辣椒种质遗传多样性的RAPD和ISSR及其表型数据分析

用RAPDI、SSR分子标记及28个表型性状数据对辣椒属5个栽培种的13份材料进行了分析,结果表明:23条RAPD引物共扩增出209条带,平均每个引物扩增出9.09条,多态性位点比率为83.73%;16条ISSR引物共扩增出94条带,平均每个引物扩增出5.88条,多态性位点比率为79.79%.与RAPD相比,ISSR标记检测到的有效等位基因数(Ne)及Shannon多样性指数(I)、遗传离散度(Ht)和遗传分化系数(Gst)等遗传多样性参数都较大,多态性位点比例在亲缘关系较近的一年生辣椒(Capsicum annuum)种内较高,说明ISSR有更高的多态性检测效率,并且适合亲缘关系较近的种群间遗传多样性分析.基于RAPDI、SSR的聚类与基于表型数据的聚类之间存在极显著正相关,且都能将C.annuum与其它栽培种区分开来.     

大戟属植物亲缘关系的ISSR分析

利用ISSR标记对9种大戟属植物的亲缘关系进行了分析.结果表明:筛选出的24条ISSR引物共扩增251条带,其中多态性条带248条,多态率为98.8%.9种大戟属材料的遗传相似系数在0.51～0.69之间,平均为0.62.利用UPGMA法将9种大戟属植物分别聚为3个类群,第一类群包含地锦亚属与一品红亚属的5个种及大戟亚属的绿玉树,第二、第三类群分别包含大戟亚属中的3个种.ISSR标记获得的聚类结果与根据形态特征获得的分类结果有很大的差异.     

应用ISSR标记研究仲彬草属植物的遗传变异

利用ISSR标记对仲彬草属14个种和1个变种共32份材料进行了研究。结果表明:仲彬草属材料间ISSR标记多态性较高,变异较大。12个引物共扩增出593条带,其中535条(90.2%)具有多态性,每个引物可扩增出11~80条多态性带,平均44.6条。ISSR标记遗传相似系数在种间变化范围为0.430~0.866,平均值为0.620。从聚类分析得知,ISSR标记能将32份仲彬草属材料完全分开,32份材料聚为4类。同种不同居群的材料分别聚在一起,亲缘关系较近,同时,在分子水平上种内不同居群间也存在分化;种间存在明显的遗传差异;形态相似、地理分布一致的物种倾向于聚类在一起,有一定的亲缘关系。因此,ISSR分子标记能有效地评价仲彬草属物种的亲缘关系。     

7种川产淫羊藿属植物遗传多样性及亲缘关系的ISSR分析

利用ISSR分子标记对7种12居群的川产淫羊藿属植物进行遗传多样性及亲缘关系分析。结果发现23个ISSR随机引物共扩增194条清晰条带,其中169条具多态性,平均多态性位点比率为87.11%,有效等位基因数Ne=1.534 2,Nei基因多样性指数H=0.314 4,Shannon多样性指数I=0.469 7,表明物种间遗传多样性丰富。7种川产淫羊藿属植物间的遗传相似系数为0.653 2～0.748 9,聚类分析和主成分分析直观的显示出12份供试材料间的亲缘关系,并将其分为两类。表明ISSR分子标记技术可以用于川产淫羊藿属植物的遗传多样性和亲缘关系分析。     

兰属Cymbidium植物ISSR遗传多样性分析

将简单重复序列区间扩增多态标记(inter-simple sequence repeats, ISSR)技术应用于16种兰属的遗传多样性分析, 15个引物共扩增出836条带, 其中有227条多态带, 多态百分比为27.2%。UPGMA聚类结果显示: 春兰与春剑的亲缘关系最近, 而兔耳兰与其他15种兰属植物的距离最远。ISSR聚类结果与传统分类结果基本相似, 表明该技术能在分子水平上对传统兰属分类进行必要补充。     

