一株嗜麦芽寡养单胞菌的分离及其生物学特性
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摘要
近年来,基于高通量测序技术对植物根际土壤微生物菌群结构的测定结果表明,土壤中绝大多数微生物难以通过传统的富营养培养基实现培养和分离。为进一步探索分离土壤中寡营养微生物的方法,结合多种培养因素,以土壤浸出液为基础培养基设计寡营养培养基分离土壤细菌菌株。并通过菌株的细胞形态、生理生化特征、16S rRNA和gyrB基因序列等对其中一株菌进行鉴定,同时对其蛋白酶与铁载体的产生能力进行检测以评价其在植物根际促生方面的应用潜力。利用土壤浸出液做培养基是有效分离土壤寡营养细菌的方法 ;编号为S35的菌株鉴定为嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia),具有蛋白酶和铁载体产生能力。采用的寡营养培养基可以分离获得在富营养培养基上不易分离的细菌,得到S. maltophilia S35菌株在植物促生方面具有一定的应用潜力。
In recent years,based on high-throughput sequencing technology,the determination of microbial flora structure in plant rhizosphere soil shows that most microorganisms in soil are difficult to culture and separate via traditional eutrophic medium. It is aimed to further explore an effective method for isolating oligotrophic microorganisms from soil. A variety of factors were combined to design an oligotrophic medium to isolate soil bacteria based on soil leaching solution. One of the strains was identified by cell morphology,physiological and biochemical characteristics,16 S rRNA,and gyrB gene sequences;meanwhile,its function of proteases-producing and siderophoreproducing was detected to assess the potential in promoting plant growth. Using soil leaching solution as a medium was a method for effectively isolating soil oligotrophic bacteria,and the strain numbered S35 was identified as Stenotrophomonas maltophilia,it had the ability to produce protease and siderophore. The oligotrophic medium used in this study can be used to obtain bacteria that are difficult to isolate through nutrientrich medium. In addition,S. maltophilia S35 has certain application potential in plant growth promotion.
In recent years,based on high-throughput sequencing technology,the determination of microbial flora structure in plant rhizosphere soil shows that most microorganisms in soil are difficult to culture and separate via traditional eutrophic medium. It is aimed to further explore an effective method for isolating oligotrophic microorganisms from soil. A variety of factors were combined to design an oligotrophic medium to isolate soil bacteria based on soil leaching solution. One of the strains was identified by cell morphology,physiological and biochemical characteristics,16 S rRNA,and gyrB gene sequences;meanwhile,its function of proteases-producing and siderophoreproducing was detected to assess the potential in promoting plant growth. Using soil leaching solution as a medium was a method for effectively isolating soil oligotrophic bacteria,and the strain numbered S35 was identified as Stenotrophomonas maltophilia,it had the ability to produce protease and siderophore. The oligotrophic medium used in this study can be used to obtain bacteria that are difficult to isolate through nutrientrich medium. In addition,S. maltophilia S35 has certain application potential in plant growth promotion.
引文
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[20]Singh RP, Jha RN. The PGPR Stenotrophomonas maltophilia SBP-9 augments resistance against biotic and abiotic stress in wheat plants[J]. Frontiers in Microbiology, 2017, 8:1945.
[2]Barer MR, Harwood CR. Bacterial viability and culturability[J].Advances in Microbial Physiology, 1999, 41:93-137.
[3]王保军,刘双江.环境微生物培养新技术的研究进展[J].微生物学通报, 2013, 40(1):6-17.
[4]Button DK, Schut F, Quang P, et al. Viability and isolation of marine bacteria by dilution culture:theory, procedures, and initial results[J]. Appl environ Microbiol, 1993, 59(3):881-891.
[5]周德庆.微生物学教程[M].北京:高等教育出版社, 2002:247-248.
[6]李阜棣.农业微生物学实验技术[M].北京:中国农业出版社,1996.
[7]张伟.纯培养和土壤中微生物形成的腐殖质研究[D].长春:吉林农业大学, 2014.
[8]Olson JB, Lord CC, McCarthy PJ. Improve recoverability of microbial colonies from the marine sponge samples[J]. MicrobEcol, 2000,40:139-147.
[9]刘丹丹,李敏,刘润进.我国植物根围促生细菌研究进展[J].生态学杂志, 2016, 35(3):815-824.
[10]Kloepper JW, Zablotowicz RM, Tipping EM, et al. Plant growth promotion mediated by bacterial rhizosphere colonizers[M]//Keister KL, Cregan PB. The rhizosphere and plant growth.Dordecht:Kluwer Academic Publishers, 1991:315-326.
[11]田方,丁延芹,朱辉,等.烟草根际铁载体产生菌G-229-21T的筛选、鉴定及拮抗机理[J].微生物学报, 2008(5):631-637.
[12]阮继生.“伯杰氏系统细菌学手册(第二版)”第5卷与我国的放线菌系统学研究[J].微生物学报, 2013, 53(6):521-530.
[13]东秀珠.常见细菌系统鉴定手册[M].北京:科学出版社,2001:371.
[14]王君.蓝莓根际促生细菌的筛选、鉴定及其促生效果[D].泰安:山东农业大学, 2016.
[15]沈萍,陈向东.微生物学实验[M].北京:高等教育出版社,2007:189.
[16]Palleroni NJ, Bradbury JF. Stenotrophomonas, a new bacterial genus for Xanthomonas maltophilia(Hugh 1980)Swings et al.1983[J]. International Journal of Systematic Bacteriology, 1993,43(3):606-609.
[17]Willems A, Falsen E, Pot B, et al. Acidovorax, a new genus for Pseudomonas facilis, Pseudomonas delafieldii, E. Falsen(EF)group 13, EF group 16, and several clinical isolates, with the species Acidovorax facilis comb. nov. Acidovorax delafieldii comb.nov. and Acidovorax temperans sp.[J]. International Journal of Systematic Bacteriology, 1990, 40(4):384-398.
[18]AdjidéCC, De Meyer A, Weyer M, et al. A sensitive, specific and predictive isolation medium developed for Stenotrophomonas maltophilia study in healthcare settings[J]. Pathology Biology(Paris), 2010, 58(1):11-17.
[19]岳秀娟,余利岩,李秋萍,等.自然界中难分离培养微生物的分离和应用[J].微生物学通报, 2006, 33(3):77-81.
[20]Singh RP, Jha RN. The PGPR Stenotrophomonas maltophilia SBP-9 augments resistance against biotic and abiotic stress in wheat plants[J]. Frontiers in Microbiology, 2017, 8:1945.