液泡分选蛋白CfVps17参与调控果生刺盘孢的生长发育和致病力  

The putative vacuolar sorting protein CfVps17 is involved in regulating development and pathogenicity of Colletotrichum fructicola

作　　者：李玲玲[1] 张盛培 李河[1] LI Ling-Ling;ZHANG Sheng-Pei;LI He(Key Laboratory for Non-Wood Forest Cultivation and Conservation of Ministry of Education,Central South University of Forestry and Technology,Changsha,Hunan 410004,China)

机构地区：[1]经济林培育与保护省部共建教育部重点实验室,中南林业科技大学,湖南长沙410004

出　　处：《菌物学报》2021年第8期2102-2113,共12页Mycosystema

基　　金：国家自然科学基金(32071765)。

摘　　要：果生刺盘孢是油茶炭疽病的主要流行致病菌。本文研究果生刺盘孢中液泡分选蛋白CfVps17的生物学功能,为阐明该蛋白调控致病分子机制提供依据。采用over-lap方法构建CfVPS17基因敲除载体片段,使用PEG介导法将敲除载体片段转化至果生刺盘孢原生质体中,通过验证筛选获得突变体菌株ΔCfvps17。构建目的基因CfVPS17回补载体pYF11::CfVPS17,经转化获得ΔCfvps17-C回补菌株。测定野生型、突变体ΔCfvps17及回补菌株ΔCfvps17-C的生物学表型,结果表明,与野生型和回补菌株相比,突变体ΔCfvps17菌丝生长速率、产孢量和附着胞形成率显著降低,突变体菌丝中糖原积累减少,对氯化钠和刚果红胁迫耐受性增强;致病力测定结果表明,突变体ΔCfvps17丧失了对油茶叶片的致病力。液泡分选蛋白CfVps17参与调控果生刺盘孢糖原积累、生长发育、外界胁迫应答和致病力。Colletotrichum fructicola is a major pathogen causing anthracnose on Camellia oleifera.This experiment aims at revealing the biological functions of the putative vacuolar sorting protein CfVps17,providing a basis for illuminating the pathogenic molecular mechanism of C.fructicola regulated by this protein.The over-lap method was used to construct the CfVPS17 gene-deleted fragment which was transformed into the protoplasts of wild-type strain CFLH16 by PEG-mediated method for obtaining mutant strainΔCfvps17 through validation screening.A complement vector pYF11::CfVPS17 of the CfVPS17 gene was constructed and transformed into the mutant strain’s protoplast by PEG-mediated method.The complementary strainΔCfvps17-C was obtained by fluorescence screening.The biological phenotypes of the wild-type strain CFLH16,the mutant strainΔCfvps17,and the complementary strainΔCfvps17-C including vegetative growth,appressorium formation,stress response,accumulation of glycogen,and pathogenicity were further tested.The result indicated that the mycelial growth rate,conidiation and appressorium formation rate of theΔCfvps17 were significantly reduced,glycogen storage in mycelium declined,and tolerance to Congo red and NaCl increased,in comparison with CFLH16 andΔCfvps17-C.Pathogenicity test showed that theΔCfvps17 lost the ability to infect Ca.oleifera leaves.Facts prove that the vacuoler sorting protein CfVps17 is involved in regulating the growth,conidiation,appressorium formation,response to stress,accumulation of glycogen and pathogenicity of C.fructicola.

关 键 词：油茶 果生刺盘孢 CfVps17 致病力 

分 类 号：S763.7[农业科学—森林保护学]