Fine mapping of the <Emphasis Type="Italic">Ht2 (Helminthosporium turcicum</Emphasis> resistance 2) gene in maize

Fine mapping of Helminthosporium turcicum resistance gene Ht2 is extremely valuable for map-based cloning of the Ht2 gene,gaining a better knowledge of the distribution of resistance genes in maize genome and marker-assisted selection in maize breeding.An F2 mapping population was developed from a cross between a resistant inbred line 77Ht2 and a susceptible inbred line Huobai.With the aid of RFLP marker analyses,the Ht2 gene was mapped between the RFLP markers UMC89 and BNL2.369on chromosome 8,with a genetic distance of 0.9cM to BNL2.369.There was a linkage between SSR markers UMC1202,BNLG1152,UMC1149 and the Ht2 gene by SSR assay,Among the SSR markers,the genetic distance between UMC1149 and the Ht2 gene was 7.2cM,By bulked segregant analysis 7 RAPD-amplified products which were probably linked to the Ht2 gene were selected after screening 450 RAPD primers and converted the single-copy ones into SCAR markers.Linkage analysis showed that the genetic distance between the SCAR marker SD-06633 and the Ht2 gene was 0.4cM.From these results,a part of linkage map around the Ht2 gene was constructed.     

与美洲黑杨抗黑斑病基因连锁的SCAR标记的开发

通过测序和引物设计,将与美洲黑杨抗黑斑病基因相连锁的RAPD标记OPAI17-1550和OPAI13-900成功地转化成显性SCAR标记(SAI17-1570)和共显性SCAR标记(SAI13-292),并对感、抗病池和F1代91个无性系进行了SCAR标记检测.SAI17-1570在抗病池和OPAI17-1550阳性的F1代植株中扩增出1条与RAPD扩增结果相符的特异性条带,而在感病池和OPAI17-1550阴性的F1代植株中没有相应的带,SAI13-292在抗病池和OPAI13-900阳性的F1代植株中扩增出2条带,而在感病池和OPAI13-900阴性的F1代植株中只有其中1条带.     

不同生态型麦冬特异分子标记的选择与鉴定

采用RAPD技术对四川、湖北、浙江及上海等地 9个不同生态型的麦冬进行遗传多样性研究 ,筛选出 8个可以扩增出清晰、稳定、重复性好和多态性高的随机引物 ,通过对 8个有效引物的指纹图谱分析 ,发现 5个特异的RAPD分子标记 ,可以对 5个不同生态型的麦冬进行区别鉴定     

Phenotypic and genetic variation between two populations of the Chinese yellow pond turtle, Mauremys mutica （Cantor, 1842）

Mauremys mutica (Cantor, 1842) is an endangered species in China. Main phenotypic variations in body color, body weight, body shape, clutch size, egg size, and hatchling size were revealed between the southern and northern populations. Both populations have the phenomenon of "larger male" sexual size dimorphism (SSD), especially in the southern population. Furthermore, genetic variations between the two populations were analyzed by RAPD band patterns of 30 random individuals in each population. The average genetic distance was 0.299±0.108 among the samples of two populations. The average genetic distance between southern and northern populations was 0.305±0.046. Cluster analysis indicated that all the individuals from the southern and northern populations were clustered among themselves to form two distinct clades. A total of 20 population-specific RAPD fragments were scored from 16 primers, and could be used as RAPD markers for distinguishing the southern and the northern population. Based on the nucleotide sequences of two RAPD markers, two pairs of SCAR primers (SC1-S and SC2-S) were designed, which could be used as SCAR markers for the southern population. According to the significant phenotypic and genetic variations, we suggested that the northern population and southern population might be considered as two separate taxa, the "northern taxon" and the "southern taxon", and the conservation should be respectively conducted on the two taxa.     

