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来自印度列城拉达克寒冷沙漠的嗜冷细菌的可培养多样性及功能注释

Culturable diversity and functional annotation of psychrotrophic bacteria from cold desert of Leh Ladakh (India).

作者信息

Yadav Ajar Nath, Sachan Shashwati Ghosh, Verma Priyanka, Tyagi Satya Prakash, Kaushik Rajeev, Saxena Anil K

机构信息

Division of Microbiology, Indian Agricultural Research Institute, New Delhi, 110012, India.

出版信息

World J Microbiol Biotechnol. 2015 Jan;31(1):95-108. doi: 10.1007/s11274-014-1768-z. Epub 2014 Nov 5.

Abstract

To study culturable bacterial diversity under subzero temperature conditions and their possible functional annotation, soil and water samples from Leh Ladakh region were analysed. Ten different nutrient combinations were used to isolate the maximum possible culturable morphotypes. A total of 325 bacterial isolates were characterized employing 16S rDNA-Amplified Ribosomal DNA Restriction Analysis with three restriction endonucleases AluI, MspI and HaeIII, which led to formation of 23-40 groups for the different sites at 75 % similarity index, adding up to 175 groups. Phylogenetic analysis based on 16S rRNA gene sequencing led to the identification of 175 bacteria, grouped in four phyla, Firmicutes (54 %), Proteobacteria (28 %), Actinobacteria (16 %) and Bacteroidetes (3 %), and included 29 different genera with 57 distinct species. Overall 39 % of the total morphotypes belonged to the Bacillus and Bacillus derived genera (BBDG) followed by Pseudomonas (14 %), Arthrobacter (9 %), Exiguobacterium (8 %), Alishewanella (4 %), Brachybacterium, Providencia, Planococcus (3 %), Janthinobacterium, Sphingobacterium, Kocuria (2 %) and Aurantimonas, Citricoccus, Cellulosimicrobium, Brevundimonas, Desemzia, Flavobacterium, Klebsiella, Paracoccus, Psychrobacter, Sporosarcina, Staphylococcus, Sinobaca, Stenotrophomonas, Sanguibacter, Vibrio (1 %). The representative isolates from each cluster were screened for their plant growth promoting characteristics at low temperature (5-15 °C). Variations were observed among strains for production of ammonia, hydrogen cyanide, indole-3-acetic acid and siderophore, solubilisation of phosphate, 1-aminocyclopropane-1-carboxylate deaminase activity and biocontrol activity against Rhizoctonia solani and Macrophomina phaseolina. Cold adapted microbes may have application as inoculants and biocontrol agents in crops growing at high altitudes under cold climate condition.

摘要

为了研究零下温度条件下可培养细菌的多样性及其可能的功能注释,对来自列城拉达克地区的土壤和水样进行了分析。使用了十种不同的营养组合来分离尽可能多的可培养形态类型。采用16S rDNA-扩增核糖体DNA限制性分析,使用三种限制性内切酶AluI、MspI和HaeIII对总共325株细菌分离株进行了表征,这导致在75%相似性指数下不同位点形成23 - 40个组,总计175个组。基于16S rRNA基因测序的系统发育分析鉴定出175种细菌,分为四个门,厚壁菌门(54%)、变形菌门(28%)、放线菌门(16%)和拟杆菌门(3%),包括29个不同的属和57个不同的种。总体而言,总形态类型的39%属于芽孢杆菌属和芽孢杆菌衍生属(BBDG),其次是假单胞菌属(14%)、节杆菌属(9%)、嗜冷栖热菌属(8%)、阿氏杆菌属(4%)、短杆菌属、普罗威登斯菌属、动性球菌属(3%)、詹氏菌属、鞘氨醇杆菌属、考克氏菌属(2%)以及橙色单胞菌属、柠檬酸球菌属、纤维单胞菌属、短波单胞菌属、德氏菌属、黄杆菌属、克雷伯菌属、副球菌属、嗜冷杆菌属、芽孢八叠球菌属、葡萄球菌属、中华芽孢杆菌属、嗜麦芽窄食单胞菌属、血杆菌属、弧菌属(1%)。对每个聚类的代表性分离株进行了低温(5 - 15°C)下促进植物生长特性的筛选。观察到菌株在氨、氰化氢、吲哚 - 3 - 乙酸和铁载体的产生、磷酸盐溶解、1 - 氨基环丙烷 - 1 - 羧酸脱氨酶活性以及对立枯丝核菌和菜豆壳球孢菌的生物防治活性方面存在差异。适应寒冷的微生物可能在寒冷气候条件下高海拔地区生长的作物中作为接种剂和生物防治剂应用。

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