‘无籽瓯柑’CsRAD51基因的克隆及在花粉发育过程中的表达分析  

Cloning and Expression of Cs RAD51 Gene of Citrus suavissima 'Wuzi Ougan'

作　　者：吴莹莹[1] 张迟[1] 陈高峰[1] 孙靳俭 朱咪咪[1] 张敏[1] 

机构地区：[1]浙江农林大学省部共建亚热带森林培育国家重点实验室,浙江临安311300

出　　处：《浙江林业科技》2017年第2期1-9,共9页Journal of Zhejiang Forestry Science and Technology

基　　金：浙江省大学生科技创新活动计划项目(2016R412003);国家自然科学基金资助项目(31000897);浙江省团队科技特派员服务计划项目(浙科发农[2013]215-122)

摘　　要：‘无籽瓯柑’的雄性不育主要表现为花粉败育,起始于小孢子母细胞减数分裂时期。本研究克隆了‘无籽瓯柑’减数分裂相关基因Cs RAD51,并采用荧光定量PCR检测其在瓯柑及‘无籽瓯柑’中的表达差异。结果显示,Cs RAD51基因的c DNA全长1210 bp,含有一个1029 bp的ORF区;其编码的蛋白质属于疏水性蛋白,分子量45740.1D,氨基酸数388,原子总数6 555,等电点10.20。同源性比对发现,‘无籽瓯柑’Cs RAD51的氨基酸序列与甜橙、麻疯树、油棕、黄瓜、黑杨等的RAD51蛋白高度同源,分别达99%,98%,98%,98%,98%。RT-PCR结果显示‘无籽瓯柑’Cs RAD51在小孢子母细胞、四分体以及单核花粉粒时期的相对表达量显著高于瓯柑。Cs RAD51的过量表达可能影响了DSBs的同源重组修复,致使‘无籽瓯柑’小孢子母细胞时期减数分裂异常。Citrus suavissima ‘Wuzi Ougan＇ is a seedless bud variation of C. suavissima with male sterility and pollen abortion from the tetrad stage. RNA of C. suavissima and C. suavissima ‘Wuzi Ougan＇ was extracted and the full-length c DNA was cloned and bioinformatics analysis of amino acid sequences was conducted. The results showed that the sequence length of Cs RAD51 was 1210 bp, containing an ORF of 1029 bp. The deduced protein was hydrophobic one, encoded 388 amino acids. Its molecular weight was 45740.1D, with atom number of 6555 and isoelectric point of 10.20. Homologous alignment demonstrated the homology coefficient of C. suavissima ‘Wuzi Ougan＇ with C. sinensis, Jatropha curcas, Elaeis guineensis,Cucumis sativus and Populus nigra was 99%,98%,98%,98%,98% respectively. The Real-time PCR results showed that the relative expression of Cs RAD51 in C. suavissima ‘Wuzi Ougan＇ was significantly higher than C. suavissima at microsporocyte stage, tetrad stage and uninucleus microspore stage, indicating that overexpression of Cs RAD51 inhibited double-strand break-induced homologous recombination and led to DNA damage and abnormal meiosis.

关 键 词：柑橘 雄性不育 减数分裂 Cs RAD51 实时荧光定量PCR. 

分 类 号：S666.1[农业科学—果树学]