用差减抑制杂交方法鉴定太谷核不育小麦花药败育过程中的特异表达基因

Identification of Differentially Expressed Genes during Anther Abortion of Taigu Genic Sterile Wheat Using SSH

以太谷核不育小麦败育花药为对照群体 ,可育花药为目标群体 ,进行抑制差减杂交。用经过败育花药差减的可育花药cDNA群体构建了一个差减抑制杂交文库。结果显示 ,插入片段的大小主要集中在 30 0～ 5 0 0bp之间。随机选取 8个克隆进行Northern杂交分析 ,结果其中 7个克隆在可育花药和不育花药之间的表达存在差异。对其中 16个插入cDNA片段测序后与GenBank同源性比较表明 ,共获得未重复序列 13条 ,占81% ,其中与已知功能基因同源的 3条 ,与EST库中的序列同源的 7条 ,新的cDNA片段 3条。

Suppression subtractive hybridization (SSH) was made with sterile anther of Taigu genic sterile wheat as driver, and fertile anther of Taigu genic sterile wheat as tester and the suppression subtracted cDNA library has been constructed. Analysis showed that most of the inserted fragments were 300～500bp. Northern blotting showsd that 7 of 8 clones selected randomly were specially expressed in fertile anther. 16 clones were selected randomly and sequenced. The BLASTN homology search analysis revealed that 13 nonredundant clones were classified into the following groups: 3 known function genes, 7 wheat ESTs of unknown function, and 3 with no match in the databases.