Assessment of genetic diversity of wild soybean (Glycine soja Siebold et Zucc.) in the far eastern region of Russia

Polymorphism of RAPD markers was analyzed in the wild soybean populations from the Far East region of Russia. The level of RAPD marker polymorphism was significantly higher in the wild than in the cultivated soybean. The results obtained suggest active development of genetically different groups of wild soybean. Geographically isolated subpopulations showed maximum distance from the main population of wild soybean.     

Investigation of genetically modified soybean biosafety in the center of the origin and diversity of Russian Far East

Wild soybean (Glycine soja Sieb. & Zucc.) is the nearest relative of a soybean crop (Glycine max (L.) Merr.). Study of population genetic structure of wild-growing relatives ofgenetically modified (GM) plants in the centers of their origin is one of the main procedures before introduction of GM crops in these areas. We have studied genetic variability of nine wild growing soya populations of Primorye Territory using RAPD analysis. The level of G. soja genetic variability was considerably higher than that of G. max. We have analyzed phylogenetic relationships in the genus Glycine subgenus Soja using RAPD markers. Our data confirm validity of allocation G. gracilis in a rank of a species.     

Detection of Polymorphic DNA and Taxonomic Relationships Among 10 Wild Perennial Soybean Species Using Specific and Arbitrary Nucleotide Primers

Polymorphic DNA in complex genomes of agronomic crops can be detected using specific nucleotide and arbitrary primers and the polymerase chain reaction (PCR). Nineteen accessions representing 10 species of the wild perennial soybean were evaluated using 4 sets of specific primers and 3 sets of random amplified polymorphic DNAs (RAPD) primers. The potential of the RAPD assays was further increased by combining two primers in a single PCR. The fragments generated by the two assays discriminated 10 wild species by banding profiles. The size of the amplified DNA fragments ranged from 100 to 2100 base pairs. The resolved PCR products yielded highly characteristic and homogeneous DNA fingerprints. The fingerprints were useful not only for investigating genetic variability but also for further characterizing the wild soybean species by detecting inter- and intra-specific polymorphisms, constructing dendrograms defining the phylogenetic relationships among these species, and identifying molecular markers for the construction of genetic linkage maps. Furthermore, unique markers distinguishing particular species were also identified. Thus, it is expected that PCR will have great relevance for taxonomic studies. This revised version was published online in July 2006 with corrections to the Cover Date.     

不同尺度下野大豆种群的遗传分化

为了阐明不同尺度范围内野大豆种群的遗传分化情况,应用随机扩增多态性DNA（RAPD）方法,分别对我国5个纬度8个不同地点的野大豆（Glycine soja)种群及浙江金华地区5个野大豆种群,进行了分子生态学研究。根据RAPD数据计算相似系数及遗传距离并进行聚类分析,发现无论是不同纬度野大豆种群还是金华地区野大豆小种群均存在较高的遗传变异,且不同纬度野大豆种群间的遗传变异与地理纬度有一定正相关。在对金华地区野大豆种群遗传多样怀的研究,利用Shannon指数估算了5个野大豆种群的遗传多样性,发现大部分的遗传变异存在于野大豆种群间（78．5％）,只有少部分的遗传变异存在于种群内。本就此探讨了不同尺度下野大豆种群的遗传多样性与环境因子的关系,并对其成因及维持机制进行了讨论。     

陕西大豆资源遗传多样性及变异特点研究

利用42个PAPD引物对75份陕西大豆种质进行遗传多样性分析,共扩增出310个条带,平均每个引物扩增7.3个条带,多态性比率为96%;田间试验考察了13个农艺性状。陕西大豆的遗传多样性在秦岭南、北两个地区有所不同,秦岭北品种遗传多样性指数较高的性状数目和性状遗传多样性指数都大于秦岭南品种,RAPD分子标记遗传多样性指数也是秦岭北品种大于秦岭南品种,但秦岭南品种RAPD分子标记的遗传多样性指数较高的个数大于秦岭北品种。聚类分析将参试大豆材料分为三大类,基本上反映了材料的地理来源。主成分分析结果显示,前两个主成分反映了10.95%的遗传变异,基于前两个主成分值的二维散点图可以将两个地区的材料基本区分开来。AMOVA分析显示,陕西大豆品种个体间的遗传变异占总变异的92.06%,地区间的遗传变异占总变异的7.94%,二者都达到了极显著水平。研究结果表明,陕西大豆资源存在丰富的遗传多样性,秦岭北品种遗传多样性较高,但秦岭南品种有着广泛的微小变异。     

