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曹阳1, 邢启凯2, 李铃仙2, 徐品三1*.可可毛色二孢效应子LT_397基因的克隆及转录分析[J].植物保护,2021,47(2):62-69.
可可毛色二孢效应子LT_397基因的克隆及转录分析
Cloning and transcription analysis of LT_397 effector in Lasiodiplodia theobromae
投稿时间:2019-11-28  修订日期:2020-01-09
DOI:10.16688/j.zwbh.2019658
中文关键词:  可可毛色二孢  LT_397基因  生物信息学  逆境胁迫  过敏反应
英文关键词:Lasiodiplodia theobromae  LT_397 gene  bioinformatics  adversity stress  hypersensitive response
基金项目:北京市自然科学基金(6184041);国家自然科学基金(31801686);国家葡萄产业技术体系(CARS-30)
作者单位E-mail
曹阳1, 邢启凯2, 李铃仙2, 徐品三1* 1. 大连理工大学生物工程学院, 大连116024
2. 北京市农林科学院植物保护环境保护研究所, 北方果树病虫害绿色防控北京市重点实验室, 北京100097 
xupinsan@dlut.edu.cn 
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中文摘要:
      可可毛色二孢Lasiodiplodia theobromae是引起葡萄溃疡病的主要致病菌之一。病原菌会分泌许多效应子抑制植物的防御反应。本试验对可可毛色二孢特有的效应子LT_397基因进行克隆, 开放阅读框(open reading frame, ORF)全长1 140 bp, 含有11个半胱氨酸。生物信息学软件预测显示:效应子LT_397信号肽为位于N端的1~16个氨基酸。qRT-PCR结果表明:LT_397受可可毛色二孢诱导下, 在侵染24 h的基因相对转录水平最高; 在逆境胁迫处理方面:LT_397在营养、氧化、离子、酸碱和UV胁迫下基因的相对转录水平都上升。烟草瞬时表达表明, 效应子LT_397可抑制水稻细菌性谷枯病菌Burkholderia glumae引发的过敏反应(hypersensitive response, HR)。以上结果为进一步分析效应子对寄主的致病机制奠定了基础。
英文摘要:
      Lasiodiplodia theobromae is one of the important pathogens causing grapevine canker. The effectors produced by the pathogens usually act as inhibitors of plant defense responses and promote successful colonization of the pathogen in plants cells. In this study, the full open reading frame of LT_397 (1 140 bp) was cloned, which included 11 cysteine residues. Bioinformatics analysis predicted that a signal peptide of 1-16 amino acids was located at the N terminus. The results of qRT-PCR showed that LT_397 was induced by L.theobromae and its relative transcription level was the highest at 24 h after infection. Under stress conditions, the transcription level of LT_397 gene was increased under nutritional stress, oxidative stress, high-temperature stress and UV treatment. The transient expression of Nicotiana benthamiana indicated that the effector LT_397 could inhibit the hypersensitive response (HR) induced by Burkholderia glumae. These results provide a reference for further analysis of pathogenesis and the effector interaction with the host.
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