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Cloning and mapping of chromosome-specific DNA markers in crops

作物染色体特异性 DNA 标记的克隆和作图

基本信息

批准号：
04454040
负责人：
YASUMURO Yoshimasa
金额：
$ 4.1万
依托单位：
Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454040/
关键词：
repeated DNA sequences Agropyron intermedium tandem repeated family Southern hybridization in situ hybridization chromosome addition line added chromosome DNA marker Ag.intermedium特異的反復配列 Xゲノム染色体マーカー Aegilops squarrosa(タハキコムギ)Secale

项目摘要

Repeated DNA sequences cloned from Agropyron intermedium (2n=42, E1E1E2E2XX) enabled to identify the Ag.intermedium chromosomes in the common wheat genome by Southern and in situ hybridization.Two repetitive DNA clones not hybridized to the total genomic DNA of Triticum aestivum cv.Chinese Spring (CS) were screened from the MboI-digested genomic DNA library of Ag.intermedium. The clone pTA100 hybridized to the 350bp tandem repeated family defined by digestion with EcoO109I.The clone pTA100 showed homology with the EcoO109I-380 bp tandem repeated family of rye (TOMITA et al.1993) which belonged to the 350 bp family or rye (BEDBROOK et al.1980, APPELS et al.1986) . the clone pTA28 hybridized to four kinds of repeated fragments, 6800 bp, 6200 bp, 3600 bp and 1850 bp defined by digestion with EcoO109I.The clone pTA100 used as a probe did not exhibit hybridization signals based on both genomic Southern and chromosomal in situ analysis for the amphiploid AgCS (2n=56, AABBDDEE) between CS (2n … More =42, AABBDD) and Ag.elongatum (2n=14, EE) . This finding indicated that the pTA100 family was absent in the E genome. However, the clone pTA100 hybridized to the terminal regions of the ten pairs of chromosomes in Ag.intermedium. These results indicated that at least three pairs of chromosomes belonged to the E1 or E2 genomes and the segments of the hybrid region in these chromosomes must have originated from the X genome.The clone pTA100 was used as a probe for Southern hybridization to seven kinds of Wheat-Ag.intermedium chromosome addition lines. The 350bp tandem patterns were observed in the addition lines A,B,C and D,while no hybridization was observed in the remaining three addition lines. Moreover, based on chromosomal in situ analysis the clone pTA100 hybridized to each terminal region of the short arm of added chromosome A,the long arm of added chromosome B and both arms of added chromosomes C and D.As the added chromosomes C and D differed in their arm rotios, it was possible to distinguish these four added chromosomes from each other by the ISH site of the clone pTA100. The three kinds of added chromosomes A,B and D showing the ISH site of the clone pTA100 and faint C-bands may have originated from the X genome of Ag.intermedium. Less

从中间偃麦草（Agropyronintermedium）基因组DNA文库中筛选出两个与中国春小麦（Triticumaestivumcv. ChineseSpring，CS）基因组DNA不杂交的重复DNA克隆，并通过Southern杂交和原位杂交鉴定了中间偃麦草（Agropyronintermedium，2n=42，E1 E1 E2 E2 XX）的染色体。克隆pTA 100与EcoO 109 Ⅰ-380 bp串联重复家族（TOMITA et al.1993）同源，属于黑麦350 bp串联重复家族（BEDBROOK et al.1980，APPELS et al.1986）。用EcoO 109 Ⅰ酶切鉴定，pTA 28与6800 bp、6200 bp、3600 bp和1850 bp四种重复片段杂交;用pTA 100作探针，对双倍体AgCS（2n=56，AABBDDEE）进行基因组Southern杂交和染色体原位杂交分析，均未显示杂交信号 ...更多信息 =42，AABBDD）和长穗银胶（2n=14，EE）。这一发现表明pTA 100家族在E基因组中不存在。然而，克隆pTA 100杂交的10对染色体的末端区域在银。中间。这些结果表明，至少有3对染色体属于E_1或E_2染色体组，这些染色体中的杂种区片段一定来自X染色体组。在附加系A、B、C和D中观察到350 bp的串联模式，而在其余三个附加系中没有观察到杂交。此外，基于染色体原位分析，克隆pTA 100与添加的染色体A的短臂、添加的染色体B的长臂以及添加的染色体C和D的两臂的每个末端区域杂交。由于添加的染色体C和D的臂旋转不同，因此可以通过克隆pTA 100的ISH位点将这四个添加的染色体彼此区分开。显示pTA 100的ISH位点和微弱C带的A、B和D三种附加染色体可能来自中间银藻的X染色体组。少