水稻白叶枯病菌1个毒性相关基因的鉴定

我们曾报道了来自水稻白叶枯病菌真基文库的重级质粒ｐＧＸＮ３０００中含有一个能互补甘蓝黑腐病菌ｒｐｆＧ致病因子调控基因突变全的基因，并通过转座子诱变获得子转座子插在ｐＧＸＮ３０００中的这一基因的转座子插重组质粒，本研究利用这些转座子插入重组质粒，通过标记置换方法构建了不稻白叶枯病菌这一基因的突变菌株，植株试验结果表明，突变株虽然在水稻中仍能正常生长繁殖，但毒性严重降低，说明水稻白叶枯病菌的这一基因在     

枯萎病尖孢镰刀菌的RAPD-PCR多态性分析

利用随机扩增多态性DNA(RAPD PCR),对采自我省不同地区、不同寄主和同一寄主不同植株的瓜类枯萎病原菌尖孢镰刀菌进行遗传多样性分析,筛选到6个多态性随机引物,共产生了72条RAPD带,其中86.11%具有多态性;通过聚类分析,将供试菌株分为4个RAPD组,确定了菌株间的亲缘关系,为瓜类枯萎病原菌尖孢镰刀菌的分类提供有利的分子证据.     

基因聚合品种对云南水稻白叶枯病的抗性分析

研究含多个抗性基因的聚合品种和单个抗性基因的近等基因系对云南省水稻白叶枯病菌的抗性差异.基因聚合品种对水稻白叶枯病菌的抗性达到高抗至免疫水平,病斑长度比单基因品系明显缩短,表明聚合抗病基因提高了水稻品种的抗性.单个抗性基因的近等基因系对这些菌株的抗性均不高,对我省生产中的主要白叶枯病菌株多表现为中感;这一研究结果为培育具有持久抗性的品种提供了新思路,它在实践应用方面将具有重要的意义.     

Molecular markers linked to Rsa resistant to soybean mosaic virus

Soybean mosaic virus (SMV) is a severe disease in worldwide soybean production. A cross was made between Kefeng No. 1 with broad spectrum resistance to SMV and Nannong 1138–2, a susceptible cultivar. The inheritance of resistance to SMV strain Sa prevailing in southern China was analyzed. Results of x² test from inoculation experiment on parents F1, F2 and F3 lines showed that the resistance to strain Sa was controlled by a single dominant gene Rsa. BSA method was adopted and 900 random 10-mer primers were used to amplify total DNA from resistant pool and susceptible pool in order to obtain polymorphic bands in two bulks. 16 primers could generate polymorphic bands, of which OPW-05 and OPAS-06 could generate the most stable RAPD patterns. RAPD markers OPW-05₆₆₀ and OPAS-06₁₈₀₀ were found to be linked to Rsa. Their order and genetic distance were OPAS-06₁₈₀₀22.2cM Rsa10.1 cM OPW-05₆₆₀. Southern blotting showed that both OPAS-06₁₈₀₀ and 0PW-05₆₆₀ were low copy DNA in genomic DNA. 0PW-05₆₆₀ has been converted into an RFLP marker successfully. Additionally, pK644H, an RFLP marker, has been identified to be linked to 0PW-05₆₆₀, and their genetic distance was 37.4 cM.     

Fine mapping of an incomplete recessive gene for leaf rolling in rice （Oryza sativa L.）

Genetic analysis and fine mapping of genes controlling leaf rolling were conducted using two backcrossed generations （BC4F2, BC4F3） derived from a cross between QMX, a non-rolled leaf cultivar as a recurrent parent, and JZB, a rolled leaf NIL of ZB as a donor parent. Results indicated that leaf rolling was mainly controlled by an incompletely recessive major gene, namely rl（t）, and at the same time, affected by quantitative trait loci （QTLs） and/or the environment. A genetic linkage map was constructed using MAPMAKER/EXP3.0 with eight polymorphic markers on chromosome 2, which were screened by BAS method from 500 SSR markers and 15 newly developed insertion/deletion （InDel） markers. The position of rl（t） was estimated with composite interval mapping （CIM） method using WinQTLcart2.5. Gene rl（t） was mapped between markers InDel 112 and RM3763, and 1.0 cM away from InDel 112 using 241 plants in BC4F2 population. To fine map r（t）, one BC4F3 line with 855 plants was generated from one semi-rolled leaf plant in BC4F2. Four new polymorphic InDel markers were developed, including InDel 112.6 and InDel 113 located between markers InDe1112 and RM3763. Based on the information of recombination offered by 191 rolled leaf plants and 185 non-rolled leaf plants from the BC4F3 line ,we mapped r（t） to a 137-kb region between markers InDel 112.6 and InDel 113. Homologous gene analysis suggested that r（t）was probably related to the process of leaf development regulated by microRNA.     

