ISSR Analysis of Chlorophytum Treated by Three Kinds of Chemical Mutagen

Chlorophytum leaves were treated with three kinds of chemical mutagen, EMS, DES and NaN3 with different concentration to obtain the variation materials with excellent properties. The results showed that the genetic similarity coefficient of variation plants with EMS treatment was between 0.648 and 0.868, with NaN3 treatment was between 0.598 and 0.859, and with DES treatment was between 0.668 and 0.904, of which the mutagenic effects with 0.8% EMS, 250 mg. L-1 NaN3 and 0.3% DES on chloro- phytum were the best. ISSR molecular marker technique was used to analyze their genetic diversities. Total 392 polymorphic bands were obtained through 18 ISSR primers. Polymorphic ratio was 72.4%, which showed that DNA mutation took place in various degrees     

ISSR molecular characterization and leaf volatiles analysis of Pittosporum undulatum Vent. naturalized in the Azores archipelago (Portugal)

Pittosporum undulatum from the Azores archipelago was characterized by both molecular and volatiles analyses. Inter simple sequence repeat (ISSR) evaluation was performed based on a random sampling of 77 individuals from a total of 123 samples collected on all the Azorean islands. Molecular studies grouped the samples mainly according to the geographical collection site, with some exceptions. Leaf volatiles were isolated by distillation-extraction and analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) from all the 123 samples. Cluster analysis based on the leaf volatiles chemical composition defined three main clusters, not related to sample site collection, and was based mainly on the relative amounts of limonene (3-89%), sabinene (0.1-64%) and terpinen-4-ol (traces-43%). Clusters obtained from both molecular studies and volatiles composition were not superimposable and no correlation between ISSR markers and volatile oils could be drawn. The results suggest that there is a high genetic variability among individuals of P. undulatum that can explain the invasive ability of this species in the Azores archipelago. The detailed characterization of this alien weed species may assist finding potential commercial uses as well as management strategies to help its eradication and/or control.     

Molecular analyses of genetic variability in soap nut (Sapindus mukorossi Gaertn.)

Sapindus mukorossi Gaertn. (Sapindaceae) is commercially an important plant much valued for its fruits commonly known as soap nut as well as for the seed oil used as a feed-stock for biodiesel production. Molecular polymorphism, determined, using three single primer amplification reaction (SPAR) methods (RAPD, DAMD and ISSR) is used to assess the genetic variability in 69 accessions of S. mukorossi Gaertn., encompassing different geographical regions in India. Cumulative band data generated using the three SPAR methods resulted in 82.49% polymorphism across all genotypes of S. mukorossi Gaertn. UPGMA tree showed two major clusters that were in accordance with the geographical diversity. The intra-population genetic diversity (0.16), Shannon information index (0.24) and polymorphic loci (45.62%) were highest in AS population, while the corresponding lowest values were recorded in HP population. The inter-population average distance ranged from 0.05 (GA and HP) to 0.26 (AP and HP). Based on these distances, the UPGMA tree for the populations was computed, which showed the presence of two major clusters. Cluster I contained all the populations from Himachal Pradesh (HP) and Uttarakhand (GA and KU) and cluster II grouped the populations from North Eastern region (AS, AP, MG). The present study showed that SPAR methods are informative and useful to unravel the diversity among different populations of the soap nut plants and is a prelude for further utilization of promising and genetically divergent materials in the breeding programmes.     

芒果ISSR反应体系的建立与优化

运用L16(45)正交设计对芒果ISSR反应的5个因素,即DNA模板浓度、Mg2+浓度、dNTPs浓度、引物浓度、Taq DNA聚合酶浓度在4个水平上进行优化试验,通过不同反应体系扩增效果比较,建立优化的芒果ISSR反应体系,即25μL的PCR体系中含有DNA模板25 ng、Mg2+浓度为1.5 mmol/L、dNTPs 0.3 mmol/L、引物0.32μmol/L、Taq DNA聚合酶1.25 U。这为进一步运用ISSR标记在DNA分子水平上对芒果种质资源进行分析奠定基础。     

中国油棕种质材料的遗传多样性研究

为开发利用中国油棕种质资源,从DNA分子水平上对海南和云南油棕种质资源进行遗传多样性研究。应用ISSR技术对海南和云南的85份油棕种质材料进行遗传多样性分析,用23个引物对所有85个新引进油棕种质及原有油棕种质材料进行ISSR扩增,23个引物共扩增出201条带,其中多态性条带占78.6%。聚类分析的结果显示,采自海南、云南的新选育油棕种质及原有的油棕种质材料基本上能按不同的采集地点或不同的来源途径而分别聚类在一起。     

