一株甘蓝内生枯草芽孢杆菌16S rDNA序列的克隆与分析
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摘要
台州学院生命科学学院实验室前期从甘蓝健康植株中分离到一株内生细菌GL06,为甘蓝内生枯草芽孢杆菌GL06的功能研究和生产应用奠定基础,利用PCR法克隆其16S rDNA序列,并进行生物信息学分析。结果表明:以27F/1492R为PCR引物,扩增到长度为1 510bp的16S rDNA序列,序列的GC含量为54%;GL06 16S与枯草芽孢杆菌16S的相似性最高,达99%,仅存在3个碱基差异;GL06与枯草芽孢杆菌、解淀粉芽孢杆菌的系统发育树聚为一组。
The 16 SrDNA sequence and biological information of an endophytic Bacillus subtilis strain GL06 isolated from healthy cabbage plants by College of Life Sciences,Taizhou University was cloned and analyzed by PCR to lay the foundation for its function research and production application.Result:The length and GC content of the 16 SrDNA sequence amplified by using 27 F/1492 Ras a primer are 1 510 bp and 54%respectively.The 16 Ssimilarity between GL06 strain and B.subtilis is up to 99%and there is a three-base difference only.The phylogenetic tree of GL06 strain,B.subtilis and Bacillus amyloliquefaciens can be divided into the same group.
The 16 SrDNA sequence and biological information of an endophytic Bacillus subtilis strain GL06 isolated from healthy cabbage plants by College of Life Sciences,Taizhou University was cloned and analyzed by PCR to lay the foundation for its function research and production application.Result:The length and GC content of the 16 SrDNA sequence amplified by using 27 F/1492 Ras a primer are 1 510 bp and 54%respectively.The 16 Ssimilarity between GL06 strain and B.subtilis is up to 99%and there is a three-base difference only.The phylogenetic tree of GL06 strain,B.subtilis and Bacillus amyloliquefaciens can be divided into the same group.
引文
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[25]张伟信,邬晓凤,喻华英.16SrDNA对四种常见细菌中的鉴定与应用[J].塔里木大学学报,2013,25(4):7-11.
[26]WATTS G S,YOUENS-CLARK K,SLEPIAN M J,et al.16SrRNA gene sequencing on a benchtop sequencer:accuracy for identification of clinically important bacteria[J].Journal of Applied Microbiology,2017,123:1584-1596.
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[28]陈美琪,王利.鲈鱼枯草芽孢杆菌的分离及16SrD-NA基因分析[J].黑龙江畜牧兽医,2017(10):197-199.
[29]吕美云,刘紫英.豆豉中5株产纤溶酶菌的筛选与鉴定[J].大豆科学,2016(1):165-170.
[30]严婉荣,肖敏,陈圆,等.芽孢杆菌基本特征、16SrRNA对比分析及特异性基因挖掘[J].基因组学与应用生物学,2017(11):4686-4692.
[2]MA K,CHEN X,GUO X,et al.Bacillus vini sp.nov.isolated from alcohol fermentation pit mud[J].Archives of Microbiology,2016,198:559-564.
[3]HASSAN MA,HAROUN BM,AMARA AA,et al.Production and characterization of keratinolytic protease from new wool-degrading Bacillus species isolated from Egyptian ecosystem[J].Biomed Research International,2013,2013(6):175012.
[4]AFOUDA P,DUBOURG G,CADORET F,et al.‘Bacillus massiliglaciei’,a new bacterial species isolated from Siberian permafrost[J].New Microbes and New Infections,2017,15:92-93.
[5]刘国红,林乃铨,林营志,等.芽孢杆菌分类与应用研究进展[J].福建农业学报,2008,23(1):92-99.
[6]李云龙.芽孢杆菌发展史综述及用途[J].农业开发与装备,2015(5):37.
[7]KIMURA K,YOKOYAMA S.Trends in the application of Bacillus in fermented foods[J].Current Opinion in Biotechnology,2018,56:36-42.
[8]TAHIR M,KHALID U,IJAZ M,et al.Combined application of bio-organic phosphate and phosphorus solubilizing bacteria(Bacillus strain MWT 14)improve the performance of bread wheat with low fertilizer input under an arid climate[J].Brazilian Journal of Microbiology,2018,49(1):15-24.
[9]BEN KHEDHER S,KILANI-FEKI O,DAMMAKM,et al.Efficacy of Bacillus subtilis V26as a biological control agent against Rhizoctonia solani on potato[J].Comptes Rendus Biologies,2015,338:784-792.
