RAPD技术对东北地区大豆的初步研究

采用ＲＡＰＤ技术对东北地区不同类型大豆共３２份材料进行了分析，以野生种０１６５０，半野生种０１７７１和栽培种吉林２１号大豆的基因组ＤＮＡ为模板，对Ｏｐｅｒｏｎ公司的Ａ，Ｆ，ＧＨ和Ｋ组共１００个随机引物进行了筛选，依据扩增结果选用了１９个引物对３２份大豆材料进行了扩增，所获得的８２个多态片段可将供试材料全部区分开。     

Characterization and molecular marker screening of a rice bacteria-resistant gene Xa-min(t)

To test the resistant spectrum of the Xa-min(t) gene introgressed from Oryza minuta, thirty-four isolates of different bacterial blight pathogen, Xanthomonas oryzae pv. oryzae (Xoo), from 11 countries were used to inoculate the Xa-min(t) introgression line 78-15. Four rice cultivars, IR24, C64 (IRBB21), Nipponbare and Zhonghua 11 were used as controls. The results showed that the Xa-min(t) gene was broad-spectrum and highly resistant to diverse Xoo isolates. The methods of bulk segregant analysis (BSA), randomly amplified polymorphic DNA (RAPD) and sequence characterized amplified regions (SCAR) were used to analyze F2 individuals of the hybrid IR24×78-15 and molecular genetic markers linked to Xa-min(t) gene were identified. A total of 800 arbitrary decamer oligonucleotide primers were used for RAPD analysis. Two RAPD markers, BE05300 and BE061400, produced by primers BE05 and BE06 respectively, were closely linked to the Xa-min(t) gene. Based on the sequences of these two markers, sequence specific primers were designed and used to screen all F2 plants. One RAPD marker, BE05300, was converted into a stable SCAR marker (ScBE05300). Linkage analysis was carried out using markers ScBE05300 and BE061400 on 948 and 719 F2 individuals of the hybrid IR24×78-15. Our results indicate that the genetic distances from Xa-min(t) to ScBE05300 and BE061400 are 2.2 cM and 3.7 cM respectively on the same side. This study may facilitate the construction of the fine physical map of the Xa-min(t) gene.     

唐山奶山羊随机扩增多态DNA与体重体尺相关性研究

利用１２种随机引物研究了唐山奶山羊的随机扩增多态ＤＮＡ,并利用ＳＰＳＳ程序对多态标记进行了方差分析。结果表明,唐山奶山羊基因组ＤＮＡ多态频率为５１．９２％。多态标记Ｋ０２Ａ、Ｋ０３Ａ、Ｑ０５Ｃ、Ｑ１４Ｄ及其互作效应对体重和体尺有显著影响。     

HFJ和MIJ近交系大鼠RAPD特征与比较研究

目的 利用人工合成的PCR随机引物,对本单位培育的HFJ和MIJ近交系大鼠基因组DNA进行扩增,建立 HFJ和MIJ大鼠RAPD扩增特征图谱;选择常用近交系大鼠Lewis和F344作为对照品系,观察HFJ和MIJ大鼠与 对照品系大鼠的RAPD多态性。方法 用酚-氯仿法分别提取4个不同品系大鼠的基因组DNA,从60条随机引 物中筛选能够得到清晰扩增条带的30条引物,对HFJ和MIJ大鼠DNA进行扩增。再从30条引物中随机选择12 条,对4个品系大鼠基因组DNA扩增,并对扩增条带多态性比较分析。结果 30条引物对HFJ和MIJ大鼠DNA 进行扩增,均能获得1～7个较清晰条带,除引物0～08外,条带的分子大小均在200～1 100 bp之间,两品系的引 物0-08扩增结果,均在2 000 bp以上的近原点处扩增出1条清晰的条带。HFJ和MU大鼠品系内个体间和两个品 系之间,扩增的结果均完全一致;用12条随机引物对4个品系大鼠同时扩增,HFJ和MIJ大鼠扩增结果一致,与 Lewis和F344比较,有5个引物出现差异条带。结论 用30条随机引物建立了HFJ和MIJ大鼠RAPD扩增特征性 图谱。由于两品系来源于同一对Wistar封闭群大鼠,虽然遗传特性有显著差异,但是在所选30条引物扩增时,未 发现多态性。两品系与Lewis和F344之间存在多态性条带     

甜菜抗根腐病基因ISSR分子标记的初步研究

以感病×抗病的甜菜种间杂交组合KWS9419×ZD204的F1代17个单株及ZD自交一代16个单株为试材,采用BSA法和ISSR技术,通过对155个随机引物的筛选,获得了一个与甜菜抗根腐病基因连锁的ISSR标记OPP0760,可以用作抗根腐病基因的分子辅助选择的依据。     

