桑褐斑病病原菌的分离鉴定及高通量测序分析  

Isolation,Identification and High-Throughput Sequencing Analysis of the Pathogenic Fungus of Mulberry Brown Spot Disease

作　　者：张兴楠 罗龙辉 黄裕鑫 刘吉平[1,2] Zhang Xingnan;Luo Longhui;Huang Yuxin;Liu Jiping(College of Animal Science,South China Agricultural University,Guangzhou 510642,China;Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding,Guangzhou 510642,China;Guangxi Key Labora-tory of Sericulture Ecology and Applied Intelligent Technology,School of Chemistry and Bioengineering,Hechi University,Hechi Guangxi 546300,China)

机构地区：[1]华南农业大学动物科学学院,广州510642 [2]广东省农业动物基因组学与分子育种重点实验室,广州510642 [3]广西蚕桑生态学与智能化技术应用重点实验室,河池学院化学与生物工程学院,广西河池546300

出　　处：《蚕业科学》2024年第1期9-16,共8页ACTA SERICOLOGICA SINICA

基　　金：现代农业产业技术体系建设专项(CARS-18)。

摘　　要：桑褐斑病是一种发生较为普遍的真菌性病害。从陕西安康蚕桑主产区收集桑褐斑病病样,结合科赫氏法则,采用形态学及分子生物学的方法对桑褐斑病病原菌进行分离鉴定;使用高通量测序技术,确定桑褐斑病病原菌的主要类群;通过水合氯醛透明反应观察桑褐斑病病原菌在宿主桑树叶片中的定植状态。结果表明,侵染陕西安康桑树的桑褐斑病病原菌为桑新褐斑壳丰孢Neophloeospora maculans;病原菌在PDA培养基上呈白色,生长速度缓慢,分生孢子为透明圆柱形,两端逐渐变细,有隔膜;菌丝主要寄生在桑叶的气孔与叶脉周边;病原菌rDNA序列全长5512 bp,GC含量51.16%。上述结果确定了侵染陕西安康蚕桑主产区桑褐斑病病原菌的种类,为桑褐斑病的防治提供了理论依据。Mulberry brown spot disease(MBSD)is a common fungal disease.In this study,samples of MBSD were collected from the main sericultural area in Ankang City,Shaanxi Province.Combined with Koch’s postula-tes,morphological and molecular biology methods were used to isolate and identify the pathogenic fungus,and high-throughput sequencing technology was used to determine the main groups of pathogenic fungi.Furthermore,the colonization state of pathogenic fungi in mulberry leaves was observed by chloral hydrate clearing reaction.The results showed that the pathogen of MBSD infecting mulberry trees in Ankang,Shaanxi Province was Neophloeospora maculans.The pathogenic fungi grow slowly on PDA medium and produce white conidia,which are transparent cylindrical with tapering at both ends and septum.The hyphae mainly parasitizes around the stomates and leaf veins of mulberry leaf.The rDNA sequence of the pathogen has a total length of 5512 bp and a GC ratio of 51.16%.Based on above results,the mulberry brown spot pathogens infecting the main sericultural area of Ankang,Shaanxi Province were determined.The research results provide a theoretical basis for the prevention and control of MBSD.

关 键 词：桑树真菌病 桑褐斑病 桑新褐斑壳丰孢 高通量测序 

分 类 号：S888.71[农业科学—特种经济动物饲养]