费氏中华根瘤菌共生质粒扩增对结瘤因子组分和共生固氮能力的影响

应用生态学报 ›› 2003, Vol. ›› Issue (9): 1517-1520.

费氏中华根瘤菌共生质粒扩增对结瘤因子组分和共生固氮能力的影响

缪礼鸿, 周俊初

华中农业大学教育部微生物重点实验室, 武汉 430070

收稿日期:2001-10-29 修回日期:2002-04-01 出版日期:2003-09-15
通讯作者: 周俊初
基金资助:
欧盟INCO-DC国际合作项目(2001AA214021);国家基础研究发展规划资助项目(001CB1089)

Effect of Sinorhizobium fredii YC4 symbiotic plasmid amplification on nod factors and symbiotic N-fixation

MIAO Lihong, ZHOU Junchu

Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China

Received:2001-10-29 Revised:2002-04-01 Online:2003-09-15

摘要/Abstract

摘要： 费氏中华根瘤菌(Sinorhizobium fredii)YC4能在大豆(Glycine max)和野大豆(G.soia)上形成正常固氮的根瘤。人工培养条件下用¹⁴C标记的薄层层析(TLC)法检测根瘤菌产生的结瘤因子(LCOs)的结果表明,与其它4株费氏中华根瘤菌相比,YC4产生的LCOs含有较多的疏水性基因。从YC4菌株中分离到1株共生质粒发生了扩增的自发突变株YSC3,其产生的LCOs中含有较野生型菌株多的1个疏水性组分,28℃培养条件下产生的LCOs量亦较YC4显著增加。结瘤试验结果表明,YSC3菌株只能在大豆和野大豆上形成无效的根瘤。

Abstract: Sinorhizobium fredii YC4 can form nitrogen-fixing nodules on soybean (Glycine max) and wild soybean (G.soja).It can produce unique lipochitooligosaccharide nod factors (LCOs), in comparison with the other four strains of S.fredii.The constitution of LCOs produced by YC4 contained more hydrophobic substitutions detected by Thin-layer chromatography (TLC) of ¹⁴C-labeled nod factors.Aspontaneous mutant termed YSC3 amplified in the symbiotic plasmid was isolated from YC4, which can produce more amount of LCOs than its parental strain at 28 ℃,and showed a difference in the construction of LCOs.Nodulation test indicated that YSC3 only formed ineffective nodules on soybean G.soja.

中图分类号:

Q939.113

缪礼鸿, 周俊初. 费氏中华根瘤菌共生质粒扩增对结瘤因子组分和共生固氮能力的影响[J]. 应用生态学报, 2003, (9): 1517-1520.

MIAO Lihong, ZHOU Junchu . Effect of Sinorhizobium fredii YC4 symbiotic plasmid amplification on nod factors and symbiotic N-fixation[J]. Chinese Journal of Applied Ecology, 2003, (9): 1517-1520.

参考文献

[1] Bec-Ferte MP, Krishnan HB, Savagnac A, et al. 1996. Rhizobium fredii synthesizes an array of lipooligosaccharides, including a novel compound with glucose inserted into the backbone of molecule.FEBS Lett, 339: 273～279
[2] Brom S, Garcia de los Santos A, Girard ML, et al. 1991. Highfrequency rearrangements in Rhizobium leguminosarum by.phaseoli plasmids. JBacterial, 173:1344～1346
[3] Buendia-Claveria, Chamber AM, Ruiz-Sainz JE. 1989. A comparative study of physiological characteristics, plasmid content and symbiotic properties of different Rhizobium fredii strains. Syst Appl Microbiol, 12:210～215
[4] Cren M, Kondorosi A, Kondorosi E. 1995. NolR controls expression of the Rhizobium meliloti nodulation genes involved in the core nod factor synthesis. Mol Microbiol, 15: 733～747
[5] Eckhardt T. 1978. A rapid method for the identification of plasmid deoxyribonucleic acid in bacteria. Plasmid, 1:581～588
[6] Fellay R, Hanin M, Montorzi G, et al. 1998. nodD2 of Rhizobium sp. NGR²34 is involved in the repression of the nodABC operon. Mol Microbiol, 27:1039～1050
[7] Frelberg C, Fellay R, Balroch A, et al. 1997.Molecular basis of symbiosis between Rhizobium and legumes.Nature, 387:394～4014
[8] Hynes MF, Jurgen Q, O' Connell MP, et al. 1989. Direct selection for curing and deletion of Rhizobium plasmids using transposom carrying the Bacillus subtilis sacB gene. Gene, 78:111～120
[9] Kim CH, Tull RE, Keister DL. 1989. Exopolysaccharide-deficient mutants of Rhizobium fredii HH303 which are symbiotically effecitive. Appl Environ Microbiol, 55: 1852～1854
[10] Kiss E, Mergaert P, Olah B, et al. 1998. Conservation of nolR in the Sinorhizobium and Rhizobium genera of the Rhizobiaceae family. Mol Plant-Microbe Interact, 11:1186～1195
[11] Mavingui P, Laeremans T, Flores M, et al. 1998. Genes essential for nod factor production and nodulation are located on a symbiotic amplicon (AMPRtr CFN299pc60) in Rhizobium tropici. J Bacteriol, 180: 2866～2874
[12] Mergaert P, Montagu MV, Holsters M. 1997. Molecular mechanisms of nod factor diversity. Mol Microbiol,25 : 811～817
[13] Olsthoorn MMA, Stokvis E, Haverkamp J, et al. 2000. Growth temperature regulation of host-specific modifications of rhizobial lipo-chintin oligosaccharides: the function of nodx is temperature regulated. Mol Plant-Microbe Interact, 13: 808～820
[14] Perret A, Staehelin C, Broughton WJ. 2000. Molecular basis of symbiotic promiscuity. Microbiol Mol Biol Rev, 14:180～201
[15] Pueppke SG, Broughton WJ. 1999. Rhizobium sp. strain NGR²34and R. fredii USDA257 share exceptionally broad, nested host range. Mol Plant-Microbe Interact, 12:293～318
[16] Pueppke SG, Ruiz-Sainz JE, Spaink HP, et al. 1999. Mutation in GDP-Fucose synthesis genes of Sinorhizobium fredii alters nod factors and significantly decreases competitiveness to nodulate soybeans. Mol Plant-Microbe Interact, 12:207～217
[17] Romero D, Martinez-Salazar J, Girara L, et al. 1995. Discrete amplifiable region(amplicons)in the symbiotic plasmid of Rhizobium etli CFN42. J Bacteriol, 177: 973～980
[18] Romero D, Brom S, Martinez-Salazar J, et al. 1991. Amplification and deletion of nod-nif region in symbiotic plasmid of Rhizobium phaseoli. J Bacteriol, 173: 2435～2441
[19] Spaink HP, Aarts A, Stacey G, et al. 1992. Detection and separation of Rhizobium and Bradyrhizobium nod metabolites using thin-layer chromatography. Mol Plant-Microbe Interact, 5: 72～80
[20] Vincent JMA. 1970. Manual for the Practical Study of Root-nodule Bacteria. Oxford: Blace Well Scientific Publication.
[21] Zhang XX, Kosier B, Pricfer UB. 2001. Symbiotic plasmid rearrangement in R hizobium leguminosarum by. viciae VF39SM. J Bacteriol, 183:2141～2144