Inheritance of resistance to race 7 of Cercospora sojina in soybeans and RAPD tagging of the resistance gene

Frog-eye leaf spot of soybean caused by Cercospora sojina Hara is a kind of worldwide disease. Resistance to race 7 of C. sojina was found to be due to a single dominant gene through analyzing resistant behavior of the cross NEAU91212 (susceptible to race 7) × NEAU9674 (resistant to all races). 3 RAPD markers linked to the resistant gene Rcsc 7 were identified using the BSA method. 2 fragments of OPS03620 and OPS03580 amplified with primer OPS03 co-dominantly segregated in the F2 individuals. The genetic distance between OPS03620 and the resistant gene is 8.7 cM. According to the co-dominant marker, the accuracy of predicting the homozygous and heterozygous resistant F2 individuals were 100% and 97.6% , respectively.     

水稻苯达松敏感致死基因的RAPD标记的克隆及测序

用3S Spin DNA Agarose Gel Purification Kit试剂盒将与水稻苯达松敏感致死基因相连锁的RAPD遗传标记S20—420和S316—600回收纯化，连接于pGEM—T载体并克隆测序，得到了S20—420和S316—600的全序列，其长度分别为423bp、606bp．将两端序列设计特异PCR扩增引物可用于检测水稻苯达松敏感致死基因和标记辅助育种．     

山羊与岩羊间特异RAPD片段分析

在两组40个随机引物中，筛选出16个重复性好的多态引物，对山羊与岩羊闻RAPD片段分析表明，16个引物扩增出67条带，其中54条带表现多态，多态率80．60％，山羊各群体共有条带为23条，山羊和岩羊共有条带为13条，岩羊有4条特异带，不同引物所扩增出的片段在各群体中分布频率不同．岩羊特异RAPD片段，OPA-10724的序列与人类全基因组比较，同源的短序列较多，最太长度为86bp，唯有30bp的长度与绵羊ZFZ基因和牛ZFY基因内含子同源。     

Genetic analysis and gene mapping of leafy head （lhd）, a mutant blocking the differen-tiation of rachis branches in rice （Oryza sativa L.）

Flowers, fruits and seeds are products of plant re- productive development and provide the important sources of foods for humans. Therefore, the moleculargenetic mechanisms of floral development have been ahotspot of research of plant developmental biology[1]. Rice is one of the most important staple food crops. Theoutcome of its reproductive development would determine the yield and quality of grains. Rice is also a model plantof cereals. Hence, the study of rice reproductivedevelopment, esp…     

基于RAPD标记的我国水青冈属植物的分类研究

应用随机引物扩增多态性DNA(RAPD)方法,对我国水青冈属(Fagus L.)6种植物,即巴山水青冈(F.pashanica)、长柄水青冈(F.longipetiolata)、米心水青冈(F.engleriana)、亮叶水青冈(F.lucida)、台湾水青冈(F.hayatae)和浙江水青冈(F.hayatae var.zhejiangensis)进行基因组多态性分析,选用80个随机引物进行扩增,其中56个引物扩增出清晰可重复的条带,片段大小在250～3050 bp之间.采用UPGMA法对求出的遗传片段进行聚类分析.结果显示:台湾水青冈、巴山水青冈和浙江水青冈的遗传距离较近,表明它们可归并为一种.这与我国水青冈属的生态解剖学研究结果完全一致.根据结果将我国水青冈植物归并分为4种:长柄水青冈、米心水青冈、亮叶水青冈、台湾水青冈(包括巴山水青冈和浙江水青冈).     

