大豆疫霉菌遗传多样性的RAPD分析

利用随机扩增DNA (RAPD)技术分析了15个大豆疫霉菌( Phytophthora sojae)的遗传多样性,并同其它7 个疫霉种的8个菌株进行了比较。运用筛选出的12个引物共获得152个RAPD标记,其中84. 2%具有多态性。通 过聚类分析结果表明,疫霉属种间的遗传相似系数的变化幅度为0. 533～0. 783,大豆疫霉菌与其它几种疫霉的遗 传距离相对较远,用引物OPB06扩增到P. sojae种特有的一个RAPD标记; P. sojae种内菌株间遗传相似系数的变化 幅度为0. 855～0. 967,亦存在较丰富的遗传多样性,该遗传多样性与菌株毒力之间有一定相关性,而与菌株地理来 源没有相关性。     

红掌细菌性疫病病原菌遗传多样性的 RAPD 和 Rep-PCR分析

由地毯草黄单胞菌花叶万年青致病变种引起的细菌性疫病是红掌生产中最具破坏性的病害。本研究对来自全国 8 个省市主产区的 93 个红掌细菌性疫病菌株，采用 RAPD 和 Rep-PCR（ERIC、BOX、REP）分子标记进行了遗传多样性分析。筛选出的 13 个 RAPD 引物共扩增出 131 个清晰、重复性好的条带，而 Rep-PCR 共扩增出65 个条带，所有位点多态性比率为 100%。基于 RAPD 和 Rep-PCR 结果计算的群体遗传距离显著相关，综合 2 种标记类型的分型结果进行 UPGMA 聚类分析，表明我国红掌细菌性疫病具有高度的遗传多样性，在遗传相似性系数范围 0.519 1~0.984 7，在 0.63 水平时可以将所有菌株划分为 3 个组群，分别包括 58、28、7 个个体，病菌群体的遗传多样性与其寄主品种、地理来源没有明显相关性。本研究为红掌细菌性疫病监测和抗病育种提供了科学依据。     

三个小麦赤霉病抗源的抗性QTL定位

为寻找小麦赤霉病抗性基因及可用于分子标记辅助育种的抗性连锁标记．对中国的三个小麦赤霉病抗源苏麦3号、望水白和宁894037进行了抗性QTL的定位研究。SSR、AFLP分析与QTL分析结果表明，尽管三个抗源的来源和遗传背景并不同，但均在3B染色体短臂上发现抗性主效QTL，不同遗传群体所获得的QTL位点所处的染色体区段略有差异，位于QTL两翼的SSR标记也有所不同。苏麦3号的赤霉病抗性主效QTL位于3B染色体上的标记区间Xgwm533～Xgwm493内；宁894037的抗性位点分布于3B和6B染色体上。分别定位于标记Xgwm493～Xbarcl33和Xgwm644-Xgwm518之间；望水白的抗性主效QTL也位于3B染色体上．定位于标记Xgwm493～Xbarc147之间。微效QTL由于遗传群体的不同，分别住于1B、3B和2A染色体上。研究还表明，寻找抗性QTL在3B染色体以外的新抗源十分必要。     

橡胶树多主棒孢病菌遗传多态性分析

利用ISSR（简单序列重复区间扩增多态性）和RAPD（随机扩增多态DNA）技术对供试的18个多主棒孢菌株进行了遗传多态性分析,研究结果表明,2种分子标记技术均能揭示橡胶多主棒孢病菌的遗传多态性。同时利用P距离模型构建了18个多主棒孢菌株的UPGMA聚类分析树状图,根据2种技术构建的UPG—MA聚类树状图可将供试菌株分为3个主要类群。ISSR类群和RAPD类群与地理来源均不存在明显的关系。     

湖南烟溪病圃稻瘟病菌的有性态及微卫星标记的遗传多样性分析

 用2对稻瘟病菌的两性能育菌株（交配型1.1和交配型1.2）测定了来自湖南烟溪病圃上的4年124个稻瘟病菌株的交配型，用微卫星分子标记分析其中105个菌株的遗传多样性及其亲缘关系。 124个菌株中， 28个为交配型1.1, 39个为交配型1.2， 57个不育, 分别占供试菌株的22.58%、31.45%和45.97%；在0.67相似性的基础上，7对微卫星引物扩增出的标记可将105个菌株区分为6个系谱群。     

