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鉴定一个短 rDNA 间隔序列,该序列高度特异于含有从 Lycopersicon hirsutum 导入的 Tm-1 基因的番茄品系。

Identification of a short rDNA spacer sequence highly specific of a tomato line containing Tm-1 gene introgressed from Lycopersicon hirsutum.

机构信息

Plant Molecular Genetics, UA 115, Bâtiment 360, University of Paris-Sud, F-91405, Orsay Cedex, France.

出版信息

Theor Appl Genet. 1990 Nov;80(5):602-8. doi: 10.1007/BF00224218.

Abstract

We studied rDNA restriction fragment length polymorphism between two tomato lines used for F1 hybrid seed production: line A, containing the Tm-1 gene responsible for tobacco mosaic virus tolerance introgressed from the wild species Lycopersicon hirsutum, and line B, a tobacco mosaic virus sensitive line. Hybridization patterns led to distinct rDNA maps with two size classes, 10.4 and 10.7 kb, in line A and a single, 8.9-kb class in line B. Size differences were located in the intergenie sequence (IGS). A highly specific 54-bp TaqI fragment was cloned from the line A IGS and used in dot blot experiments to probe total DNA from line A, line B, and their F1 hybrid. It proved capable of discriminating B from A and the hybrid. This probe could thus serve to screen inbreds in commercial seed lots where line A is used as male. This fragment showed 80-90% sequence homology with the 53-bp subrepeats previously characterized in a region of the tomato IGS close to the 25S rRNA gene. Preliminary comparison of rDNA in line A and several wild related species indicated that the L. hirsutum H2 genotype was the closest to line A. rDNA variations between line A and this wild genotype could be explained by recombination during the introgression process involving numerous backcrosses or by an important intraspecific polymorphism. Our results strongly suggest that Tm-1 and the rDNA were introgressed together into tomato from L. hirsutum through linkage drag.

摘要

我们研究了用于 F1 杂交种子生产的两个番茄品系之间的 rDNA 限制片段长度多态性:A 系,含有从野生种 Lycopersicon hirsutum 中导入的烟草花叶病毒耐受性基因 Tm-1;B 系,是烟草花叶病毒敏感系。杂交模式导致 A 系存在两种大小类别的 rDNA 图谱,分别为 10.4 和 10.7kb,而 B 系则只有一种大小为 8.9kb 的类。大小差异位于基因间序列(IGS)中。我们从 A 系的 IGS 中克隆了一个高度特异的 54-bp TaqI 片段,并在斑点杂交实验中使用该片段来探测 A 系、B 系及其 F1 杂种的总 DNA。结果表明,该探针能够区分 B 系和 A 系及其杂种。因此,该探针可用于筛选含有 A 系作为雄性的商业种子中的自交系。该片段与先前在靠近 25S rRNA 基因的番茄 IGS 区域中表征的 53-bp 亚重复序列具有 80-90%的序列同源性。A 系和几种野生相关种的 rDNA 初步比较表明,L. hirsutum H2 基因型与 A 系最为接近。A 系与该野生基因型之间的 rDNA 变异可以通过涉及多次回交的导入过程中的重组来解释,或者通过重要的种内多态性来解释。我们的结果强烈表明,Tm-1 和 rDNA 是通过连锁拖拽从 L. hirsutum 中共同导入番茄的。

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