中国栽培白灵侧耳的RAPD和IGS分析

以RAPD和IGS为分子标记,研究了中国栽培白灵侧耳Pleurotus nebrodensis菌株的遗传多样性,RAPD结果表明,供试菌株间存在多态性;对供试菌株的IGS分析表明,白灵侧耳P. nebrodensis的IGS1区域较保守,菌株间没有多态性;IGS2区域进化较快,菌株间具丰富的多态性。IGS2-RFLP可应用于白灵侧耳的菌株鉴定鉴别,较RAPD更稳定,更具可操作性。     

新疆野生阿魏蘑的IGS2-RFLP及培养特征多样性分析

对28个新疆野生阿魏蘑样本进行IGS2-RFLP分析和双核体培养特征多样性分析。IGS2-RFLP分析结果表明,3种限制性内切酶共产生的多态性条带比率（PPB）为96.4%,总样本的Shannon信息指数（I）和Nei’s基因多样性指数（H）分别为0.364和0.230,平均相似系数0.665,表明其具有丰富的遗传多样性。在相似系数0.36水平上,UPGMA聚类分析将供试样本分为白灵侧耳和刺芹侧耳阿魏变种两个种群,且白灵侧耳的遗传多样性更为丰富。双核体培养特征多样性分析结果表明,供试样本间在最适生长温度、温度敏感性以及菌落形态等培养特征上具明显差异,基于UPGMA聚类分析大致将供试的野生阿魏蘑样本分为菌丝生长快、菌落形态舒展和菌丝生长慢、菌落局限两大组。双核体培养特征聚类与IGS2-RFLP分析聚类存在一定相关关系,培养特征有可能作为野生阿魏蘑种质鉴定的白灵侧耳的检出标志。     

抗风桐(Pisonia grandis)的生态生物学特征

对收集于广西桂林的17份野生毛葡萄种质和24份栽培葡萄种质,分别使用12条ISSR和12条SCoT引物进行了遗传多样性和亲缘关系检测。结果表明：两种分子标记均能产生较丰富的多态性片段,可有效应用于葡萄的遗传多样性检测,但在聚类分析结果上表现出一定的差异性,SCoT分子标记能更好地区分野生种质和栽培品种,说明SCoT分子标记在葡萄遗传多样性检测和系统进化研究上可能更有优势。从SCoT聚类结果上看,广西植物研究所收集的3个野生毛葡萄种质zws1、zws2和zws3相对其它野生种质而言,更偏向于与栽培种质聚为一类,说明这一类野生毛葡萄可能是这些栽培品种的原始亲本来源之一。不同的野生种质聚为多个类群,并表现出明显的地域特性,但遗传距离相对较远,说明桂林野生毛葡萄资源具有丰富的遗传变异。栽培品种没有明显的聚类特点,可能因为所选用的栽培品种的地域代表性并不是很强,也可能是因为栽培品种在不断的人工杂交选育过程中,遗传背景趋向一致,遗传多样性降低。该研究证明SCoT分子标记在葡萄遗传多样性研究上具有一定的优势。该研究结果为桂林毛葡萄资源的保护、利用和品种选育提供了理论依据,也为葡萄的系统进化研究提供了参考。     

41份葡萄种质遗传多样性的ISSR和SCoT对比分析

对收集于广西桂林的17份野生毛葡萄种质和24份栽培葡萄种质,分别使用12条ISSR和12条SCoT引物进行了遗传多样性和亲缘关系检测。结果表明:两种分子标记均能产生较丰富的多态性片段,可有效应用于葡萄的遗传多样性检测,但在聚类分析结果上表现出一定的差异性,SCoT分子标记能更好地区分野生种质和栽培品种,说明SCoT分子标记在葡萄遗传多样性检测和系统进化研究上可能更有优势。从SCoT聚类结果上看,广西植物研究所收集的3个野生毛葡萄种质zws1、zws2和zws3相对其它野生种质而言,更偏向于与栽培种质聚为一类,说明这一类野生毛葡萄可能是这些栽培品种的原始亲本来源之一。不同的野生种质聚为多个类群,并表现出明显的地域特性,但遗传距离相对较远,说明桂林野生毛葡萄资源具有丰富的遗传变异。栽培品种没有明显的聚类特点,可能因为所选用的栽培品种的地域代表性并不是很强,也可能是因为栽培品种在不断的人工杂交选育过程中,遗传背景趋向一致,遗传多样性降低。该研究证明SCoT分子标记在葡萄遗传多样性研究上具有一定的优势。该研究结果为桂林毛葡萄资源的保护、利用和品种选育提供了理论依据,也为葡萄的系统进化研究提供了参考。     

