水稻OsCatB敲除突变体的构建及耐逆性初步分析  

Construction of Rice OsCatB Knockout Mutants and Preliminary Analysis of Their Stress Tolerance

作　　者：刘珊 赵李剑[1] 刘聪 邓勇 黄剑 唐冬英[1] 刘选明[1] 林建中[1] LIU Shan;ZHAO Li-jian;LIU Cong;DENG Yong;HUANG Jian;TANG Dong-ying;LIU Xuan-ming;LIN Jian-zhong(Hunan Province Key Laboratory of Plant Functional Genomics and Developmental Regulation,College of Biology,Hunan University,Changsha 410082,Hunan,China)

机构地区：[1]湖南大学生物学院植物功能基因组学和发育调控湖南省重点实验室,中国湖南长沙410082

出　　处：《生命科学研究》2020年第4期301-309,共9页Life Science Research

基　　金：国家自然科学基金资助项目(31571635,31871595);湖南省科技重大专项(2018NK1010);湖南省自然科学基金资助项目(2017-JJ2042);长沙市科技计划项目(kq1801001,kq1701028);杂交水稻国家重点实验室(湖南杂交水稻研究中心)开放课题(2019KF02);广西水稻遗传育种重点实验室开放课题(2018-05-Z06-KF05)。

摘　　要：水稻过氧化氢酶B (catalase B, CatB)主要在非光合组织、愈伤组织、根和种子中表达,参与调控活性氧动态水平的平衡和根系的生长。目前, CatB参与非生物胁迫响应的功能在很大程度上是未知的。本研究利用基因编辑技术CRISPR/Cas9获得OsCatB基因敲除的纯合突变体catb,并分析了catb在盐、高温和氧化等胁迫处理下的生理和生化表型。研究结果发现,在盐处理下,突变体catb的生理和生化表型与野生型植株无显著差异;在热胁迫下, catb的过氧化氢酶活性和存活率与野生型植株相比显著降低,而H2O2含量显著升高;在氧化胁迫下, catb的幼苗高度低于野生型。这些结果说明,水稻CatB参与盐胁迫响应的调控不明显,而主要参与了高温和氧化胁迫响应,并正调控水稻的高温和氧化胁迫耐受性。该研究为进一步探究Os CatB参与逆境响应的分子机制和培育抗逆水稻品种提供了一定的理论指导和遗传材料。Catalase B(CatB) is mainly expressed in non-photosynthetic tissues, callus, roots and seeds, and is involved in the regulation of homeostasis of reactive oxygen species(ROS) and root growth. So far, its function involved in abiotic stress response is largely unknown. In present study, the homozygous OsCatB knockout rice mutants were generated by CRISPR/Cas9, a gene editing technology, and their physiological and bio-chemical phenotypes under salt, high temperature and oxidative stresses were analyzed, respectively. The re-sults showed that the physiological and biochemical phenotypes of catb mutants were not significantly different from those of wild type(WT) plants under salinity condition. Whereas, under heat stress condition, the catalase(CAT) activities and survival rates in catb mutants were significantly lower than those in WT plants,but their H2 O2 contents were markedly higher than that of WT plants. Meanwhile, catb mutants exhibited much shorter seedlings than WT plants under oxidative condition. These results indicate that OsCatB partici-pating in the regulation of response to salt stress is not obvious, and that the enzyme is mainly involved in the modulation of response to high temperature and oxidative stresses, playing a positive role in the regula-tion of tolerance to these two factors. This study might provide a certain theoretical guidance and genetic materials for further studying molecular mechanism of OsCatB in response to abiotic stresses and breeding stress-tolerance rice varieties.

关 键 词：过氧化氢酶B(CatB) 活性氧(ROS) CRISPR/Cas9技术 非生物胁迫 水稻 

分 类 号：Q945.78[生物学—植物学]