高粱抗丝黑穗病基因的分子标记RAPD分析

丝黑穗病是影响我国高粱生产发展的主要病害之一,在高粱产区每年都有发生,发病率有时高达70 %,给生产造成很大损失,而选育抗病品种是最为有效的抗病策略.分别以高粱2381恢复系(抗病)、矮四恢复系(感病)的叶片为试材,采用CTAB法提取高粱基因组DNA,并应用RAPD筛选技术对高粱基因进行分子标记.在最佳的RAPD反应体系中共筛选了100个RAPD随机引物,筛选出来27个适宜引物,获得了稳定的RAPD扩增结果及多态性较好的DNA谱带.其中有6个引物对DNA扩增产生了差异谱带.     

应用RAPD标记构建马尾松单株树遗传连锁图谱

利用马尾松（Ｐｉｎｕｓｍａｓｓｏｎｉａｎａ）崇义群体Ｃ－１６号单株上４０粒种子的胚乳为作图群体材料，用随机扩增多态性ＤＮＡ（ＲＡＰＤ）方法构建马尾松单株遗传连锁图谱，从被步筛选的８０个引物中，得到１６个具有多态性产物的引物，对这１６种引物进行Ｍｅｎｄｅｌｉａｎ１：１分离检测（卡方检验，Ｐ＜０．０５），我们得到符合Ｍｅｎｄｅｌｉａｎ１：１分离比例的ＲＡＰＤｓ标记４９个．通过对４９个标记的连锁关系进行分析，其中２９个标记分布在１２个连锁群上，总图距为４８３．３ｃＭ，平均图距为２８．４２ｃＭ，２０个标记没有连锁到任何连锁群上，需要继续进行大量标记的连续分析，构建完善的高密度的遗传图谱，使之能够对数量性状位点ＱＴＬｓ进行定位，从而进行分子标记辅助选择育种（ＭＡＳ）．     

Genetic and physical mapping of AvrPi7, a novel avirulence gene of Magnaporthe oryzae using physical position-ready markers

Rice blast, caused by the fungal pathogen Magnaporthe oryzae, is one of the most devastating crop diseases worldwide. The avirulence gene corresponding to rice blast resistance gene Pi7 in field isolate CHL346 was inherited as a single gene, designated AvrPi7, in a segregating population consisting of 189 ascospore progenies derived from a cross between field isolates CHL346 and CHL42. In order to determine the chromosomal location of the AvrPi7 locus, a total of 121 simple sequence repeat (SSR) markers were developed based on the whole-genome sequence of reference isolate 70-15 of M. oryzae. Linkage analysis of the locus with these SSR markers showed that eight SSR markers on chromosome 1 were linked to the locus, among which the closest flanking markers MS1-9 and MS1-15 were 3.2 and 16.4 cM from the locus, respectively. For fine mapping, additional PCR-based makers including eight SSR markers and three candidate avirulence gene (CAG) markers were developed in the region flanking both markers. The AvrPi7 locus was genetically delimited within a 1.6-cM region flanked by markers MS1-21 and MS1-22, and co-segregated with the marker CAG2. To construct a physical map of the AvrPi7 locus, molecular markers linked to the Avr gene were mapped on the supercontigs of the ref-erence isolate 70-15 through bioinformation analysis (BIA). Consequently, the AvrPi7 locus was delim-ited to a 75-kb interval flanked by markers MS1-21 and MS1-22 based on the reference sequence. Merodiploids observed in this study are also discussed.     

Construction of a genetic linkage map for cotton based on SRAP

DNA markers have been widely used in construction of molecular genetic linkage maps in plants. The first genetic linkage map of cotton was constructed by Reinish in 1994 using RFLP (restriction fragment length polymorphism)[1], which included 705 polymorphic loci on 41 linkage groups with a total length of 4675 cM. Afterwards, several genetic linkage maps were constructed[2—7], but no map is comparable to this one in marker density. A high-density genetic linkage map could be applied effec…     

罗汉果性别相关的RAPD标记

应用随机扩增多态性DNA(RAPD)技术寻找与罗汉果性别相关的分子标记,筛选了130个10 bp的随机引物,发现有4个引物(S60、S90、S100、S343)能在雌、雄株DNA间扩增出5条差异性片段,大小在300~1 300 bp之间,表明这些特异带可被用来作为性别鉴别的特异性分子遗传标记。