绿色荧光蛋白基因在小麦禾谷镰刀菌中的表达与鉴定

为了获得稳定表达绿色荧光蛋白（GFP）的我国代表性小麦禾谷镰刀菌菌株,用于小麦抗赤霉病接种鉴定与分析,以禾谷镰刀菌武昌菌株Fg175原生质体为受体,经PEG介导将序列优化的水母GFP基因转入该菌株中,分析鉴定了转基因菌株GFP基因的表达。PCR分析表明,Fg175基因组中整合了GFP基因,Western blot检测到高效表达的GFP特异蛋白;激光共聚焦显微镜及体视镜观察进一步证实,转基因菌株Fg175-GFP的分生孢子、菌丝及接种小麦麦穗后,均可发出清晰的绿色荧光。     

东北地区玉米弯孢菌的致病性分化与RAPD分析

在东北三省15个地区采集分离26个玉米弯孢菌菌株,通过鉴别寄主技术与RAPD分子标记技术进行致病性与遗传分化的研究。结果表明,东北地区玉米弯孢菌在致病性与遗传上都存在明显的变异,二者有一定的相关性,未发现变异程度与菌株地理来源有明显的直接关系。26个菌株的致病性分为5种类型,其中中等致病类型分布遍及东北三省,为优势致病类群;强致病类型分布在吉林省的白城市;弱致病类型主要分布在吉林省的白山和梨树。用8个引物对供试菌株进行RAPD扩增,共获得77个RAPD标记,其中多态性标记71个,多态性比例92.2%。供试菌株在相似系数约0.71处共聚为4组,大多数致病性较强的菌株聚在一起,而弱致病株在相似系数相对较低时和其他菌株聚在一起。     

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。     

Occurrence, pathogenicity and distribution of Fusarium spp. in stored wheat seeds Kermanshah Province, Iran

Fusarium is one of the most important pathogenic and toxigenic fungi widely distributed all over the world, including Iran. Fusarium species are found frequently in stored agriculture products especially wheat. The objective of this study was to identify Fusarium species associated with stored wheat seeds and their pathogenicity on root and head of wheat in Kermanshah, the leading province in wheat production in Iran. In this survey 75 seed samples of stored wheat were collected from 10 different regions during 2006-2008 and tested for the presence of Fusarium. Fusarium spp. were found in 51 (68%) of 75 samples. A total of 580 Fusarium strains were isolated, identified and preserved. All these strains belong to 20 Fusarium spp. according to morphological characters. Each conidial suspension of selected strains representing all species was evaluated for their pathogenicity on roots and spikes of healthy wheat var. Fallat in the greenhouse. F. graminearum, F. crookwellense, F. trichothecioides, F. culmorum and F. verticillioides were the most pathogenic to wheat's head. Foot rot assessment revealed that F. pseudograminearum and F. culmorum were the most damaging species. Of the Fusarium isolates, F. graminearum was the most prevalent followed by F. verticillioides and F. proliferatum. This is the first comprehensive report on identity and distribution of Fusarium spp. from stored wheat seeds in Iran while F. nelsonii was reported for the first time from wheat seeds in Iran.     

安徽省小麦赤霉病菌群体遗传多样性AFLP分析

为了解安徽省小麦赤霉病菌的群体分化和遗传变异规律,利用AFLP技术对安徽省4个地理群体的68个供试菌株进行了群体遗传多样性分析。结果表明,8对引物共扩增出245条带,供试菌株间的遗传距离为0.014~0.021。4个小麦赤霉病菌群体之间的遗传距离与地理分布没有明显的相关性。4个赤霉病菌群体的Shannon信息指数I为0.321,基因多样性指数H为0.193,表明具有一定的遗传多样性。方差分析表明,被测小麦赤霉病菌群体的遗传变异主要存在于群体内(99.80%),群体间的遗传变异仅占0.20%。4个赤霉病菌群体间存在较低的遗传分化(Gst=0.049)和频繁的基因交流(Nm=9.648)。相比较而言,北部群体和中部群体亲缘关系较近,而南部群体和中、北部群体亲缘关系较远。     

