与西瓜野生种质抗枯萎病基因连锁的RAPD标记

运用ＲＡＰＤ技术,采用混合分组分析（ｂｕｌｋｅｄｓｅｇｒｅｇａｎｔａｎａｌｙｓｉｓ,ＢＳＡ）方法进行了西瓜（Ｃｉｔｒｕｌｌｕｓｌａｎａｔｕｓ（Ｔｈｕｎｂ．）Ｍａｎｓｆｅｌｄｖａｒ． ｃｉｔｒｏｉｄｅｓ） 野生种质ＰＩ２９６３４１ 抗枯萎病基因连锁的分子标记研究。研究结果表明：西瓜野生种质Ｐ１２９６３４１ 抗枯萎病生理小种１ 的抗性由单显性基因控制,ＲＡＰＤ标记ＯＰＰＯＬ／７００ 与其抗病基因连锁,其遗传距离为３０ ｃＭ（ｃｅｎｔｉｍｏｒｇａｎ）。这为进行抗病分子标记辅助选择,以及最终定位与克隆其抗病基因打下了良好基础。     

小麦抗白粉病基因Pm21的分子鉴定和标记辅助选择

利用小麦抗白粉病基因Ｐｍ２１的ＲＡＰＤ标记、ＳＣＡＲ标记和荧光源位杂交技术对小麦抗病育种材料中的抗白粉病Ｐｍ２１基因进行了分子鉴定和标记辅助选择。     

应用荧光原位杂交和RFLP标记检测多小穗小麦新种质10—A中的黑麦染 …

利用ＡＰＡＧＥ,荧光原位杂交技术和ＲＦＬＰ标记,对导入黑麦（ＳｅｃａｌｅｃｅｒｅａｌｅＬ．）多小穗等性状创制的小麦新种质１０－Ａ进行了分子标记检测,ＡＰＡＧＥ分析发现,１０－Ａ与其他１ＲＳ／１ＢＬ易位系一样,含有１ＲＳ的醇溶蛋白标记位点Ｇｌｄ１Ｂ３。以黑麦基因组总ＤＮＡ作探针,用中国春（Ｔｒｉｔｉｃｕｍａｅｓｔｉｕｍｃｖ．ＣｈｉｎｅｓｅＳｐｒｉｎｇ）基因组ＤＮＡ作封阻,与１０－Ａ根尖细胞有丝分裂染     

利用RAPD技术进行植物性状标记及辅助选择

近等基因系、混合分离群体法是ＲＡＰＤ 标记的主要策略。目前,ＲＡＰＤ标记广泛用于抗线虫、抗病、雄性不育等辅助选择的研究中,取得了可喜的成绩。由于遗传距离的不同,使ＲＡＰＤ 标记具有基因型的差异。寻找无重组的ＲＡＰＤ 标记或将ＲＡＰＤ标记转化为ＲＦＬＰ标记,可以解决这一问题。随着连锁程度的降低选择效率也随着降低。相斥相的ＲＡＰＤ标注可提高选择效率将ＲＡＰＤ标记转化为ＳＣＡＲｓ、ＡＰＳＰｓ 标记,可以解决ＲＡＰＤ 标记稳定性差的问题。来源于ＲＡＰＤ 标记的ＳＣＡＲｓ 标记将在辅助选择中发挥巨大的作用。     

