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Bacteria in root nodules of legumes play important roles in promoting plant growth. In this study, we investigated root nodule-associated bacteria isolated from leguminous plants along an elevation gradient on the northern slope of the Kunlun Mountains, China, using a cultivation approach. In total, 300 isolates were obtained from seven legume species within six ecological zones. Isolates were identified based on 16S rRNA gene phylogenetic analysis and potential rhizobia were further identified using a  rec A gene phylogeny. Among the isolates,  Bacillales  (particularly  Bacillus ) were the dominant isolates from all host legumes and all elevations (63.5%), followed by  Rhizobiales  (13%) and  Pseudomonadales  (11.7%). Less than 3% of the isolates belonged to  Burkholderiales ,  Paenibacillales ,  Enterobacteriales ,  Actinomycetales ,  Sphingomonadales ,  Xanthomonadales ,  Chitinophagales ,  Brevibacillales ,  Staphylococcales , or  Mycobacteriales . A few elevation-specific patterns emerged within the  Bacillales  and  Pseudomonadales . For example, isolates related to the psychrotroph  Bacillus psychrosaccharolyticus  were only isolated from the highest elevation sites (>3,500 m) whereas those related to the mesophile  Bacillus endophyticus  were only isolated from lowest elevation sites (1,350 m), suggestive of a role of soil temperature in their distribution. Similarly, isolates related to  Pseudomonas brassicacearum  were the dominant  Pseudomonadales  isolates, but they were only isolated from middle and low elevations (<3,200 m). A total of 39 isolates belonged to the  Rhizobiales , 36 of which were confirmed to the genus level using the  recA  gene. In all,  Rhizobiales  isolates were obtained from five different host legumes spanning the entire elevation gradient. Those from the low-elevation Qira Desert-Oasis Transition Zone (1,350–1,960 m) suggested some patterns of host preference. For example, most isolates from  Albizia julibrissin  formed a monophyletic group related to  Rhizobium lemnae  and most from  Alhagi sparsifolia  were closely related to  Ensifer kummerowiae . In general, this study shows that most bacteria associated with root nodules of legumes are widely distributed in distinct ecological zones within a single geographic region but suggests that both climate and host interactions may influence their distributions.

Diversity of Root Nodule-Associated Bacteria of Diverse Legumes Along an Elevation Gradient in the Kunlun Mountains, China