白菜紫色性状RAPD连锁标记的筛选与染色体定位研究

以紫菜薹自交系95T2-5、大白菜自交系94S17-1及其F1、F2、BC1植株为材料,进行RAPD分析,从640个随机引物中筛选出2个引物S79和S123,分别能扩增出与紫色性状连锁的条带S79-934和S123-750。连锁分析发现,标记S79-934和S123-750与紫色基因间的遗传距离分别为13.73cM和18.65cM,并且位于紫色基因的两边。回收S79-934特异带,克隆转化并测序,比较分析表明其与大白菜1号染色体上已知克隆KBrH077A05的全序列(113253bp)有99%的相似性,初步推断控制紫色性状的主基因位于大白菜1号染色体上。     

茎瘤芥(榨菜)瘤茎形状的遗传研究

利用瘤茎形状有较大差异的4个茎瘤芥(Brassica juncea var.tumida Tsen et Lee)自交系作为亲本配置了2个杂交组合,并以瘤茎形状指数作为度量指标,应用主基因+多基因混合遗传模型对其衍生后代家系群体P1、P2、F1、B1、B2和F2瘤茎形状进行了多世代联合遗传分析。杂交组合y203×b145的瘤茎形状受2对加性-显性-上位性主基因控制,其遗传率在B1、B2和F2群体中分别为59.89%、26.18%和54.14%;而瘤茎形状在杂交组合y92×b146中受2对加性-显性-上位性主基因+加性-显性多基因控制,在B1、B2和F2家系群体中,其主基因遗传率分别为22.44%、58.06%和63.14%,多基因遗传率分别为40.42%、4.36%和1.26%。这些结果表明,对瘤茎形状改良时要以主基因利用为主,且宜在中高世代选择。     

小麦品系5R618抗叶锈基因的初步定位

5R618是高抗叶锈病小麦品系。为了确定该品系所携带的抗叶锈基因,以5R618与感病小麦品种郑州5389杂交获得F1,自交获得F2分离群体以及F2∶3家系,用叶锈菌生理小种THJP对亲本、F2分离群体以及F2∶3家系进行叶锈抗性鉴定,然后进行分子标记分析。结果显示,5R618对生理小种THJP的抗病性由1对显性基因控制,该基因暂命名为Lr5R。经过亲本和抗感池间分子标记筛选以及F2∶3家系的标记检测,Lr5R定位于染色体3DL上,barc71和STS24-16是Lr5R最近的2个标记,遗传距离分别为0.9 c M和2.1 c M。     

人工合成小麦抗白粉病未知基因的SSR标记

YAV-2/TEZ//A.SQ(895)是硬粒小麦与粗山羊草杂交获得的抗白粉病人工合成小麦。本研究利用人工合成小麦YAV-2/TEZ//A.SQ(895)与感白粉病的普通小麦品系品资50098杂交和自交获得的F2代群体及F3家系,在温室条件下鉴定群体的白粉病抗性。遗传分析结果表明,该抗白粉病基因为显性单基因遗传。利用647对小麦SSR引物进行了白粉病抗性基因的分子标记分析,结果表明该白粉病抗性基因与2A染色体的6个SSR标记连锁,与标记Xcfa2086的遗传距离最近,为11.8cM。     

小麦农家品种红麦(京2747)主效抗条锈病基因的RAPD标记

小麦农家品种红麦(京2747)可抗中国小麦条锈菌多个生理小种。遗传分析表明,该品种对于小麦条锈菌条中19号生理小种的抗性由1对显性基因控制。本研究采用铭贤169×红麦的F2分离群体建立抗、感DNA池,用RAPD方法进行DNA多态性分析。共筛选236个10碱基随机引物,其中引物S1167所扩增出的1条约245 bp的多态性DNA片段只出现在抗病DNA池和红麦中,而不出现在感病DNA池和感病品种铭贤169中。经用201株杂交F2植株对多态性DNA片段S1167245与目的基因的遗传连锁性进行分析,在164株抗病单株中有156株可稳定扩增出该特异DNA片段,而在37株感病单株中则有34株不能扩增出该特异DNA片段,经统计共有11株发生了交换,标记S1167245与目的抗病基因间的遗传距离为6.1cM。本研究得到的RAPD标记S1167245表现稳定、重复性强,可用于小麦抗锈育种中的标记辅助选择,促进红麦的抗条锈基因的利用。     

