Bioinformatic analysis and functional characterization of CFEM proteins in Setosphaeria turcica

doi:10.1016/S2095-3119(20)63342-1

Journal of Integrative Agriculture ›› 2021, Vol. 20 ›› Issue (9): 2438-2449.DOI: 10.1016/S2095-3119(20)63342-1

所属专题：动物科学合辑Animal Science；植物抗病遗传合辑Plant Disease-resistance Genetics

收稿日期:2020-05-21 出版日期:2021-09-01 发布日期:2021-07-20

Bioinformatic analysis and functional characterization of CFEM proteins in Setosphaeria turcica

WANG Jian-xia^1*, LONG Feng^1*, ZHU Hang¹, ZHANG Yan¹, WU Jian-ying¹, SHEN Shen¹, DONG Jin-gao^{1, 2}, HAO Zhi-min¹

¹ State Key Laboratory of North China Crop Improvement and Regulation/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology, College of Life Sciences, Hebei Agricultural University, Baoding 071001, P.R.China
² College of Plant Protection, Hebei Agricultural University, Baoding 071001, P.R.China

Received:2020-05-21 Online:2021-09-01 Published:2021-07-20
Contact: Correspondence SHEN Shen, E-mail: shenshen0428@163.com; DONG Jin-gao, E-mail: shmdjg@hebau.edu.cn; HAO Zhi-min, E-mail: haozhimin@hebau.edu.cn
About author:WANG Jian-xia, E-mail: 746514180@qq.com; * These authors contributed equally to this study.
Supported by:
This work was supported by the grants from the Natural Science Foundation of Hebei Province, China (C2020204172 and C2018204120), the China Agriculture Research System of MOF and MARA (CARS-02-25) and the National Natural Science Foundation of China (31601598 and 31901827).

摘要/Abstract

摘要：

本研究在引起北方玉米叶枯病的半活体性营养真菌——玉米大斑病菌（Setosphaeria turcica）基因组中鉴定了13个StCFEM蛋白。CFEM结构域的序列比对和WebLogo分析表明，StCFEM氨基酸高度保守，除StCFEM1、2、3和6外，整体上含有8个典型的半胱氨酸；系统发育分析表明，根据有无跨膜结构域的存在，13个蛋白（StCFEM1-13）可分为2个分支，其中6个有信号肽且无跨膜结构域的StCFEM蛋白（StCFEM3, 4, 5, 10, 12, 13）被假定为候选效应蛋白；对蛋白三级结构进行预测，发现候选效应蛋白的CFEM结构域可以形成螺旋结构，与白色念珠菌（Candida albicans）Csa2同源；进一步利用pSuc2t7M13or酵母分泌系统验证效应蛋白的分泌功能，结果发现6个候选效应蛋白均具有分泌能力，鉴定为分泌蛋白；转录组分析表明，6个候选基因在真菌侵染过程中不同时期均表达，其中SCFEM3、4、5和12在附着胞形成时期高度表达；我们还发现StCFEM3、4和5对BAX/INF诱导的本氏烟草程序性细胞死亡（PCD）没有影响，而StCFEM12可以抑制INF诱导的PCD，但对BAX诱导的PCD没有影响。本研究发现玉米大斑病菌（S. turcica ）CFEM蛋白家族共有13个成员，鉴定StCFEM12为候选效应子，为阐明CFEM蛋白在植物原发病过程中的作用奠定了基础。

Abstract:

Common in Fungal Extracellular Membrane (CFEM) domains are uniquely found in fungal extracellular membrane proteins which are important for pathogens. This study identified 13 StCFEM proteins in the genome of Setosphaeria turcica, the hemibiotrophic fungus that causes northern corn leaf blight. Sequence alignment and WebLogo analysis of their CFEM domains indicated that the amino acids were highly conserved and that, with the exception of StCFEM1, 2, 3, and 6, they contained eight cysteines. Phylogenic analysis suggested that these 13 proteins (StCFEM1–13) could be divided into 2 clades based on the presence of the trans-membrane domain. Six StCFEM proteins with a signal peptide and without a trans-membrane domain were considered as candidate effector proteins. The CFEM domain in the candidate effector proteins could form a helical-basket structure homologous to Csa2 in Candida albicans. Transcriptome analysis suggested that the 13 genes were expressed during fungal infection and a yeast secretion assay revealed that these candidate effectors were secreted proteins. It was also found that StCFEM3, 4, and 5 couldn’t affect BAX/INF1-induced programmed cell death (PCD) in Nicotiana benthamiana and while StCFEM12 could suppress INF1-induced PCD, it showed no effect on BAX-induced PCD. This study found that there were 13 members of the S. turcica CFEM protein family and that StCFEM12 was a candidate effector. This study laid the foundation for illustrating the roles of CFEM proteins during the pathogenic processes of phytopathogens.

Key words: CFEM domain , effector , Setosphaeria turcica , signal peptide , PCD

. [J]. Journal of Integrative Agriculture, 2021, 20(9): 2438-2449.

WANG Jian-xia, LONG Feng, ZHU Hang, ZHANG Yan, WU Jian-ying, SHEN Shen, DONG Jin-gao, HAO Zhi-min. Bioinformatic analysis and functional characterization of CFEM proteins in Setosphaeria turcica[J]. Journal of Integrative Agriculture, 2021, 20(9): 2438-2449.

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Bioinformatic analysis and functional characterization of CFEM proteins in Setosphaeria turcica

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