应用荧光原位杂交和RFLP标记检测多小穗小麦新种质10-A中的黑麦染色质

利用ＡＰＡＧＥ、荧光原位杂交技术和ＲＦＬＰ标记,对导入黑麦（ＳｅｃａｌｅｃｅｒｅａｌｅＬ．）多小穗等性状创制的小麦新种质１０＿Ａ进行了分子标记检测。ＡＰＡＧＥ分析发现,１０＿Ａ与其他１ＲＳ／１ＢＬ易位系一样,含有１ＲＳ的醇溶蛋白标记位点Ｇｌｄ１Ｂ３。以黑麦基因组总ＤＮＡ作探针,用中国春（Ｔｒｉｔｉｃｕｍａｅｓｔｉｖｕｍｃｖ．ＣｈｉｎｅｓｅＳｐｒｉｎｇ）基因组ＤＮＡ作封阻,与１０＿Ａ根尖细胞有丝分裂染色体进行荧光原位杂交。结果表明,黑麦的１ＲＳ易位到１０＿Ａ中。用２５个ＲＦＬＰ探针进行Ｓｏｕｔｈｅｒｎ分析,进一步发现１０＿Ａ的１ＢＳ特异限制性片段发生丢失,代之以黑麦１ＲＳ的特异限制性片段,而位于其他染色体上的特异限制性片段未发生缺失。据此认为,多小穗小麦新种质１０＿Ａ属于１ＲＳ／１ＢＬ易位系。同时还讨论了１０＿Ａ在小麦遗传改良中的利用情况。     

应用RAPD技术对不同基因组组合生物型遗传多态性的研究(英文)

利用ＲＡＰＤ技术对不同基因组合的鱼类进行了基因组指纹图谱构建,在ＤＮＡ水平上对基因组成分进行了分析,探讨了其遗传多态性。ＲＡＰＤ结果发现,在２６个随机引物扩增的产物中,平均每个个体观察到约１４２个ＲＡＰＤ标记,单个引物获得的标记平均为５．４。其中４个引物扩增的图谱可将不同的生物型区分开：Ｓ－２６引物的扩增图谱（Ｆｉｇ．１）可将红鲫（ＲＡ）与其它组合区分开,还可将鲤鲫杂种一倍体（ＣＡ）与鲫鲤杂种三倍体（ＣＡＡ）和人工复合三倍体鲤（ＣＣＡ）区分开;Ｓ－８引物（Ｆｉｇ．２）可区分开红鲤（ＲＣ）和镜鲤（ＭＣ）;Ｓ－４５引物（Ｆｉｇ．３）可区分开ＲＣ和ＣＡ;Ｓ－２２引物则可区分开ＣＡＡ和ＣＣＡ。六种生物型均存在基因组特异性的图谱即各自独特的“诊断性”图谱,作者由此建立了详细的分子标记检索表（Ｔａｂｌｅ１）。通过对ＲＡＰＤ图谱的量化分析,利用ＵＰＧＭＡ构建了不同生物型的遗传关系树图;反映了鲤鲫及各种组合生物型之间的遗传相似关系：ＲＣ和ＭＣ属同一种系,聚为一族;ＣＡＡ和ＣＡ基因组类型相同,聚为一族;ＣＣＡ虽自成一体,但可与ＣＡＡ和ＣＡ聚为一族;而ＲＡ与其它组合遗传距离较远,自成一族。ＲＡＰＤ的结果也表明各种生物型内个体间     

黑麦1R染色体的微切微克隆研究

显微分离出黑麦（ＳｅｃａｌｅｃｅｒｅａｌｅＬ．）１Ｒ染色体,用ＣｏｈｅｓｉｖｅａｄａｐｔｅｒｓｓｉｎｇｌｅｐｒｉｍｅｒＰＣＲ（ＣＡＳＰＰＣＲ）方法进行体外扩增,以ＤＩＧ１１ｄＵＴＰ标记扩增产物为探针,进行Ｓｏｕｔｈｅｒｎ分子杂交,结果表明扩增产物来自黑麦１Ｒ染色体。用１／１０体积的连接物转化Ｅ．ｃｏｌｉＤＨ５α,获得１００００多个重组菌落。经酶切分析,克隆子的插入片段为２５０～５００ｂｐ,为进一步筛选１Ｒ染色体的分子标记打下了基础     