葡萄感霜霉病基因的分子标记(英文)

 在葡萄抗病育种中 ,幼苗期排除感霜霉病的后代具有特别重要的意义 .用 BSA,RAPD和SCAR方法研究了葡萄感霜霉病基因的分子标记 .分析了两个种间杂交组合 [毛葡萄 (抗病 )×欧洲葡萄 (感病 ) ]88- 1 1 0和 88- 84与 88- 1 1 0的 F1代自交或互交所得的 3个 F2 代 ,以及欧洲葡萄品种和中国野生葡萄种 .共筛选了 2 80个随机引物 .引物 OPO1 0产生了一个 RAPD标记 OPO1 0 - 80 0与葡萄感霜霉病主效基因紧密联锁 .将该 DNA片段克隆并测序 .OPO1 0 - 80 0的实际长度为 835bp,所以 OPO1 0 - 80 0应为 OPO1 0 - 835.据其两端序列 ,设计了一对长度为 2 6bp和 2 8bp的特异引物分别扩增上述试材 ,获得了与该 RAPD标记相同大小的一条带 ,将 RAPD标记转化为 SCAR标记SCO1 0 - 835.并证实了此 SCAR标记的通用性 ,该 SCAR标记可用于葡萄抗病育种中杂种后代对霜霉病的抗病与感病性鉴定 .     

水稻苯达松敏感致死基因的RAPD标记和SCAR标记

利用RAPD技术对水稻品种农林 8号 (含苯达松抗性基因Ben)和其突变体农林 8号m (含苯达松敏感致死基因ben)进行标记 ,从 36 0个 10bp寡核苷酸随机引物中筛选出 5个引物产生的 7个RAPD标记。经对多态性标记的克隆和序列分析 ,再设计PCR引物 ,将其中 4个RAPD标记OPG18/ 94 3、OPG18/ 972、OPD10 / 12 4 8和OPF0 3/ 1198转化成SCAR标记SCAR/G18/ 883、SCAR/G18/ 890、SCAR/G18/ 919/ 94 8、SCAR/D10 / 12 37、SCAR/F0 3/ 1186。通过对农林 8号×农林 8号mF2 分离群体 32 0个单株的连锁分析及在 1对含ben基因的近等基因系H12 1和Hben12 1中验证 ,标记SCAR/G18/ 883、SCAR/G18/ 890、SCAR/G18/ 919/ 94 8与Ben或ben基因共分离 ,SCAR/D10 / 12 37与Ben基因的遗传距离为 (14 .8± 2 .1)cM。经Southernblotting分析并结合F2 代分离比例表明 ,标记OPG18/ 94 3、OPG18/ 972及其转化的SCAR标记在基因组中为单拷贝序列 ,且OPG18/ 94 3和OPG18/ 972为一对等位STS位点。这是首次报道与ben或Ben基因相连锁的分子标记。本研究为利用分子标记辅助ben基因的转育及利用图位克隆技术分离ben基因提供了有用的分子标记。     

人工合成甘蓝型油菜中花色与芥酸含量的遗传连锁分析

利用人工合成的甘蓝型油菜品系．No．2127-17(白花、有芥酸)与加拿大双低甘蓝型油菜品种Quantum(黄花、低芥酸)配制杂交组合。对亲本、F1、BC1、F2和DH(doubled haploid)5个世代的花色及芥酸含量进行分析,结果表明：花色受单基因控制,且白花对黄花为显性;芥酸含量仅表现出一对基因的差异且具有加性效应的遗传模式。花色和芥酸含量的连锁分析表明：白花与高芥酸紧密连锁．在DH群体中重组频率为5．8％。采用集团分离分析法(Bulked Segregant Analysis,BSA),从685条10个碱基的随机引物筛选到一个与黄花和低芥酸含量紧密连锁的RAPD标记S92-1400。在遗传图谱上黄花基因和低芥酸基因距离S92-1400标记的图距分别为2．2cM和5．4cM。     