[10]RYU H,PARK H,SUH D S,et al.Biological control of Colletotrichum panacicola on Panax ginseng by Bacillus subtilis HK-CSM-1[J].Journal of Ginseng Research,2014,38:215-219.
[11]李元召,孙俊良,李东霄,等.一株产淀粉酶枯草芽孢杆菌16SrDNA的克隆及序列分析[J].河南科技学院学报,2013,41(3):37-41.
[12]CLARRIDGE J E.Impact of 16SrRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases[J].Clinical Microbiology Reviews,2004,17(4):840-862.
[13]MATSUMOTO T,SUGANO M.16SrRNA gene sequence analysis for bacterial identification in the clinical laboratory[J].The Japanese Journal of Clinical Pathology,2013,61(12):1107-1115.
[14]WOO P C,LAU S K,TENG J L,et al.Then and now:use of 16SrDNA gene sequencing for bacterial identification and discovery of novel bacteria in clinical microbiology laboratories[J].Clinical Microbiology and Infection,2008,14(10):908-934.
[15]RAMPINI S K,BLOEMBERG G V,KELLER P M,et al.Broad-range 16SrRNA gene polymerase chain reaction for diagnosis of culture-negative bacterial infections[J].Clinical Infectious Diseases,2011,53(12):1245-1251.
[16]LOTFY W A,MOSTAFA S W,ADEL A A,et al.Production of di-(2-ethylhexyl)phthalate by Bacillus subtilis AD35:Isolation,purification,characterization and biological activities[J].Microbial Pathogenesis,2018,124:89-100.
[17]SUBBA REDDY GANGIREDDYGARI V,KANDERID,GOLLA R,et al.Biodegradation of quinalphos by a soil bacterium-Bacillus subtilis[J].Pakistan Journal of Biological Sciences,2017,20(8):410-422.
[18]杜昕波,赵耘,李伟杰.菌种保藏中的细菌鉴定方法[J].中国兽药杂志,2009,43(3):50-52.
[19]安然,易图永,肖启明,等.gyrB基因在细菌分类和检测中的应用[J].江西农业学报,20l0,22(4):18-20.
[20]盛强,罗明,管晶晶,等.新疆加工型辣椒一种新的细菌性叶斑病病原的鉴定[J].植物保护学报,2017,44(2):260-268.
[21]PUAH S M,KHOR W C,KEE B P,et al.Development of a species-specific PCR-RFLP targeting rpoDgene fragment for discrimination of Aeromonas species[J].Journal of Medical Microbiology,2018,67:1271-1278.
[22]MARONICHE G A,GARCIA J E,SALCEDO F,et al.Molecular identification of Azospirillum spp.:Limitations of 16SrRNA and qualities of rpoD as genetic markers[J].Microbiological Research,2017,195:1-10.
[23]TEIXEIRA A B,BARIN J,HERMES D M,et al.PCRAssay based on the gyrB gene for rapid identification of Acinetobacter baumannii-calcoaceticus complex at specie level[J].Journal of Clinical Laboratory Analysis,2017,31(3):e22046.
[24]卢佳琦,李市场,王大红,等.6SrDNA与gyrB序列联合法鉴定一株蜡样芽孢杆菌[J].河南科技大学学报(自然科学版),2018,39(1):73-77.
[25]张伟信,邬晓凤,喻华英.16SrDNA对四种常见细菌中的鉴定与应用[J].塔里木大学学报,2013,25(4):7-11.
[26]WATTS G S,YOUENS-CLARK K,SLEPIAN M J,et al.16SrRNA gene sequencing on a benchtop sequencer:accuracy for identification of clinically important bacteria[J].Journal of Applied Microbiology,2017,123:1584-1596.
[27]BOSSHARD P P,ABELS S,ZBINDEN R,et al.Ribosomal DNA sequencing for identification of aerobic gram-positive rods in the clinical laboratory(an 18-month evaluation)[J].Journal of Clinical Microbiology,2003,41(9):4134-4140.
[28]陈美琪,王利.鲈鱼枯草芽孢杆菌的分离及16SrD-NA基因分析[J].黑龙江畜牧兽医,2017(10):197-199.
[29]吕美云,刘紫英.豆豉中5株产纤溶酶菌的筛选与鉴定[J].大豆科学,2016(1):165-170.
[30]严婉荣,肖敏,陈圆,等.芽孢杆菌基本特征、16SrRNA对比分析及特异性基因挖掘[J].基因组学与应用生物学,2017(11):4686-4692.
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