不同龙眼资源遗传多样性的SCoT和ISSR 比较分析

应用SCoT和ISSR标记对36份龙眼资源和1份近缘种龙荔的遗传多样性进行分析。结果表明:12对SCoT引物共扩增出127条带,平均每条引物扩增10.58条带;15条ISSR引物共扩增出117个条带,平均扩增7.8条带。UPGMA聚类结果表明:SCoT标记和ISSR标记分别在相似系数0.672和0.685水平上,均可将37份材料分成6大类群,SCoT和ISSR标记均适用于龙眼材料的遗传多样性分析,如果将两种标记的数据进行综合分析,可以缩小单一标记的误差。研究结果为龙眼种质资源的保存和利用提供了重要的依据。     

棉花遗传多样性SCoT和SRAP标记的研究及比较分析

利用SCoT和SRAP两种分子标记技术对30份彩色棉与白色棉种质资源,进行遗传多样性研究。用29对SRAP引物组合和26个SCoT引物分别对供试棉花的基因组DNA进行扩增。SCoT引物共扩增出163条带,多态性比率为61.96%,遗传相似系数GS值变化范围为0.5405～0.9972。SRAP引物组合共扩增条带1067条,多态性比率仅为14.1%,遗传相似系数GS值变化范围为0.5415～0.9109。两种标记系统得到了相似但并不完全相同的聚类图,2种标记方法间存在显著相关性(r=0.5518,P<0.05)。结果表明,SRAP与SCoT标记均适用于棉花种质的遗传多样性分析,且SCoT的标记指数MI高于SRAP标记,为SCoT这种新兴的标记技术在棉花育种中的应用提供了重要的依据。     

安徽野生香菇遗传多样性及杂种优势的ISSR分析

采用简单序列重复区间扩增多态性(Inter–simplesequencerepeat,ISSR)技术,利用13个引物对26个安徽野生香菇Lentinulaedodes菌株和6个香菇栽培品种进行了遗传多样性分析,构建了遗传相关聚类图,并根据ISSR分析选择遗传距离远近不同的亲本进行组合杂交,评价了其杂种优势。在78个ISSR标记中,多态性标记为61个,占78.2%。聚类分析显示,当以相异距离0.23为阈值,32个菌株被划为5个ISSR遗传组,多数菌株之间遗传相似性较低。这表明供试菌株在DNA水平上存在比较显著的遗传变异,具有较丰富的遗传多样性。杂种优势的检测表明亲本间遗传距离较大的组合其杂种优势也较强。     

云南黑籽南瓜种质遗传多样性的RAPD和ISSR分析

摘要: 采用RAPD和ISSR分子标记技术对来源于云南省6个地州13份的黑籽南瓜种质进行遗传多样性分析。结果表明：6个RAPD和6个ISSR引物分别扩增出43条和41条带,多态性比率分别为90.70%和51.21%;RAPD和ISSR标记检测供试材的遗传相似性系数(Gs)范围,分别为0.340-0.895和0.162-0.941,ISSR(平均GS值0.698)检测多态性效果高于RAPD(平均GS值0.481)。RAPD标记聚类分析将供试种质分为3个类群5组;ISSR标记聚类分析将供试种质分为4个类群6组,RAPD和ISSR标记的遗传相似性系数呈显著相关(r=0.536)。基于UPGMA聚类结果,可为黑籽南瓜的引种栽培或品种改良提供参考。     

SRAP结合SCoT标记分析番木瓜种质的遗传多样性

利用SRAP和SCoT两种分子标记相结合对中国22个番木瓜主要栽培品种(系)进行遗传多样性研究。SCoT标记检测获得的遗传多样性参数值和遗传相似系数范围均高于SRAP标记检测的结果,表明SCoT标记检测多态性的能力高于SRAP标记。基于两种标记数据合并后的UPGMA聚类结果显示,番木瓜种质间的遗传相似系数为0.65~0.90,种质间遗传多样性水平较低;在遗传相似系数为0.82时,将所有参试材料划分为3个类群。应用Mantel检测对SRAP和SCoT及两种标记合并进行相关性分析,表明三者之间具有显著的相关性,且相关性很高。聚类结果从分子水平反映出中国番木瓜主要栽培品种(系)的遗传基础狭窄。     