甘蓝型油菜含油量的主基因+多基因遗传效应分析

应用多世代联合分析数量性状主基因和多基因混合遗传的统计方法,分析了甘蓝型油菜两个组合的5个世代——亲本P1、P2、F1、F2和F2：3家系材料含油量的遗传效应。结果表明,分离世代F2及F2：3家系含油量次数分布均呈混合的正态分布,符合主基因＋多基因的遗传特征。D-2模型是该项研究两个甘蓝型油菜杂交组合含油量的最适遗传模型,含油量的遗传是由一对加性主基因和加-显性多基因共同控制的。组合1（1141Bx垦C1-1）主基因加性效应值为-1．74,表明亲本1141B中主基因位点上的等位基因降低含油量,而亲本垦C1-1中的等位基因增加含油量。多基因加性效应值和显性效应值分别为1．20和-1．93;F2的主基因遗传力和多基因遗传力分别为68．21％和27．17％;F2：3的主基因遗传力和多基因遗传力分别为81．70％和16．80％。组合2（32Bx垦C1-2）主基因加性效应值为-3．74,表明亲本32B中主基因位点上的等位基因降低含油量,而亲本垦C1-2中的等位基因增加含油量。多基因加性效应值和显性效应值分别为-1．99和0．93;F2的主基因遗传力和多基因遗传力分别为66．20％,和28．10％;F2：3的主基因遗传力和多基因遗传力为81．00％和14．90％。两组合在F2：3家系世代含油量的主基因遗传力均较F2高,因此认为高含油量育种中在F2：3家系进行选择效率较高。     

小麦耐盐基因的标记和标记的克隆

普通小麦（Triticum aesticum L）耐盐农家品种茶淀红与盐敏感品种农大85021杂交,对杂交后代141株F2分离个体进行耐盐性鉴定。通过遗传分析和卡方检验证实小麦耐盐农家品种茶淀红含有一个主效耐盐基因,杂交后代符合1:2:1的分离规律。依据混合群体分组分析法（Bulked Segregate Analysis,BSA）建立耐盐基因池和盐敏感基因池,通过RAPD实验,从520个引物中筛选出了一个在两池间具有多态性的引物OPZ09,用双亲、F1、F2单株DNA进行RAPD实验证明了该引物扩增出的特异性片段OPZ09-590是一个与茶淀红耐盐基因连锁的RAPD标记,用JOINMAP Version1.4计算基因与标记间的重组值为5.674%,连锁距离为6.557cM。从琼脂糖凝胶回收OPZ09-590与载体pUCm-T连接,并转入受体菌JM109,对克隆片段测序表明其实际大小为591bp,故此小麦茶淀红耐盐基因的RAPD标记为OPZ09-591。     

Analysis of genetic heterogeneity among five gynogenetic clones of silver crucian carp, Carassius auratus gibelio Bloch, based on detection of RAPD molecular markers

The gynogenetic silver crucian carp, Carassius auratus gibelio, is a unique model system for studying evolutionary genetics and selective breeding, owing to its specific genetic background and reproductive modes. Five gynogenetic clones were analyzed by the random amplified polymorphic DNA (RAPD) technique, using 30 10-nucleotide-long primers. Twenty-six primers produced well-amplified DNA fragments with reproducible banding patterns, and 24 primers were polymorphic. Nearly identical banding patterns were observed among individuals within each clone, suggesting that each clone might possess a specific pattern owing to its gynogenesis. In contrast, the RAPD patterns of the five clones differed from each other. A phylogenetic tree was constructed using UPGMA cluster analysis based on a total of 3,744 distinguishable fragments (156 per individual). Average genetic distances within and among the five clones clearly indicated their intraclonal homogeneity, interclonal heterogeneity, and phylogenetic relationships. Clones A and P were the most closely related, whereas the most divergence was seen between clone D and clone E or F. A total of 88 polymorphic fragments were scored from 24 primers after excluding bands that were monomorphic for the five clones. Most primers corresponding to the polymorphic fragments amplified reproducible markers specific for one clone or that were shared by two, three, or four clones. Several primers (e.g., Opj-1, Opj-7, and Opp-10) produced abundant banding patterns that could be used to discriminate between the five clones. Markers specific for one or two clones were also identified. The RAPD markers identified in this study will likely benefit evolutionary genetics and selective breeding studies.     