大豆遗传图谱的构建和分析

分子标记连锁图的构建为植物基因组的结构和功能分析提供了有力的工具。较高密度的遗传图谱在数量性状基因定位、图位克隆重要农艺性状基因等研究中发挥了巨大作用。应用栽培大豆长农４和半野生大豆新民６杂交得到的Ｆ８代重组自交系,构建了一张较高密度的遗传图谱。该图谱共有２４０个标记,其中包括２个形态标记、１００个ＲＦＬＰ标记、３３个ＳＳＲ标记、４２个ＡＦＬＰ标记、６２个ＲＡＰＤ标记和１个ＳＣＡＲ标记,分布在２２     

非同位素PCR－单链构象多态性技术的建立和应用

ＰＣＲ－单链构象多态性技术问世以来,成为研究基因突变的工具,特别是在分子肿瘤学研究中,广泛应用于癌基因,抑癌基因突变的研究,常规ＰＣＲ－ＳＳＣＰ采用同位素标记ＰＣＲ产物,测序板电泳分离突变,在操作和费用上有种种局限,文章建立了一种非同位素ＰＣＲ－ＳＳＣＰ技术;通过不对称ＰＣＲ获得单链,普通ＰＡＧＥ分离,经银染检出突变,用这种方法,还研究了四株鼻咽癌细胞株ＣＮＥ１,ＣＮＥ２,ＨＫ１和ＳＵＮＥ１中肿瘤     

DNA分子标记在柑桔中的应用

ＤＮＡ分子标记是最为理想的遗传标记 ,依其多态性检出所用的分子生物学技术 ,大致可分为Ｓｏｕｔｈｅｍ杂交技术为核心的分子标记和ＰＣＲ技术为核心的分子标记。前者的代表性技术有ＲＥＬＰ(ｒｅｓｔｒｉｃｔｉｏｎｆｒａｇｍｅｎｔｌｅｎｇｔｈｐｏｌｙｍｏｒｐｈｉｓｍ)和ＤＮＡ指纹技术 (ＤＮＡｆｉｎｇｅｒｐｒｉｎｔｉｎｇｔｅｃｈｎｉｑｕｅｓ)。后者的代表性技术有ＲＡＰＤ(ｒａｎｄｏｍａｍｐｌｉｆｉｅｄｐｏｌｙｍｏｒｐｈｉｃＤＮＡ)、ＳＣＡＲ(ｓｅｑｕｅｎｃｅｃｈａｒａｃ ｔｅｒｉｓｔｉｃａｍｐｌｉｆｉｅｄｒｅｇｉｏｎ)…     

Development of SRAP, SRAP-RGA, RAPD and SCAR markers linked with a Fusarium wilt resistance gene in eggplant

Fusarium wilt (Fusarium oxysporum Schlecht. f. sp. melongenae) is a vascular disease of eggplant (Solanum melongena L.). The objectives of this work were (1) to confirm the monogenic inheritance of fusarium wilt resistance in eggplant, (2) to identify molecular markers linked to this resistance, and (3) to develop SCAR markers from most informative markers. We report the tagging of the gene for resistance to fusarium wilt (FOM) in eggplant using SRAP, RGA, SRAP-RGA and RAPD markers. Analysis of segregation data confirmed the monogenic inheritance of resistance. DNA from F₂ and BC₁ populations of eggplant segregating for fusarium wilt resistance was screened with 2,316 primer combinations to detect polymorphism. Three markers were linked within 2.6 cM of the gene. The codominant SRAP marker Me8/Em5 and dominant SRAP-RGA marker Em12/GLPL2 were tightly linked to each other and mapped 1.2 cM from the resistance gene, whereas RAPD marker H12 mapped 2.6 cM from the gene and on the same side as the other two markers. The SRAP marker was converted into two dominant SCAR markers that were confirmed to be linked to the resistance gene in the F_2, BC₁ and F₂ of BC₃ generations of the same cross. These markers provide a starting point for mapping the eggplant FOM resistance gene in eggplant and for exploring the synteny between solanaceous crops for fusarium wilt resistance genes. The SCAR markers will be useful for identifying fusarium wilt-resistant genotypes in marker-assisted selection breeding programs using segregating progenies of the resistant eggplant progenitor used in this study.     