Characterization of Natural and Simulated Herbivory on Wild Soybean (Glycine soja Seib. et Zucc.) for Use in Ecological Risk Assessment of Insect Protected Soybean

Insect-protected soybean (Glycine max (L.) Merr.) was developed to protect against foliage feeding by certain Lepidopteran insects. The assessment of potential consequences of transgene introgression from soybean to wild soybean (Glycine soja Seib. et Zucc.) is required as one aspect of the environmental risk assessment (ERA) in Japan. A potential hazard of insect-protected soybean may be hypothesized as transfer of a trait by gene flow to wild soybean and subsequent reduction in foliage feeding by Lepidopteran insects that result in increased weediness of wild soybean in Japan. To assess this potential hazard two studies were conducted. A three-year survey of wild soybean populations in Japan was conducted to establish basic information on foliage damage caused by different herbivores. When assessed across all populations and years within each prefecture, the total foliage from different herbivores was ≤ 30%, with the lowest levels of defoliation (< 2%) caused by Lepidopteran insects. A separate experiment using five levels of simulated defoliation (0%, 10%, 25%, 50% and 100%) was conducted to assess the impact on pod and seed production and time to maturity of wild soybean. The results indicated that there was no decrease in wild soybean plants pod or seed number or time to maturity at defoliation rates up to 50%. The results from these experiments indicate that wild soybean is not limited by lepidopteran feeding and has an ability to compensate for defoliation levels observed in nature. Therefore, the potential hazard to wild soybean from the importation of insect-protected soybean for food and feed into Japan is negligible.     

Identification of wild and cultivated Hordeum species using two-primer RAPD fragments

Random amplified polymorphic DNAs (RAPD) analysis has been adapted to assess the degree of RAPD polymorphism within the genus Hordeum to determine if this approach can distinguish wild and cultivated species. Nineteen wild and seven cultivated accessions were evaluated using 4 random 10-mer primers. The potential of the RAPD assay was further increased by combining two primers in a single polymerase chain reaction (PCR). RAPD fragments generated by two pairs of arbitrary 10-mer primers discriminated six wild species and one cultivated species by banding profiles. The size of the amplified DNA fragments ranged from 150 to 2300 base pairs. 33 %percent of the fragments were common to both wild and cultivated species; 67% were specific to either wild or cultivated species. The average difference in fragments was less within the species than among the species. By comparing RAPD fingerprints of wild and cultivated barley, markers were identified among the set of amplified DNA fragments which could be used to distinguish wild and cultivated Hordeum species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Random amplified polymorphic DNA analysis of genetically modified organisms

Randomly amplified polymorphic DNA (RAPD) was used to analyzed 78 samples comprises of certified reference materials (soya and maize powder), raw seeds (soybean and maize), processed food and animal feed. Combination assay of two arbitrary primers in the RAPD analysis enable to distinguish genetically modified organism (GMO) reference materials from the samples tested. Dendrogram analysis revealed 13 clusters at 45% similarity from the RAPD. RAPD analysis showed that the maize and soybean samples were clustered differently besides the GMO and non-GMO products.     

盐渍条件下野大豆群体的遗传分化和生理适应:同工酶和随机扩增多态DNA研究

黄河三角洲野大豆(Glycine soja Sieb. et Zucc.)盐渍群体的耐盐性高于附近的正常群体。群体内个体间耐盐能力差别很大。盐渍群体有比最耐盐的栽培大豆(G.max (L.)Merr.)品种耐盐能力高得多的个体,也有对盐相当敏感的植株。同工酶分析表明群体内高水半多态性,但酶谱与抗性没有相关性。盐渍与正常群体间的遗传一致性高达0.96。用改良的随机扩增多态DNA(RAPD)方法,10个引物扩增得出群体内多态位点百分数为68/188=0.36。看来,绝大多数位点与耐盐能力无关。上述资料说明,盐渍条件下野大豆自然群体的高度遗传多样性和发育变通性,可能足对盐胁迫强度随时随地变化的环境的适应。     

中国部分地区一年生野生大豆资源考察、收集及分布现状分析

2002-2004年对中国10省(市)221个县(市)的一年生野生大豆资源进行了考察,新发现有野生大豆分布的县(市)45个,收集野生大豆资源814份.通过考察发现野生大豆在中国分布面积锐减,濒危状况严重.     

