西北部分地区苦马豆根瘤菌的遗传多样性

doi:10.3724/SP.J.1003.2009.08218

摘要/Abstract

摘要：

苦马豆(Sphaerophysa salsula)是荒漠区重要的豆科植物。为了研究其共生根瘤菌的多样性, 本试验采用16S rDNA PCR-RFLP和16S rDNA全序列分析方法, 对西北部分地区的苦马豆根瘤菌进行了遗传多样性及系统发育分析。结果表明, 57株供试菌株共产生了9种遗传图谱类型, 对每种图谱类型的代表性菌株进行16S rDNA全序列分析的结果表明, 它们分别归属于中慢生根瘤菌属(Mesorhizobium)、根瘤菌属(Rhizobium)、中华根瘤菌属(Sinorhizobium)、土壤杆菌属(Agrobacterium)、叶杆菌属(Phyllobacterium)和Shinella kummerowiae。不同地域的菌株在多样性方面也有明显差异: 分离自银川的苦马豆根瘤菌的Jaccard相似性系数较低; 而来自民乐县和临泽县的菌株有着非常丰富的遗传多样性, 其Simpson指数分别为0.826和0.710, Shannon-Wiener指数分别为1.831和1.530。以上结果为进一步确定西北地区豆科植物根瘤菌的系统分类地位提供了依据。

关键词: Sphaerophysa salsula, 系统发育, 16S rDNA PCR-RFLP, 16S rDNA全序列, 遗传多样性

Abstract

Sphaerophysa salsula is a highly nutritive and drought-tolerant perennial grass distributed in mid-Asia and northwestern China. This legume plant is highly prized for the revegetation of drought and high alkali soils. Our objective was to better understand the diversity and phylogeny of rhizobia collected from nodules of Sphaerophsa salsula in different regions of northwestern China. We estimated genetic diversity using 16S rDNA PCR-RFLP and 16S rDNA sequencing. Nine genotypes were tested from 57 strains. 16S rDNA sequencing revealed that these rhizobial strains belonged to the following genera: Mesorhizobium, Rhizobium, Sinorhizobium, Agrobacterium, Phyllobacterium and Shinella kummerowiae. The Jaccard similarity coefficient of the strains from Yinchuan was low while the strains isolated from Minle and Linze had rich diversity, with Simpson indices of 0.826 and 0.710, respectively, and Shannon-Wiener indices of 1.831 and 1.530, respectively. This study provides basic information for the phylogeny of rhizobia in northwestern China.

Key words: Sphaerophysa salsula, phylogeny, 16S rDNA PCR-RFLP, 16S rDNA sequencing, genetic diversity

徐琳, 徐佳洁, 刘巧莉, 谢瑞美, 韦革宏 (2009) 西北部分地区苦马豆根瘤菌的遗传多样性. 生物多样性, 17, 69-75. DOI: 10.3724/SP.J.1003.2009.08218.

Lin Xu, Jiajie Xu, Qiaoli Liu, Ruimei Xie, Gehong Wei (2009) Genetic diversity in rhizobia isolated from Sphaerophysa salsula in several regions of northwestern China. Biodiversity Science, 17, 69-75. DOI: 10.3724/SP.J.1003.2009.08218.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2009.08218