Identification of a RAPD Marker Linked to Fusarium Wilt Resistant Gene in Wild Watermelon Germplasm (Citrullus lanatus var. citroides

Random amplified polymorphic DNA(RAPD) was employed to detect a molecular marker linked to Fusarium wilt resistant gene in the wild watermelon ( Citrullus lanatus (Thunb.) Mansfeld var. citroides ) germplasm P1296341. The resistance to race 1 Fusarium wilt of PI296341 was controlled by one dominant gene. A RAPD marker OPPO1/700 was proved to be linked to the resistant gene. The genetic distance is 3.0 cM (centimorgan). This work has provided a solid basis for molecular marker-assisted selection (MAS) for disease resistance, and made location and cloning of disease resistant genes possible.     

单核苷酸多态性与甜瓜抗枯萎病分子育种研究

目的:结合单核苷酸多态性标记技术,利用甜瓜本身的抗病性以解决新疆甜瓜病害问题。方法:对新疆甜瓜抗枯萎病基因Fom-2基因进行克隆分析,并根据Fom-2基因在不同抗性甜瓜亲本的单核苷酸多态性,设计检测SNP标记的PCR扩增引物,验证其多态性;并利用F2代分析该标记与筛选获得的甜瓜抗枯萎病基因连锁的SSR标记的遗传关系。结果:在抗病与感病甜瓜品种中均扩增获得PCR条带,试验中设计单核苷酸多态性分子标记在抗病品种为显性,与筛选的和抗枯萎病基因紧密连锁的共显性标记SSR430共分离。结论:不同抗性甜瓜品种均含有Fom-2基因或其高度同源序列,SNP显性标记和共显性标记SSR430均可用于甜瓜抗枯萎病分子标记辅助育种。     

Identification of molecular markers for the detection of the yellow rust resistance gene Yr17 in wheat

The Yr17 gene, which is present in many European wheat cultivars, displays yellow rust resistance at the seedling stage. The gene introduced into chromosome 2A from Aegilops ventricosa was previously found to be closely linked (0.5 cM) to leaf and stem rust resistance genes Lr37 and Sr38, respectively. The objective of this study was to identify molecular markers linked to the Yr17 gene. We screened with RAPD primers, for polymorphism, the DNAs of cv. Thatcher and the leaf rust-resistant near-isogenic line (NIL) RL 6081 of cv. Thatcher carrying the Lr37 gene. Using a F2 progeny of the cross between VPM1 (resistant) and Thésée (susceptible), the RAPD marker OP-Y15580 was found to be closely linked to the Yr17 gene. We converted the OP- Y15580 RAPD marker into a sequence characterized amplified region (SCAR). This SCAR marker (SC-Y15) was linked at 0.8 ± 0.7 cM to the Yr17 resistance gene. We tested the SC-Y15 marker over a survey of 37 wheat cultivars in order to verify its consistency in different genetic backgrounds and to explain the resistance of some cultivars against yellow rust. Moreover, we showed that the Xpsr150-2Mv locus marker of Lr gene described by Bonhomme et al. [6] which possesses A. ventricosa introgression on the 2A chromosome was also closely linked to the Yr17 gene. Both the SCAR SC-Y15 and Xpsr150-2Mv markers should be used in breeding programmes in order to detect the cluster of the three genes Yr17, Lr37 and Sr38 in cross progenies. This revised version was published online in June 2006 with corrections to the Cover Date.     

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 .     

DNA markers linked to a T10 loose smut resistance gene in wheat (Triticum aestivum L.).

