A Preliminary Study on Screening Adversity Resistance of Rhizobium Strains in Jianfengling Reserve and the 16S rDNA Determination

PENG Yu-hong; JIAO Ru-zhen; MOU Xin-tao

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A Preliminary Study on Screening Adversity Resistance of Rhizobium Strains in Jianfengling Reserve and the 16S rDNA Determination

1.
Research Institute of Forestry, Chinese Academy of Forestry
2.
Key Laboratory of Tree Breeding and Cultivation, State Forestry Administration, Beijing 100091, China

Abstract

In order to identify the resistance abilities of woody Legume Rhizobium, 9 strains isolated from roots nodules of different legume plants in the Jianfengling National Nature Reserve, Hainan Province, were preliminary studied. Results show that the optimal pH and NaCl concentration for growth are respectively 7 and 10.0 g·kg-1 for all the 9 isolates, but the temperatures are varied, the optimal growth temperature for CAF224 is 37 ℃, for CAF416, CAF438 and CAF279 is between 20-28 ℃, and for the other 5 strains is 28 ℃. All the 9 isolates can grow under pH 4-11, 20.0 g·kg-1 NaCl, and at 37 ℃, of which the isolates CAF224 and CAF276 could tolerate pH 3, while CAF226 could grow at 40.0 g·kg-1 NaCl; CAF276 could survive the high temperature of 50 ℃ for 20 min. and 60 ℃ for 10 min. The Rhizobium strains from different hosts and under different ecological conditions demonstrate different adversity resistances. 4 strains have higher resistances, which are CAF226, CAF276, CAF224 and CAF414. A phylogenetic analysis based on 16S rDNA gene sequences reveals that the 9 strains gather a cluster with Rhizobium tropici Clone H12 (GenBank EF054889) with a similarity of 99.4%. The 9 strains presented here are tentatively identified as Rhizobium tropici Martinez-Romero et al.
- Jianfengling,
- salt tolerance,
- pH scope,
- adversity resistance,
- 16S rDNA sequences,
- Rhizobium tropici Martinez-Romero et al.

References

[1]	张炎周,周立江.豆科植物在退耕还林中的应用[J]. 四川林勘设计,2002(2):1-7
[2]	de Lajudie P, Willems A. Polyphasic taxonomy of rhizobia: emendation of the genus Sinorhizobium and Description of Sinorhizobium meliloti comb. nov., Sinorhizobium saheli sp. nov., and Sinorhizobium teranga sp. nov.[J]. Int J Syst Bacteriol ,1994,44: 715-733
[3]	de Lajudie P, Anne Willems, Giselle Nick.Characterization of tropical tree rhizobia and description of Mesorhizobium plurifarium sp.nov[J]. International Journal of Systematic Bacteriology, 1998, 48: 369-382
[4]	康丽华,李素翠.相思根瘤菌耐酸性研究及耐酸菌株的筛选[J].林业科学研究,1998,11(6):581-585
[5]	韦原莲,黄宝灵,吕成群,等.马占相思根瘤菌生物学特性的研究[J].西北林学院学报,2006,21(5):120-124
[6]	黄宝灵,吕成群,韦原莲,等.相思树种根瘤菌的若干抗逆特性[J].南京林业大学学报:自然科学版,2004, 28(1):29-32
[7]	关桂兰,王卫卫,杨玉锁.新疆干旱地区固氮生物资源[M].北京: 科学出版社,1991
[8]	杨文权,郭军康,冯春生,等.宁夏豆科植物根瘤菌资源调查及其生态分布[J].干旱地区农业研究,25(5):176-181
[9]	张执欣,陈卫民,韦革宏,等.甘肃省部分地区豆科植物根瘤菌资源调查[J].西北农林科技大学学报:自然科学版, 2006, 34(2):77-82
[10]	韦革宏,龚明福,吕双庆.中国帕米尔高原根瘤菌一豆科植物共生资源调查[J].西北植物学,2005,25(8):1618-1622
[11]	师尚礼,刘建荣,张勃,等.甘肃寒旱区苜蓿根瘤菌抗逆性评价[J].草地学报,2007,15 (1):1-6
[12]	李东,黄莉莉,韩素芬.23株豆科木本植物根瘤菌16S rDNA序列分析[J].南京林业大学学报,2007,31(6): 117-120
[13]	陈文峰,陈文新.豆科树种刺槐、黄檀、合欢根瘤菌的数值分类及16S rDNA-PCR RFLP研究[J].应用与环境生物学报,2003,9(1) :53-58
[14]	刘杰.我国中东部地区紫穗槐、紫荆、紫藤根瘤菌的数值分类及16S rDNA PCR-RFLP研究[J].中国农业科学,2003, 36(1) :17-25
[15]	Odee D W, Sutherland JM, Makatiani E T,et al. Phenotypic characteristics and composition of rhizobia associated with woody legumes growing in diverse Kenyan conditions[J]. Plant and Soil, 1997(188): 65-75
[16]	Doignon-Bourcier R, Willems A, Coopman R, et al. Genotypic characterization of Bradyrhizobium strains undulating small Senegalese legumes by 16S-23s rRNA intergenic gene spacers and amplified fragment length polymorphism fingerprint analyse[J]. Applied and Environmental Microbiology,2000, 66: 3987-3997
[17]	周宇光.中国菌种目录[M].北京:化学工业出版社,2007
[18]	黄成敏,龚子同.海南岛尖峰岭地区山地土壤发生特性[J].山地学报,2000, l8(3):193-200
[19]	Michiels J, Verreth C, Vanderleyden J. Effects of temperature stress onbean nodulating Rhizobium strains[J]. Appl Environ Microbiol,1994,60:1206-1212
[20]	Graham P H. Stress tolerance in Rhizobium and Bradyrhizobium,and nodulation under adverse soil conditions[J]. Can J Microbiol, 1992,38: 475-484
[21]	Bordeleau L M,Prevost D.Nodulation and nitrogen fixation in extreme environments [J]. Plant Soil, 1994, 161: 115-124
[22]	Brockwell J,Pilka A,Holliday R A. Soil pH is a major determinant of the numbers of naturally-occurring Rhizobium meliloti in non-cultivated soils of New South Wales [J]. Aust J Exp Agric, 1991(31): 211-219
[23]	Hungria M, Franco A A. Effects of high temperature on nodulation and nitrogen fixation by Phaseolus vulgaris L.[J]. Plant Soil, 1993,149: 95-102

