《植物生理学报》 2009, 45(12): 1191-1196
通信作者:张凤兰;E-mail: zhangfenglan@nercv.org;Tel: 010-51503038
摘 要:
以易出胚大白菜‘09C3’ 为材料, 克隆到7 个小孢子胚胎发生相关候选基因的cDNA核心片段, 并采用半定量RT-PCR方法研究了这些基因在游离小孢子热激处理和培养期间的表达。结果表明, 获得的 7 个基因的核心片段长度在 213 bp 和417 bp 之间, 克隆号分别为 C31、C33、99H6、99H8、EV14、EX01 和 C35。NCBI 进行 Blastn 和 Blastx 分析结果显示, 这些片段与源基因 HSP70、BcHSP、Lec 2、Fus 3、PAP14、HSP17.6 和 HSP18.2 的同源性均超过 80%。半定量 RT-PCR 结果表明, HSP70 和 Lec 2 在热激 24 h 后上调表达, 而常温培养下 24 h 后没有变化; BcHSP、HSP18.2、Fus 3 和 HSP17.6 在热 激 24 h 后和常温下 24 h 都上调表达; PAP14 在热激 24 h 后和常温下 24 h 表达下调。关键词:大白菜; 小孢子胚胎发生; 热激; 半定量RT-PCR
收稿:2009-10-26 修定:2009-11-17
资助:国家农业行业计划(nyhyzx07-007)、北京市科技计划项目(Z09090501040902)和现代农业产业技术体系建设专项资金。
Corresponding author: ZHANG Feng-Lan; E-mail: zhangfenglan@nercv.org; Tel: 010-51503038
Abstract:
The highly embryogenic inbred line ‘09C3’ was used to clone cDNA fragments of seven candidate genes involving in microspore embryogenesis of Chinese cabbage, and the expression of these genes were studied by semi-quantitative RT-PCR to monitor the effect of 24 h heat-shock and gene behavior after 24 h nonheat-shock culture. The length of seven obtained fragments ranged from 213 bp to 417 bp (the cloning IDs were C31, C33, 99H6, 99H8, EV14, EX01 and C35, respectively). By Blastn and Blastx on the NCBI, these fragments showed over 80% in amino acid identity against HSP70, BcHSP, Lec 2, Fus 3, PAP14, HSP17.6 and HSP18.2, respectively. The results from semi-quantitative RT-PCR showed that HSP70 and Lec 2 were upregulated after 24 h heat shock, while no changes were found in normal culture. BcHSP, HSP18.2, Fus 3 and HSP17.6 were up-regulated under both treatments, while PAP14 was down-regulated.Key words: Chinese cabbage; microspore embryogenesis; heat shock; semi-quantitative RT-PCR
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