Identification of RAPD marker linked with fertility-restoring gene of cytoplasmic male sterile lines in upland cotton

Bulked segregant analysis was employed to construct two mixed DNA pools to screen the RAPd marker linked with the fertility-restoring gene(Rf _i) of upland cotton. A total of 425 arbitrary 10-mer oligonucleotide primers were screened on two DNA pools, bulked male fertile and sterile DNAs isolated from BC₃ segregating population of (0-613-2R X Simian No. 3). Three primers produced repeatable polymorphisms between the paired bulks and their parents. DNA was extracted and amplified with these three primers for 92 plants of (Zhong 12A-1 × 0-613-2R)F₂. Based on the male fertility scoring and RAPD amplification, it is found that one RAPD marker fragment designated OPV-15₃₀₀ was linked with the fertility-restoring gene (Rf₁) with a recombination value of 13.0±2.57%.     

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.     

Identification of an AFLP marker linked to the stripe rust resistance geneYr10 in wheat

AFLP analysis of near-isogenic lines of the stripe rust resistance gene Yr10 was carried out with 6 PstⅠ- primers and 10 TaqⅠ-primers with the donor parent of Yr10 gene as the check. A total of about 4200 distinguishable bands were amplified, of which 5 were stable. The genetic linkage of the 5 polymorphic DNA fragments with the target gene were tested preliminarily on a segregating F₂ population derived from a cross between the gene donor parent “Moro” and susceptible cultivar “Mingxian 169”. The DNA fragment PT0502 was found closely linked to the Yr10 gene and cloned and sequenced. Based on the sequence specific primers for PCR were designed and synthesized. Genetic linkage analysis with 195 segregating F₂ plants indicated that the genetic distance was 0.5 cM between the main product SC₂₀₀ fragment produced by PCR with the primers and the Yr10 gene. The primers can be used to detect the Yr10 gene quickly, effectively and exactly.     

Genetic relationships among Chinese and American isolates of Phytophthora sojae assessed by RAPD markers

The genetic diversity of three geographic populations of Phytophthora sojae from China and the United States was determined using random amplified polymorphic DNA （RAPD）. The purpose was to explore genetic relationships among Chinese and American isolates of the organism. 21 random primers were selected among 200 random primers screened. A total of 223 reproducible RAPD fragments were scored among 111 individuals, of which 199 （89.23%） were polymorphic. Analysis of genetic variation showed that there existed higher genetic variation in the United States population in comparison to the Chinese populations. Nei＇s genetic identity and principal component analysis indicated that the populations of Fujian and United States are closer to each other than to Heilongjiang populations. Shannon-Wiener diversity index revealed that the United States populations have a higher genetic di- versity than that of Chinese populations. These data are in support of the hypothesis that P. sojae in the United States might not have been introduced from China.     

Genetic diversity of reared N.miichthioides population

The genetic diversity of 30 rearedNibea miichthioides individuals was analyzed by random amplified polymorphic DNA (RAPD) with 20 random primers. The result showed that the genetic diversity of reared individuals was relatively low with 15.31% polymorphism and 0.031 9 of the average difference (AD). The result also indicated that RAPD is a useful way in genetic diversity analysis of fish population.     

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).     

丁(鱼岁)遗传多样性的随机扩增多态性DNA分析

利用RAPD分子标记技术对15个丁鱼岁养殖个体的遗传多样性进行分析.选用20个随机引物对其基因组DNA进行PCR扩增,有16个引物可以扩增出稳定且清晰的条带,其中S103、S104和S105为多态引物.16个引物共扩增出65个DNA位点,片段大小在200~3 000 bp之间,其中多态性位点6个,多态位点比例为9.23%.个体间遗传距离在0~0.067 4之间,平均遗传距离为0.028 6.结果显示,丁鱼岁养殖群体的遗传多样性水平较低.     

淤泥湖和梁子湖团头鲂遗传多样性的研究

采用随机扩增多态DNA（RAPD）技术对淤泥湖和梁子湖的团头鲂进行了遗传分析,在所用20个10碱基随机引物中,除OPP－1、OPP－13外,其余18个引物都能对团头鲂扩增出稳定的RAPD谱带,其中OPP－7、8、9、12、15、17等6个引物在团头鲂不同个体间扩增出多态性片段,用这些引物对团头鲂进行遗传分析,结果表明,淤泥湖团头鲂个体间的遗传相似度在0．9543－0．9718之间,平均值为0．9617;梁子湖团头鲂个体间的遗传相似度在0．9541－0．9622之间,平均值为0．9589;淤泥湖和梁子湖团头鲂个体间的遗传相似度在0．9395－0．9614之间,平均值为0．9541。     

