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可可毛色二孢β-1,4-葡聚糖内切酶LtEg1信号肽的鉴定及其酶活性分析
引用本文:彭军波,李兴红,张玮,刘梅,邢启凯,燕继晔.可可毛色二孢β-1,4-葡聚糖内切酶LtEg1信号肽的鉴定及其酶活性分析[J].植物保护学报,2020,47(1):119-126.
作者姓名:彭军波  李兴红  张玮  刘梅  邢启凯  燕继晔
作者单位: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,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,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,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,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,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
基金项目:国家现代农业(葡萄)产业技术体系(CARS-29),北京市农林科学院院储备项目(KJCX20170412)
摘    要:为明确可可毛色二孢β-1,4-葡聚糖内切酶LtEg1是否能够分泌至胞外及其活性,采用酵母互补试验分析了内切酶LtEg1的信号肽活性,利用3,5-二硝基水杨酸法检测了内切酶LtEg1活性,通过实时荧光定量PCR测定了LtEg1基因在不同侵染阶段的表达量。内切酶LtEg1的信号肽和酵母蔗糖酶的融合表达能够引起酵母蔗糖酶的外泌,使得酵母转化子能够在以棉子糖为唯一碳源的培养基上生长。内切酶LtEg1能够以羧甲基纤维素钠为底物,发挥其酶活功能。与营养菌丝阶段相比,LtEg1基因在病原菌侵染阶段的表达水平显著升高;且在接种后48 h,LtEg1基因的表达量达到高峰,约为营养菌丝阶段的12倍,表明内切酶LtEg1作为重要的外泌蛋白酶,在可可毛色二孢侵染致病过程发挥了重要作用。

关 键 词:可可毛色二孢  β-1  4-葡聚糖内切酶  信号肽  酶活性
收稿时间:2019/2/28 0:00:00

Identification of the signal peptide and enzymatic activity analysis of the endo-β-1,4-glucanase LtEg1 of fungal pathogen Lasiodiplodia theobromae
PENG Junbo,LI Xinghong,ZHANG Wei,LIU Mei,XING Qikai and YAN Jiye.Identification of the signal peptide and enzymatic activity analysis of the endo-β-1,4-glucanase LtEg1 of fungal pathogen Lasiodiplodia theobromae[J].Acta Phytophylacica Sinica,2020,47(1):119-126.
Authors:PENG Junbo  LI Xinghong  ZHANG Wei  LIU Mei  XING Qikai and YAN Jiye
Institution:Beijing Key Laboratory of Environment-Friendly Management of Diseases and Pests of North China Fruits, Institute of Plant and Environmental Protection, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory of Environment-Friendly Management of Diseases and Pests of North China Fruits, Institute of Plant and Environmental Protection, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory of Environment-Friendly Management of Diseases and Pests of North China Fruits, Institute of Plant and Environmental Protection, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory of Environment-Friendly Management of Diseases and Pests of North China Fruits, Institute of Plant and Environmental Protection, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China,Beijing Key Laboratory of Environment-Friendly Management of Diseases and Pests of North China Fruits, Institute of Plant and Environmental Protection, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China and Beijing Key Laboratory of Environment-Friendly Management of Diseases and Pests of North China Fruits, Institute of Plant and Environmental Protection, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China
Abstract:
Keywords:Lasiodiplodia theobromae  endo-β-1  4-glucanase  signal peptide  enzymatic activity
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