https://www.biodiversity-science.net/CN/Y2009/V17/I1/69

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参考文献 22

[1]	Beringer JE (1974) R-factor transfer in Rhizobium leguminosarum. Journal of General Microbiology, 84, 188-198.
[2]	Chen WX (陈文新), Wang ET (汪恩涛), Chen WF (陈文峰) (2004) The relationship between the symbiotic promiscuity of rhizobia and legumes and their geographical environments. Scientia Agricultura Sinica (中国农业科学), 37, 81-86. (in Chinese with English abstract)
[3]	Gao LF (高丽锋), Deng X (邓馨), Wang HX (王洪新) Hu ZA (胡志昂) (2004) Diversity and resistance of rhizobia isolated from Caragana intermedia in Maowusu sandland. Chinese Journal of Applied Ecology (应用生态学报), 15, 44-48. (in Chinese with English abstract)
[4]	Gaunt MW, Turner SL, Rigottier GL, Rigottier-Gois L, Lloyd-Macgilp SA, Young JPW (2001) Phylogenies of atpD and recA support the small subunit rRNA-based classification of rhizobia. International Journal of Systematic and Evolutionary Microbiology, 51, 2037-2048.
[5]	Ma KP (马克平) (1994) The measurement of community diversity. In: Principles and Methodologies of Biodiversity Studies (生物多样性研究的原理与方法), pp. 141-165, China Science and Technology Press, Beijing.
[6]	Mhamdi R, Mrabet M, Laguerre G, Tiwari R, Aouani ME (2005) Colonization of Phaseolus vulgaris nodules by Agrobacterium-like strains. Canadian Journal of Microbiology, 51, 105-111. URL PMID
[7]	Martens M, Dawyndt P, Coopman R, Gillis M, De Vos P, Willems A (2008) Advantages of multilocus sequence analysis for taxonomic studies: a case study using 10 housekeeping genes in the genus Ensifer (including former Sinorhizobium). International Journal of Systematic and Evolutionary Microbiology, 58, 200-214.
[8]	Peng GX, Tan ZY, Wang ET, Reinhold-Hurek B, Chen WF and Chen WX (2002) Identification of isolates from soybean nodules in Xinjiang region as Sinorhizobium xinjiangense and genetic differentiation of S. xinjiangense from Sinorhizobium fredii. International Journal of Systematic and Evolutionary Microbiology, 52, 457-462.
[9]	Selenska-Pobell S, Evguenieva-Hackenberg E, Radeva G, Squartini A (1996) Characterization of Rhizobium‘hedysari’by RFLP analysis of PCR amplified rDNA and by genomic PCR fingerprinting. Journal of Applied Bacteriology, 80, 517-528.
[10]	Stackebrandt E, Frederiksen W, Garrity GM, Grimont PAD, Kampfer P, Maiden MCJ, Nesme X, Rossello-Mora R, Swings J, Truper HG, Vauterin L, Ward AC, Whitman WB (2002) Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology. International Journal of Systematic and Evolutionary Microbiology, 52, 1043-1047. URL PMID
[11]	Tan ZY (谭志远), Chen WX (陈文新) (1997) Sequencing the 16S rDNA of representative strain of new rhizobial group and determining of its phylogenetic relationship. Acta Microbiologica Sinica (微生物学报), 37, 411-416. (in Chinese with English abstract)
[12]	Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25, 4876-4882. URL PMID
[13]	van de Peer Y, De Wachter R (1994) TREECON for Windows: a software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Computer Applications in the Bioscience, 10, 569-570.
[14]	Vandamme P, Pot B, Gillis M, de Vos P, Kersters K, Swings J (1996) Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiological Reviews, 60, 407-438.
[15]	Vincent JM (1970) A Manual for the Practical Study of Rroot-Nodule Bacteria. IBP Handbook 15. Blackwell Scientific Publications, Oxford.
[16]	Wang LL, Wang ET, Liu J, Li Y, Chen WX (2006) Endophytic occupation of root nodules and roots of Melilotus dentatus by Agrobacterium tumefaciens. Microbial Ecology, 52, 436-443.
[17]	Wu D (吴达), Shi YP (师彦平), Liang B (梁冰), Wang JH (王建华) (2003) Advances in studies on swainsonine. Chinese Traditional and Herbal Drugs (中草药), 34, A5-7. (in Chinese)
[18]	Xu L (徐莉), Yang JK (杨江科), Zhang WT (张伟涛), Yuan TY (袁天英), Zhou JC (周俊初) (2007) Phenotypic and genetic diversity of cowpea rhizobia. Acta Ecologica Sinina (生态学报), 27, 3655-3662. (in Chinese with English abstract)
[19]	Yang J, Hu Z, Guo GQ, Zheng GC (2001) In vitro plant regeneration from cotyledon explants of Swainsona salsula Taubert. Plant Cell, Tissue and Organ Culture, 66, 35-39.
[20]	Yao RC (姚荣成) (2003) Advances in studies on Swainsona salsula. The Journal of Pharmaceutical Practice (药学实践杂志), 21, 43-44. (in Chinese)
[21]	Zhang Y (张英) (1999) The nutrition and feed value of Sphaerophysa salsula Tauberts. Feed Research (饲料研究), (7), 22. (in Chinese)
[22]	Zou L (邹林), Yang MQ (杨鸣琦) (2006) Progress on anti-tumor activity of swainsonine. Progress in Veterinary Medicine (动物医学进展), 27(4), 36-38. (in Chinese with English abstract)

