Screening On Rhizobia Strains From Woody Legumes In Jianfengling Nature Reserve Of Hainan

227 strains isolated from 24 species woody legumes that 9 genus 14 kinds blongs to Papilionoids, 3 genus 3 kinds blongs to Caesalpinioideae and 5 genus 7 kinds are Mimosoideae, in Jianfengling National Nature Reserve, Hainan province. 117 strains which had the colony characters of rhizobia were carried on 16S rDNA appraisal, 27 strains which were rhizobia had carried on further research, physiological biochemistry tests, the BIOLOG carbon source utility, dissolving phosphors and potassium,9 Rhizobium tropici strains had carried on preliminary stress resistance. As follows are the results.The results of 16S rDNA sequential analysis indicated that woody leguminous rhizobia had the multiplity heredity. 16S rDNA cloned sequences blasted with that of the GenBank, from the Phylogenetic tree, twenty two strains were R.tropici,one strain was Rhizobium rhizogenes, two strains were Ensifer mexicanum, two strains were Burkholderia sp.; the others 90 strains were endophyte. The hosts of the 27 rhizobia strains were different, the hosts of the 22 R.tropici were Ormosia balansae Drake, Ormosia semicastrata, Acacia mangium Willd., Indigofera suffruticosa Mill., Desmodium heterocarpon(L.)DC., lagenirisa siceraria, the hosts of the Rhizobium rhizogenes was the Erythrophleum fordii Oliv., the hosts of the Ensifer mexicanum were the Gliricidia sepium (Jacq. ) Walp., the hosts of the Burkholderia sp. were the Dalbergia hancei Benth. The similarities of 16S rDNA were different of each other, for that of caf 443 and caf436 was as low as 98.8%, but that of the others all above 99.0%.The results of physiology and biochemistry tests show that, its cluster results were similar with that of the 16S rDNA. Rhizobium rhizogenes caf448 strain clusted near with the standard Rhizobium rhizogenes strain CGMCC1.2556, Ensifer mexicanum caf249 and caf254 clusted near with the standard Sinorhizobium fredii strain CGMCC1.2556, Burkholderia sp. caf420 and caf421 clusted near with the standard Burkholderia cepacia strain. CGMCC1.1813. 22 Rhizobium tropici strains clusted with standard Rhizobium tropici and Sinorhizobium teranga, with large difference in characters of physiology and biochemistry.The strains had abilities in utilizing 95 kinds of carbon sources for different rhizobia, for that all strains could used only pentite,L-arabinose andα-D-glucose,could not useα-cyclodextrin and Glycogen. The cluster results of BIOLOG was different from that of the physiological and biochemical tests, for that Ensifer mexicanum caf249 clustered with Rhizobium tropici, but not with the same Sinorhizobium fredii which blonged to same genus.The results of dissolving phosphors indicated that, rhizobia isolated from Jiangfengling distrct had strong ability of dissolving phosphors. 24 strains could dissolving inorganic phosphorus Ca3(PO4)2, not include the lecithin. caf446 D/d ratio is most greatly 2.037,the D/d ratio of caf415(1.854), caf437(1.570), caf226(1.529), caf276(1.880), caf278 (1.610), caf334(1.678), caf341(1.630),caf344(1.596), caf414(1.611), caf420(1.543) and caf436 (1.743) are bigger than 1.5. The results of dissolving phosphors screening on Colormetry method indicated that, all except caf438 the available phosphorus to be higher than the comparison (CK), the caf341 effective phosphorus content as high as 629.63mg/kg, caf436, caf276, caf444, caf278, caf336 and the caf344 are above 400mg/kg.The results of dissolving Potassium indicated that the soluble potassium's in 26 rhizobia nutrient fluid content were obviously higher than that of the comparison, and the soluble potassium contents of caf224(1.617), caf416(1.605), caf420(1.498), caf278(1.459), caf440(1.466), caf341(1.478), caf344(1.497), caf414(1.572) and caf415(1.532) was higher than that of the standard strain CGMCC1.2540(1.445).Generally analysed, there are 23 strains simultaneously dissolving phosphors and potassium ability, caf224, caf225, caf226, caf414, caf415, caf416, caf341, caf344, caf333, caf334, caf336, caf276, caf278, caf279, caf437, caf438, caf439, caf440, caf443, caf444, caf446, caf436 and caf420. And caf278, caf341, caf344, caf414, caf420 had stronger dissolving phosphors and potassium ability than the others.The Rhizobium tropici resistance tests discovered that: the pH growth scope was wide, from pH4 to pH11, caf224 and caf276 could endure pH3; bears salty ability, could endure 2%NaCl, caf226 could endure 4%NaCl; temperature compatible, all strains grew well from 20℃to 37℃,three strains caf224, caf414, caf336 could grow in 10℃, six strains caf224, caf226, caf414, caf415, caf438, caf276 could endure 45℃high temperature, moreover the strains had the tolerance to short time high temperature, caf414, caf276 could endure 50℃20min, caf224, caf226, caf415, caf416, caf438, caf279 could endure 60℃10min.From all above, the rhizobia, isolated from woody legume tree of Jianfengling Nature Reserve District, had diverisity in heredity and phenotype , for the 16S rDNA sequences, physiology and biochemistry characters and carbon utility abilities. It's found that the rhizobia, and obtained 23 strains which could dissolving Phosphors and Potassium, 4 of them also had many kinds of resistance, caf224, caf226, caf276 and caf414.