基因组原位杂交中封阻DNA制备方法研究   总被引：3，自引：0，他引：3  

顾爱侠  李雪姣  冯大领  王彦华  陈雪平  申书兴 《河北农业大学学报》2010,33(5)

在基因组原位杂交中,适当的封阻可以大大提高基因组原位杂交的效率。本研究采用煮沸法、超声波剪切法对大白菜基因组DNA进行剪切,研究了大白菜封阻DNA的制备方法。结果表明:珠沸法效率高,操作简单,当煮沸70 min DNA片段大小主要集中在200～500 bp,适于在基因组原位杂交中作为封阻。研究结果为基因组原位杂交的应用奠定了基础。  相似文献   

茄子抗病导入系的抗青枯病性及GISH鉴定     

曹必好  雷建军  陈国菊  陈清华  肖熙鸥 《吉林蔬菜》2010,(4):25-27

利用栽培茄与野生茄(Solanumtorvum)的种间杂种为供体,以两个栽培茄的自交系为轮回亲本进行多代回交,得到2个导入系"BILR-1","BILR-2"。经过抗青枯病及GISH鉴定,表明所得两个导入系对青枯病有很强的抗性,且它们含有野生茄(Solanumtorvum)的遗传物质,成功把野生茄(Solanumtorvum)的抗青枯病性状转到栽培茄子中。所得抗病导入系的染色体数量为24条,它们可以作为一种新的抗源未来在茄子抗病育种中加以应用。  相似文献   

小麦 黑麦大穗型衍生后代的分子细胞学鉴定     

杜万里  武军  赵继新  刘淑会  杨群慧  庞玉辉  陈林刚  陈新宏 《麦类作物学报》2009,29(4):565-570

为创制大穗型小麦种质材料,利用具有大穗多小穗性状的小麦-黑麦双二倍体材料"兰小黑"和普通小麦杂交得到一批大穗型衍生后代.综合采用基因组原位杂交(GISH)、SCAR标记、微卫星(SSR)和醇溶蛋白(A-PAGE)技术对这些大穗型后代中的8个单株进行分子细胞学鉴定.结果表明,GISH检测后代含2个外源信号;1RS特异SCAR标记检测后代均含有黑麦1.5 kb的1RS特征条带;醇溶蛋白检测后代都出现了黑麦碱基因Sec-1特征条带.筛选小麦21条染色体长短臂上各6对引物,结果发现只有1BS上的3对引物未扩增出1BS的条带,其余引物均扩增出了各自的相应条带.由此确定这8株小麦-黑麦大穗型衍生后代为1BL/1RS易位材料.  相似文献   

甘薯近缘野生种Ipomoea trifida(4x) GISH分析     

向素琼  汪卫星  李晓林  陈瑶  郭启高  何桥  梁国鲁 《作物学报》2008,34(2):341-343

以甘薯近缘野生种I. trifida (2x)为探针, 与I. trifida (4x) 2个株系“695104”和“697288”的体细胞染色体进行基因组荧光原位杂交, 结果显示, 2株系都与I. trifida (2x)有很近的亲缘关系, 但2株系的信号存在差异。“695104”几乎所有染色体整条都有均匀明亮的信号, 应为I. trifida (2x)基因组直接加倍而来;而 “697288”与“695104”不同, 虽然各条染色体也均有杂交信号, 但信号的区域与亮度有差异, 较为复杂, 可分为三种情况。第1种是整条染色体有均匀明亮的信号, 亮度与分布区域同“695104” , 有41条;第2种是几乎整条染色体有信号, 但亮度较第一种暗, 有14条;第3种为染色体部分区域有信号, 亮度较前二者更暗, 有5条。推测 “697288”是在加倍同时或之后又发生了基因组重组与部分变异。  相似文献   

Creation of new maize germplasm using alien introgression from <Emphasis Type="Italic">Zea mays</Emphasis> ssp. <Emphasis Type="Italic">mexicana</Emphasis>     

