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Molecular Studies on Nuclear Gene(s) Interacting with Rye Cytoplasm

核基因与黑麦细胞质相互作用的分子研究

基本信息

批准号：
02660007
负责人：
MURATA Minoru
金额：
$ 1.09万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

In a common wheat strain with rye cytoplasm, at least one diminutive chromosome, called "midget" always exists. The midget chromosome does not pair with any of the wheat chromosomes at meiosis, and the seeds of this strain without this chromosome are inviable because of their endosperm abortion. These results indicate that the midget chromosome was derived from certain segment(s) of rye chromosome(s), having the gene(s)essential for rye cytoplasmic function. To analyze the gene(s), in this study, we attempted to isolate the midget chromosome-specific DNA sequences by using a genomic subtraction method.Three different rye-specific sequences were isolated. Dot and Southern blot analyses showed that the inserted sequence of one clone(pScm-6.39)was derived from chromosome 5R of rye, and that clone pScm-6.44 contained a common sequence among rye chromosomes 1R, 2R, 3R, 5R and 7R. The insert of pScm-2.3 was shown to be derived from chloroplast DNA of rye.The origin and molecular structure of the midget chromosome were studied by using in situ hybridization. In situ hybridization with biotinylated rye genomic DNA as a probe clearly showed that the midget chromosome had originated from certain part(s)of rye chromosome(s). The midget chromosome did not possess sequences similar to wheat rDNA nor to rye telomeric sequence with 350-bp repeat unit. However, another repetitive sequence(120-bp family)of rye was found to occur in one distal end of the midget chromosome. The telomere repeat sequences from Arabidopsis thaliana cross-hybridized to both ends of the midget chromosome as well as wheat chromosomes.

在具有黑麦细胞质的普通小麦品系中，至少存在一条称为“侏儒”的小染色体。在减数分裂时，小染色体不与任何一条小麦染色体配对，没有这条染色体的小麦种子由于胚乳流产而不能存活。这些结果表明，小染色体来源于黑麦染色体的某些片段，具有黑麦细胞质功能所必需的基因。为了分析基因，在本研究中，我们试图通过基因组减法分离出侏儒染色体特异性DNA序列。分离出三种不同的黑麦特异性序列。Dot和Southern blot分析表明，其中一个克隆（pScm-6.39）的插入序列来源于黑麦的5R染色体，pScm-6.44在黑麦的1R、2R、3R、5R和7R染色体中含有一个共同的序列。pScm-2.3插入物来源于黑麦叶绿体DNA。用原位杂交技术研究了小染色体的起源和分子结构。以生物素化黑麦基因组DNA为探针，原位杂交结果清楚地表明，小染色体起源于黑麦染色体的某些部分。小染色体不具有与小麦rDNA相似的序列，也不具有与黑麦350 bp重复单元相似的端粒序列。然而，在黑麦小染色体的一个远端发现了另一个重复序列（120bp家族）。拟南芥的端粒重复序列与小麦染色体和侏儒染色体的两端交叉杂交。