Osa-miR166i-3p介导活性氧积累途径正调控水稻纹枯病抗性
#共同第一作者
收稿日期: 2024-01-14
修回日期: 2024-02-28
网络出版日期: 2025-03-19
基金资助
江苏省重点研发计划现代农业项目(BE2022335);扬州大学大学生创新创业项目(XCX20230650)
Osa-miR166i-3 Positively Regulates Resistance to Sheath Blight Through Mediating the Accumulation of Reactive Oxygen Species
#These authors contributed equally to this work
Received date: 2024-01-14
Revised date: 2024-02-28
Online published: 2025-03-19
【目的】纹枯病是水稻上的重要病害，严重影响稻米品质和产量。在感病的粳型常规水稻品种徐稻3号和高抗籼粳交后代群体YSBR1中，Osa-miR166i-3p响应立枯丝核菌（Rhizoctonia solani）的侵染。明确Osa-miR166i-3p在水稻抗纹枯病过程中发挥的作用及其分子机制，同时探究下游基因可能涉及的通路具有重要意义。【方法】通过构建Osa-miR166i-3p的敲除和过表达载体，使用农杆菌转化法创制徐稻3号背景下的转基因植株，通过测序及检测Osa-miR166i-3p表达水平验证转基因植株真实性。在温室环境对筛选后的植株进行纹枯病菌接种，统计病斑长度，同时对大田环境下正常生长的转基因植株及对照进行主要农艺性状考察。选取立枯丝核菌接种后0 h、8 h、16 h的水稻叶鞘组织构建文库进行RNA-seq分析，对Osa-miR166i-3p的生物学功能进行研究。【结果】与徐稻3号相比，在敲除植株中Osa-miR166i-3p表达水平明显降低，在过表达植株中则明显升高。接种立枯丝核菌后，Osa-miR166i-3p敲除植株病斑长度增加，对纹枯病的抗性下降；过表达植株病斑长度减小，对纹枯病的抗性增强。大田中转基因植株及对照的株高、穗长、每穗枝梗数、千粒重的统计结果没有显著差异，表明过表达和敲除Osa-miR166i-3p不影响水稻的农艺性状。富集分析结果显示，在接种立枯丝核菌8 h后的Osa-miR166i-3p过表达植株中，多个过氧化物酶基因被诱导表达。【结论】综上所述，Osa-miR166i-3p主要通过调节植物第三类过氧化物酶相关基因的表达，影响水稻中活性氧的积累，正调控水稻纹枯病抗性，可为提高水稻抗病性提供新思路。
关键词： 水稻; 纹枯病; Osa-miR166i-3p; 生物学功能; RNA-seq
冯涛, 张朝阳, 黄新妮, 王月, 钟旭志, 冯志明, 刘欣, 左示敏, 欧阳寿强 . Osa-miR166i-3p介导活性氧积累途径正调控水稻纹枯病抗性[J]. 中国水稻科学, 2025 , 39(2) : 187 -196 . DOI: 10.16819/j.1001-7216.2025.240107
【Objective】Sheath blight is a major disease in rice, leading to significant yield and quality losses. Osa-miR166i-3p responds to Rhizoctonia solani infection in both the susceptible japonica rice variety Xudao 3 and the resistant indica-japonica hybrid variety YSBR1. This study aimed to clarify the role and molecular mechanism of Osa-miR166i-3p in rice resistance to sheath blight and to explore the potential pathways involving downstream genes. 【Method】Vectors containing Osa-miR166i-3p were constructed, and transgenic plants were generated in the susceptible variety Xudao 3 using Agrobacterium-mediated transformation. The authenticity of the transgenic plants was confirmed by sequencing and detecting the expression level of Osa-miR166i-3p. The plants were inoculated with R. solani in a greenhouse, and lesion lengths were measured. Additionally, the main agronomic traits of the transgenic and control plants grown under field conditions were investigated. Leaf sheath tissues collected at 0 h, 8 h, and 16 h post-inoculation were used for RNA-seq analysis to study the biological function of Osa-miR166i-3p. 【Results】Compared with Xudao 3, the expression level of Osa-miR166i-3p was significantly reduced in knockout plants and significantly increased in overexpressed plants. After inoculation with R. solani, the lesion length of Osa-miR166i-3p knockout plants increased, indicating enhanced susceptibility, while the lesion length of overexpression plants decreased, indicating enhanced resistance. Statistical analysis of agronomic traits, including plant height, panicle length, number of branches per panicle, and 1000-grain weight, showed no significant differences between transgenic and control plants, suggesting that overexpression or knockout of Osa-miR166i-3p did not affect rice agronomic traits. Enrichment analysis revealed that multiple peroxidase genes were induced in Osa-miR166i-3p overexpression plants 8 hours after inoculation with R. solani. 【Conclusion】This study demonstrates that Osa-miR166i-3p positively regulates rice resistance to sheath blight by modulating the expression of class III peroxidase-related genes and influencing reactive oxygen species (ROS) accumulation. These findings provide new insights for improving rice disease resistance.
Key words： rice; sheath blight; Osa-miR166i-3p; biological function; RNA-seq
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