Lingzhi Wang  Aifang Yang  Chunmei He  Mingli Qu  Juren Zhang 《Euphytica》2008,164(3):789-801

Zea mays ssp. mexicana, an annual wild relative of maize, has many desirable characteristics for maize improvement. To transfer alien genetic germplasm into maize background, F₁ hybrids were generated by using Z. mays ssp. mexicana as the female parent and cultivated maize inbred line Ye515 as the male parent. Alien introgression lines, with a large range of genetic diversity, were produced by backcross and successive self-pollinations. A number of alien introgression lines with the predominant traits of cultivated maize were selected. Genomic in situ hybridization (GISH) proved that small chromosome segments of Z. mays ssp. mexicana had been integrated into the maize genome. Some outstanding alien introgression lines were evaluated in performance trials which showed 54.6% hybrids had grain yield greater than that of hybrid check Yedan12 which possessed 50% Ye515 parentage, and 17.1, 9.9% hybrids had grain yield competitive or greater than those of Nongda108 and Zheng958, which were elite commercial hybrids in China, respectively. The results indicated that some of the introgression lines had excellent agronomic traits and combining ability for maize cultivar, and demonstrated that Z. mays ssp. mexicana was a valuable source for maize breeding, and could be used to broaden and enrich the maize germplasm.  相似文献   

小麦-滨麦2D/2Ns异代换系DM2411的分子细胞遗传学鉴定     

刘洋  赵继新  谈宏斌  王秀娟  李家创  梁邦平  刁慧珊  白宇皓  武军  陈新宏 《麦类作物学报》2017,(5):571-577

为进一步挖掘利用滨麦优异基因,并丰富小麦遗传种质资源,利用形态学、细胞遗传学、基因组原位杂交(Genomic in situ hybridization,GISH)、EST-STS分子标记、SSR分子标记等技术,对从八倍体小滨麦M842-16和硬粒小麦D4286杂交F_7代材料中筛选出的1个遗传稳定的小滨麦异代换系DM2411进行了鉴定。细胞遗传学观察表明,DM2411的染色体主要构型为2n=42=21Ⅱ,遗传稳定。根尖体细胞和花粉母细胞的原位杂交研究表明,DM2411含有1对滨麦Ns基因组。SSR分析表明,DM2411可能缺失了小麦2D染色体。EST分析表明,DM2411可能含有滨麦2Ns染色体。形态学调查表明,DM2411的株高极显著降低。  相似文献   

Detection of wheat-barley translocations by genomic in situ hybridization in derivatives of hybrids multiplied in vitro     

Márta Molnár-Láng  Gabriella Linc  Bernd R. Friebe  József Sutka 《Euphytica》2000,112(2):117-123

Wheat-barley translocations were identified by genomicin situ hybridization (GISH) in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) × barley (Hordeum vulgare L. cv. Betzes) hybrids. The regenerated hybrids were pollinated with the wheat line Martonvásári 9 kr1. Five translocated wheat-barley chromosomes were recovered among 51 BC₂F₂ progeny from the in vitro regenerated wheat × barley hybrids. All were single breakpoint translocations with the relative positions of the breakpoints ranging from the centromere to about 0.8 of the relative arm length. Of the four translocations with intercalary breakpoints, three were transfers of terminal barley segments to wheat chromosomes; one was a transfer of a terminal wheat segment to a barley chromosome. Because of the absence of diagnostic N-bands, the identity of three barley segments could not be determined; in one translocation the barley chromosome involved had a NOR so it must have been 5H or 6H, and the centric translocation was 4HS.2BL. Following selfing, homozygotes of four translocations were selected. The experiment suggests that in vitro culture conditions are conducive for major genome rearrangements in wheat-barley hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

Characterization of wheat-Thinopyrum partial amphiploids for resistance to barley yellow dwarf virus     

