山西省褐腐病菌种群结构及致病性研究

发布时间：2019-05-13 12:38

【摘要】：褐腐病是水果上的重要病害之一,能引起多种水果果实腐烂,影响水果生产,带来经济损失。该病害主要由链核盘菌属(Monilinia)中关系较近的三个种-美澳型核果链核盘菌Monilinia fructicola、果生链核盘菌Monilinia fructigena和核果链核盘菌Monilinia laxa引起,本研究对山西省褐腐病病原菌种群结构及致病性进行了研究。从2011年至2014年,采集褐腐病病果。采样过程中发现褐腐病在山西省主要发生在管理粗放的果园,在管理精细的果园鲜有发生,在套袋果上极少发生。通过分离共获得93株褐腐病菌菌株的纯培养物,对其进行了形态学和分子鉴定,并对其中的81株菌的致病性进行了测定。1.形态学鉴定结果表明：果生链核盘菌M. fructigena有69株,核果链核盘菌M. laxa有15株,美澳型核果链核盘菌M. fructicola有3株,其中6株不能通过形态进行准确判断。2.分别用三对褐腐病菌的特异引物ITS 1 Mfcl/ITS4Mfcl、ITS1Mfgn/ITS4Mfgn和ITS1Mlx/ITS4Mlx对供试褐腐病菌菌株进行PCR扩增,93个供试菌株中,有79株扩增出了特异性单一条带,7株扩增出两条条带,7株不能有效扩增。利用真菌ITS通用引物对7株未能进行种特异性扩增的菌株进行扩增和测序,测序结果在GeneBank核酸数据库中进行BLAST比对和同源性分析。形态学鉴定和分子生物学鉴定结果表明,供试菌株中,M. fructigena有75株,M. laxa有15株,M. fructicola有3株,分别占供试菌株数的80.7%、16.1%、3.2%。3.81株褐腐病菌在接种苹果和梨果实后,均能引起发病,但病斑大小有显著差异。对病斑直径进行方差分析,将褐腐病菌菌株划分为弱、中、强三种致病性类型。接种苹果时弱、中、强三种类型分别占菌株总数的29.6%、46.9%、23.5%,接种梨时三种类型分别占菌株总数的18.5%、50.6%、30.9%。总体来说,不同地区来源的菌株间致病力存在分化,同一致病力类群在各地区所占比例不同。
[Abstract]:Brown rot is one of the important diseases on fruit, which can cause many kinds of fruit decay, affect fruit production and bring economic loss. The disease was mainly caused by three species of Sclerotinia (Monilinia), Monilinia fructicola, Monilinia fructigena and Monilinia laxa, which are closely related to Sclerotinia sclerotinia Sclerotinia and Sclerotinia pyrenoides Monilinia laxa. In this study, the population structure and pathogenicity of brown rot pathogens in Shanxi Province were studied. From 2011 to 2014, the fruit of brown rot was collected. In the process of sampling, it was found that brown rot mainly occurred in orchards with extensive management, rarely occurred in orchards with fine management, and rarely occurred on bagged fruits. A total of 93 strains of brown rot bacteria were isolated and purified, and their morphology and molecular identification were carried out, and the pathogenicity of 81 of them was determined. The results of morphological identification showed that there were 69 strains of M. fructigena, 15 strains of M. laxa and 3 strains of M. fructicola, 6 of which could not be accurately judged by morphology. Three pairs of specific primers ITS1 Mfcl/ITS4Mfcl,ITS1Mfgn/ITS4Mfgn and ITS1Mlx/ITS4Mlx were used to amplify the tested strains of brown rot bacteria by PCR. Among the 93 tested strains, 79 strains amplified specific single bands and 7 strains amplified two bands. 7 strains could not be amplified effectively. Seven strains which could not be specifically amplified by species were amplified and sequenced by using universal primers of fungal ITS. BLAST comparison and homology analysis were carried out in GeneBank nucleic acid database. The results of morphological identification and molecular biological identification showed that 75 strains of, M. fructigena, 15 strains of, M. fructicola and 3 strains of, M. fructicola accounted for 80.7% and 16.1% of the tested strains, respectively. 3.2%. 3.81 strains of brown rot bacteria could cause the disease after inoculating apple and pear fruit, but the size of disease spot was significantly different. According to the variance analysis of the diameter of the disease spot, the strains of brown rot bacteria were divided into three pathogenic types: weak, medium and strong. When inoculated with apple, the three types accounted for 29.6%, 46.9% and 23.5% respectively, and the three types accounted for 18.5%, 50.6% and 30.9%, respectively. Generally speaking, there was differentiation of pathogenicity among strains from different regions, and the proportion of pathogenicity groups was different from that of consistent pathogenicity groups in different regions.
【学位授予单位】：山西农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S432.4

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