酶切类型 Fingerprint pattern	分离地 Location
酶切类型 Fingerprint pattern	甘肃民乐县 Minle County, Gansu	甘肃张掖市 Zhangye City, Gansu	甘肃临泽县 Linze County, Gansu	甘肃玉门市 Yumen City, Gansu	宁夏银川市 Yinchuan City, Ningxia	新疆伊犁市 Yili City, Xinjiang
AAB	GS0181, GS0185, GS0188, GS0192, GS0198	GS0215	GS0240
BBD	GS0184		GS0218	GS0237
CCB	GS0170, GS0171 GS0168		GS0219, GS0223, GS0227, GS0230, GS0231, GS0232		NX0171, NX0172, NX0173	XJ0216
CEB	GS0176, GS0196, GS0199, GS0200, GS0201, GS0204	GS0206, GS0209 GS0210, GS0214	GS0216, GS0233 GS0234
DDB			GS0228
DFB	GS0172, GS0202
EDB	GS0177, GS0178, GS0187			GS0238
EDC	GS0166, GS0169, GS0173, GS0180	GS0207, GS0211, GS0212, GS0213	GS0220			XJ0217, XJ0218, XJ0219, XJ0221
FDC						XJ0229

酶切类型 Fingerprint pattern	分离地 Location
酶切类型 Fingerprint pattern	甘肃民乐县 Minle County, Gansu	甘肃张掖市 Zhangye City, Gansu	甘肃临泽县 Linze County, Gansu	甘肃玉门市 Yumen City, Gansu	宁夏银川市 Yinchuan City, Ningxia	新疆伊犁市 Yili City, Xinjiang
AAB	GS0181, GS0185, GS0188, GS0192, GS0198	GS0215	GS0240
BBD	GS0184		GS0218	GS0237
CCB	GS0170, GS0171 GS0168		GS0219, GS0223, GS0227, GS0230, GS0231, GS0232		NX0171, NX0172, NX0173	XJ0216
CEB	GS0176, GS0196, GS0199, GS0200, GS0201, GS0204	GS0206, GS0209 GS0210, GS0214	GS0216, GS0233 GS0234
DDB			GS0228
DFB	GS0172, GS0202
EDB	GS0177, GS0178, GS0187			GS0238
EDC	GS0166, GS0169, GS0173, GS0180	GS0207, GS0211, GS0212, GS0213	GS0220			XJ0217, XJ0218, XJ0219, XJ0221
FDC						XJ0229

来源 Location	Jaccard相似性系数 Jaccard similarity coefficient					Simpson 指数 Simpson index	Shannon-Wiener指数 Shannon-Wiener index
来源 Location	甘肃张掖市 Zhangye City, Gansu	甘肃临泽县 Linze County, Gansu	甘肃玉门市 Yumen City, Gansu	宁夏银川市 Yinchuan City, Ningxia	新疆伊犁市 Yili City, Xinjiang	Simpson 指数 Simpson index	Shannon-Wiener指数 Shannon-Wiener index
甘肃张掖市 Zhangye City, Gansu						0.593	0. 965
甘肃临泽县 Linze County, Gansu	0.50					0.710	1.530
甘肃玉门市 Yumen City, Gansu	0.25	0.14				0.500	0.693
宁夏银川市 Yinchuan City, Ningxia	0.00	0.17	0.00			0.000	0.000
新疆伊犁市 Yili City, Xinjiang	0.20	0.30	0.00	0.33		0.500	0.868
甘肃民乐县 Minle County, Gansu	0.38	0.56	0.25	0.13	0.22	0.826	1.831

来源 Location	Jaccard相似性系数 Jaccard similarity coefficient					Simpson 指数 Simpson index	Shannon-Wiener指数 Shannon-Wiener index
来源 Location	甘肃张掖市 Zhangye City, Gansu	甘肃临泽县 Linze County, Gansu	甘肃玉门市 Yumen City, Gansu	宁夏银川市 Yinchuan City, Ningxia	新疆伊犁市 Yili City, Xinjiang	Simpson 指数 Simpson index	Shannon-Wiener指数 Shannon-Wiener index
甘肃张掖市 Zhangye City, Gansu						0.593	0. 965
甘肃临泽县 Linze County, Gansu	0.50					0.710	1.530
甘肃玉门市 Yumen City, Gansu	0.25	0.14				0.500	0.693
宁夏银川市 Yinchuan City, Ningxia	0.00	0.17	0.00			0.000	0.000
新疆伊犁市 Yili City, Xinjiang	0.20	0.30	0.00	0.33		0.500	0.868
甘肃民乐县 Minle County, Gansu	0.38	0.56	0.25	0.13	0.22	0.826	1.831