果腐病是导致采后橄榄果实腐烂的主要病害。为明确导致采后橄榄果实果腐病的病原菌种类,本文以福建省主栽橄榄品种‘长营’果实为研究对象,试验采用组织分离法和传统纯化培养法分离采后橄榄果实果腐病病原菌,并用柯赫氏法则测定其致病性;利用真菌基因组DNA提取试剂盒法提取橄榄果实果腐病病原菌DNA,运用PCR技术对其rDNA—ITS基因序列进行扩增,并测定扩增产物的核苷酸序列,再用局部序列比对基本检索工具(BLAST)方法对测序结果进行同源性分析;采用MEGA6.0软件的邻接法(Neighbor-joining,NJ)构建橄榄果实果腐病病原菌系统发育树,结合形态学、分子生物学及系统发育分析法鉴定橄榄果实果腐病病原菌种类。结果表明,导致采后橄榄果实果腐病的病原菌为小孢拟盘多毛孢(Pestalotiopsi smicrosporao本研究为进一步控制采后橄榄果实果腐病、延长橄榄果实贮藏保鲜期提供科学依据。
Fruit rot is a major disease symptom in harvested Chinese olive fruit. In order to clarify the chief pathogens causing fruit rot in harvested Chinese olives, the fi'uit of 'Changying' Chinese olive [Canarium album (Lour.) Raeusch 'Changying'], a main Chinese olive cultivar in Fujian Province, was used for isolating pathogens causing fruit rot via tissue isolation and traditional pure culture, followed by pathogenicity verification of the isolated pathogen by Koch's postulates. The DNA of the isolated pathogen was extracted by fungal genomic DNA-isolation methods, polymerase chain reaction (PCR) was used to amplify rDNA internal transcribed spacer-region sequences, and the amplified product was sequenced. The homologous nucleotide sequences of the pathogen were further analyzed using the basic local alignment search tool (http://blast.ncbi.nlm.nih.gov/Blast.cgi). The corresponding phylogenetic tree for the isolated pathogen was constructed using the neighbor-joining method combined with morphological, molecular biology, and phylogenetic analysis to identify the pathogenic bacterial species causing 'Changying' Chinese olive fruit rot. The results showed that Pestalotiopsis microspora was the pathogen causing fruit rot in harvested Chinese olives. These findings provide a scientific basis for fruit-rot control and extension of the storage life of Chinese olives.