Screening for loose smut resistance in wheat is difficult. Selecting lines with DNA markers linked to loose smut resistance would be more reliable and less costly. Molecular markers linked to a race T10 loose smut resistance gene were identified using a F6 single seed descent segregating population. A RAPD marker and a RFLP marker were located on opposite flanks of the resistance gene and were shown to be loosely linked. The RAPD marker was converted to a user friendly polymorphic SCAR marker that represented a single genetically defined locus in hexaploid wheat. Using these two bracketing markers simultaneously, the error rate for T10 resistance selection due to crossing-over was reduced to 4%. These markers can be used for a faster and more reliable selection of T10 resistant plants than previous conventional loose smut ratings.     

Inheritance of resistance to covered smut in barley and development of a tightly linked SCAR marker

Inheritance of resistance to covered smut in the barley line Q21861 was studied using a doubled-haploid population produced by crossing Q21861 with the line SM89010. Based on 3 years of screening in the field and two seasons in the greenhouse, segregation for resistance/susceptibility fits a one-gene ratio, indicating a single major gene for resistance in Q21861. Of 440 random 10-mer primers tested using bulked segregant analysis, one primer (OPJ10) resulted in a reproducible polymorphic band. RAPD marker OPJ10₄₅₀ co-segregated in repulsion with the covered smut resistance. This marker was converted to a sequence-characterized amplified region (SCAR) marker linked in coupling (5.5 cM) with the covered smut resistant gene in Q21861. The SCAR marker was amplified in the line TR640 which is also resistant to covered smut, but not in the other resistant lines. The SCAR marker will be useful for marker-assisted selection for covered smut in barley breeding programs. Received: 9 January 2001 / Accepted: 31 May 2001     

Identification and mapping on chromosome 9 of RAPD markers linked to Sw-5 in tomato by bulked segregant analysis

Bulked segregant analysis was used to identify random amplified polymorphic DNA (RAPD) markers linked to the Sw-5 gene for resistance to tomato spotted wilt virus (TSWV) in tomato. Using two pools of phenotyped individuals from one segregating population, we identified four RAPD markers linked to the gene of interest. Two of these appeared tightly linked to Sw-5, whereas another, linked in repulsion phase, enabled the identification of heterozygous and susceptible plants. After linkage analysis of an F₂ population, the RAPD markers were shown to be linked to Sw-5 within a distance of 10.5 cM. One of the RAPD markers close to Sw-5 was used to develop a SCAR (sequence characterized amplified region) marker. Another RAPD marker was stabilized into a pseudo-SCAR marker by enhancing the specificity of its primer sequence without cloning and sequencing. RAPD markers were mapped to chromosome 9 on the RFLP tomato map developed by Tanksley et al. (1992). The analysis of 13 F₃ families and eight BC₂ populations segregating for resistance to TSWV confirmed the linkage of the RAPD markers found. These markers are presently being used in marker-assisted plant breeding.     

Development of RAPD and SCAR markers linked to the Pvr4 locus for resistance to PVY in pepper ( Capsicum annuum L.)

Potato Virus Y (PVY) is the only potyvirus infecting pepper ( Capsicum annuum L.) in Europe. Currently, the development of pepper varieties resistant to PVY seems to be the most-efficient method to control PVY damage. Among the sources of resistance, a monogenic dominant gene Pvr4 confers resistance against all known PVY pathotypes. In this work, bulked segregant analysis (BSA) was used to search for randomly amplified polymorphic DNA (RAPD) markers linked to the Pvr4 gene, using segregating progenies obtained by crossing a homozygous resistant ('Serrano Criollo de Morelos-334') with a homozygous susceptible ('Yolo Wonder') cultivar. Eight hundred decamer primers were screened to identify one RAPD marker (UBC19(1432)) linked in repulsion phase to Pvr4. This marker was converted into a dominant sequence characterised amplified region (SCAR) marker (SCUBC19(1423)). This marker was mapped into a dense Capsicum genetic map in a region where several genes for resistance to different diseases are located. This marker can be useful to identify PVY-resistant genotypes in segregating progenies of pepper in marker-assisted selection (MAS) breeding programs.