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从油菜(甘蓝型油菜)中分离的花器官特异性多聚半乳糖醛酸酶基因的分子分析与表达。

Molecular analysis and expression of a floral organ-specific polygalacturonase gene isolated from rapeseed (Brassica napus L.).

机构信息

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, China.

出版信息

Mol Biol Rep. 2010 Dec;37(8):3851-62. doi: 10.1007/s11033-010-0041-2. Epub 2010 Mar 7.

Abstract

High throughput screening of stage-specific differentially expressed genes in a Brassica napus two-line Rs1046A/B subtractive library was used to identify the BnQRT3 gene associated with cell wall metabolism. Phylogenetic analysis indicates the protein product of BnQRT3 is polygalacturonase. According to cytological comparisons of Rs1046 sterile and fertile anthers, RT-PCR studies and in situ hybridizations, BnQRT3 is expressed most strongly in floral organs and may play an essential role in pollen maturation. Analysis of the histological staining pattern of BnQRT3 promoter-GUS constructs in transgenic Arabidopsis and Brassica napus revealed that proximal part of 5'-flanking region directed expression in the vascular tissue of filaments, veins in sepal and petals, stigma, branch connective and the floral organ abscission zone during the open flower stage. In the meanwhile, Activity of BnQRT3 was detected in the anthers, which commences at the microsporocyte stage and persists as anther approaches dehiscence. Strong GUS expression also can be observed in the vascular tissue of leaves and stem by compression with forceps or excision, suggesting that the BnQRT3 promoter is responsive to wounding.

摘要

利用甘蓝型油菜二系恢复系 Rs1046A/B 消减文库进行差异表达基因的高通量筛选,鉴定与细胞壁代谢相关的 BnQRT3 基因。系统进化分析表明,BnQRT3 蛋白产物是多聚半乳糖醛酸酶。根据 Rs1046 不育和可育花药的细胞学比较、RT-PCR 研究和原位杂交,BnQRT3 在花器官中表达最强,可能在花粉成熟中发挥重要作用。对转 BnQRT3 启动子-GUS 构建体的拟南芥和油菜的组织化学染色模式进行分析表明,5'-侧翼区近端部分在开花期定向表达于花丝、萼片和花瓣中的维管束、柱头、分枝连接组织和花器官离层。同时,在花粉母细胞阶段开始并在花药接近开裂时持续存在的花药中检测到 BnQRT3 的活性。用镊子或切除对叶片和茎的维管束进行压缩,也可以观察到强烈的 GUS 表达,这表明 BnQRT3 启动子对创伤有反应。

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