22种木莲属植物亲缘关系的ISSR分析

利用ISSR标记分析技术探讨了22种木莲属植物的遗传亲缘关系。选用10对ISSR引物组合对木莲属22个种的基因组进行分析。结果表明：22个种共扩增出595条DNA带,其中多态性条带占94.45%;根据Nei Li遗传相似性系数在0.68处,UPGMA聚类结果将22个种划分为6个类群,在0.70处进一步可分为12个亚类群;22种木莲属植物的Nei-Li遗传相似性系数变化范围为0.605(乳源木莲与球果木莲之间)～0.956(中缅木莲与滇南木莲之间),平均遗传相似性系数为0.698。说明木莲属植物种间差异性大。其中中缅木莲、滇南木莲之间遗传相似性系数为0.956,两者亲缘关系最近。该结果同时支持将巴东木莲、乳源木莲、滇桂木莲分别作为独立的种。     

木霉肽类代谢产物对豇豆土著根瘤菌遗传性状的影响

利用RAPD与ISSR分子标记检测手段,分析了哈茨木霉T2-16肽类代谢产物处理豇豆土著根瘤菌,对其遗传性状的影响,同时,比较了RAPD和ISSR两种不同分子标记在检测根瘤菌种间的遗传相似性以及遗传变异性的分辨力.实验中,从100条引物中筛选到具有多态性的ISSR引物5条,从80条引物中筛选到具有多态性的RAPD引物6条,用5条ISSR引物扩增出54条带,多态性条带比率为75.93 %;6条RAPD引物扩增出61条带,多态性条带比率为68.85 %.两种分子标记均能揭示出处理前后根瘤菌间的遗传差异,但ISSR标记比RAPD标记可检测到更大的遗传变异.根据两种标记的结果,对供试的根瘤菌进行聚类分析,结果表明,土著根瘤菌经木霉肽类代谢产物处理后,与出发菌株相比,表现出一定程度的遗传分化和遗传差异性.     

利用RAPD和ISSR标记分析青麻种质遗传多样性

利用RAPD和ISSR分子标记检测来自全国11个省(市)的48份青麻种质资源的遗传多样性,为青麻资源利用和育种提供分子生物学依据.在48份青麻种质资源中,17条RAPD引物扩增出191条带,多态条带比率为87.43%;9条ISSR引物扩增出82条带,多态条带比率为88.89%,扩增产物片段大小都在0.1～3.0kb之间.两种分子标记的结果呈显著正相关(r=0.80).基于UPGMA聚类,野生种和栽培种各自聚为相应的类别.     

云南普通野生稻遗传多样性和亲缘关系

野生稻（Oryza rufipogon）是稻属的重要组成部分,具有许多优良性状,是水稻遗传改良的天然基因库。本研究通过对形态学性状的观测,及ISSR和RAPDUPGMA聚类分析,将云南普通野生稻划分为4个类型,即元江类型、景洪紫杆直立型、景洪绿杆直立型和景洪匍匐型。在供试材料中筛选到具有多态性的ISSR和RAPD引物各11个,ISSR引物扩增出多态带113条,多态性条带比率（PPB）为82．26％,RAPD引物共扩增出多态性条带76条,PPB值为76．77％,两种分子标记的分析结果呈极显著正相关（r=0.951）。此外UPGMA聚类结果表明,云南普通野生稻不同类型与其它地区普通野生稻之间的遗传亲缘关系差异明显。     

云南普通野生稻遗传多样性和亲缘关系

野生稻(Oryza rufipogon)是稻属的重要组成部分, 具有许多优良性状, 是水稻遗传改良的天然基因库。本研究通过对形态学性状的观测, 及ISSR和RAPD UPGMA聚类分析, 将云南普通野生稻划分为4个类型, 即元江类型、景洪紫杆直立型、景洪绿杆直立型和景洪匍匐型。在供试材料中筛选到具有多态性的ISSR和RAPD引物各11个, ISSR引物扩增出多态带113条, 多态性条带比率(PPB)为82.26%, RAPD引物共扩增出多态性条带76条, PPB值为76.77%, 两种分子标记的分析结果呈极显著正相关(r = 0.951)。此外UPGMA聚类结果表明, 云南普通野生稻不同类型与其它地区普通野生稻之间的遗传亲缘关系差异明显。     