中国白灵侧耳自然群体的交配型因子分析

为了研究我国新疆白灵侧耳自然种群的遗传多样性,估测自然种群的遗传丰度,对51株新疆野生白灵侧耳的84个原生质体单核体的交配型因子进行分析。在84个单核体中存在54个不同的A因子和59个不同的B因子。x2检测表明交配型因子A和B的系列因子均为等概率分布。据此估算我国白灵侧耳自然群体中有79个A因子,100个B因子,自然群体中的单核交配型总数79×100=7,900个,双核交配型总数31,201,050个。可见,新疆白灵侧耳遗传多样性丰富,新疆是我国白灵侧耳的野生种质资源宝库。     

Genetic diversity and population structure detection in sponge gourd ( Luffa cylindrica ) using ISSR,SCoT and morphological markers

Investigation of genetic diversity is essential for the selection of parents for crop breeding and conservation of genetic resources. To estimate the genetic variability and population structure in the midst of 45 accessions of sponge gourd brought together from different geographical areas of India, morphological traits and two molecular markers, ISSR and SCoT markers were compared. Principal components analysis of 20 morphological traits showed 72.70% variability and significant positive correlations between fruit traits. All three marker techniques clustered all accessions into two groups with few outgroups. High level of polymorphism was observed among ISSR (74.6%) and SCoT (71.5%) primers. The Bayesian model revealed the hidden grouping and showed admixture type of population. The diversity pattern is influenced by genetic marker used, as different molecular markers have different polymorphism evaluation efficiency. This study can be helpful in amplifying the genetic base and selection of specific traits for breeding. Thus, ISSR and SCoT markers are potential marker for identification in sponge gourd and provide valuable data on its genetic correlation and structure.     

一种新的DNA分子标记技术——起始密码子-微卫星扩增多态性

综合SCoT和ISSR分子标记技术开发了一种既能将标记位点与表达序列紧密联系,又具有相对较高的多态性的新的分子标记技术——起始密码子一微卫星扩增多态性（start codon-simple sequence repeat, SC-SSR）。SC—SSR标记是基于PCR的目的基因标记系统．上游引物用SCoT标记引物,瞄准基因区域,下游引物用ISSR标记引物,上下游引物间可自由组配。引物设计原则同SCoT标记和ISSR标记。使用50℃的退火温度,保证了扩增结果的稳定性。PCR结果采用琼脂糖凝胶电冰和聚丙烯酰胺凝胶电泳检测。SC—SSR分子标记结合了ISSR标记和SCoT标记的优点,具有操作简单、成本低廉、多态性丰富、重复性好、引物设计简单且通用性良好、同时与表达序列紧密连锁等诸多优点,可用于种质资源的鉴定评价、遗传图谱的构建、重要性状基因标记、gDNA与cDNA指纹分析乃至图位克隆等方面。     

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.     

Genetic diversity of Indian jujube cultivars using SCoT,ISSR, and rDNA markers

Genetic variation and relationships among 37 cultivars of Ziziphus mauritiana (Lamk.) native of India were analyzed using start codon targeted (SCoT), inter-simple sequence repeats (ISSR), and ribosomal DNA (rDNA) markers. High level of polymorphism among SCoT (61.6%) and ISSR (61%) primers with higher PIC values ranging from 63.1 to 90.4% of SCoT and 47.3 to 88.8% of ISSR primers was recorded. SCoT and ISSR dendrograms revealed similarity coefficients ranging from 0.80 to 0.92 and 0.79 to 0.96, respectively, and clearly delineated all the cultivars of Z. mauritiana into well-supported distinct clusters. Greater Gst signifies higher amount of differentiation observed over multiple loci among seven Z. mauritiana populations. On the other hand, higher gene flow demonstrating a very high migration rate between Z. mauritiana populations indicated higher rates of transfer of alleles or genes from one population to another. The genetic diversity of population 1 (Rajasthan) was the richest among all the seven populations. The largest genetic distance was measured between Maharashtra and West Bengal and the least between Rajasthan and Punjab cultivars. Most of the genetic diversity exists within population rather than among populations. Substantial variation in the ITS-1 region signifies its phylogenetic utility specifically in assessing genetic diversity in Z. mauritiana. The clustering patterns using three molecular marker systems vis-à-vis place of origin exhibited no consistency in grouping of Z. mauritiana cultivars as cultivars from the same place of origin were genetically cataloged into different SCoT, ISSR, and ITS phylogram clusters indicating wide genetic diversity and distribution across agro-climatic zones validating the robustness of marker systems tested.     