Gramineous and non-gramineous weed species as alternative hosts of Fusarium graminearum,causal agent of Fusarium head blight of wheat,in Argentina

Weeds and wild plants around and within crops could serve as alternative hosts of fungal pathogens. In this work we describe the isolation of Fusarium graminearum, the main causal agent of Fusarium head blight (FHB) in Argentina from the inflorescences of healthy weed plants belonging to sixty seven gramineous and non-gramineous species, which showed no symptoms of Fusarium infection, sampled throughout a year. Fifty four of the weed species considered, belonging to 19 botanical families, were first identified as alternative hosts of F. graminearum in the present work. Furthermore, the trichothecene chemotype of a group of isolates was analysed and strains belonging to 15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol and nivalenol chemotypes were found. The information provided could prove valuable to study further the epidemiological role of weeds in FHB epidemics, which might help to improve management of the disease in wheat growing areas.     

利用ISSR技术分析禾谷镰孢菌群体遗传多样性的研究

采用ISSR标记对采自我国11个省的玉米茎腐病相关禾谷镰孢的遗传多样性进行分析。利用筛选出的14个ISSR引物对供试的115株禾谷镰孢菌株进行扩增,共获得63条扩增清晰、重复性高的条带,其中多态性条带为60条,占95.2%。扩增条带分子量为150～2000bp,平均每个引物扩增出4.5条带。遗传多样性分析表明,在地理种群水平上,基因多样性指数在0～0.2919之间,平均为0.1591;Shannon‘s多样性指数在0～0.4252之间,平均为0.2360,表明不同地理种群间存在一定的遗传变异。多样性指数、等位基因数的增大与各种群内样本数量增加有关。遗传相似性分析证明,山东省种群与河南省种群间的遗传相似性最高,内蒙古种群与河北省种群间遗传相似性最低。在相似性系数为0.682时,可将115个菌株区分为2个聚类组,各组下又可分为3个亚组,分组结果与菌株的地理来源有一定相关性,表明禾谷镰孢的遗传分化与生态地理有关。     

Genetic diversity of accessions of Iranian Aloe vera based on horticultural traits and RAPD markers

In this study, genetic diversity of 10 accessions of Aloe vera, collected from different parts of Iran, were evaluated using horticultural and random amplified polymorphic DNA (RAPD) data. Statistical analysis showed significant differences for all horticultural characteristics among the accessions, suggesting that selection for relevant characteristics could be possible. The analysis of molecular diversity was used the RAPD technique, with 10 random primers of 10-mer oligonucleotides. Out of the 10 primers, five were polymorphic, producing 269 DNA bands, 189 of which were polymorphic among accessions. A dendrogram was prepared on the basis of a similarity matrix using the Unweighted Pair Group Method Arithmetic Mean (UPGMA) algorithm, separating the 10 accessions into two groups. Results show that both environmental and genetic factors are effective in observing variations. Our results also indicate that the RAPD approach, along with horticultural analysis, seemed to be best-suited for assessing with high accuracy the genetic relationships among distinct A. vera accessions.     

Evaluation of genetic diversity in turmeric (Curcuma longa L.) using RAPD and ISSR markers