墨天牛属三个近缘种的RAPD分析

应用ＲＡＰＤ（随机扩增的多态性ＤＮＡ）技术,分析了墨天牛属三个近缘种－松墨天牛、云杉大墨天牛和云杉小墨天牛的ＤＮＡ多态性。通过采用多种引物对同种天牛幼虫的浸渍标本和新鲜标本的ＤＮＡ多态性研究,发现浸渍标本与新鲜标本ＤＮＡ扩增后的电泳指纹图谱完全一致;同种害虫不同虫态间的ＤＮＡ指纹图谱完全一致;三种随机引物可明显鉴别三种天牛幼虫。这为天牛近缘种的鉴定,尤其是天牛幼虫的鉴定提供了一种新方法。     

Molecular characterization of RAPD and SCAR marker linked to the frog-eye leaf spot resistance gene in soybean

Two fragments SCS3₆₂₀ and SCS3₅₈₀ of the co-dominant marker OPS03_{620 & 580} that were linked to the resistance gene of soybean frog-eye leaf spot have been completely sequenced. A significant insertion of 30 bp is the main reason of the polymorphism between the two fragments. The results of Southern hybridization indicate that SCS3₆₂₀ derives from a single-or low-copy sequence and can be used as an RFLP probe. A co-dominant SCAR marker SCS3_{620 & 580} has been developed based on the sequences. The segregation of SCS3_{620 & 580} is similar to that of RAPD marker OPS03_{620 & 580}-Significant polymorphism has been shown between resistant and susceptible genotypes when 62 soybean genotypes were surveyed for the SCAR marker. Therefore, the marker can be used in the resistance breeding of soybean frog-eye leaf spot by marker-assisted selection.     

松枯梢病菌RAPD分子水平遗传距离分析

对来自中国、美国、英国、南非和智利的松枯梢病菌23个菌株进行RAPD分析。用12个引物共扩增出135个RAPD标记,其中多态性标记占96.3%。各菌株间的Nei相似系数UPGMA法聚类结果表明,国内外23个菌株可大致分为3个类群:来自智利的CWS41与所有菌株的遗传关系最远;来自中国的F2和J2次之;其他的国外菌株与国内菌株的遗传关系则较近。此次实验未将来自美国的B型菌株与其他菌株区分开。     

滩羊体大品系遗传标记的研究

采用RAPD(Rondom Amplified Polymorphic DNA)技术,利用混合基因池(DNA pool)法,对滩羊体大品系、普通品系进行了DNA多态性分析,从100种具有10个碱基的随机引物中,筛选出84种引物在滩羊群体基因组中共扩增出358条带,其中22种引物的扩增产物表现为多态(占22％),且扩增出32条有差异的条带,占总带数的8.94％;62种引物的扩增产物表现为单态(占62％)。滩羊体大品系的特异性条带有5条,而普通品系的特异性条带有7条,这些特异标记可以用来鉴定滩羊的两个品系;滩羊体大品系与普通品系间的遗传距离为0.136±0.087,表明两品系之间的亲缘关系很近。     

5种菊头蝠的随机扩增DNA多态(RAPD)的分析

以5种菊头蝠为材料,用随机扩增DNA多态(RAPD)方法分析它们的亲缘关系。结果显示:5种菊头蝠均表现出各自不同的多态性RAPD标记,6条单个引物分别扩增出2~10条的RAPD条带,其分子量约在700~2 600bp之间。依据这些多态性标记计算其种间遗传相似性,建立亲缘关系树形图。     

青龙本地山羊随机扩增多态DNA与体重体尺相关性研究

利用９ 种随机引物研究了青龙本地山羊的随机扩增多态ＤＮＡ,并利用ＳＰＳＳ程序对多态标记进行了方差分析。结果表明,青龙本地山羊基因组ＤＮＡ 多态频率为４８．６５％ 。多态标记Ｋ０９Ｂ、Ｐ１４Ｄ和Ｑ１４Ｄ及其互作效应与体重和体尺有显著相关     

马氏珠母贝与解氏珠母贝的随机扩增多态DNA分析

用OPM组20种碱基随机引物对采自海南洋浦的马氏珠母贝（Pincada martensii）和解氏珠母贝（P.chemmitri）天然群体进行RAPD分析,其中6种引物扩增有效,这6种引物对马氏珠母贝和解氏珠母贝的总扩增带数分别为52条和58条。每一引物的扩增带数为3～16条,同一引物对两种贝类的扩增带数不同（OPM19B除外）,对两贝类的扩增带型也有较大差异。群体内的各个个体（实验个体为10个）RAPD扩增带谱全部呈多态,表明在同一群体中,各个体存在着明显的个体特征带,由于样品数量及引物数均少,2种贝类各自种群的共同特征尚未能确定。