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水稻穗顶部退化突变体tutou3的基因精细定位
邢欣欣, 郭秀平, 李帅, 常艳琪, 寻子琦, 张欣
0
(中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程)
摘要:
水稻穗顶部退化在水稻生产中时有发生,影响水稻的穗粒数,严重时会导致水稻产量下降。目前,虽然从水稻穗顶部退化突变体中已经鉴定出了一些基因如PAA1、PAA2、TUT1等,但是对穗顶部退化发生的分子机制的认知仍十分有限。本研究报道了一个穗顶部退化的隐性突变体tutou3的基因定位及克隆结果。该突变体在营养生长时期表现为株高变矮,穗发育后期穗顶部发生严重退化,穗粒数、穗长、千粒重显著降低。遗传分析表明,tutou3受1对隐性基因控制。利用tutou3 × IRAT129杂交组合F2分离群体中的856个穗顶部退化极端个体,将基因TUTOU3定位在了4号染色体长臂端的29kb区间内,测序研究发现,区间内一个编码质膜H+-ATP酶基因LOC_Os04g56160的第8外显子存在1个C→A单碱基替换,导致第608个氨基酸从丙氨酸突变为天冬氨酸。qRT-PCR分析发现,TUTOU3基因在水稻中呈组成型表达,在突变体中该基因表达量下降。诱发穗顶部细胞程序性死亡的基因OsVPE2和OsVPE3在突变体tutou3中表达量显著上升。对TUTOU3基因的深入研究将会揭示H+-ATP 酶在穗顶部细胞的生长发育中的作用,有助于深入阐明穗顶部退化在细胞水平上发生的机制。
关键词:  水稻(Oryza sativa L.)  顶部退化  精细定位  H+-ATP酶
DOI:10.13430/j.cnki.jpgr.20201223001
投稿时间:2020-12-23修订日期:2021-02-25
基金项目:国家自然科学基金项目(31871603);转基因生物新品种培育重大专项(2016ZX08010004)
Fine Mapping of Panicle Apical Abortion Mutant tutou3 in Rice
XING Xin-xin, GUO Xiu-ping, LI Shuai, CHANG Yan-qi, XUN Zi-qi, ZHANG Xin
(Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement)
Abstract:
Panicle apical abortion occurs frequently in rice production, which can affect grain number per panicle and when serious may lead to the reduction of grain yield. Although some rice panicle apical abortion genes such as PAA1, PAA2, and TUT1 have been identified, their underling molecular mechanism are still poorly understood. In this study, we report a gene mapping and cloning of a recessive PAA tutou3 mutant in rice. The mutant had shorter plant height during vegetative development stage and serious panicle apical abortion at the later stage of panicle development, which resulted in significantly reduced grain number per panicle, panicle length and 1000-grain weight. Genetic analysis demonstrated that the phenotype of tutou3 was determined by a single recessive gene. Using 856 individuals with serious panicle apical abortion in the F2 segregating population derived from tutou3 × IRAT129, TUTOU3 locus was mapped in a 29 kb interval flanked by markers A-2 and A-17 on chromosome 4 containing 3 putative open reading frames (ORFs). Sequence analysis revealed a single nucleotide substitution of C to A at the eighth exon of LOC_Os04g56160, a plasma membrane H+-ATPase encoding gene, in tutou3, resulting in an amino acid change from alanine to aspartic acid. qRT-PCR analysis showed that TUTOU3 gene was constitutively expressed in rice, and its expression level decreased in the mutants. The mRNA expression levels of OsVPE2 and OsVPE3, which induce programmed cell death of apical panicle, increased significantly in the degraded spikelets in tutou3. Further study of TUTOU3 gene will reveal the role of H+-ATPase in the growth and development of panicle apical cells and help to elucidate the mechanism of panicle apical abortion at the cellular level.
Key words:  rice (Oryza sativa L.)  apical abortion  fine mapping  H+-ATPase

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