Genetic diversity of mango cultivars estimated using SCoT and ISSR markers

Two molecular marker systems, SCoT and ISSR were used for identification and genetic comparison analysis of 23 mango germplasm accessions collected within Guangxi province of China. Using 18 selected SCoT primers 158 bands were generated, of which 104 (65.82%) were polymorphic. Eighteen selected ISSR primers amplified 156 bands with 87 (55.77%) being polymorphic. The cultivars of Xiang Ya Mango type and their progeny have high genetic similarity with each other. The 23 cultivars were clustered into two major groups based on the SCoT analysis and three major groups based on the ISSR analysis with UPGMA. These clusters are in accordance with their known origins and main phenotypic characteristics. Our results indicated that the SCoT analysis better represents the actual relationships than ISSR analysis, although both analyses give similar results. The results also demonstrate that the SCoT marker system is useful for identification and genetic diversity analysis of mango cultivars.     

Evaluation of genetic diversity and population structure of polygonati rhizoma germplasms: implications for better crop development and conservation of a traditional Chinese medicine

Polygonati rhizoma (PR) is an important and widely used product in Chinese traditional medicine and edible goods. The time-consuming nature of breaking dormancy in both rhizomes and seeds means that improving variety selection is limited to collection, identification, and selection of germplasms. In this study, we used two DNA-based molecular marker techniques—inter simple sequence repeat (ISSR) and start codon targeted (SCoT)—to assess the genetic diversity and population structure among PR source plants collected from 47 different regions and belonging to 12 populations (P1–P12). For molecular markers analysis, 15 ISSR and 10 SCoT markers were tested. Total number of 159 fragments (150–4000 bp) were amplified based on ISSR analysis with a range from 6 to 17 bands, 153 of them were polymorphic, ranging from 97.27 to 100%. For SCoT analysis, 164 polymorphic bands (150–5000 bp) were observed, varying from 14 to 19 bands for each primer. Nei’s genetic diversity analysis showed that the highest value was found in P11 for ISSR and P4 for SCoT markers. The highest Nei’s genetic diversity value was observed in P5 for combined markers and the low in P2. Nei’s dendrogram constructed with combined markers indicated a 75–89% of genetic similarity coefficient among populations. Population structure analysis revealed an optimum number of three groups, the same as their geographical distribution. This knowledge on PR genetic diversity can be used in future breeding programs, genetic improvement, product enhancement, and germplasms conservation.     

Comparative assessment of ISSR, DAMD and SCoT markers for evaluation of genetic diversity and conservation of landrace chickpea (Cicer arietinum L.) genotypes collected from north-west of Iran

Morphological traits and three molecular markers techniques: start codon targeted (SCoT), inter-simple sequence repeat (ISSR) and directly amplified minisatellite DNA (DAMD) markers were compared for fingerprinting of 40 landraces chickpea genotypes collected from different geographical locations of north-west of Iran. Variance analysis of ten measured morphological traits showed significant differences existed between genotypes. Cluster analysis based on morphological traits, divided genotypes in three distinct clusters. Average polymorphism information content (PIC) for ISSR, DAMD and SCoT markers was 0.216, 0.232 and 0.232, respectively, and this revealed that SCoT markers were more informative, followed by ISSRs marker, than other markers for the assessment of diversity amongst genotypes. Cluster analysis for three different molecular types revealed that genotypes taken for the analysis can be divided in three and four distinct clusters. Accessions from same geographical regions mostly showed more genetic similarities than those from origins far isolated apart. These results suggest that efficiency of SCOT, DAMD and ISSR markers was relatively the same in fingerprinting of genotypes but SCOT and DAMD analysis are more effective in fingerprinting of chickpea genotypes. To our knowledge, this is the first detailed report of a comparison of performance among two targeted DNA region molecular markers (SCoT and DAMD) and the ISSR technique on a set of samples of chickpea. Overall, our results indicate that SCOT, ISSR and DAMD fingerprinting could be used to detect polymorphism for genotypes of chickpea.