摘要
为明确IDD家族IDD4基因在拟南芥Arabidopsis thaliana抵抗灰葡萄孢菌Botrytis cinerea侵染过程中的作用,通过统计病情指数检测拟南芥野生型(wild type,WT)植株、过表达植株IDD4-OE和缺失突变体idd4植株感染灰葡萄孢菌情况,利用组织染色检测叶片细胞死亡和H2O2的积累情况,采用实时荧光定量PCR(real-time quantitative-PT-PCR,q RT-PCR)技术分析灰葡萄孢菌肌动蛋白基因Bc.ACTIN在3种植株叶片中的表达情况,并施加0.1 mmol/L外源水杨酸(salicylic acid,SA)后测定IDD4-OE植株的病情指数。结果显示,不同株系对灰葡萄孢菌的抗性由高到低依次为idd4>WT>IDD4-OE,IDD4-OE植株中病原菌感染部位的寄主细胞死亡程度比idd4植株严重。染色结果表明,病原菌侵染拟南芥后4 h,接种部位已有H2O2积累。qRT-PCR反应结果显示,Bc.ACTIN在IDD4-OE中比在idd4植株中的表达水平更高,表明灰葡萄孢菌在IDD4-OE植株中的繁殖速率更快。对IDD4-OE植株外源施加SA后,其病情指数、Bc.ACTIN表达量与WT植株间均无显著差异,说明SA能将感病植株的抗性提高至WT植株的水平,表明IDD4作为负调控因子参与了拟南芥对灰葡萄孢菌的抗性调控,SA在其中发挥着重要作用。
To determine the role of IDD family gene IDD4 in Arabidopsis thaliana in resistance against fungal pathogen Botrytis cinerea infection,the disease indices of over-expressing lines IDD4-OE,knock-out mutant idd4 and wild type(WT)plants were determined after inoculation with the pathogen,leaf cell death and accumulation of H2O2 were measured with tissue staining,and the expression level of Bc.ACTIN gene which encodes actin of pathogen was analyzed by real-time quantitative RT-PCR(qRT-PCR).The disease indices and Bc.ACTIN expression in leaves of IDD4-OE were determined after applying 0.1 mmol/L of exogenous salicylic acid(SA).The results showed that IDD4-OE plant was susceptible and idd4 plant was resistant to this pathogen comparing with wild type.The cell death at pathogen infection site in IDD4-OE plants was more severe than that in idd4 plants.Staining results showed that H_(2)O_(2) was accumulated in the infection site four hours post inoculation with this pathogen.The Bc.ACTIN was expressed at higher levels in IDD4-OE than in idd4 plants,indicating that B.cinerea reproduced at a faster rate in IDD4-OE.With SA pretreatment,the disease index and Bc.ACTIN expression in IDD4-OE plants were not significantly different from those of WT plants upon B.cinerea attack,suggesting that SA could increase the resistance of susceptible plants to the level of WT plants.The results indicated that IDD4 was acting as a negative regulator of SA-mediated resistance to B.cinerea in A.thaliana.
作者
李林倩
张豪
徐晓东
于德嘉
张国斌
鲁旭蓉
陈健鑫
王云月
杨红玉
Li Linqian;Zhang Hao;Xu Xiaodong;Yu Dejia;Zhang Guobin;Lu Xurong;Chen Jianxin;Wang Yunyue;Yang Hongyu(Ministry of Education Key Laboratory for Agricultural Biodiversity and Pest Management,College of Plant Protection,Yunnan Agricultural University,Kunming 650201,Yunnan Province,China;College of Agriculture and Life Sciences,Kunming University,Kunming 650214,Yunnan Province,China;College of Life Sciences,Xishuangbanna Vocational and Technical College,Jinghong 666100,Yunnan Province,China;Key Laboratory of Forest Disaster Warning and Control in Universities of Yunnan Province,College of Biodiversity Conservation,Southwest Forestry University,Kunming 650224,Yunnan Province,China)
出处
《植物保护学报》
CAS
CSCD
北大核心
2024年第1期261-268,共8页
Journal of Plant Protection
基金
国家自然科学基金(31760078,31860208)
昆明学院校级人才引进项目(YJL18002)。
关键词
IDD4转录因子
灰葡萄孢菌
水杨酸
抗性鉴定
拟南芥
IDD4 transcription factor
Botrytis cinerea
salicylic acid
resistance identification
Arabidopsis thaliana
