Selvakumar Govindan, Joshi Piyush, Mishra Pankaj K, Bisht Jaideep K, Gupta Hari S
Vivekananda Institute of Hill Agriculture, Indian Council of Agricultural Research, Almora, India.
Curr Microbiol. 2009 Oct;59(4):432-8. doi: 10.1007/s00284-009-9456-1. Epub 2009 Jul 28.
Fourteen cold tolerant phosphate solubilizing bacteria isolated from high altitude representative locations of the two major mountain aspects of the Uttarakhand Himalayas (cooler north and warmer south facing slopes) were selected for this study. The tricalcium phosphate (TCP) solubilizing abilities of the isolates were estimated at three different incubation temperatures viz., 4, 15, and 30 degrees C under in vitro conditions. Irrespective of their geographical origin, all the isolates recorded maximum P release values at 30 degrees C. At 4 degrees C, the isolates from the north facing slope were found to release significantly higher levels of P, as compared to the isolates from the south facing slopes. Alternatively at 15 degrees C, the isolates from the south facing slope were found to release significantly higher levels of P. Initial confirmation of their genus level identity as Pseudomonads was arrived by amplification of a 990 bp fragment of the 16S rRNA gene using genus specific primers. Further putative species level identification was arrived by sequencing of the 16S rRNA gene. The diversity among the isolates was determined by rep-PCR using the primers BOX, ERIC, and ERIC2. A composite dendrogram constructed using the rep-PCR profiles revealed that the isolates from the north and south mountain aspects formed separate major clusters. The extent of diversity was greater among the isolates from the south mountain aspect. This study reveals the potential of rep-PCR in determining the genetic diversity among Pseudomonads selected for a single functional trait, but varying in their geographical origin.
从北阿坎德邦喜马拉雅山脉两个主要坡面(较凉爽的北坡和较温暖的南坡)的高海拔代表性地点分离出14株耐冷解磷细菌用于本研究。在体外条件下,于三种不同培养温度(即4、15和30摄氏度)下评估分离株的磷酸三钙(TCP)溶解能力。无论其地理来源如何,所有分离株在30摄氏度时的磷释放值均最高。在4摄氏度时,发现来自北坡的分离株比来自南坡的分离株释放出显著更高水平的磷。相反,在15摄氏度时,发现来自南坡的分离株释放出显著更高水平的磷。通过使用属特异性引物扩增16S rRNA基因的990 bp片段,初步确认它们在属水平上属于假单胞菌属。通过对16S rRNA基因进行测序进一步进行假定的种水平鉴定。使用BOX、ERIC和ERIC2引物通过rep-PCR确定分离株之间的多样性。使用rep-PCR图谱构建的复合树状图显示,来自北坡和南坡的分离株形成了单独的主要聚类。来自南山坡的分离株之间的多样性程度更大。本研究揭示了rep-PCR在确定因单一功能性状而选择但地理来源不同的假单胞菌之间遗传多样性方面的潜力。