[1]吴莹莹,张迟,陈高峰,等.‘无籽瓯柑’CsRAD51 基因的克隆及在花粉发育过程中的表达分析[J].浙江林业科技,2017,37(02):1-9.[doi:10.3969/j.issn.1001-3776.2017.02.001]
 WU Ying-ying,ZHANG Chi,CHEN Gao-feng,et al.Cloning and Expression of CsRAD51 Gene of Citrus suavissima ’Wuzi Ougan’[J].Journal of Zhejiang Forestry Science and Technology,2017,37(02):1-9.[doi:10.3969/j.issn.1001-3776.2017.02.001]
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‘无籽瓯柑’CsRAD51 基因的克隆及在花粉发育过程中的表达分析()
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《浙江林业科技》[ISSN:1001-3776/CN:33-1112/S]

卷:
37
期数:
2017年02期
页码:
1-9
栏目:
出版日期:
2017-06-30

文章信息/Info

Title:
Cloning and Expression of CsRAD51 Gene of Citrus suavissima ’Wuzi Ougan’
作者:
吴莹莹张迟陈高峰孙靳俭朱咪咪张敏
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
Author(s):
WU Ying-yingZHANG ChiCHEN Gao-fengSUN Jin-jianZHU Mi-miZHANG Min
State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, Lin’an 311300, China
关键词:
柑橘雄性不育减数分裂CsRAD51实时荧光定量PCR.
Keywords:
Citrus male sterility meiosis CsRAD51 Real-time PCR.
分类号:
S666.1
DOI:
10.3969/j.issn.1001-3776.2017.02.001
文献标志码:
A
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CsRAD51 , 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,CsRAD51 基因的cDNA 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’CsRAD51 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’CsRAD51 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。CsRAD51 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。
Abstract:
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 cDNA was cloned and bioinformatics analysis of aminoacid sequences was conducted. The results showed that the sequence length of CsRAD51 was 1210bp, containing an ORF of 1029bp. The deducedprotein 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 ofCsRAD51 in C. suavissima ‘Wuzi Ougan’ was significantly higher than C. suavissima at microsporocyte stage, tetrad stage and uninucleusmicrospore stage, indicating that overexpression of CsRAD51 inhibited double-strand break-induced homologous recombination and led to DNAdamage and abnormal meiosis.

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备注/Memo

备注/Memo:
收稿日期:2016-11-21 ;修回日期:2017-02-28
基金项目: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)
作者简介:吴莹莹,本科生,从事经济林栽培与利用研究;E-mail:1009069852@qq.com。通信作者:张敏,副教授,博士,从事经济林遗传育种研究;E-mail:mzhang@zafu.edu.cn。
更新日期/Last Update: 2017-04-20