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杨树PtrWRKY89在转基因拟南芥中的过表达通过调控水杨酸和茉莉酸依赖信号通路中的防御相关基因导致抗病性降低。

Overexpression of Poplar PtrWRKY89 in Transgenic Arabidopsis Leads to a Reduction of Disease Resistance by Regulating Defense-Related Genes in Salicylate- and Jasmonate-Dependent Signaling.

作者信息

Jiang Yuanzhong, Guo Li, Liu Rui, Jiao Bo, Zhao Xin, Ling Zhengyi, Luo Keming

机构信息

Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing, 400715, China.

State Key Laboratory of Forest Genetics and Tree Breeding, Chinese Academy of Forestry, Beijing, 100091, PR China.

出版信息

PLoS One. 2016 Mar 28;11(3):e0149137. doi: 10.1371/journal.pone.0149137. eCollection 2016.

Abstract

The plant hormones jasmonic acid (JA) and salicylic acid (SA) play key roles in plant defenses against pathogens and several WRKY transcription factors have been shown to have a role in SA/JA crosstalk. In a previous study, overexpression of the poplar WRKY gene PtrWRKY89 enhanced resistance to pathogens in transgenic poplars. In this study, the promoter of PtrWRKY89 (ProPtrWRKY89) was isolated and used to drive GUS reporter gene. High GUS activity was observed in old leaves of transgenic Arabidopsis containing ProPtrWRKY89-GUS construct and GUS expression was extremely induced by SA solution and SA+MeJA mixture but not by MeJA treatment. Subcellular localization and transactivation assays showed that PtrWRKY89 acted as a transcription activator in the nucleus. Constitutive expression of PtrWRKY89 in Arabidopsis resulted in more susceptible to Pseudomonas syringae and Botrytis cinerea compared to wild-type plants. Quantitative real-time PCR (qRT-PCR) analysis confirmed that marker genes of SA and JA pathways were down-regulated in transgenic Arabidopsis after pathogen inoculations. Overall, our results indicated that PtrWRKY89 modulates a cross talk in resistance to P. syringe and B. cinerea by negatively regulating both SA and JA pathways in Arabidopsis.

摘要

植物激素茉莉酸(JA)和水杨酸(SA)在植物抵御病原体的过程中发挥关键作用,并且已有研究表明几种WRKY转录因子在SA/JA信号转导途径中发挥作用。在之前的一项研究中,杨树WRKY基因PtrWRKY89的过表达增强了转基因杨树对病原体的抗性。在本研究中,PtrWRKY89的启动子(ProPtrWRKY89)被分离出来并用于驱动GUS报告基因。在含有ProPtrWRKY89-GUS构建体的转基因拟南芥的老叶中观察到高GUS活性,并且GUS表达受到SA溶液和SA+MeJA混合物的强烈诱导,但不受MeJA处理的诱导。亚细胞定位和反式激活分析表明,PtrWRKY89在细胞核中作为转录激活因子发挥作用。与野生型植物相比,PtrWRKY89在拟南芥中的组成型表达导致对丁香假单胞菌和灰霉病菌更敏感。定量实时PCR(qRT-PCR)分析证实,在接种病原体后,转基因拟南芥中SA和JA途径的标记基因被下调。总体而言,我们的结果表明,PtrWRKY89通过负向调节拟南芥中的SA和JA途径,调节对丁香假单胞菌和灰霉病菌抗性的信号转导途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ed69/4809744/543d176d4136/pone.0149137.g001.jpg

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