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水稻雄性不育突变体tip2-zh的鉴定与基因定位
彭泽群, 龚 柯, 陈代波, 李延慧, 孙志豪, 陈 驰, 于 萍, 张迎信, 周正平, 廖飞飞, 程式华, 曹立勇, 占小登, 孙廉平
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(中国水稻研究所/国家水稻改良中心/浙江省超级稻研究重点实验室/水稻生物学国家重点实验室)
摘要:
本研究从籼稻中恢8015经甲基磺乙酯(Ethylmethylsulfone,EMS)诱变的突变体库中分离鉴定出了一个无花粉型雄性不育突变体tip2-zh。突变体tip2-zh与野生型中恢8015在株高、分蘖数、分裂角度、抽穗期等农艺性状上无显著差异,但其花药瘦小、细长且呈半透明状,花药中无成熟花粉细胞;花药半薄切片观察结果显示,tip2-zh减数分裂异常,花粉母细胞不能生成小孢子细胞,绒毡层空泡化、降解延迟,最终不能形成有活力的花粉粒。扫描电镜结果显示,突变体花药外壁的蜡质和角质等线状物质更加浓缩、内壁呈光滑圆润但不规则分层状态、花粉细胞最终降解破碎。遗传分析表明,该突变性状受一对隐性核基因控制,且对目的基因的图位克隆证实,该突变位点位于水稻第1号染色体长臂JX21和JX26之间27 kb区间内;测序分析发现该区间内bHLH转录因子TIP2/bHLH142(LOC_Os01g18870)的第1个外显子区存在1处单碱基替换,由野生型的C替换成T,使得其编码的氨基酸直接替换为终止密码子,转录和翻译提前终止,导致出现该不育表型。qPCR检测结果表明,tip2-zh突变体中许多雄性不育相关基因的表达量受到了该突变位点的影响,进一步证明TIP2-ZH在水稻花药发育的脂质合成、绒毡层降解以及花粉壁形成等生物学过程中具有重要作用。
关键词:  水稻  隐形核不育  基因定位  tip2-zh
DOI:10.13430/j.cnki.jpgr.20201203001
投稿时间:2020-12-03修订日期:2021-02-17
基金项目:国家自然科学基金(31801440,31961143016);浙江省自然科学基金(LY17C130003,LY21C130003,LY18C130008);国家重点研发计划(2018YFD0100806);国家转基因重大专项(2014ZX08001-002);中国农业科学院创新工程 (CAAS-ASTIP-2013-CNRRI)
Characterization and Genetic Mapping of a No-pollen Male Sterility Mutant tip2-zh in Rice (Oryza sativa L.)
PENG Ze-qun, GONG Ke, CHEN Dai-bo, LI Yan-hui, SUN Zhi-hao, CHEN Chi, YU Ping, ZHANG Ying-xin, ZHOU Zheng-ping, LIAO Fei-fei, CHENG Shi-hua, CAO Li-yong, ZHAN Xiao-deng, SUN Lian-ping
(China National Rice Research Institute/National Center for Rice Improvement/ Key Laboratory for Zhejiang Super Rice Research/ State Key Laboratory of Rice Biology)
Abstract:
A no-pollen male sterile mutant tip2-zh was isolated from the EMS-soaked?progeny?of Zhonghui 8015 in this study. No significant difference in agronomic traits such as plant height, tiller number, tiller angle and heading date was observed between the tip2-zh mutant and the wild type, whereas tip2-zh mutant showed thinner, slender and translucent anthers as well as no mature pollen grains in the anther locule. The semi-thin sections of anthers during different development stages showed that the tip2-zh exhibited abnormal meiosis vacuolated tapetum with delayed degradation, the meiocyte mother cell could not generate microspore cells and finally resulting in failure of pollen grains formation. SEM observation also found that the linear substances of the outer surface in the mutant anther such as wax and cutin were more concentrated, the inner wall of the mutant anther exhibits irregular stratified and the microspores were finally broken and degraded. Genetic analysis showed that the mutation of tip2-zh was controlled by a recessive single gene in a 27 kb interval flanked by JX21 and JX26 on chromosome 1. Sequencing analysis revealed a C-to-T substitution in the first exon of the bHLH transcription factor TIP2/bHLH142 (LOC_Os01g18870), which resulted in a stop codon and encoded for an early-maturity protein, thus causing the male sterility. The transcriptional analysis using qPCR suggested that the expression levels on a series of GMS genes were changed significantly in the tip2-zh mutant. Collectively, these results proved that the gene TIP2-ZH played an important role in many biological processes during rice anther development such as the lipid synthesis of anthers, tapetum degradation and pollen wall formation.
Key words:  rice  recessive genic male sterility  gene mapping  tip2-zh.

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