{"title":"胡萝卜果乳杆菌亚种的特性研究。波斯尼亚和黑塞哥维那最近在卷心菜上爆发的胡萝卜素分离株","authors":"T. Popović, A. Jelušić, S. Markovic, R. Iličić","doi":"10.2298/pif1904211p","DOIUrl":null,"url":null,"abstract":"The causal agent of soft rot disease associated with a cabbage outbreak in Semberija region, Bosnia and Herzegovina, in 2018 was identified and characterized. Symptoms appeared in the form of water-soaked lesions on leaves and specific odour. Disease incidence ranged from 20% to 30%. The causal pathogen was isolated on nutrient agar (NA), King?s B and crystal violet pectate (CVP) media. Eight creamy-white, round and convex bacterial isolates, which produced characteristic pits on CVP medium were taken as representative. They were gram negative, facultative anaerobe, oxidase negative, catalase positive, nonfluorescent on King?s B medium, levan and arginine dehydrolase negative. The isolates were able to cause soft rot on cabbage and potato tuber slices 24 h after inoculation under conditions of high relative humidity. Polymerase chain reaction (PCR) was performed for preliminary identification by using three specific primer sets: F0145/E2477 (specific for Pectobacterium carotovorum subsp. carotovorum), Br1f/L1r (specific for P. carotovorum subsp. brasiliensis) and ECA1f/ECA2r (specific for P. atrosepticum). All isolates produced the band size of 666 bp with F0145/E2477 primer pair, indicating that they belong to the species P. carotovorum subsp. carotovorum. Further genetic characterization was based on sequence analysis of the gapA and mdh housekeeping genes. BLAST analysis confirmed 99.39% (Q. cover 100%, E. value 0.0) and 100% (Q. cover 100%, E. value 0.0) identity of the isolates with P. carotovorum subsp. carotovorum strains deposited in the NCBI database as M34 (KY047594) for gapA and Pcc t0437 (KC337296) for mdh genes, respectively. Phylogenetic analysis showed genetic homogeneity among the cabbage isolates.","PeriodicalId":19801,"journal":{"name":"Pesticidi I Fitomedicina","volume":"1 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"2","resultStr":"{\"title\":\"Characterization of Pectobacterium carotovorum subsp. carotovorum isolates from a recent outbreak on cabbage in Bosnia and Herzegovina\",\"authors\":\"T. Popović, A. Jelušić, S. Markovic, R. Iličić\",\"doi\":\"10.2298/pif1904211p\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"The causal agent of soft rot disease associated with a cabbage outbreak in Semberija region, Bosnia and Herzegovina, in 2018 was identified and characterized. Symptoms appeared in the form of water-soaked lesions on leaves and specific odour. Disease incidence ranged from 20% to 30%. The causal pathogen was isolated on nutrient agar (NA), King?s B and crystal violet pectate (CVP) media. Eight creamy-white, round and convex bacterial isolates, which produced characteristic pits on CVP medium were taken as representative. They were gram negative, facultative anaerobe, oxidase negative, catalase positive, nonfluorescent on King?s B medium, levan and arginine dehydrolase negative. The isolates were able to cause soft rot on cabbage and potato tuber slices 24 h after inoculation under conditions of high relative humidity. Polymerase chain reaction (PCR) was performed for preliminary identification by using three specific primer sets: F0145/E2477 (specific for Pectobacterium carotovorum subsp. carotovorum), Br1f/L1r (specific for P. carotovorum subsp. brasiliensis) and ECA1f/ECA2r (specific for P. atrosepticum). All isolates produced the band size of 666 bp with F0145/E2477 primer pair, indicating that they belong to the species P. carotovorum subsp. carotovorum. Further genetic characterization was based on sequence analysis of the gapA and mdh housekeeping genes. BLAST analysis confirmed 99.39% (Q. cover 100%, E. value 0.0) and 100% (Q. cover 100%, E. value 0.0) identity of the isolates with P. carotovorum subsp. carotovorum strains deposited in the NCBI database as M34 (KY047594) for gapA and Pcc t0437 (KC337296) for mdh genes, respectively. 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引用次数: 2
摘要
对2018年波斯尼亚和黑塞哥维那Semberija地区卷心菜暴发相关的软腐病病原进行了鉴定和鉴定。症状表现为叶子上被水浸透的病变和特殊气味。发病率从20%到30%不等。在营养琼脂(NA)上分离病原菌,King?s B和结晶紫果胶(CVP)介质。以8株乳白色、圆形和凸形的分离菌为代表,在CVP培养基上产生特征坑。革兰氏阴性,兼性厌氧菌,氧化酶阴性,过氧化氢酶阳性,金?s B培养基,levan和精氨酸脱氢酶阴性。在较高的相对湿度条件下,接种24 h后可对白菜和马铃薯块茎片造成软腐病。采用三组特异性引物F0145/E2477 (Pectobacterium carotovorum subsp特异性引物)进行聚合酶链反应(PCR)初步鉴定。cartovorum), Br1f/L1r(特异于p.a rotovorum亚种)。brasiliensis)和ECA1f/ECA2r (P. atrosepticum特异性)。F0145/E2477引物对的条带大小均为666 bp,表明该菌株属于胡萝卜芽孢杆菌亚种。carotovorum。进一步的遗传特征是基于gapA和mdh管家基因的序列分析。BLAST分析证实,分离株与胡萝卜假单胞菌亚种的同源性分别为99.39% (q - cover 100%, e - value 0.0)和100% (q - cover 100%, e - value 0.0)。在NCBI数据库中,cartovorum菌株的gapA基因为M34 (KY047594), mdh基因为Pcc t0437 (KC337296)。系统发育分析表明,各分离株遗传同源性较好。
Characterization of Pectobacterium carotovorum subsp. carotovorum isolates from a recent outbreak on cabbage in Bosnia and Herzegovina
The causal agent of soft rot disease associated with a cabbage outbreak in Semberija region, Bosnia and Herzegovina, in 2018 was identified and characterized. Symptoms appeared in the form of water-soaked lesions on leaves and specific odour. Disease incidence ranged from 20% to 30%. The causal pathogen was isolated on nutrient agar (NA), King?s B and crystal violet pectate (CVP) media. Eight creamy-white, round and convex bacterial isolates, which produced characteristic pits on CVP medium were taken as representative. They were gram negative, facultative anaerobe, oxidase negative, catalase positive, nonfluorescent on King?s B medium, levan and arginine dehydrolase negative. The isolates were able to cause soft rot on cabbage and potato tuber slices 24 h after inoculation under conditions of high relative humidity. Polymerase chain reaction (PCR) was performed for preliminary identification by using three specific primer sets: F0145/E2477 (specific for Pectobacterium carotovorum subsp. carotovorum), Br1f/L1r (specific for P. carotovorum subsp. brasiliensis) and ECA1f/ECA2r (specific for P. atrosepticum). All isolates produced the band size of 666 bp with F0145/E2477 primer pair, indicating that they belong to the species P. carotovorum subsp. carotovorum. Further genetic characterization was based on sequence analysis of the gapA and mdh housekeeping genes. BLAST analysis confirmed 99.39% (Q. cover 100%, E. value 0.0) and 100% (Q. cover 100%, E. value 0.0) identity of the isolates with P. carotovorum subsp. carotovorum strains deposited in the NCBI database as M34 (KY047594) for gapA and Pcc t0437 (KC337296) for mdh genes, respectively. Phylogenetic analysis showed genetic homogeneity among the cabbage isolates.