与葡萄抗霜霉病基因紧密连锁的分子遗传标记

以种间杂交组合88-110［83-4-96（毛葡萄,抗霜霉病）×粉红玫瑰（欧洲葡萄,感霜霉病）］的F     

Identification of the Molecular Markers Linked to the Salt-resistance Locus in the Wheat Using RAPD-BSA Technique

Calli from mature embryo of “Jimai-24” wheat ( Triticum aestivum L.) were induced on medium containing Zhengdingmycin then continuously cultured on medium containing 0.5% NaCl till to regenerate plants named 8901-17 salt-tolerant mutant. “Jimai-24” was compared with 8901-17 by using the technique of RAPD. Thirty-five out of 280 random primers could detect DNA polymorphism. The similarity index was 0.978, indicating that they were NILs (near-isogenic lines). Two F2 populations (“Jimai-24”×8901-17 and 8901-17דZhongmai-9”) had been constructed using the method of half-division. The two relative DNA pools (salt tolerant DNA pool and susceptible DNA pool) which come from the two F2 populations, respectively, had been made according to the method of BSA (bulked segregant analysis). RAPD analysis between the two DNA relative pools was carried on with above 35 random primers which could detect DNA polymorphism definitely. The identical polymorphism between the two sets of DNA pools come from the two F2 populations could be determined only by OperonQ4 primer. This result implied that the polymorphic fragment amplified by OperonQ4 primer was the molecular marker of RAPD closely linked to the salt tolerant mutation locus.     

Pathogenic and molecular characterisations of pigeonpea wilt pathogen Fusarium udum

Thirty two pathogenic isolates of Fusarium udum from different pigeonpea growing areas in India were studied for pathogenic and molecular variability. Pathogenic variability was tested on 12 pigeonpea differential genotypes, which revealed prevalence of five variants in F. udum. The amount of genetic variation was evaluated by Polymerase Chain Reaction (PCR) amplification with 20 random amplified polymorphic DNA (RAPD) markers and nine microsatellite markers. All amplifications revealed scorable polymorphisms among the isolates, and a total of 137 polymorphic fragments were scored for the RAPD markers and 16 alleles for the simple sequence repeat (SSR) markers. RAPD primers showed 86% polymorphism. Genetic similarity was calculated using Jaccard's similarity coefficient and cluster analysis was used to generate a dendrogram showing relationships between them. Isolates could be grouped into three subpopulations based on molecular analysis. Results indicated that there is high genetic variability among a subpopulation of F. udum as identified by RAPD and SSR markers and pathogenicity on differential genotypes.     

人工雌核发育鲢的遗传多样性及异源遗传物质整入的RAPD分析

运用RAPD技术对连续二代人工雌核发育鲢的遗传多样性及异源遗传物质的整入进行了分析 ,结果表明 :一代雌核发育鲢 ,个体间遗传相似度为 0 94 5— 0 995 6 ,多样性指数为 0 175 ;二代雌核发育鲢 ,个体间遗传相似度为0 96 15— 1 0 0 ,平均为 0 985 2 ,多样性指数为 0 0 6 2。研究揭示经过连续二代人工雌核发育后 ,其遗传多样性明显减少 ,种质进一步纯化。通过对雌核发育鲢二代、亲本鲢和雄鲤的RAPD扩增比较 ,发现雌核发育鲢含有少数与父本相同的特异DNA扩增带 ,而亲本鲢没有 ,在基因水平上表明雌核发育鲢整入了雄鲤的遗传物质     