Genetic variation of the genus Kengyilia by ISSR markers

We investigated the genetic variation within 32 accessions distributed to 14 species and one variety by using ISSR (inter-simple sequence repeat) markers. The results showed that genetic variation was relatively higher among the accessions. A total of 593 bands were amplified by 12 ISSR primers, of which 535 bands (90.2%) were polymorphic. Eleven to 80 polymorphic bands were amplified from each prime, with an average of 44.6 bands. The interspecies GS (genetic similarity) value ranged from 0.430 to 0.866, and the average was 0.620. Cluster analysis showed that all accessions could be classified into 4 groups by ISSR markers. The different accessions in a species were clustered together, but they had genetic variation in molecular levels. There was obvious interspecies genetic variation. Species with similar morphological characteristics and from the same areas or neighboring geographical regions were clustered together and had close relationships. ISSR markers are useful in analyzing interspecies variation in Kengyilia. __________ Translated from Guihaia, 2006, 26 (4): 375–380 [译自: 广西植物]     

Genetic similarity among cultivars of Phyllostachys pubescens

Phyllostachys pubescens is the most important economic bamboo species in China, which grows widely in the South of China. There are more than ten cultivars in this species but their genetic relationship still remains unknown. We used both amplified fragment length polymorphism (AFLP) and inter-simple sequence repeat (ISSR) techniques to determine genetic similarity among ten cultivars of P. pubescens and two related species. Eight hundred and twenty seven bands, in which 495 are polymorphic, were detected using 15 pairs of AFLP primers whereas total 231 bands, in which 154 bands are polymorphic, were scored using 16 ISSR primers. Statistic analysis showed that the genetic similarity matrices obtained from these two sets of molecular markers had a significant correlation (R = 0.959, P = 0.013). The dendrogram generated with AFLP and ISSR markers could clearly genetically identify ten cultivars of P. pubescens that had high similarity with genetic distances ranging from 0.023 to 0.108, and could be divided into three groups based on their genetic variation and similarity. Our results suggest that these molecular markers are useful to genetically classify cultivars or varieties of a species, particularly a bamboo species.     

Differentiation of commercial strains of <Emphasis Type="Italic">Agaricus</Emphasis> species in China with inter-simple sequence repeat marker

In recent years, Agaricus mushroom production has undergone an important increase in the world. However, there is no consensus over the taxonomy of some commercial strains. So rapid and reliable methods to identify Agaricus strains are urgently needed. In this paper, six inter-simple sequence repeat (ISSR) primers were screened from 20 primers to analyse the genetic diversities of the 12 main commercial strains of Agaricus in China. The results showed that a total of 124 bands were produced, among which 80.6% was polymorphic, the similarity coefficients ranged from 0.69 to 0.98. Twelve tested strains were divided into 2 groups on the similarity coefficient of 0.75. Three Agaricus bitorquis strains were clustered in one group, and seven Agaricus bisporus strains were clustered with Agaricus strains S1 and S2 in the other group. As comparison with ISSR, intergenic spacers combined with restriction fragment length polymorphisms (IGS-RFLP) was employed to analyse genetic diversities of the tested strains by digesting IGS amplified products with restriction enzymes Hae III, Alu I, Rsa I, respectively, and IGS-RFLP cluster results were similar with those of ISSR. However, ISSR was superior to IGS-RFLP in differentiating closely related strains. It is suggested that the ISSR marker is an alternative method to differentiate Agaricus strains.     

