CfNop12参与调控果生刺盘孢生长发育、低温胁迫响应和致病力  

CfNop12 regulates the development,cold stress response and pathogenicity of Colletotrichum fructicola

作　　者：姚权 李司政 王成玉 李河[1] YAO Quan;LI Sizheng;WANG Chengyu;LI He(Key Laboratory of National Forestry and Grassland Administration on Control of Artificial Forest Diseases and Pests in South China,Hunan Provincial Key Laboratory for Control of Forest Diseases and Pests,Key Laboratory of Forest Bio-resources and Integrated Pest Management for Higher Education in Hunan Province,Central South University of Forestry and Technology,Changsha 410004,Hunan,China)

机构地区：[1]中南林业科技大学、南方人工林病虫害防控国家林业和草原局重点实验室森林有害生物防控湖南省重点实验室森林生物资源与有害生物综合管理湖南省普通高等学校重点实验室,湖南长沙410004

出　　处：《菌物学报》2023年第11期2257-2268,共12页Mycosystema

基　　金：国家自然科学基金(32071765);湖南省研究生科研创新项目(QL20220175)。

摘　　要：果生刺盘孢是油茶炭疽病的主要致病菌。研究果生刺盘孢RNA结合蛋白基因CfNOP12的生物学功能,阐述果生刺盘孢致病的分子机制,为油茶炭疽病的防治提供理论基础。根据同源重组原理,在果生刺盘孢中敲除目标基因CfNOP12,PCR验证获得正确的突变体ΔCfnop12;进一步构建PYF11::CfNOP12回补质粒,导入突变体原生质体中,筛选成功互补菌株ΔCfnop12-C。对这些菌株进行生物学表型测定,发现突变体ΔCfnop12营养生长速率显著下降,分生孢子的产量及附着胞形成率显著降低;在含有细胞壁胁迫剂的PDA培养基上,突变体ΔCfnop12的抑制率相较于野生型和回补菌株显著升高,在低温条件下,突变体的生长速率表现出明显下降;野生型和回补菌株几丁质聚集在菌丝顶端,突变体ΔCfnop12尖端几丁质分布不正常;相比于野生型和回补菌株,突变体致病力显著降低。综上所述,潜在RNA结合蛋白基因CfNOP12参与调控了果生刺盘孢生长发育、低温胁迫响应和致病力。Colletotrichum fructicola is a major causal pathogen of oil-tea tree.The purpose of this study is to investigate the biological function of the RNA-binding proteins CfNop12 of C.fructicola and provide a theoretical basis for the prevention and control of oil-tea tree anthracnose.Based on the principle of homologous recombination,the CfNOP12 gene knockout vector fragment was constructed and transformed into the protoplast of C.fructicola to get the mutant strain ofDCfnop12.The mutant strain was verified by PCR amplification.The PCR-amplified CfNOP12 gene-containing complement of the promoter was taken to construct a complementary vector fragment which was transformed into the mutant protoplasts.The complemented strain was screened by fluorescence.The biological phenotypes of wild-type strain,theDCfnop12 mutant and the gene complementationDCfnop12-C were measured.The results showed that the vegetative growth,conidiation,and appressorium formation rate ofDCfnop12 was significantly lower than those of the wild-type strain and the complemented strain.Compared with the wild-type strain,the CfNOP12 gene deletion mutant was more sensitive to osmotic stress and cell wall-damaging agents.At the low temperature,the growth rate of the mutant was markedly slowed down.In addition,the distribution of chitin of the mutant was not aggregated at the tip of mycelium.Taken together,our investigation reveals that the RNA-binding protein CfNop12 plays a critical role in development,cold stress response and pathogenicity of C.fructicola.

关 键 词：果生刺盘孢 CfNOP12基因 低温胁迫响应 致病力 

分 类 号：S763.7[农业科学—森林保护学]