进境美国苹果牛眼果腐病病原真菌(Neofabraea perennans)的鉴定  

Identification of the causal fungal agent of bull's-eye rot on apple(Neofabraea perennans)imported from US

作　　者：王卫芳[1,2] 胡佳[3] 温庆秋[3] 何瑞芳[1,2] 于璇[1,2] 胡学难[1,2] 何日荣[1,2] 

机构地区：[1]广东出入境检验检疫局检验检疫技术中心,广州510623 [2]广东省动植物与食品进出口技术措施研究重点实验室,广州510623 [3]广州出入境检验检疫局,广州510623

出　　处：《果树学报》2016年第4期457-465,共9页Journal of Fruit Science

基　　金：国家质检总局科技计划项目(2014IK004);广东省科技计划项目(2011B020308011)

摘　　要：【目的】探明进境截获的美国苹果牛眼果腐病的病原真菌种类,为评估病菌的入侵风险提供科学依据。【方法】对苹果（Malus domestica）腐烂果实的症状特点、病菌的形态特征与培养特性进行观察,对分离物的致病性和DNA序列进行分析。【结果】32批次腐烂果实均表现果梗凹腐烂症状,病部较坚硬,病斑圆形褐色,中央色浅形似牛眼状。累计从病果上分离46株菌株,菌落圆形,气生菌丝稀少,白色至乳白色,加深至黄褐色至黑褐色,并形成深浅褐色相间同心轮纹,菌丝体最适生长温度15~20℃,经柯赫法则证实为病原菌。在病果及PDA上,病菌形成奶油色胶质状分生孢子盘,分生孢子梗瓶梗状,大型分生孢子及小型分生孢子圆柱形、直或下部稍弯曲、无色透明、单胞。基于β-微管蛋白基因序列的系统发育分析,8个代表性的分离物与已知的苹果牛眼果腐病菌Neofabraea perennans聚在同一簇。【结论】根据症状学、形态学与致病性及系统发育分析,将屡次从进境美国苹果中截获的苹果牛眼果腐病病原真菌鉴定为Neofabraea perennans,是我国高度关注的进境植物检疫性有害生物,对我国果树的生产存在安全隐患。【Objective】Bull＇s-eye rot of apple caused by species of Neofabraea can infect a variety of fruit trees and ornamental trees in the family of Rosaceae. This is an important branch and postharvest disease of apples and pears producing cankers on branches and twigs and/or decay on postharvest fruits, which lead to more than 40% postharvest losses in severe year. So far, this disease distributes in Europe, America and Oceania while there is no record of occurrence in China. Up to September 2015, in Guangdong port, decayed fruits of apple with suspicious symptom of bull＇s-eye rot had already been intercepted from32 shipments of apples imported from Washington state, USA and 46 fungal strains were isolated from decayed fruits. Up to now, four species of the genus Neofabraea are known to be associated with bull＇s-eye rot of apple, i.e. N. malicorticis, N. perennans, N. alba and N. kienholzii. It was reported that the geographical distribution of the four bull＇s-eye rot pathogens described above often overlap, morphological an molecular difference between N. malicorticis and N. perennans are considered to be subtle. To identify the species of the causal agent associated with decayed apple fruits intercepted from US and provide scientific basis to assess the risk of the invasion of the pathogen, fungal strains were isolated from fruit samples and subjected to DNA sequence, morphological and cultural analysis.【Methods】Suspicious decayed fruits of apple（Malus domestica）were collected in Guangdong port from imported fruits, the symptoms were photographed and main features of decayed spots were observed under microscope. Isolates were obtained from decayed fruits. After spraying with 75% ethanol, the skin of decayed fruit was peeled from the marginal area between decayed and healthy tissues and small fragments（4×4 mm） were excised and plated on potatodextrose agar（PDA）, or mycelium or conidia were picked from the skin surface of decayed fruits onto PDA plate and incubated under the condition of 24

关 键 词：苹果牛眼果腐病 蔷薇科 Neofabraea perennans 形态学 分子 Β-微管蛋白 系统发育分析 

分 类 号：S661.1[农业科学—果树学]