陕西等地区豆科植物根瘤内生细菌遗传多样性和系统发育研究Studies on the Diversity and Phylogeny of Endophytes Isolated from Root Nodules of Legumes in Shaanxi and Other Regions
齐雅琳;王金平;李正;韩素贞;
摘要(Abstract):
采用16S r DNA PCR-RFLP、16S r DNA全序列分析、BOX-PCR等多相分类技术,对采集自陕西神木、甘肃天水、青海泽库地区的豆科植物根瘤内生细菌资源的多样性进行了研究。从采集的豆科植物根瘤中共分离到50株供试菌株,对这50株供试菌株和8株参比菌株进行了16S r DNA PCR-RFLP聚类分析。结果显示,全部供试菌株在81.5%的相似性水平上分成4个分支(在86%相似性水平上,分支I包括3个群,分支III包括3个群)。选取各群代表菌株进行16S r DNA测序,确定了50株供试菌株中,1株属于慢生根瘤菌属(Bradyrhizobium),1株属于土壤杆菌属(Agrobacterium),6株属于中华根瘤菌属(Sinorhizobium),27株属于芽孢杆菌属(Bacillus),2株属于类芽孢杆菌属(Paenibacillus),其余13株则分别归属于寡养单胞菌属(Stenotrophomonas)、假单胞菌属(Pseudomonas)和泛菌属(Pantoea)。表明16S r DNA PCR-RFLP和16S r DNA全序列分析结果具有较好的一致性。对部分16Sr DNA序列相似性达到100%的菌株进行BOXPCR来确定是否为同一克隆。研究结果表明,陕西神木等地区豆科植物根瘤内生菌具有很丰富的遗传多样性。
关键词(KeyWords): 根瘤内生菌;根瘤菌系统发育;多样性
基金项目(Foundation):
作者(Author): 齐雅琳;王金平;李正;韩素贞;
Email:
DOI: 10.13989/j.cnki.0517-6611.2015.18.019
参考文献(References):
- [1]刘杰,汪恩涛,陈文新.豆科植物根瘤内生细菌的发现及其研究进展[J].微生物学报,2011,51(8):1001-1006.
- [2]WILSON D.Endophyte:The evolution of a term,and clarification of its use and definition[J].Plant Soil,1995,73:274-276.
- [3]ARKHIPOVA T N,VESELOV S U,MELENTIEV A I,et al.Ability of bacterium Bacillus subtilis to produce cytokinins and to influence the growth and endogenous hormone content of lettuce plants[J].Plant Soil,2005,272:201-209.
- [4]HALLMANN J,QUADT-HALLMANN A,MAHAFFEE W F,et al.Bacterial endophytes in agricultural crops[J].Canadian Journal of Microbiology,1997,43:895-914.
- [5]冯永君,宋未.植物内生细菌[J].自然杂志,2001,23(5):249-252.
- [6]VINCENT J.The cultivation,isolation and maintenance of rhizobia[M]//VINCENT J M.A manual for the practical study of the root-nodule bacteria.Oxford:Blackwell Scientific,1970:1-13.
- [7]SMIBERT R,KRIEG N.Phenotypic characterization[M]//GERHARDT P,MURRAY R G E,WOOD W A,et al.Methods for general and molecular bacteriology.Washington DC:American Society for Microbiology,1994:607-654.
- [8]CHEN W P,KUO T T.A simple and rapid method for the preparation of gram-negative bacterial genomic DNA[J].Nucleic Acids Research,1993,21(9):2260.
- [9]WEISBURG W G,BARNS S M,PELLETIER D A,et al.16S Ribosomal DNA amplification for phylogenetic study[J].Journal of Bacteriology,1991,173(2):697-703.
- [10]毕江涛,贺达汉,韦革宏,等.沙冬青根瘤菌遗传多样性和系统发育分析[J].西北植物学报,2009,29(4):695-703.
- [11]LAGUERRE G,MAVINGUI P,ALLARD M,et al.Typing of rhizobia by PCR DNA fingerprinting and PCR-restriction fragment length polymorphism analysis of chromosomal and symbiotic gene regions:Application to Rhizobium legu minosarum and its different biovars[J].Applied and Environmental Microbiology,1996,62:2029-2036.