梨属植物ISSR技术体系的建立与优化

本研究以雪花梨为材料,对ISSR反应体系中的模板用量、引物浓度、dNTPs浓度、TaqDNA聚合酶用量等一些重要参数进行探索和优化,初步建立了稳定的具有广泛适用性的梨属植物ISSR技术的反应体系及扩增程序,即20 μL体系中含1×buffer (内含1.5 mmol? L-1 MgCl2)、模板DNA 40～60 ng、引物浓度为0.2 μmol?L-1、 dNTP 200 μmol?L-1、Taq DNA聚合酶 1U。扩增程序为：94℃ 5 min、退火60s、72℃延伸120 s,然后连续39个循环为94℃ 60s、退火温度(52℃左右)60s 、72℃延伸120s,完成最后一个循环后,在72℃继续延伸10 min,然后在4℃保温,最后通过1.4%的琼脂糖凝胶电泳检测其扩增产物。     

饲用型小黑麦遗传图谱构建及草产量相关性状QTLs初步定位

本研究以饲用型小黑麦杂交F₂代为作图群体,利用ISSR分子标记构建了遗传连锁图谱,并对小黑麦草产量相关性状（株高,分蘖数,单株生物量）进行了QTLs定位,可为小黑麦草产量相关主效QTL挖掘、功能基因定位以及分子标记辅助育种奠定基础。结果表明：521个F₂代单株草产量相关性状的田间表型数据呈连续变异,分布频率大致接近正态分布,可用于QTLs定位。该遗传图谱包含7个连锁群,图谱总长度为542.9 cM,标记间平均距离为5.90 cM,共有92个位点。对草产量相关性状基因进行QTL定位分析,共检测到17个QTLs,分布在6个连锁群上,控制株高的QTLs有5个,贡献率为6.7%~13.2%;控制分蘖数的QTLs有7个,贡献率为5.4%~15.4%;控制单株生物量的QTLs有5个,贡献率为7.4%~12.4%。其中QPH7-2,QNT2-2和QBS2分别对小黑麦株高、分蘖数和单株生物量的贡献率最大,为主效QTLs,可对其进行进一步克隆、转化及利用。     

美洲黑杨新无性系的ISSR鉴别及SCAR标记转化

为了从分子水平上对美洲黑杨新无性系材料进行准确快速鉴定,从100条ISSR引物中筛选出10条具多态性的引物对包括钻天杨和69杨在内的6个黑杨无性系开展指纹图谱的初步构建,并将钻天杨和03-北-1-15中扩增出的特异ISSR条带转为可用于快速鉴别黑杨无性系的SCAR标记。结果表明,ISSR指纹分析结合SCAR标记可以对供试黑杨无性系进行区分与鉴定,进而为杨树林木品种鉴定以及黑杨育种研究提供理论依据。     

应用ISSR和线粒体COI序列对不同地理种群双齿围沙蚕（Perinereis aibuhitensis）遗传多样性的分析

采用COI基因序列和ISSR分子标记对分布于我国大连（DL）、乳山（RS）、盘锦（PJ）、青岛（QD）、盐城（YC）、晋江（JJ）和朝鲜西海岸（CX） 7个地理种群双齿围沙蚕（Perinereis aibuhitensis）的遗传多样性和遗传结构进行了研究。10个ISSR引物在7个种群中共扩增出116条条带,其中多态性条带109条,总多态位点比率为93.97%,Neis基因多样性指数为0.365 2,Shannons信息指数为0.518 6,群体间遗传分化系数Gst值为0246 0。数据表明24.60%的遗传分化发生在不同地理种群间,76.40%的遗传分化发生在整个种群内,群体内分化大于群体间分化。对7个群体70个个体COI基因序列进行分析,对比长度包括469 个位点,其中变异位点68个,简约信息位点47个。在68个变异位点中,62个位点发生转换,6个位点发生颠换,简约信息位点率达9.611%,系统树分析表明供试群体内具有较高的遗传多样性,与ISSR分析结果相一致。该结果为科学有效的利用和保护沙蚕种质资源提供了理论依据。     

云南椰子种质资源遗传多样性的ISSR分析

为摸清云南椰子种质资源遗传多样性现状,促进资源的有效利用,以在云南收集的60份椰子种质为试材,利用ISSR分子标记技术对其遗传多样性进行了分析。结果表明：27条ISSR引物共扩增出272个位点,其中230个为多态性位点,多态性比率为84.56%,引物多态性指数为0.85,SM遗传相似系数平均值为0.664。根据各种质的SM,运用NTSYSpc2.10e软件的UPGMA方法构建了种质的系统聚类图,60份椰子被聚为3个类群(A、B、C)。其中,A类群为全部由本地种质组成的一个独立类群;C类群既包括部分本地种质,也包括来源于本地以外的其他种质。研究结果显示出云南椰子种质具有较高的遗传多样性;这些种质与外地种质具有独立性和关联性的特点。该研究结果可为云南椰子种质资源的进一步研究和利用提供参考。