水稻糖苷水解酶基因OsINV3影响花粉育性的研究  

Effects of Rice Glycoside Hydrolase Gene OsINV3 on Pollen Fertility

作　　者：夏思奇 杨汉树 邱牡丹 李磊 李懿星[2] 宋书锋[2] 李莉[2] 王建龙[1] XIA Siqi;YANG Hanshu;QIU Mudan;LI Lei;LI Yixing;SONG Shufeng;LI Li;WANG Jianlong(College of Agronomy,Hunan Agricultural University,Changsha,Hunan 410128,China;State Key Laboratory of Hybrid Rice,Hunan Hybrid Rice Research Center,Changsha,Hunan 410125,China)

机构地区：[1]湖南农业大学农学院,湖南长沙410128 [2]湖南杂交水稻研究中心杂交水稻全国重点实验室,湖南长沙410125

出　　处：《杂交水稻》2024年第1期35-43,共9页Hybrid Rice

基　　金：湖南省科技创新计划岳麓山种业创新项目(2021NK 1002);长沙市科技计划项目(kq2004053)。

摘　　要：花粉育性是水稻生殖发育中的重要性状,直接影响水稻的产量。本研究鉴定到一个包含Glyco_32核心结构域的糖苷水解酶编码基因OsINV3。通过实时定量PCR对OsINV3的时空表达模式分析发现,该基因在穗发育各阶段均有表达且在幼穗分化第6期具有较高的表达水平。通过亚细胞定位分析,发现OsINV3定位于细胞质。利用CRISPR/Cas9技术获得了粳稻9522和籼稻五山丝苗背景下的功能缺失突变体952201590-1和952201590-2、WSSM01590-1和WSSM01590-2。通过对表型分析及农艺性状统计发现,2种不同背景的突变体材料均表现为株高矮化、花粉育性下降、结实率降低。进一步分析发现,不同背景的突变材料,对育性下降的程度存在差异,在籼稻五山丝苗背景下,OsINV3的功能缺失导致花粉完全败育。通过半薄切片超显微观察发现,OsINV3的功能缺陷导致花药壁和小孢子发育异常。本研究为水稻花药发育提供新的研究思路,为水稻遗传育种提供遗传材料。Pollen fertility is an important character in rice reproduction and development,which directly affects rice yield.A glycoside hydrolase-encoding gene OsINV3 containing Glyco_32 core domain was identified.Temporal and spatial expression pattern analysis of OsINV3 by real-time quantitative PCR showed that OsINV3 gene was expressed at all stages of panicle development and had a high expression level in the 6th stage of young panicle differentiation.Subcellular localization analysis showed that OsINV3 was localized in cytoplasm.The non-functional mutants 952201590-1 and 952201590-2,WSSM01590-1 and WSSM01590-2 in the background of japonica rice 9522 and indica rice Wushansimiao were obtained by CRISPR/Cas9 technique.Through phenotypic analysis and statistical analysis of agronomic traits,it was found that the mutant materials of two different backgrounds showed dwarfing of plant height,decreased pollen fertility and decreased seed setting rate.Further analysis showed that there were differences in the degree of fertility decline among the mutant materials in different backgrounds.In the background of indica rice Wushansimiao,the functional loss of OsINV3 led to complete pollen abortion.By ultramicroscopic observation of semi-thin section,the functional defect of OsINV3 resulted in abnormal development of anther wall and microspore.This study provides a new idea for research of rice anther development and genetic materials for rice genetic breeding.

关 键 词：水稻 花粉育性 OsINV3基因 糖苷水解酶 CRISPR/Cas9 

分 类 号：S511.01[农业科学—作物学] S311