Turmeric (Curcuma longa L.) is an industrially important plant used for production of curcumin, oleoresin and essential oil. In the present study we examined the genetic diversity among turmeric accessions from 10 different agro-climatic regions comprising 5 cultivars and 55 accessions. Two DNA-based molecular marker techniques, viz., random amplified polymorphism DNA (RAPD) and inter simple sequence repeat (ISSR) were used to assess the genetic diversity in turmeric genotypes. A total of 17 polymorphic primers (11 RAPDs and 6 ISSRs) were used in this study. RAPD analysis of 60 genotypes yielded 94 fragments of which 75 were polymorphic with an average of 6.83 polymorphic fragments per primer. Number of amplified fragments with RAPD primers ranged from 3 to 13 with the size of amplicons ranging from 230 to 3000 bp in size. The polymorphism ranged from 45 to 100 with an average of 91.4%. The 6 ISSR primers produced 66 bands across 60 genotypes of which 52 were polymorphic with an average of 8.6 polymorphic fragments per primer. The number of amplified bands varied from 1 to 14 with size of amplicons ranging from 200 to 2000 bp. The percentage of polymorphism using ISSR primers ranged from 83 to 100 with an average of 95.4%. Nei's dendrogram for 60 samples using both RAPD and ISSR markers demonstrated an extent of 62% correlation between the genetic similarity and geographical location. The result of Nei's genetic diversity (H) generated from the POP gene analysis shows relatively low genetic diversity in turmeric accessions of South eastern ghat (P7), Western undulating zone (P8) with 0.181 and 0.199 value whereas highest genetic diversity (0.257) has been observed in Western central table land (P9). Knowledge on the genetic diversity of turmeric from different agro-climatic regions can be used to future breeding programs for increased curcumin, oleoresin and essential oil production to meet the ever-increasing demand of turmeric for industrial and pharmaceutical uses.     

小麦新品种生选6号的丰产稳产性和赤霉病抗性分析

生选6号是以半矮秆、中感赤霉病的宁麦8号为母本,多穗、中抗赤霉病的宁麦9号为父本杂交,F1代利用花培育种技术育成的高产稳产高抗赤霉病的小麦新品种。为了给该品种的合理利用提供依据,利用国家长江中下游冬麦组区域试验和生产试验以及多年多点赤霉病抗性鉴定的实验数据分析了其丰产性、稳定性及赤霉病抗性。此外,选用与赤霉病抗性相关的9对SSR引物对生选6号及其亲本宁麦8号和宁麦9号进行了基因型分析。结果表明,生选6号在两年国家区域试验和一年的生产试验中均比对照增产,适应性较广,静态和动态稳定性较好;其赤霉病抗性相当于苏麦3号,是目前通过审定并大面积推广应用的品种中唯一高抗赤霉病的小麦新品种。生选6号既存在来自双亲的赤霉病抗性位点,还存在双亲均没有而苏麦3号和望水白都存在的赤霉病抗性位点,这可能是生选6号赤霉病抗性超亲的主要遗传基础。     

中国北方玉蜀黍黑粉菌交配型a位点鉴别及遗传多样性分析

利用特异性引物对2017年采自7个省(自治区)的84份玉米瘤黑粉病菌进行交配型a位点鉴别,采用相关序列扩增多态性(ISSR)分子标记技术对供试菌株进行遗传多样性分析,分析我国北方玉米主要产区玉蜀黍黑粉菌交配型a位点基因型种类及其遗传多样性水平。结果表明,玉蜀黍黑粉菌交配型a位点基因型主要有mfa1、mfa2和mfa1+mfa2三个种类,84株菌株中,mfa1基因型和mfa1+mfa2基因型均有31株,占鉴定株数的36.9%;22株为mfa2位点,占鉴定株数的26.2%。通过筛选出的8条ISSR引物共扩增出61条清晰条带,特异性条带55条,多态性位点所占比例为90.2%。UPGMA聚类分析结果表明,供试菌株遗传相似系数介于0.59～0.92,在遗传相似系数为0.65时供试菌株被分为6个类群,表现出丰富的遗传多样性。ISSR揭示的玉蜀黍黑粉菌遗传多样性、交配型a位点基因型以及地理来源之间的关系分析结果表明,菌株的遗传类群和交配型a位点基因型之间无密切关系,并且两者与菌株地理来源也无明显相关性。     

