Wang Meiling, Yang Dongyong, Ma Feilong, Zhu Mulan, Shi Zhenying, Miao Xuexia
Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
University of Chinese Academy of Sciences, Beijing, China.
Rice (N Y). 2019 Feb 22;12(1):9. doi: 10.1186/s12284-019-0267-0.
In plants, basic helix-loop-helix (bHLH) proteins form the largest transcription factor (TF) family. Among them, HLH proteins are a small group of atypical members that lack the basic domain, and form dimers with bHLH proteins. Although bHLH proteins have been proved to play important roles in plant development and physiology, the function of HLH proteins is rarely studied, not to mention in plant biotic resistance. Brown planthopper (BPH) is a kind of rice-specific insect that causes devastating yield losses each year.
In this study, we identified OsHLH61 gene that encodes HLH protein. OsHLH61 gene could be highly induced by BPH infestation. Furthermore, Methyl Jasmonic acid (Me-JA) and cis-12-oxo- phytodienoic acid (OPDA) induced expression of OsHLH61, while SA repressed it. We knocked down expression of OsHLH61 by RNA interference (RNAi), the transgenic plants were susceptible to BPH infestation. RNA-seq analysis revealed that some pathogen-related (PR) genes in the Salicylic acid (SA) signaling pathway that mediate plant immunity were obviously down-regulated in the OsHLH61 RNAi plants. Meanwhile, yeast two-hybrid assay and bimolecular luciferase complementation (BiLC) analysis identified bHLH096 to be an interacting factor of OsHLH61. Also, some PR genes were down-regulated in the OsbHLH96 over expressing lines. Expression of OsbHLH96 was inhibited. Besides, OsbHLH96 might interact with Jasmonate Zim-Domain3 (OsJAZ3).
Altogether, we identified an OsHLH61-OsbHLH96 complex that might mediate defense to BPH through regulating PR genes. And OsHLH61-OsbHLH96 might be important in mediating SA and JA signaling crosstalk.
在植物中,碱性螺旋-环-螺旋(bHLH)蛋白构成了最大的转录因子(TF)家族。其中,HLH蛋白是一小群缺乏碱性结构域的非典型成员,它们与bHLH蛋白形成二聚体。尽管bHLH蛋白已被证明在植物发育和生理过程中发挥重要作用,但HLH蛋白的功能却鲜有研究,更不用说在植物生物抗性方面了。褐飞虱(BPH)是一种专食水稻的昆虫,每年都会造成毁灭性的产量损失。
在本研究中,我们鉴定了编码HLH蛋白的OsHLH61基因。OsHLH61基因可被褐飞虱侵害高度诱导。此外,茉莉酸甲酯(Me-JA)和顺式-12-氧-植物二烯酸(OPDA)可诱导OsHLH61的表达,而水杨酸(SA)则抑制其表达。我们通过RNA干扰(RNAi)敲低了OsHLH61的表达,转基因植物对褐飞虱侵害敏感。RNA测序分析表明,水杨酸(SA)信号通路中一些介导植物免疫的病程相关(PR)基因在OsHLH61 RNAi植物中明显下调。同时,酵母双杂交试验和双分子荧光素酶互补(BiLC)分析确定bHLH096是OsHLH61的一个相互作用因子。此外,一些PR基因在过表达OsbHLH96的株系中下调。OsbHLH96的表达受到抑制。此外,OsbHLH96可能与茉莉酸锌指结构域3(OsJAZ3)相互作用。
总之,我们鉴定了一个OsHLH61-OsbHLH96复合物,该复合物可能通过调节PR基因介导对褐飞虱的防御。并且OsHLH61-OsbHLH96在介导SA和JA信号转导的相互作用中可能很重要。