- [12]SAITOU N,NEI M.The neighbor-joining method:A new method for reconstructing phylogenetic trees[J].Molecular Biology and Evolution,1987,4:406-425.
- [13]KIMURA M.A simple method for estimationg evolutionary rates of base substitutions through comparative studies of nucleotide sequences[J].Journal of Molecular Biology,1980,16:111-120.
- [14]JUKES T H,CANTOR C R.Evolution of protein molecule[M]//MUNTO H N.Mammalian protein metabolism.New York:Academic Press,1969:21-132.
- [15]FELSENSTEIN J.Confidence limits on phylogenies:An approach using the bootstrap[J].Evolution,1985,39:783-791.
- [16]VERSALOVIC J,SCHNEIDER M,DE BRUIJIN F J,et al.Genomic fingerprinting of bacteria using repetitive sequence based polymerase chain reaction[J].Methods in Molecular and Cellular Biology,1994,5:25-40.
- [17]杨凤环,李正楠,姬惜珠,等.BOX-PCR技术在微生物多样性研究中的应用[J].微生物学通报,2008,35(8):1282-1286.
- [18]SCHOLLA M H,ELKAN G H.Rhizobium fredii sp.nov.a fast-growing species that effectively nodulates soybeans[J].International Journal of Systematic Bacteriology,1984,34:484-486.
- [19]CHEN W X,YAN G H,LI J L.Numerical taxonomic study of fast-growing soybean rhizobia and a proposal that Rhizobium fredii be assigned to Sinorhzobium gen.nov[J].International Journal of Systematic Bacteriology,1988,38:392-397.
- [20]HAN L L,WANG E T,LU Y L,et al.Bradyrhizobium spp.and Sinorhizobium fredii are predo minant in root nodules of Vigna angularis,a native legume crop in the subtropical region of China[J].The Journal of Microbiology,2009,47:287-296.
- [21]KHBAYA B,NORMAND P.Genetic diversity and phylogeny of rhizobia that nodulate Aacacia spp.in morocco assessed by analysis of rRNA genes[J].Applied and Environmental Microbiology,1998,64:4912-4917.
- [22]AGUILAR O M,LPEZ M V,RICCILLO P M.The diversity of rhizobia nodulating beans in Northwest Argentina as a source of more efficient inoculants strains[J].Journal of Biotechnology,2001,91:181-188.
- [23]SALDANA G,MARTINEZ-ALCNTARA V,VINARDELL J M,et al.Genetic diversity of fast-growing rhizobia that nodulate soybean(Glycine max L.Merr)[J].Archives of Microbiology,2003,180:45-52.
- [24]TOLEDO I,LLORET L,MARTHEZ-ROMERO E.Sinorhizobium americanus sp.nov.,a new Sinorhizobium species nodulating native Acacia spp.in Mexico[J].Systematic and Applied Microbiology,2003,26:54-64.
- [25]陈强,陈文新,张小平,等.四川省部分豆科植物根瘤菌的遗传多样性[J].应用与环境生物学报,2008(14):83-89.
- [26]陈文新,汪恩涛.中国根瘤菌[M].北京:科学出版社,2011.
- [27]TAN Z Y,XU X D,WANG E T,et al.Phylogenetic and genetic relationships of Mesorhizobium tianshanense and related rhizobia[J].International Journal of Systematic Bacteriology,1997,47:874-879.
- [28]DE LAJUDIE P,WILLEMS A,NICK G,et al.Agrobacterium bv strains isolated from nodules of tropical legumes[J].Systematic and Applied Microbiology,1999,22:119-132.
- [29]LIU J,WANG E T,CHEN W X.Diverse rhizobia associated with woody legumes Wisteria sinensis,Cercis racemosa and Amoroha fruticosa grown in the temoerate zone of China[J].Systematic and Applied Microbiology,2005,28(5):465-477.
- [30]KAN F L,CHEN Z Y,WANG E T,et al.Characterization of symbiotic and endophytic bacteria isolated from root nodules of herbaceous legumes grown in Qinghai-Tibet Plateau and on other zones of China[J].Archives of Microbiology,2007,188:103-115.
- [31]LIU X Y,WANG E T,L I Y,et al.Diverse bacteria isolated from root nodules of Trifolium,Crotalaria and Mimosa grown in the subtropical regions of China[J].Archives of Microbiology,2007,188:1-14.