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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A Preliminary Study on Screening Adversity Resistance of Rhizobium Strains in Jianfengling Reserve and the 16S rDNA Determination

1. Research Institute of Forestry, Chinese Academy of Forestry

2. Key Laboratory of Tree Breeding and Cultivation, State Forestry Administration, Beijing 100091, China

Abstract: In order to identify the resistance abilities of woody Legume Rhizobium, 9 strains isolated from roots nodules of different legume plants in the Jianfengling National Nature Reserve, Hainan Province, were preliminary studied. Results show that the optimal pH and NaCl concentration for growth are respectively 7 and 10.0 g·kg-1 for all the 9 isolates, but the temperatures are varied, the optimal growth temperature for CAF224 is 37 ℃, for CAF416, CAF438 and CAF279 is between 20-28 ℃, and for the other 5 strains is 28 ℃. All the 9 isolates can grow under pH 4-11, 20.0 g·kg-1 NaCl, and at 37 ℃, of which the isolates CAF224 and CAF276 could tolerate pH 3, while CAF226 could grow at 40.0 g·kg-1 NaCl; CAF276 could survive the high temperature of 50 ℃ for 20 min. and 60 ℃ for 10 min. The Rhizobium strains from different hosts and under different ecological conditions demonstrate different adversity resistances. 4 strains have higher resistances, which are CAF226, CAF276, CAF224 and CAF414. A phylogenetic analysis based on 16S rDNA gene sequences reveals that the 9 strains gather a cluster with Rhizobium tropici Clone H12 (GenBank EF054889) with a similarity of 99.4%. The 9 strains presented here are tentatively identified as Rhizobium tropici Martinez-Romero et al.

