State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Rice Research Institute, Sichuan Agricultural University, Chengdu 611130, China.
Int J Mol Sci. 2022 Mar 30;23(7):3827. doi: 10.3390/ijms23073827.
Catalases (CATs) are important self-originating enzymes and are involved in many of the biological functions of plants. Multiple forms of CATs suggest their versatile role in lesion mimic mutants (LMMs), H2O2 homeostasis and abiotic and biotic stress tolerance. In the current study, we identified a large lesion mimic mutant9428 (llm9428) from Ethyl-methane-sulfonate (EMS) mutagenized population. The llm9428 showed a typical phenotype of LMMs including decreased agronomic yield traits. The histochemical assays showed decreased cell viability and increased reactive oxygen species (ROS) in the leaves of llm9428 compared to its wild type (WT). The llm9428 showed enhanced blast disease resistance and increased relative expression of pathogenesis-related (PR) genes. Studies of the sub-cellular structure of the leaf and quantification of starch contents revealed a significant decrease in starch granule formation in llm9428. Genetic analysis revealed a single nucleotide change (C > T) that altered an amino acid (Ala > Val) in the candidate gene (Os03g0131200) encoding a CATALASE C in llm9428. CRISPR-Cas9 targetted knockout lines of LLM9428/OsCATC showed the phenotype of LMMs and reduced starch metabolism. Taken together, the current study results revealed a novel role of OsCATC in starch metabolism in addition to validating previously studied functions of CATs.
过氧化氢酶(CATs)是重要的内源性酶,参与植物的许多生物学功能。多种形式的 CATs 表明它们在病变模拟突变体(LMMs)、H2O2 动态平衡以及非生物和生物胁迫耐受中的多功能作用。在本研究中,我们从乙磺酸(EMS)诱变群体中鉴定出一个大的病变模拟突变体 9428(llm9428)。llm9428 表现出典型的 LMMs 表型,包括农艺产量性状下降。组织化学分析显示,与野生型(WT)相比,llm9428 叶片中的细胞活力降低,活性氧(ROS)增加。llm9428 表现出增强的稻瘟病抗性和增加的病程相关(PR)基因的相对表达。对叶片亚细胞结构的研究和淀粉含量的定量表明,llm9428 中淀粉颗粒形成明显减少。遗传分析显示候选基因(编码 CATALASE C 的 Os03g0131200)中的一个单核苷酸变化(C > T)改变了一个氨基酸(Ala > Val)。CRISPR-Cas9 靶向敲除 llm9428/OsCATC 的突变体显示出 LMMs 的表型和减少的淀粉代谢。总之,本研究结果揭示了 OsCATC 在淀粉代谢中的新作用,除了验证 CATs 的先前研究功能。