云南主要核桃品种的ISSR分子鉴别

以核桃当年萌发的新叶为材料,在建立了良好的反应体系基础上,筛选出8条引物对云南省11个主要栽培和推广的核桃品种进行了ISSR分子标记研究。结果表明:8个ISSR引物共检测出位点102个,其中多态性位点62个,占60.78%,参试样品具有较高的多态比率;用其中的两条引物建立了11个品种的指纹图谱,并可用之鉴别参试品种;POPGENE 32软件的计算结果有效等位基因数、基因的多样性和Shannon信息指数分别为1.370 7,0.214 3和0.321 6,供试的云南核桃品种在分子水平上存在比较丰富的遗传变异。     

‘兰州百合’ISSR-PCR体系的优化

以‘兰州百合’为试材,采用CTAB法提取百合嫩叶DNA,对‘兰州百合’ISSR-PCR反应体系的Taq酶、dNTP、引物和模板DNA进行4个因素4个水平优化试验,通过聚丙烯酰胺凝胶电泳结果分析,最终获得的最佳反应体系为:25μL反应体系中含Taq DNA聚合酶0.5U,dNTP 0.3mmol/L,引物0.5μmol/L,模板DNA 60ng,10×Buffer(20mmol/L Mg2+plus)2.5μL,不足部分用ddH2O补足.在获得最佳反应体系的基础上确定了引物最佳退火温度为53℃,最佳循环次数45次.该优化体系以UBC873作为引物对18份‘兰州百合’材料DNA进行ISSR-PCR扩增,结果显示该优化体系具有较高的扩增稳定性.     

ISSR-Based Molecular Characterization of an Elite Germplasm Collection of Sweet Potato （Ipomoea batatas L.） in China

To determine the genetic diversity and population structure of sweet potato accessions cultivated in China, and to establish the genetic relationships among their germplasm types, a representative collection of 240 accessions was analyzed using inter-simple sequence repeat （ISSR） markers. The mean genetic similarity coefifcient, Nei’s gene diversity, and shared allele distance of tested sweet potato accessions were 0.7302, 0.3167 and 0.2698, respectively. The 240 accessions could be divided into six subgroups and ifve subpopulations based on neighbor-joining （NJ） clustering and STRUCTURE results, and obvious genetic relationships among the tested sweet potato accessions were identiifed. The marker-based NJ clustering and population structure showed no distinct assignment pattern corresponding to lfesh color or geographical ecotype of the tested sweet potato germplasm. Analysis of molecular variance （AMOVA） revealed small but signiifcant difference between white and orange-lfeshed sweet potato accessions. Small but signiifcant difference were also observed among sweet potato accessions from the Southern summer-autumn sweet potato region, the Yellow River Basin spring and summer sweet potato region and the Yangtze River Basin summer sweet potato region. This study demonstrates that genetic diversity in the tested sweet potato germplasm collection in China is lower than that in some reported sweet potato germplasm collections from other regions. Pedigree investigations suggest that more diverse Chinese sweet potato varieties should be formed by broadening the selection scope of breeding parents and incorporating the introduced varieties into future breeding programs.     

黔产金樱子变型的ISSR分析

[目的]揭示贵州境内不同变型金樱子10个居群间的遗传变异情况。[方法]采用ISSR分子标记技术,从ISSR随机引物中,筛选有效引物,进行扩增,用NTSYSpc,ver．2．02软件进行UPGMA聚类分析。[结果]10个居群不同变型金樱子的相似性系数在0．09～0．35,聚成A、B2大类。[结论]同一变型的金樱子样品并不严格为一类,不同变型金樱子样品之间的遗传变异较大。     

凤凰单丛茶树资源遗传多样性的ISSR分析

采用ISSR分子标记技术对48份凤凰单丛茶品种(系)进行了遗传多样性分析。选用10个多态性高、分辨力强的ISSR引物分别对供试材料基因组DNA进行扩增,共获得121个位点,其中多态位点带98个,多态位点比率(PPB)为81.0%。品种(系)之间的遗传相似系数变化范围在0.590 9~0.967 4之间。UPGMA聚类分析结果表明,在GS值为0.792的水平上供试材料可划分为7大类,其亲缘关系远近与地理来源关系不大且与香味类型没有必然的联系。     