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Reference (23)

[1]	张炎周,周立江.豆科植物在退耕还林中的应用[J]. 四川林勘设计,2002(2):1-7
[2]	de Lajudie P, Willems A. Polyphasic taxonomy of rhizobia: emendation of the genus Sinorhizobium and Description of Sinorhizobium meliloti comb. nov., Sinorhizobium saheli sp. nov., and Sinorhizobium teranga sp. nov.[J]. Int J Syst Bacteriol ,1994,44: 715-733
[3]	de Lajudie P, Anne Willems, Giselle Nick.Characterization of tropical tree rhizobia and description of Mesorhizobium plurifarium sp.nov[J]. International Journal of Systematic Bacteriology, 1998, 48: 369-382
[4]	康丽华,李素翠.相思根瘤菌耐酸性研究及耐酸菌株的筛选[J].林业科学研究,1998,11(6):581-585
[5]	韦原莲,黄宝灵,吕成群,等.马占相思根瘤菌生物学特性的研究[J].西北林学院学报,2006,21(5):120-124
[6]	黄宝灵,吕成群,韦原莲,等.相思树种根瘤菌的若干抗逆特性[J].南京林业大学学报:自然科学版,2004, 28(1):29-32
[7]	关桂兰,王卫卫,杨玉锁.新疆干旱地区固氮生物资源[M].北京: 科学出版社,1991
[8]	杨文权,郭军康,冯春生,等.宁夏豆科植物根瘤菌资源调查及其生态分布[J].干旱地区农业研究,25(5):176-181
[9]	张执欣,陈卫民,韦革宏,等.甘肃省部分地区豆科植物根瘤菌资源调查[J].西北农林科技大学学报:自然科学版, 2006, 34(2):77-82
[10]	韦革宏,龚明福,吕双庆.中国帕米尔高原根瘤菌一豆科植物共生资源调查[J].西北植物学,2005,25(8):1618-1622
[11]	师尚礼,刘建荣,张勃,等.甘肃寒旱区苜蓿根瘤菌抗逆性评价[J].草地学报,2007,15 (1):1-6
[12]	李东,黄莉莉,韩素芬.23株豆科木本植物根瘤菌16S rDNA序列分析[J].南京林业大学学报,2007,31(6): 117-120
[13]	陈文峰,陈文新.豆科树种刺槐、黄檀、合欢根瘤菌的数值分类及16S rDNA-PCR RFLP研究[J].应用与环境生物学报,2003,9(1) :53-58
[14]	刘杰.我国中东部地区紫穗槐、紫荆、紫藤根瘤菌的数值分类及16S rDNA PCR-RFLP研究[J].中国农业科学,2003, 36(1) :17-25
[15]	Odee D W, Sutherland JM, Makatiani E T,et al. Phenotypic characteristics and composition of rhizobia associated with woody legumes growing in diverse Kenyan conditions[J]. Plant and Soil, 1997(188): 65-75
[16]	Doignon-Bourcier R, Willems A, Coopman R, et al. Genotypic characterization of Bradyrhizobium strains undulating small Senegalese legumes by 16S-23s rRNA intergenic gene spacers and amplified fragment length polymorphism fingerprint analyse[J]. Applied and Environmental Microbiology,2000, 66: 3987-3997
[17]	周宇光.中国菌种目录[M].北京:化学工业出版社,2007
[18]	黄成敏,龚子同.海南岛尖峰岭地区山地土壤发生特性[J].山地学报,2000, l8(3):193-200
[19]	Michiels J, Verreth C, Vanderleyden J. Effects of temperature stress onbean nodulating Rhizobium strains[J]. Appl Environ Microbiol,1994,60:1206-1212
[20]	Graham P H. Stress tolerance in Rhizobium and Bradyrhizobium,and nodulation under adverse soil conditions[J]. Can J Microbiol, 1992,38: 475-484
[21]	Bordeleau L M,Prevost D.Nodulation and nitrogen fixation in extreme environments [J]. Plant Soil, 1994, 161: 115-124
[22]	Brockwell J,Pilka A,Holliday R A. Soil pH is a major determinant of the numbers of naturally-occurring Rhizobium meliloti in non-cultivated soils of New South Wales [J]. Aust J Exp Agric, 1991(31): 211-219
[23]	Hungria M, Franco A A. Effects of high temperature on nodulation and nitrogen fixation by Phaseolus vulgaris L.[J]. Plant Soil, 1993,149: 95-102

A Preliminary Study on Screening Adversity Resistance of Rhizobium Strains in Jianfengling Reserve and the 16S rDNA Determination

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通讯作者: 陈斌, bchen63@163.com

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A Preliminary Study on Screening Adversity Resistance of Rhizobium Strains in Jianfengling Reserve and the 16S rDNA Determination

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