送春与多花兰种间杂交后代的ISSR分析

该文使用简单重复序列间( ISSR)分子标记,对送春与多花兰种间杂交后代进行了研究。结果表明：从80个ISSR引物中筛选出14个扩增效果稳定的ISSR引物,对两亲本和59个F1代个体进行了ISSR扩增,得到107个扩增位点,扩增的片段大小位于90~2100 bp之间,平均每个引物扩增7.64条条带,得到11种类型的带。 ISSR标记在送春×多花兰的F1代中表现出一定的多态性,分离频率为44.86％,分离位点有83.33％符合孟德尔1︰1或3︰1的分离规律,产生偏孟德尔分离的位点占12.50％,余下的4.17％属于特殊分离带型。可能导致后代变异的位点为偏孟德尔分离的6条带、缺失的8条带或新生成的2条带。聚类图中父本和母本与F1代个体间的遗传距离较远,59个杂交后代先聚集成一组,再同母本相聚为一组,最后才同父本聚在一起,59个杂种均偏母本型。送春与多花兰的杂交后代在植株形态、染色体、遗传物质方面都具备双亲特点,61个个体间的ISSR分子量标记结果和植株形态学特征都说明,59个F1代杂种包含送春和多花兰的遗传特性是真杂种;F1代杂种既有双亲的互补特征带,又有双亲的重组片断即产生新的特异带,这说明送春与多花兰的杂交后代具有遗传变异的特点。该研究结果可以有效地对杂交后代进行定向选择,为兰花的杂交育种提供了分子依据。     

福建省野牡丹属种质资源的ISSR分析

为探讨福建省野牡丹属(Melastoma L.)植物的亲缘关系,运用ISSR分子标记技术对源自福建省的野牡丹属野生种质及部分实生后代共34份材料进行分析。结果表明,11条多态性引物对34份种质DNA进行扩增,共获得112条完整、清晰的谱带,其中多态性条带104条,多态性比率为92.9%,表明野牡丹属种质资源具有较高的遗传多样性。34份种质材料的相似系数为0.55~0.93,平均相似系数为0.71,表明这些材料间的亲缘关系较近。聚类分析表明34份材料可划分为3个类群5个亚类,主坐标散点分析可分为4个类群,这与亲缘关系分析的结果基本一致。这从分子水平上揭示了福建省野牡丹属野生种质及部分具备优良园艺性状实生后代的亲缘关系,为该属植物的良种选育工作提供了理论依据。     

A Comparative Analysis of ISSR and RAPD Markers for Study of Genetic Diversity in Shisham (<Emphasis Type="Italic">Dalbergia sissoo</Emphasis>)

Shisham (Dalbergia sissoo) is one of the most preferred timber tree species of South Asia. Two DNA-based molecular marker techniques, intersimple sequence repeat (ISSR) and random amplified polymorphism DNA (RAPD), were compared to study the genetic diversity in this species. A total of 30 polymorphic primers (15 ISSR and 15 random) were used. Amplification of genomic DNA of 22 genotypes, using ISSR analysis, yielded 117 fragments, of which 64 were polymorphic. Number of amplified fragments with ISSR primers ranged from five to ten and varied in size from 180 to 1,900 bp. Percentage polymorphism ranged from 0 to 87.5. The 15 RAPD primers produced 144 bands across 22 genotypes, of which 84 were polymorphic. The number of amplified bands varied from five to 13, with size range from 180 to 2,400 bp. Percentage polymorphism ranged from 0 to 100, with an average of 58.3 across. RAPD markers were relatively more efficient than the ISSR assay. The mental test between two Jaccard’s similarity matrices gave r ≥ 0.90, showing very good fit correlation in between ISSR- and RAPD-based similarities. Clustering of isolates remained more or less the same in RAPD and combined data of RAPD and ISSR. The similarity coefficient ranged from 0.734 to 0.939, 0.563 to 0.946, and 0.648 to 0.920 with ISSR, RAPD, and combined dendrogram, respectively.     

Genetic diversity of Alternanthera philoxeroides in China

Alternanthera philoxeroides (Mart.) Grisb was introduced into China in the 1930s, and today occurs in most regions of southern China. Techniques using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) markers were applied to analyze genetic diversity of this invasive, weedy species. The fragments amplified by both 28 RAPD primers and 23 ISSR primers showed no polymorphic bands within and among the seven populations sampled. These results might be a consequence of the short introduction history in China and the clonal propagation of this aquatic plant. Although A. philoxeroides is widely distributed in China, the molecular data indicated its genetic diversity is extremely low, which implies that the low genetic diversity did not affect the success of its expansion in China. The rapid range expansion of A. philoxeroides is most likely the result of a massive clonal propagation since its introduction.     