Genetic Differentiation and Physiological Adaptation of Wild Soybean (Glycine soja) Populations Under Saline Conditions: Isozymatic and Random Amplified Polymorphic DNA Study

A higher level of salt tolerance in saline populations of wild soybean ( Glycine soja Sieb. et Zucc. ) has been found in the Yellow River Delta. The levels of salt tolerance for individuals with in populations vary widely. In the saline populations, there are a few plants with much higher tolerant level than that of the most tolerant cultivars, such as Morgan, and others as saline sensitive plants. Isozyme analysis showed a high level of genetic diversity, however, no relationship was found between isozymatic patterns and salt tolerance. Saline and normal populations shared similar genetic structure with a genetic distance of 0. 045. By using a modified procedure of random amplified polymorphic DNA(RAPD), a higher level of DNA diversity was detected in saline populations. The authors propose that the high level of genetic diversity and developmental flexibility is responsible for adaptation of wild soybean to changing saline conditions.     

东北地区大豆种质资源的RAPD聚类分析

对东北地区不同类型大豆共３２份材料进行了ＲＡＰＤ分析,并应用ＰＳＢＡＰＣ２软件按照最远法构建了聚类图。结果表明,东北不同地区的栽培大豆各以较近的关系聚在一起,基本反映了大豆品种生态类型的地理分布。野生种和半野生种大豆可以聚为一类,但二者与栽培大豆明显分开,故初步认为与野生种的亲缘关系较近,或应归类到野生种中。     

我国野生大豆资源保护管理问题

阐述了国家重点保护野生植物野生大豆的利用价值及在我国面临的问题,介绍了湖南省的野生大豆资源保护管理的经验,科学合理地提出了保护好我国野生大豆资源保护管理的建议.     

Population structure of the wild soybean (Glycine soja) in China: implications from microsatellite analyses

Background and Aims Wild soybean (Glycine soja), a native species of East Asia, is the closest wild relative of the cultivated soybean (G. max) and supplies valuable genetic resources for cultivar breeding. Analyses of the genetic variation and population structure of wild soybean are fundamental for effective conservation studies and utilization of this valuable genetic resource. Methods In this study, 40 wild soybean populations from China were genotyped with 20 microsatellites to investigate the natural population structure and genetic diversity. These results were integrated with previous microsatellite analyses for 231 representative individuals from East Asia to investigate the genetic relationships of wild soybeans from China. Key Results Analysis of molecular variance (AMOVA) revealed that 43·92 % of the molecular variance occurred within populations, although relatively low genetic diversity was detected for natural wild soybean populations. Most of the populations exhibited significant effects of a genetic bottleneck. Principal co-ordinate analysis, construction of a Neighbor-Joining tree and Bayesian clustering indicated two main genotypic clusters of wild soybean from China. The wild soybean populations, which are distributed in north-east and south China, separated by the Huang-Huai Valley, displayed similar genotypes, whereas those populations from the Huang-Huai Valley were different. Conclusions The previously unknown population structure of the natural populations of wild soybean distributed throughout China was determined. Two evolutionarily significant units were defined and further analysed by combining genetic diversity and structure analyses from Chinese populations with representative samples from Eastern Asia. The study suggests that during the glacial period there may have been an expansion route between south-east and north-east China, via the temperate forests in the East China Sea Land Bridge, which resulted in similar genotypes of wild soybean populations from these regions. Genetic diversity and bottleneck analysis supports that both extensive collection of germplasm resources and habitat management strategies should be undertaken for effective conservation studies of these important wild soybean resources.     

RAPD Determinations on the Genetic Differentiation of Chinese Common Wild Rice

Ninety accessions which included Chinese common wild rice (Oryza rufipogon) from 8 provinces and traditional cultivars from lower and middle basins of Yangtze River, southeast of China and Yunnan Province as well as some commercial varieties were analyzed by RAPD with 24 primers. A scattered figure suggesting the indica-japonica and wild-domestication differentiations among 90 rice accessions was generated based on RAPD data. The results indicated that Chinese common wild rice, indica and japonica accessions were divided into 3 groups respectively. Chinese common wild rice were somewhat closer to the japonica type than the indica type.     

DNA amplification polymorphisms of the cultivated mushroom Agaricus bisporus.