- [32]LI J H,WANG E T,CHEN W X,et al.Genetic diversity and potential for promotion of plant growth detected in nodule endophytic bacteria of soybean grown in Heilongjiang province of China[J].Soil Biology and Biochemistry,2008,40:238-246.
- [33]AN Z Y,WANG E T,PENG G X,et al.Characterization of bacteria isolated from wild legumes in the north-western regions of China[J].International Journal of Systematic and Evolutionary Microbiology,1999,49:1457-1469.
- [34]AO J L,TURNER S L,KAN F L,et al.Mesorhizobium septentrionale sp.nov.and Mesorhizobium temperatum sp.nov.,isolated from Astragalus adsurgens growing in the northern regions of China[J].International Journal of Systematic and Evolutionary Microbiology,2004,54:2003-2012.
- [35]MRABET M,MNASRI B,ROMDHANE S B,et al.Agrobacterium strains isolated from root nodules of common bean specifically reduce nodulation by Rhizobium gallicum[J].FEMS Microbiology Ecology,2006,56:304-309.
- [36]SOHAIL H,SUMERA Y,KAUSER A M,et al.Rhizobium,Bradyrhizobium and Agrobacterium strains isolated from cultivated legumes[J].Biology and Fertility of Soils,2004,39(3):179-185.
- [37]BAI Y,D’AOUST F,SMITH D,et al.Isolution of plant-growth-promoting Bacillus strain from soybean root nodules[J].Canadian Journal of Microbiology,2002,48:230-238.
- [38]ZHANG Y Z,CHEN WF,LI M,et al.Bacillus endoradici ssp.nov.,an endophytic bacterium isolated from soybean root[J].International Journal of Systematic and Evolutionary Microbiology,2012,62:359-363.
- [39]ASIS C,ADACHI K.Isolution of endophytic diazotroph Pantoea agglomerans and nondiazotroph Enterobacter asburiae from sweet potato stem in Japan[J].Letters in Applied Microbiology,2004,38:19-23.
- [40]DI GREGORIO S,LAMPIS S,VALLINI G.Selenite precipitation by a rhizospheric strain of Stenotrophomonas sp.isolated from the root system of Astragalus bisulcatus:a biotechnological perspective[J].Environment International,2005,31:233-241.
- [41]CARRO L,FLORES-FLIX J D,CERDA-CASTILLO E,et al.Paenibacillus endophyticus sp.nov.,isolated from nodules of Cicer arietinum[J].International Journal of Systematic and Evolutionary Microbiology,2013,63:4433-4438.
- [42]ZAKHIA F,JEDER H,WILLEMS A,et al.Diverse bacteria associated with root nodules of spontaneous legumes in Tunisia and first report for nif H-like gene within the genera Microbacterium and Starkeya[J].Microbial Ecology,2006,51(3):375-393.
- [43]VALVERDE A,PEIX A,RIVAS R,et al.Paenibacillus castaneae sp.nov.,isolated from the phyllosphere of Castanea sativa Miller[J].International Journal of Systematic and Evolutionary Microbiology,2008,58:2560-2564.
- [44]VALVERDE A,FTERICH A,MAHDHI M,et al.Paenibacillus prosopidis sp.nov.,isolated from the nodules of Prosopis farcta[J].International Journal of Systematic and Evolutionary Microbiology,2010,60:2182-2186.
- [45]SON S H,KHAN Z,KIM Y H.Plant growth-promoting rhizobacteria,Paenibacillus polymyxa and Paenibacillus lentimorbus suppress disease complex caused by root-knot nematode and fusarium wilt fungus[J].Journal of Applied Microbiology,2009,107:525-532.
- [46]ZHANG J,WANG Z T,YU H M,et al.Paenibacillus catalpa sp.nov.,isolated from the rhizosphere soil of Catalpa speciosa[J].International Journal of Systematic and Evolutionary Microbiology,2013,63:1776-1781.
- [47]CARRO L,FLORES-FLIX J D,RAMIREZ BAHENA M H,et al.Paenibacillus lupini sp.nov.,isolated from nodulesof Lupinus albus[J].International Journal of Systematic and Evolutionary Microbiology,2014,64:3028-3033.