybridization to the genomic DNA of forage grasses. Genetic variation was evaluated for five varieties and one population of forage grasses using 12 polymorphic rice cDNA RFLP probes. Genetic variability within varieties as measured by Rogers’ genetic distance was considerably lower for the F. pratensis variety ‘Tomosakae’ than for the L. perenne and F. arundinacea varieties. To determine the genetic diversity between varieties of different species, cluster analysis was performed using data from the 12 RFLP probes. The two accessions of Lolium perenne were clustered more closely together than the three varieties of F. arundinacea. Two Japanese varieties of F. arundinacea were grouped in the same cluster. The variety‐specific RFLP markers were seen among six accessions of L. perenne, F. pratensis and F. arundinacea. Such variety‐specific RFLP markers would provide very useful tools for breeding programmes such as the intergeneric hybridization of Lolium and Festuca genera.     

Conformity and genetic relatedness estimation in crop species having a narrow genetic base: the case of cucumber (Cucumis sativus L.)*

A set of 155 SSR (107) and SCAR (48) markers were used to evaluate 53 cucumber (Cucumis sativus L.) accessions of diverse origin to characterize genetic relationships and to define a standard marker array that was most effective in detecting genetic differences in this germplasm array. A multivariate marker‐based analysis of diverse germplasm using this standard marker array (17 SSR and 5 SCAR markers) was compared with results from a set of 70 previously reported RAPD markers, and then used to explore the potential value of these genetic markers for plant variety protection (PVP) and the establishment of essential derivation (ED) threshold values in this species using elite lines and hybrids and backcross progeny. Diversity analysis allowed identification of distinctly different lines that were used for the construction of three sets of backcross families (BC₁‐BC₃). While general genetic relationships among accessions were similar in SSR/SCAR analyses (r_s= 0.65) using two genetic distance (GD) estimators, differences in accession relationships were detected between RAPD and SSR/SCAR marker evaluations regardless of the estimator used. The GDs among elite germplasm with known pedigrees were relatively small (0.06‐0.23 for any pairwise comparison). GD values decreased and degree of fixation (at three to seven loci depending on the mating) increased with increased backcrossing such that recurrent parent allelic fixation occurred in least one family of each of the BC₃ families. In many instances the degree of fixation of loci was not uniformly achieved in the BC₃. Although the level of genetic polymorphisms will likely restrict the use of molecular markers for PVP and the establishment of ED values, the use of single nucleotide differences will likely provide opportunities to define specific functional distances that have potential for PVP in cucumber. Nevertheless, without an expanded, genetically robust standard marker array (e.g. 50 codominant markers), ED threshold values will be difficult to define in this species, and perhaps will require the appraisal of single nucleotide polymorphisms as discriminators of difference in this species.     

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.     

不同类型啤酒大麦品种遗传多样性及遗传差异的SSR分析

利用28对SSR引物对包括22份野生材料,8份耐盐碱材料,21份优质高产甘啤系列品种(系)材料和9份日本早熟材料在内的60份啤酒大麦品种资源的遗传多样性及其亲缘关系进行分析。结果表明,所用28对引物,有25对有多态性;聚类结果表明这些材料遗传相似系数分布在0.4792~0.9948之间。在遗传相似系数0.63的水平上,这些材料聚类为2大类,下分7个亚类。从聚类结果看出,不同类型材料基本各自聚为一类,表明不同类型品种(系)内遗传差异较小;不同类型品种(系)间遗传差异较大,杂交可能会产生较强的杂种优势和较高的产量潜力。     

Phylogenetic relationships as in Ocimum revealed by RAPD markers

Genetic relationships were examined among thirty germplasm accessions belonging to five Ocimum species using RAPD markers. A very high degree of polymorphism (98.20%) was observed. UPGMA cluster analysis of genetic similarity indices grouped all the accessions into two major clusters corresponding to previously reported botanical sections. Intra-clustering within the two clusters precisely grouped the accessions belonging to one species in one sub-cluster as expected from their genetic background. Our results show that RAPD technique is a sensitive, precise and efficient tool for genomic analysis in Ocimum species, that may be useful in future studies, by assigning new unclassified germplasm accessions to specific taxonomic groups and reclassifying previously classified accessions of other Ocimum species by traditional criteria on a more objective basis. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effectiveness of different classes of molecular marker for classifying and revealing variation in rice (Oryza sativa) germplasm