Single 10-bp primers were used to generate random amplified polymorphic DNA (RAPD) markers from commercial and wild strains of the cultivated mushroom Agaricus bisporus via the polymerase chain reaction. Of 20 primers tested, 19 amplified A. bisporus DNA, each producing 5 to 15 scorable markers ranging from 0.5 to 3.0 kbp. RAPD markers identified seven distinct genotypes among eight heterokaryotic strains; two of the commercial strains were shown to be related to each other through single-spore descent. Homokaryons recovered from protoplast regenerants of heterokaryotic strains carried a subset of the RAPD markers found in the heterokaryon, and both of the haploid nuclei from two heterokaryons were distinguishable. RAPD markers also served to verify the creation of a hybrid heterokaryon and to analyze meiotic progeny from this new strain: most of the basidiospores displayed RAPD fingerprints identical to that of the parental heterokaryon, although a few selected slow growers were homoallelic at a number of loci that were heteroallelic in the parent, suggesting that they represented rare homokaryotic basidiospores; crossover events between a RAPD marker locus and its respective centromere appeared to be infrequent. These results demonstrate that RAPD markers provide an efficient alternative for strain fingerprinting and a versatile tool for genetic studies and manipulations of A. bisporus.     

应用RAPD标记研究野生荔枝种质资源

应用RAPD方法对海南60份野生荔枝进行基因组多态性分析,20个随机引物共产生165条RAPD带,DNA片段大小在200～1500bp之间,其中121条为多态性带,占总带数的73．3％,并利用遗传距离进行聚类分析。结果表明,60份野生荔枝可归为6类,表明种群存在一定的遗传变异,也为分析野生荔枝群体间及群体内的遗传多样性探索了有效方法。     

橡胶树种质资源遗传多样性研究——Ⅰ．速生种质遗传多样性RAPD分析

应用RAPD技术对8份野生种质和12份栽培种质进行遗传多样性分析,筛选到18个具有多态性扩增的引物．共扩增出128条带。据Nei-Li相似系数将20份材料分别聚为野生种质和栽培种质两大类。5个野生种质聚为野生种质类群,12个栽培种质和3个野生种质聚为栽培种质类群。研究结果表明,RAPD技术用于橡胶树种质资源研究,能够为野生种质优良特性导入栽培种质提供分子水平的参考依据。     

海南主要普通野生稻的调查及利用RAPD技术进行鉴别分析

通过对海南现存的主要普通野生稻资源进行考察,报道了海南主要普通野生稻的最新生存状况;并用RAPD技术对其进行分析。从供试材料中筛选出具有多态性的RAPD引物13条,共扩增出155条带,其中多态性条带120条,多态性条带比率(PPB)值为77.42%。UPGMA聚类结果表明:海南主要普通野生稻之间遗传关系较复杂、差异较明显,并将海南主要普通野生稻资源划分为5个小类群,可以很好地将海南本地野生稻与收集保存的野生稻鉴别开来,为海南普通野生稻资源的合理利用与有效保护提供了重要依据。     

DNA amplification polymorphisms of the cultivated mushroom Agaricus bisporus.

Single 10-bp primers were used to generate random amplified polymorphic DNA (RAPD) markers from commercial and wild strains of the cultivated mushroom Agaricus bisporus via the polymerase chain reaction. Of 20 primers tested, 19 amplified A. bisporus DNA, each producing 5 to 15 scorable markers ranging from 0.5 to 3.0 kbp. RAPD markers identified seven distinct genotypes among eight heterokaryotic strains; two of the commercial strains were shown to be related to each other through single-spore descent. Homokaryons recovered from protoplast regenerants of heterokaryotic strains carried a subset of the RAPD markers found in the heterokaryon, and both of the haploid nuclei from two heterokaryons were distinguishable. RAPD markers also served to verify the creation of a hybrid heterokaryon and to analyze meiotic progeny from this new strain: most of the basidiospores displayed RAPD fingerprints identical to that of the parental heterokaryon, although a few selected slow growers were homoallelic at a number of loci that were heteroallelic in the parent, suggesting that they represented rare homokaryotic basidiospores; crossover events between a RAPD marker locus and its respective centromere appeared to be infrequent. These results demonstrate that RAPD markers provide an efficient alternative for strain fingerprinting and a versatile tool for genetic studies and manipulations of A. bisporus.