橡胶树抗寒种质遗传多样性分析

应用RAPD和ISSR技术对20份橡胶树抗寒种质进行遗传多样性分析。结果表明,16个RAPD引物和12个ISSR引物分别扩增出条带113和101条,多态性条带指数(PPB)分别为61.1%和63.4%。据Nei-Li相似系数,ISSR标记在相似系数约0.74处,可将20份材料分为野生种质和栽培种质两大类;RAPD标记在相似系数约0.70处,也可将供试材料分为野生种质和栽培种质两类,但只有XJ002420为野生种质。对2种标记的分析结果进行相关分析,结果表明,RAPD和ISSR所检测的遗传相似系数呈极显著正相关,相关系数为0.672。以上结论表明,RAPD和ISSR标记可用于橡胶树种质资源的遗传多样性研究,但鉴于ISSR较RAPD有更强的多态性检出能力,ISSR技术应作为遗传多样性研究的首选单引物标记。     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.     

基于ISSR标记的大菊品种资源遗传多样性分析

【目的】大菊品种数量众多,形态繁杂。本研究通过探讨不同瓣类大菊品种的遗传多样性,为菊花品种分类和品种演化研究提供重要资料。【方法】选取涵盖大菊品种5个瓣类的150个品种,采用ISSR分子标记进行遗传多样性分析。【结果】从50对引物中筛选出的10对引物共检测到清晰可重复的229个位点,其中多态性位点220个,多态性位点百分率（PPB）96.07%,Nei’s基因多样性指数（He）和Shannon信息指数（I）分别为0.3485和0.5154;5个瓣类内的遗传多样性水平都比较高,其中平瓣类最高（PPB＝95.20%）,桂瓣类（PPB＝86.90%）和匙瓣类（PPB＝89.52%）最低;瓣类类群间遗传分化程度较低,遗传多样性主要来源于不同花型内品种间的变异。在瓣类的UPGMA聚类中,聚类结果与瓣类形态差异显著相关。【结论】ISSR可以有效揭示大菊品种瓣类间的遗传多样性、分化程度和亲缘关系,研究结果对于品种演化和分类鉴定研究具有重要参考价值。     

东南亚茄子种质资源ISSR遗传多样性分析

为探讨东南亚和中国华南地区茄子育种资源的遗传差异,研究采用ISSR分子标记对来自泰国和马来西亚的10份茄子材料及中国和南美的8份材料进行遗传多样性分析。结果表明,17条ISSR引物共扩增出433条谱带,其中多态性条带390条,平均多态率为92%。17条引物在18份材料中的平均观测等位基因数、平均有效等位基因数、平均Nei’s基因多样度和平均香农信息指数分别为1.8971、1.4778、0.2850、0.4336,18份材料间的遗传相似系数GS在0.55~0.81之间。运用NTSYSpc2.10软件的UPGMA方法进行聚类分析,结果表明在遗传相似系数0.67时18份材料可分为2个类群,其中来自于东南亚的10份材料在第1类群,来自于南美的3份材料在第2类群。研究结果表明,东南亚茄子材料与中国和南美茄子材料间遗传多样性丰富,可为茄子种质资源的进一步研究和利用提供理论依据。     

江苏近海银鲳群体遗传多样性的同工酶与ISSR分析

采用同工酶和ISSR分子标记技术对江苏吕泗海域银鲳群体进行了研究,通过聚丙烯酰胺凝胶电泳技术,在银鲳肌肉组织中共检测出7种酶,分别为ADH、EST、GDH、MDH、IDH、LDH、AMY,共统计了15个位点,其中9个位点呈多态,多态位点比率达60%,位点平均有效等位基因数为1.351 0,群体的平均杂合度为观测值0.140 0,平均杂合度期望值0.227 O,说明该银鲳群体具有较高的遗传多样性,资源状况良好.同时,利用现代分子标记技术,建立了银鲳ISSR分析体系,筛选出了21个可用于银鲳群体遗传状况分析的引物,并对冷冻保存和70%乙醇保存两种处理样本进行了比较研究,结果表明,两种样本处理方法都可以满足银鲳群体遗传多样性研究要求.