We have examined the effectiveness of similar numbers of markers from four molecular marker systems (AFLP, isozymes, ISSR and RAPD) for revealing genetic diversity and discriminating between infraspecific groups of Oryza sativa germplasm. Each marker system classifies the germplasm into three major groups (most effectively with isozymes and AFLPs), but with differences (primarily with ISSR) between the precise classifications generated. However, at the highest levels of genetic similarity there was only partial agreement as to relationships between individual accessions when different markers were used. When variance was partitioned among and within the three subspecific groups, although the differences were not significant, greater variation was found among than within groups using AFLP and isozymes, with the reverse for RAPD and ISSR. Measurement of polymorphism using average heterozygosity and effective number of alleles gave similar results for each marker system. These results are discussed in relation to various genetic resources conservation activities, and the advisability of extrapolating to other sets of germplasm particularly of other crop species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic variability among sugarcane genotypes based on polymorphisms in sucrose metabolism and drought tolerance genes

Target region amplification polymorphism (TRAP) markers were used to estimate the genetic similarity (GS) among 53 sugarcane varieties and five species of the Saccharum complex. Seven fixed primers designed from candidate genes involved in sucrose metabolism and three from those involved in drought response metabolism were used in combination with three arbitrary primers. The clustering of the genotypes for sucrose metabolism and drought response were similar, but the GS based on Jaccard’s coefficient changed. The GS based on polymorphism in sucrose genes estimated in a set of 46 Brazilian varieties, all of which belong to the three Brazilian breeding programs, ranged from 0.52 to 0.9, and that based on drought data ranged from 0.44 to 0.95. The results suggest that genetic variability in the evaluated genes was lower in the sucrose metabolism genes than in the drought response metabolism ones.     

Population genetic structure and genetic diversity of <Emphasis Type="Italic">Jatropha curcas</Emphasis> germplasm as investigated by 5′-anchored simple sequence repeat primers

Jatropha (Jatropha curcas L.) is widely distributed in the tropics with a promise for use as an oil crop for biodiesel. Understanding population genetic structure and diversity of J. curcas is important for germplasm collection and planning for breeding programs. Twelve 5′-anchored simple sequence repeat primers were selected and used for evaluation of population genetic structure and genetic diversity of J. curcas accessions collected from different regions within Thailand and introduced from other countries. A moderate level of genetic variation was found in all accessions studied. Cluster analysis did not reflect geographical distance in general as revealed by the Mantel test. An analysis of molecular variance showed that the majority (83.3%) of genetic diversity was within populations, indicating a high level of overlap and little population subdivision with respect to geographic area. An absence of subpopulation structure among neighboring regions may be caused by exchanging germplasm with vegetative propagation. This study provides a basis for breeders on selection of parental material to maximize genetic polymorphism in J. curcas breeding programs.     

Genetic diversity of garlic (Allium sativum L.) germplasm from China by fluorescent‐based AFLP,SSR and InDel markers

Garlic (Allium sativum L.), an asexually propagated crop, is an important vegetable and medicinal plant. China is the biggest garlic producer in the world; however, the genetic background of garlic from China is not well understood. In this study, population structure and clustering analysis of garlic germplasm was performed using amplified fragment length polymorphism (AFLP), simple sequence repeat (SSR) and insertion–deletion (InDel) markers. Among 212 accessions of garlic, genetic diversity analysis identified 546 alleles amplified by AFLP, SSR and InDel primers, and 492 of these were polymorphic. All accessions were divided into five groups by structure analysis and neighbor‐joining clustering. Most traits, including allicin content, were only slightly affected by population structure, which indicated that this germplasm can be used as populations for association mapping. The results provide a molecular basis for understanding the genetic diversity of the garlic germplasm preserved in China.