Morphological and molecular genetic variations of oat genotypes grown in Kermanshah,Iran

Morphological traits and molecular markers are two common methods for genetic variation studies. Molecular markers, morphological traits methods and relationship between the two were used to study genetic variation among 43 oat genotypes and varieties. For this purpose, an augmented design was conducted in three replicates at 2008–2009 cropping season in the experimental field of Campus of Agriculture and Natural Resources of Razi University, Kermanshah, Iran. Four wild oat accessions (Avena sterilis) were added to evaluated genotypes in molecular experiment. Results showed a significant variation among genotypes for all morphological traits and they were classified based on this variation in four groups by WARD cluster analysis. In molecular experiment, 28 inter simple sequence repeat (ISSR) primers amplified 206 polymorph bands. Based on Jaccard similarity matrix, similarity among genotypes was varied from 0.23 to 0.66 and cluster analysis classified genotypes in seven groups by complete linkage method. The correlation between ISSR marker and morphological traits classifications was not significant. ISSR showed to be a helpful marker for genotype identity and separation as it put wild accessions in a group.     

Genetic variability and structure of Quercus brantii assessed by ISSR,IRAP and SCoT markers

Persian oak (Quercus brantii Lindl.) is one of the most important woody species of the Zagros forests in Iran. Three molecular marker techniques: start codon targeted (SCoT), inter-simple sequence repeat (ISSR) and inter-retrotransposon amplified polymorphism (IRAP) markers were compared for fingerprinting of 125 individuals of this species collected from different geographical locations of north-west of Iran. A total of 233 bands were amplified by 18 ISSR primers, of which 224 (96.10%) were polymorphic, and 126 polymorphic bands (97.65%) were observed in 129 bands amplified by 10 IRAP primers. Besides, 118 bands were observed for all 10 SCoT primers, of which 113 were polymorphic (95.71%). Average polymorphism information content (PIC) for ISSR, IRAP and SCoT markers was 0.30, 0.32 and 0.38, respectively, and this revealed that SCoT markers were more informative than IRAP and ISSR for the assessment of diversity among individuals. Based on the three different molecular types, cluster analysis revealed that 125 individuals taken for the analysis can be divided into three distinct clusters. The Jaccard's genetic similarity based on the combined data ranged from 0.23 to 0.76. These results suggest that efficiency of SCoT, IRAP and ISSR markers was relatively the same in fingerprinting of individuals. All molecular marker types revealed a low genetic differentiation among populations, indicating the possibility of gene flow between the studied populations. These results have an important implication for Persian oak (Q. brantii) germplasm characterization, improvement, and conservation.     

Assessing Genetic Diversity in Mexican Husk Tomato Species

Mexico is the center of diversity of the husk tomato (Physalis L., Solanaceae), which includes a number of commercially important edible and ornamental species. Taxonomic identification is presently based on morphological characteristics, but the presence of high inter- and intraspecific morphological variation makes this task difficult. Six ISSR primers were used on eight Mexican species of Physalis to determine their utility for interspecific taxonomic discrimination and to assess their potential for inferring interspecific relationships. The six ISSR primers amplified 101 bands, with 100% polymorphism across samples. The number of bands per primer varied from 10 to 21. All primers produced different fingerprint profiles for each species, confirming the ISSR value in taxonomic discrimination. Discrimination values based on Simpson’s diversity index varied from 0.48 to 0.58. Genetic interspecific similarity values ranged from 0.20 to 0.57, and intraspecific similarity values were highest for Physalis angulata (0.71), followed by Physalis philadelphica (0.63) and Physalis lagascae (0.55). The UPGMA analysis grouped accessions of the same species together and clustered together Physalis species of similar morphological traits. Thus, ISSR markers are useful in estimating genetic relationships in Physalis.