Identification of Broad2spectrum Antagonistic Strain B962II and GFP Tagging

doi:10.7668/hbnxb.2009.05.039

Abstract

Abstract: 16S rDNA of broad-spectrum antagonistic strain B96-II was amplified by PCR and sequenced.Using BLAST software,comparison results showed B96-II was possibly identified as Bacillus subtilis combining its morphological,physiological and biochemical characteristics.In order to construct Escherichia coli-Bacillus shuttle vector containing gfp and B96-II self promoter,digested fragments of total DNA of B96-II that was digested by Sau3A I were ligated to pNW33N-gfp that was digested by BamH I,and the product was transformed into E.coli DH5αcompetent cells.Protoplast transformation method was used to gain tagged strain by gfp and GFP was well expressed in tagged B96-II under fluorescence microscope.B96-II-gfp remained strong antifungal activities against 12 kinds of plant pathogentic fungi as B96-II.The results provided fundamental possibility for the further study on the ecological behavior of broad-spectrum antagonistic bacterium B96-II.

Key words: Broad-spectrum antagonistic strain, 16S rDNA, GFP, Protoplast transformation

CLC Number:

S432.4

HAO Bian-qing, MA Li-ping, QIAO Xiong-wu. Identification of Broad2spectrum Antagonistic Strain B962II and GFP Tagging[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(5): 188-191. doi: 10.7668/hbnxb.2009.05.039.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2009.05.039

http://www.hbnxb.net/EN/Y2009/V24/I5/188

References

[1] 杨祥田,李伟龙,叶建人,等.设施栽培几种土传病害的发生为害现状及控制途径探讨[J].浙江农业科学,2005(4):313-315,288.
[2] 李宝聚.我国蔬菜病害研究现状与展望[J].中国蔬菜,2006(1):1-5.
[3] 孔建,王文夕,赵白鸽,等.枯草芽孢杆菌B-903 菌株的研究 I.对植物病原菌的抑制作用和防治试验[J].中国生物防治,1999,15(4):157-161.
[4] 檀根甲,李增智,刘淑芳,等.枯草芽孢杆菌BS8026对苹果采后炭疽病的控病效果及作用机制[J].植物保护学报,2008,38(3):227-232.
[5] 齐东梅,惠明,梁启美,等.枯草芽孢杆菌H110对苹果梨采后青霉病和黑斑病的抑制效果[J].应用与环境生物学报,2005,11 (2):171-174.
[6] 陈晓斌,张炳欣,楼兵干,等.运用生色基因标记黄瓜根围促生菌(PGPR)筛选菌株[J].微生物学报,2001,41(3):287-292.
[7] Xi C W,Lambrecht M,Vanderleyden J,et al.Bi-functional gfp and gus A-containing mini-Tn5 transposon derivatives for combined gene expression and bacterial localization studies[J].Microbiol Meth,1999,35(1):85-92.
[8] Ramos H J O,Roncato-Maccari L D B,Souza E M,et al.Monitoring Azospirillum-wheat interactions using the gfp and gus A genes constitutively expressed from a new broad-host range vector[J].Biotechnol,2002,97(3):243-252.
[9] 张昕,张炳欣,喻景权,等.生防菌ZJY-1及ZJY-116的GFP标记极其在黄瓜根围的生态适应性[J].应用生态学报,2005,16(11):2144-2148.
[10] Scott K P,Mercer D K,Glover L A,et al.The green fluorescent protein as a visible marker for lactic acid bacteria in complex ecosystems[J].Fems Microbiol Ecol,1998,26(3):219-230.
[11] 马利平,乔雄梧,高芬,等.B96-II 对3种枯萎病的防治效果及拮抗物质初步分析[J].华北农学报,2006,21(4):99-102.
[12] 马利平,郝变青,秦曙,等.芽孢杆菌B96-II对芦笋茎枯病的防治及机制研究[J].华北农学报,2008,23(2):180-184.
[13] 郝变青,马利平,乔雄梧,等.拮抗菌对黄瓜枯萎病菌的室内生物活性[J].应用与环境生物学报,2001,7(2):155-157.
[14] 马利平,郝变青,乔雄梧.山西省芦笋病害现状及无公害生产技术[J].山西农业科学,2009,37(1):69-73.
[15] Delgado S,Suárez A,Mayo B.Identification of dominant bacteria in feces and colonic mucosa from healthy spanish adults by culturing and by 16S rDNA sequence analysis[J].Digestive Diseases and Sciences,2006,51(4):744-751.
[16] Katano T,Fukui M,Watanabe Y.Identification of cultured and uncultured picocyanobacteria from a mesotrophic freshwater lake based on the partial sequences of 16S rDNA[J].Limnology,2001,2:213-218.
[17] 张艾青,刘书亮,敖灵.产广谱细菌素乳酸菌的筛选和鉴定[J].微生物学通报,2007,34(4):753-756.
[18] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001:353-399.
[19] 诸葛健.现代发酵微生物实验技术[M].北京:化学工业出版社,2005:186-187.
[20] 周延清,杨清香,张改娜.生物遗传标记与应用[M].北京:化学工业出版社,2008:240.
[21] 田宏先,崔林,王秀英,等.马铃薯内生促生菌的促生长作用[J].山西农业科学,2003,31(1):28-30.
[22] 陈中义,张杰,曹景萍,等.杀虫防病基因工程枯草芽孢杆菌的构建[J].生物工程学报,1999,15(2):215-220.
[23] 张丽珍,马利平,乔雄梧,等.一株多菌灵降解菌NY97-1的分子鉴定及GFP标记[J].应用与环境生物学报,2006,12(4):555-558.

Please choose a citation manager

Content to export