自噬相关基因PlATG12对荔枝霜疫霉生长发育和致病性的影响  

Effect of Autophagy-related Gene PlATG12 on the Growth,Development and Pathogenicity of Peronophythora litchii

作　　者：陈泰旭 杨成东 于戈 罗曼菲[1,2] 张雪 吕林 陈庆河[1,2] CHEN Tai-Xu;YANG Cheng-Dong;YU Ge;LUO Man-Fei;ZHANG Xue;LYU Lin;CHEN Qing-He(Sanya Nanfan Research Institute/School of Tropical Agriculture and Forestry,Hainan University,Sanya 572025,China;Key Laboratory of Green Prevention and Control of Tropical Agricultural and Forestry Biological Disasters,Ministry of Education,Haikou 570228,China)

机构地区：[1]海南大学三亚南繁研究院/热带农林学院,三亚572025 [2]热带农林生物灾害绿色防控教育部重点实验室,海口570228

出　　处：《农业生物技术学报》2024年第9期2137-2149,共13页Journal of Agricultural Biotechnology

基　　金：海南省自然科学基金创新研究团队项目(321CXTD437);国家自然科学基金(32160614);海南大学科研启动基金[KYQD(ZR)-20080]。

摘　　要：细胞自噬在植物抵御病原菌侵染过程中发挥重要作用,其中自噬相关蛋白12 (autophagyrelated protein 12, ATG12)主要参与自噬体双层膜的形成和延伸,然而其在植物病原菌特别是卵菌中的作用鲜有报道。为明确自噬相关基因PlATG12 (GenBank No. OR909654)在荔枝霜疫霉(Peronophythora litchii)生长发育和致病过程中的作用,本研究通过CRISPR/Cas9基因编辑和PEG介导的原生质体转化技术,获得了荔枝霜疫霉自噬相关基因PlATG12敲除突变体,并进行原位回补。表型测定结果显示,与野生型(SHS3)相比,突变体ΔPlatg12-77和ΔPlatg12-315的菌落生长速率分别下降了6.50%和7.49%,孢子囊数量分别降低了33.86%和34.02%。同时,在低温12℃处理0.5和2 h后,突变体的游动孢子释放率也受到显著抑制。此外,突变体卵孢子产量分别仅为野生型的20.25%和22.00%,且Ⅲ型和Ⅳ型卵孢子产量显著增加。致病性测定发现PlATG12敲除突变体在荔枝(Litchi chinensis)叶片上的致病力低于野生型。PlATG12原位回补后突变体的表型恢复到野生型水平。研究结果表明,自噬相关基因PlATG12参与荔枝霜疫霉的生长发育和致病性。本研究为进一步解析细胞自噬途径对卵菌致病过程的调控提供理论依据。Autophagy plays an important role in plant resistance to pathogen infection.The autophagy-related protein 12(ATG12),is mainly involved in the formation and extension of the double-layer membrane of autophagy.However,its role in plant pathogens,especially Oomycetes,has been rarely reported.In order to clarify the role of Peronophythora litchii PlATG12(GeneBank No.OR909654)in the growth and development,and pathogenesis,the PlATG12 knockout mutants were obtained by CRISPR/Cas9 gene editing and PEG mediated protoplast transformation technology,and the in situ complementation was performed.Phenotypic assay results showed that compared with the wild-type strain SHS3(WT),the average mycelial growth rate of theΔPlatg12-77 andΔPlatg12-315 mutants was decreased by 6.50%and 7.49%,respectively,and the average sporangium number was reduced by 33.86%and 34.02%,respectively.Meanwhile,the zoospore release rate of the mutants was significantly lower than those of WT after incubating at 12℃for 0.5 and 2 h.In addition,the oospore production was only 20.25%and 22.00%of the WT,respectively,and the oospore of typeⅢand typeⅣincreased significantly.The pathogenicity of PlATG12 knockout mutants on litchi(Litchi chinensis)leaves was lower than that of WT.However,the phenotypic defects of PlATG12 knockout mutants were rescued after in situ complementation of PlATG12.These results indicated that autophagy-related gene PlATG12 plays a key role in the growth and development,and pathogenesis of P.litchii.This study provides theoretical basis for the functions of autophagy pathway in the pathogenic process of oomycetes.

关 键 词：荔枝霜疫霉 细胞自噬 荔枝霜疫霉自噬相关蛋白12(PlATG12) 生长发育 致病性 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]