G. Fedak  Q. Chen  R.L. Conner  A. Laroche  A. Comeau  C.A. St.-Pierre 《Euphytica》2001,120(3):373-378

Resistance to viruses such as wheat streak mosaic virus (WSMV) and barley yellow dwarf virus(BYDV) is lacking in the primary gene pool of wheat, and therefore resistance is being introgressed from wild relatives such as Thinopyrum species. Resistance to BYDV was found in partial amphiploids (2n = 8x = 56, consisting of 42 wheat and14 alien chromosomes) obtained in hybrids between wheat and both Th. intermedium and Th.ponticum. GISH analysis revealed that the alien genomes of all but one resistant partial amphiploid were heterogeneous consisting of different ratios of St, J^s and J genome chromosomes obtained from theThinopyrum parent. Translocated chromosomes consisting of Robertsonian, interstitial and terminal translocations between the different genomes were also detected. The tissue blot immunoassay showed that partial amphiploids having resistance could be inoculated with the virus but both virus multiplication and spread were completely blocked. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

Identification for H. villosa chromatin in wheat lines using genomic in situ hybridization, C-banding and gliadin electrophoresis techniques     

M.Q. Yu  J. Chen  G.B. Deng  M. Cerbah  X.R. Ma  O. Panaud  S. Yakovlev 《Euphytica》2001,121(2):157-162

Detection of H. villosa chromosomes in telosomic addition and translocation lines of common wheat was undertaken using genomic in situ hybridization (GISH), C-banding techniques and polyacrylamide gels electrophoresis. The result of GISH on mitotic metaphase cells of the addition line `95039' indicated that the added telochromosomes originated from H. villosa, and it was probably 6VS or 7Vs of H. villosa according to the C-banding pattern. Furthermore, the analysis of gliadin profiles demonstrated that the telochromosome was 6VS. A pair of 1RS/1BL translocation chromosome was also found in `95039'. In addition, mitotic GISH analysis showed that the 6VS/6AL translocation chromosome remained unchanged after being transferred into new wheat background. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

小麦-大麦大穗大粒渐渗系WB13的分子细胞学鉴定     

昝凯  孙树贵  刘淑会  张军  鲁敏  王秀娟  武军  赵继新  陈新宏 《农业生物技术学报》2015,23(1):61-70

小麦新种质WB13是普通小麦(Triticum aestivum L.)品种7182与农家二棱大麦(Hordeum vulgare ssp.distichon Hsü.)杂交后回交多代衍生而来的大穗大粒材料。为了明确WB13的遗传基础,本研究利用形态学、细胞学、分子标记及特异片段回收测序等技术对其进行了鉴定。结果表明,采用小麦不同同源群简单重复序列(simple sequence repeat,SSR)标记对WB13和小麦7182进行遗传背景分析发现,两者遗传相似系数(genetic similarity coefficient,GS)达97.0%,田间表现为大穗、大粒,综合农艺性状较好;根尖细胞染色体数目为2n=42,以大麦基因组DNA为探针进行基因组原位杂交(genomic in situ hybridization,GISH)未出现杂交信号;利用大麦特异序列标签位点(sequence-tagged site,STS)标记对WB13和农家二棱大麦进行扩增,发现ABG054(4H)和ABC305B(7H)两个标记在WB13中扩增出了大麦特征条带(分别记为WS1和WS3),经测序及序列比对,发现其与扩增到的大麦序列分别具有100%和98%的相似性,与EMBL数据库中的序列比对,与两者有相似性的全为大麦的序列。利用大麦4H和7H染色体上的35对SSR引物对WB13及其亲本进行扩增,发现与千粒重相关的标记MGB396(4H)在WB13中扩增出了大麦的特异条带。综合以上结果确定WB13含有大麦4H和7H染色体的遗传物质,为小麦-大麦渐渗系材料,且4H染色体渗入片段中可能携带与千粒重相关的有益基因。本研究确定了WB13为小麦-大麦杂种后代,此材料的育成丰富了小麦-大麦中间材料和大穗大粒材料的种质资源。同时本研究在分子水平上初步揭示了WB13大穗大粒特性的成因,为后续构建遗传分析群体进行相关数量性状(quantitative trait loci,QTL)定位及推动WB13的研究利用积累了基础资料。  相似文献