抗赤霉病小麦优异新种质的分子标记辅助选择

赤霉病(Fusarium head blight,FHB)是危害全球小麦生产安全的重要病害。创制抗赤霉病优异新种质是培育抗病新品种的首要任务。本研究以979-5(中感赤霉病)和苏麦3号(高抗赤霉病)杂交后代为材料,采用系谱育种法,综合利用农艺性状选择、SSR标记选择和赤霉病菌单花接种鉴定等技术,进行抗赤霉病新种质选育。结果表明:(1)用13个分别与苏麦3号3BS上主效抗赤霉病基因 Fhb1、6B上微效基因 Fhb2和5AS上微效基因 Fhb5紧密连锁的SSR标记对双亲进行分析,发现其中的6个标记在979-5和苏麦3号间扩增呈单态,另7个标记在双亲间扩增呈多态,表明979-5与苏麦3号在这3个基因位点具有较高的遗传相似性;(2)创制出中抗新品系13个,高抗赤霉病新品系7个,这20个新品系均携带有苏麦3号的 Fhb1基因位点,其中7个高抗新品系还兼具 Fhb2或 Fhb5标记位点的优异等位变异;(3)20个新品系不仅越冬抗寒性好,且株高较矮,生育期与亲本979-5相近,产量性状好,蛋白质含量、出粉率、湿面筋含量、面团稳定时间等籽粒品质性状较优,其中,k15、k32和k40三个新品系的四项品质指标均达到国家中强筋品种标准。育成的新品系可用作黄淮冬麦区抗赤霉病品种改良的优异育种材料。     

利用醇溶蛋白、盐溶蛋白和RAPD标记划分玉米自交系类群的比较研究

利用醇溶蛋白、盐溶蛋白和RAPD分子标记研究了16个玉米自交系的遗传多样性,并对这16个自交系进行聚类分析。结果显示,在供试材料中筛选到多态性、重复性均比较好的随机引物13个,检测到醇溶蛋白、盐溶蛋白和RAPD标记的多态性条带分别为30条、34条和112条,分别占总带数的83.3%、85.0%和86.8%。3种标记所得遗传相似系数的相关性均达到了极显著水平,所得平均遗传相似系数(0.597、0.541、0.554)基本一致,RAPD标记的遗传相似系数标准差最小(0.080),相对较可靠。依据醇溶蛋白、盐溶蛋白和RAPD分子标记分别将16个供试自交系划分为3类、3类和4类,聚类分析结果都与系谱分析大体相似,同时也存在与所知系谱不符的现象。     

A Set of SCAR Markers Efficiently Differentiating Hybrid Rice

Molecular markers have been widely used in crop genetic improvement,seed test and genetic mapping.Of which,sequence characterized amplified region(SCAR) markers are particularly popular for its diversity,stable reproducibility,and suitability for analyzing large number of samples.In this study,500 random amplified polymorphic DNA(RAPD) primers were tested,and a set of SCAR markers comprising 37 pairs of loci-specific primers were developed from the DNA fragments ranging from 300 to 1000 bp which correspond to the stable,distinctive RAPD banding patterns.Using these SCAR markers,59 hybrid rice combinations were assessed and distinguished into 58 subgroups at the similarity coefficient of 0.97 in a genetic clustering tree based on the allele diversities of the SCAR markers.Furthermore,13 hybrid rice combinations were reassayed with 40 randomly selected simple sequence repeat(SSR) markers to evaluate the effectiveness of these SCAR markers.SSR markers produced similar results to SCAR markers as the 13 hybrid rice combinations were completely separated at the similarity coefficient of 0.91 in the clustering tree established from SSR patterns.Taken together,SCAR markers prove to be effective tools for identifying and differentiating hybrid rice combinations.     

基于转录组的剑麻SSR标记开发与筛选

本研究基于剑麻转录组测序获得的70 110条Unigene序列,采用MISA 1.0软件查找SSR位点,利用Primer 3.0设计SSR引物,通过聚丙烯酰胺凝胶电泳技术对其中的100对SSR引物有效性进行验证。总计获得了13 175个SSR位点,SSR的分布频率为15.61%。70 110条Unigene序列总计包括60种重复基元,其中单核苷酸重复为主导重复类型（37.96%）,其次是二核苷酸重复和三核苷酸重复,比例分别为32.92%和27.90%,四核苷酸、五核苷酸、六核苷酸重复所占比例总计为1.21%。A/T、AG/CT和AGG/CCT分别为单核苷酸、二核苷酸和三核苷酸重复的优势基元。100对SSR引物中有68对引物可扩增出目标产物,其中18对引物在6份剑麻种质中表现出多态性,该结果为利用SSR分子标记技术开展剑麻种质资源鉴定、遗传多样性分析等奠定基础。