珍稀濒危植物天目木兰（Magnolia amoena）遗传多样性的RAPD分析

运用随机扩增多态DNA(RAPD)技术,对天目木兰(Magnolia amoena)居群的遗传多样性进行了研究．从40个10-mer随机引物中筛选出14个能得到清晰、稳定扩增带的引物进行扩增,14个引物共检测了94个位点,其中多态性位点为23,占24．4％,计算了12个居群之间的遗传相似度和遗传距离,并运用UPGMA法进行了聚类分析,结果显示相同严地个体间(居群内)的遗传距离较小,遗传多样性水平很低;不同产地个体间(居群间)遗传距离较大,遗传多样性水平较前者高,即天目木兰个体间遗传多样性水平与它的地理分布有关,天目木兰总体较低的遗传多样性是导致它濒危的原因之一。     

Identification of Molecular Marker for Septumless Bold Pod in Brassica campestris

Two cultivars of yellow sarson (Brassica campestris), B9 and NC1 have sharp phenotypic differences: a) pubescent or glabrous leaves, b) septumed pod with less number or septumless bold pod with higher number of seeds, and c) erect or drooping pods relative to stem axis. It is established that septum less pod is related to enhancement of seed yield and also related to high shattering resistance. The character septumed pod and pubescent leaf are controlled by single genes with complete dominance and are situated on the same linkage group, as evidenced by the study of F₁, F₂, F₃ and back cross population. To identify some DNA markers associated with septumless pod, firstly, Restriction Fragment Length Polymorphism (RFLP) between the parents were searched using 30 nonrepetitive clones picked from 89 partial genomic library as probes, secondly, Randomly Amplified Polymorphic DNA (RAPD) analysis was done by using 45 random decamer primers. RFLP analysis produced 182 discrete monomorphic bands i.e., they are unable to differentiate the two parents. In RAPD analysis, six primers produced 15 polymorphic fragments out of total 430 bands amplified by 45 primers. Among them A8-350, A10-250, A10-560 RAPD bands are expected to be linked with septumless bold pod and A3-720 with pub locus as evidenced from the bulked segregant analysis (8SA). These RAPD marker fragments of DNA were subsequently used as RFLP probes. A8-350 used as a probe revealed polymorphic bands in Eco RI digested parental DNA and also showed linkage both with septumless and glabrous loci in BSA. Approximate likelihood estimator of genetic distance between septumless locus and the marker is 1.67 cM as calculated through pooled sample mapping.     

烤烟品种资源的RAPD分析

从156个随机引物中筛选出30个引物,对来自国内外的31个烤烟品种进行了遗传多样性和亲缘关系的RAPD分析。在检测的246个位点中,127个位点为多态位点（52％）。聚类分析表明,不同烤烟品种之间存在明显差异,31个品种基本上可分为4大类,品种最多的第一大类（19个）主要由来自美国的Orinoco烤烟选育而成,反映了我国现在推广的烤烟品种遗传基因比较狭窄。研究结果表明,RAPD技术可用于烤烟品种的鉴别和纯度测定。研究为烤烟杂种育种中亲本的选配提供了一定的理论依据。     

用微卫星标记定位小麦T型CMS的恢复基因

以T型细胞质雄性不育系 75 336 9A×恢复系 72 6 9 10的F2 群体作为育性调查和基因定位群体。通过育性分析 ,确定该恢复系含有 2个主效恢复基因 ;结合群分法 ,对恢复基因进行了SSR分子标记定位 ,在 2 30对微卫星引物中 ,微卫星标记Xgwm136和Xgwm5 5 0分别与 2个主效恢复基因连锁。这两个标记与Rf基因之间的遗传距离分别为 6 7cM和 5 1cM ,从而将该恢复基因定位在 1AS、1BS染色体上。