杨树(细菌性)枯萎病菌快速检测方法研究
|
摘要
本文对杨树(细菌性)枯萎病菌的培养特征、生理生化特征、分子生物学特征等方面进行了研究与描述,设计并合成了特异性PCR引物,杨树(细菌性)枯萎病菌的PCR产物出现718 bp的特异性扩增条带,而其他近似种的细菌均未出现扩增条带,说明该对引物具有杨树(细菌性)枯萎病菌鉴定特异性,将病菌DNA模板做浓度梯度稀释,灵敏度可达10 pg/μL。该方法快速、灵敏、准确,可用于杨树枯萎病菌检测。
The culture characteristics,physiological and biochemical characteristics and molecular biological characteristics of Enterobacter cancerogenus were studied and described in this paper. Specific primers were designed and synthesized,and 718 bp specific amplification bands appeared in the PCR products of Enterobacter cancerogenus,while no amplification bands were found in other similar species,which proved that the primers were specific for identification of Enterobacter cancerogenus. By diluting the DNA template of the pathogen with concentration gradient,and the sensitivity was up to 10 pg/μL.This can be used as a rapid,sensitive and accurate PCR method for identification of Enterobacter cancerogenus.
The culture characteristics,physiological and biochemical characteristics and molecular biological characteristics of Enterobacter cancerogenus were studied and described in this paper. Specific primers were designed and synthesized,and 718 bp specific amplification bands appeared in the PCR products of Enterobacter cancerogenus,while no amplification bands were found in other similar species,which proved that the primers were specific for identification of Enterobacter cancerogenus. By diluting the DNA template of the pathogen with concentration gradient,and the sensitivity was up to 10 pg/μL.This can be used as a rapid,sensitive and accurate PCR method for identification of Enterobacter cancerogenus.
引文
[1]Uro觤evic'B.Canker of polar caused by Erwinia cancerogena sp.[in Czech].Lesnicky Casopis,1966,12:493-505.
[2]Dickey R S,Zumoff C H.Emended description of Enterobacter cancerogenus comb.nov.(formerly Erwinia cancerogena).International Journal of Systematic and Evolutionary Microbiology,1988,38:371-374.
[3]王国芬.桑细菌性枯萎病病原学及其致病机理研究[D].杭州:浙江大学,2009.
[4]朱勃.桑细菌性枯萎病病原的定名、分子检测及肠杆菌基因水平转移的研究[D].杭州:浙江大学,2009.
[5]Sanders W E J,Sanders C C.Enterobacter spp.:pathogens poised to flourish at the turn of the century.Clin Microbiol Rev,1997,10:220-241.
[6]Iversen C,Forsythe S J.Risk profile of Enterobacter sakazakii,an emergent pathogen associated with infant milk formula.Trends Food Sci Technol,2003,14:443-454.
[7]Abbott S L,Janda J M.Enterobacter cancerogenus("Enterobacter taylorae")Infections Associated With Severe Trauma or Crush Injuries.American journal of clinical pathology,1997,107(3):359-361.
[8]Uro觤evic'B.Relationships between Erwinia cancerogena Ur.and Aplanobacterium populi Ride.Forest Pathology,1973,3(2):83-97.
[9]任欣正.植物病原细菌的分类和鉴定.北京:农业出版社,1994.
[10]Weisburg W G,Barns S M,Pelletier D A,et al.16S ribosomal DNA amplification for phylogenetic study.Journal of Bacteriology,1991,173(2):697-703.
[2]Dickey R S,Zumoff C H.Emended description of Enterobacter cancerogenus comb.nov.(formerly Erwinia cancerogena).International Journal of Systematic and Evolutionary Microbiology,1988,38:371-374.
[3]王国芬.桑细菌性枯萎病病原学及其致病机理研究[D].杭州:浙江大学,2009.
[4]朱勃.桑细菌性枯萎病病原的定名、分子检测及肠杆菌基因水平转移的研究[D].杭州:浙江大学,2009.
[5]Sanders W E J,Sanders C C.Enterobacter spp.:pathogens poised to flourish at the turn of the century.Clin Microbiol Rev,1997,10:220-241.
[6]Iversen C,Forsythe S J.Risk profile of Enterobacter sakazakii,an emergent pathogen associated with infant milk formula.Trends Food Sci Technol,2003,14:443-454.
[7]Abbott S L,Janda J M.Enterobacter cancerogenus("Enterobacter taylorae")Infections Associated With Severe Trauma or Crush Injuries.American journal of clinical pathology,1997,107(3):359-361.
[8]Uro觤evic'B.Relationships between Erwinia cancerogena Ur.and Aplanobacterium populi Ride.Forest Pathology,1973,3(2):83-97.
[9]任欣正.植物病原细菌的分类和鉴定.北京:农业出版社,1994.
[10]Weisburg W G,Barns S M,Pelletier D A,et al.16S ribosomal DNA amplification for phylogenetic study.Journal of Bacteriology,1991,173(2):697-703.