橄榄果实潜伏病原真菌的分离与鉴定  

Isolation and Identification of Latent Fungal Pathogens from Chinese Olive Fruit

作　　者：陈蓬莲 陈南泉[1] 林河通[1,2,3] 林育钊 陈艺晖[1,2,3] CHEN Penglian;CHEN Nanquan;LIN Hetong;LIN Yuzhao;CHEN Yihui(College of Food Science,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Key Laboratory of Postharvest Biology of Subtropical Special Agricultural Products(Fujian Agriculture and Forestry University),Fujian Province University,Fuzhou 350002,China;Institute of Postharvest Technology of Agricultural Products,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学食品科学学院,福建福州350002 [2]亚热带特色农产品采后生物学(福建农林大学)福建省高校重点实验室,福建福州350002 [3]福建农林大学农产品产后技术研究所,福建福州350002

出　　处：《食品科学》2020年第18期165-171,共7页Food Science

基　　金：福建省科技计划项目(2015N0002);福建省财政厅科技项目(KLe16H01A;KLe16002A);福建农林大学科技创新专项基金项目(CXZX2017284;CXZX2018054;CXZX2016086;KF2015051)。

摘　　要：为明确橄榄果实潜伏性病原真菌的种类,采用组织分离法对‘长营’橄榄花器、果实上的潜伏性病原真菌进行分离,并用柯赫氏法则进行致病性测定;利用十六烷基三甲基溴化铵法提取真菌基因组DNA,运用聚合酶链式反应技术对真菌的核糖体基因转录间隔区基因序列进行扩增并对扩增产物进行核苷酸序列测定,再用基本局部比对检索工具对测序结果进行同源性分析;采用MEGA6.0软件的邻接法构建系统发育树。根据形态鉴定和系统发育学鉴定分离得到的6种潜伏性病原真菌为小新壳梭孢(Neofusicoccum parvum)、小孢拟盘多毛孢(Pestalotiopsis microspora)、出芽短梗霉(Aureobasidium pullulans)、拟茎点霉属(Phomopsis)、革菌属(Phanerochaete)和球座菌属(Guignardia)。其中N.parvum和P.microspora为橄榄果实最主要的潜伏性病原真菌,在橄榄盛花期即可侵染。本研究通过对主要潜伏性病原真菌的分离鉴定,可为指导橄榄果实采后病害的综合防治提供科学依据。In order to clarify the species of latent pathogenic fungi in Chinese olive fruit,the pathogenic fungi from the flower organ and fruit of‘Changying’Chinese olive(Canarium album(Lour.)Raeusch cv.Changying)were isolated by using tissue isolation,and their pathogenicity was assayed using Koch’s postulates.The genomic(DNA) of the isolated pathogens was extracted via the cetyl trimethyl ammonium bromide(CTAB)method Polymerase chain reaction(PCR)was employed to amplify of the ribosomal DNA internal transcribed spacer(rDNA-ITS)region,and then the amplified product was sequenced and subjected to homology analysis by basic local alignment search tool(BLAST).A phylogenetic tree for each of the six isolates was established using the neighbor-joining(NJ)method with MEGA6.0 software.According to their morphological and phylogenetic characteristics,the six latent pathogenic fungi were identified as Neofusicoccum parvum,Pestalotiopsis microspora,Aureobasidium pullulans,Phomopsis,Phanerochaete,and Guignardia with N.parvum and P.microspora being the dominant ones,which could infect the plant at the blooming stage.The results of this work can provide a scientific basis for controlling postharvest disease of Chinese olive fruit.

关 键 词：橄榄 果实 采后病害 潜伏侵染 小新壳梭孢 小孢拟盘多毛孢 

分 类 号：TS255.3[轻工技术与工程—农产品加工及贮藏工程] S667.5[轻工技术与工程—食品科学与工程]