鸡毛松根瘤内生细菌的初步研究
摘要
本论文从广西十万大山海拔600米处一棵与红豆伴生上百年的鸡毛松根瘤中分离到13株内生细菌,从红豆中分离到3株内生细菌,从移栽回种植的鸡毛松小苗春季根瘤中分离到9株内生细菌,鸡毛松小苗夏季根瘤中分离到20株内生细菌。
本论文分离的42株鸡毛松根瘤内生细菌和3株红豆根瘤内生细菌菌株中,有9株菌株革兰氏染色阳性。有20%的菌株能水解淀粉,20%的菌株水解明胶,87%的菌株能利用柠檬酸盐,全部菌株都能利用葡萄糖、半乳糖、已二酸、D-果糖、阿拉伯糖、蔗糖、乳糖7种碳源和精氨酸、组氨酸、天冬氨酸、磷酸氢二氨、硝酸钾5种氮源。
采用16S rDNA序列分析和16S rDNA PCR-RFLP的方法确定这些菌株的分类地位。所有45株菌株共产生7种遗传图谱类型,聚类分析结果表明,在79%的相似水平上,45株未知菌株构成了7个遗传表观群。16S rDNA序列分析和聚类结果表明鸡毛松根瘤内生细菌与红豆根瘤内生细菌都表现出多样性。菌株个数组成在16S rDNA PCR-RFLP聚类和数值聚类结果基本一致。
16SrDNA全序列分析表明,45株菌株属于6个属,分别为不动杆菌(Acinetobacter sp)、假单胞菌(Pseudomonas sp)、类芽孢杆菌(Paenibacillussp)、寡养单胞菌属(Stenotrophomonas sp)、芽孢杆菌(Bacillus sp)和葡萄球菌属(Staphylococcus sp)。
On the 600 meters high place of Shiwandashan Mountain in Guangxi Province, we found one Podocarpus imbricatus that grown with a Taxus for hundreds of years, from which we separated 13 endogenous bacteria, 3 endogenous bacteria from the Taxus. We also separated 9 endogenous bacteria from the vernal P. imbricatus. seedlings nodule which was transplanted, and 20 endogenous bacteria from aestival P. imbricatus. seedlings nodule.
Among 42 endogenous bacteria of P. imbricatus. and 3 endogenous bacteria of Taxus nodule, 9 strains are Gram-positive. 20 percent of the strains are hydrolyzed starch positive, and 20 percent of the strains are hydrolyzed gelatin positive, 87 percent of the strains could utilize citrate. All strains are able to use 7 kinds of carbon and 5 nitrogen.
Using the 16S rDNA sequence analysis and 16S rDNA PCR-RFLP to determine the classification status of these strains. There are totally 7 kinds of genetic map type generated from these 45 strains. The cluster analysis showed that in 79 percent of the similar level, 45 unknown strains constitute 7 apparent group. There a diversity from the endogenous bacteria of P. imbricatus. and Taxus nodule based on 16S rDNA sequence analysis and the cluster tree.
The 16SrDNA sequence analysis showed that the 45 strains belong to 6 categories, which are Acinetobacter sp, Pseudomonas sp, Paenibacillus sp, Stenotrophomonas sp, Bacillus sp and Staphylococcus sp.
本论文分离的42株鸡毛松根瘤内生细菌和3株红豆根瘤内生细菌菌株中,有9株菌株革兰氏染色阳性。有20%的菌株能水解淀粉,20%的菌株水解明胶,87%的菌株能利用柠檬酸盐,全部菌株都能利用葡萄糖、半乳糖、已二酸、D-果糖、阿拉伯糖、蔗糖、乳糖7种碳源和精氨酸、组氨酸、天冬氨酸、磷酸氢二氨、硝酸钾5种氮源。
采用16S rDNA序列分析和16S rDNA PCR-RFLP的方法确定这些菌株的分类地位。所有45株菌株共产生7种遗传图谱类型,聚类分析结果表明,在79%的相似水平上,45株未知菌株构成了7个遗传表观群。16S rDNA序列分析和聚类结果表明鸡毛松根瘤内生细菌与红豆根瘤内生细菌都表现出多样性。菌株个数组成在16S rDNA PCR-RFLP聚类和数值聚类结果基本一致。
16SrDNA全序列分析表明,45株菌株属于6个属,分别为不动杆菌(Acinetobacter sp)、假单胞菌(Pseudomonas sp)、类芽孢杆菌(Paenibacillussp)、寡养单胞菌属(Stenotrophomonas sp)、芽孢杆菌(Bacillus sp)和葡萄球菌属(Staphylococcus sp)。
On the 600 meters high place of Shiwandashan Mountain in Guangxi Province, we found one Podocarpus imbricatus that grown with a Taxus for hundreds of years, from which we separated 13 endogenous bacteria, 3 endogenous bacteria from the Taxus. We also separated 9 endogenous bacteria from the vernal P. imbricatus. seedlings nodule which was transplanted, and 20 endogenous bacteria from aestival P. imbricatus. seedlings nodule.
Among 42 endogenous bacteria of P. imbricatus. and 3 endogenous bacteria of Taxus nodule, 9 strains are Gram-positive. 20 percent of the strains are hydrolyzed starch positive, and 20 percent of the strains are hydrolyzed gelatin positive, 87 percent of the strains could utilize citrate. All strains are able to use 7 kinds of carbon and 5 nitrogen.
Using the 16S rDNA sequence analysis and 16S rDNA PCR-RFLP to determine the classification status of these strains. There are totally 7 kinds of genetic map type generated from these 45 strains. The cluster analysis showed that in 79 percent of the similar level, 45 unknown strains constitute 7 apparent group. There a diversity from the endogenous bacteria of P. imbricatus. and Taxus nodule based on 16S rDNA sequence analysis and the cluster tree.
The 16SrDNA sequence analysis showed that the 45 strains belong to 6 categories, which are Acinetobacter sp, Pseudomonas sp, Paenibacillus sp, Stenotrophomonas sp, Bacillus sp and Staphylococcus sp.
引文
[1]洪国藩,宋鸿遇,固氮之光[M].长沙:湖南科学技术出版社,1997.8 120-121
[2]黄群策.生物固氮研究的前景[J].科技导报,1999,26(2)
[3]朱奇,陈彦.生物固氮在我国农业生产中的研究现状及发展对策[J].微生物学杂志,2003,23(5):40-43
[4]陈国安.重视生物固氮在我国农业持续发展中的地位[J].科技导报,1998,2:58-59
[5]窦新田.生物固氮[M].北京:农业出版社,1989,129-133
[6]Kuykcndall L D,Saxcna B,Dcvimc T E,ctal.Genetic diversity in Bradyrhyizobium japonicum Jordan 1982 and a Propcsal for Bradyhizobium clkanii sp.now[J].Can J Microbiol,1992,38:501-505
[7]陈丹明,曾昭海,隋新华,等.紫花首稽高效共生根瘤菌的筛选[J].草业科学,2002,19(6):27-31
[8]Denarie J,Roche P.R hizbitun nodulaton signals[M].Vcnma DPS.Molecular signals in plant-mirobe communications.London,CRC Press,1991:296-324
[9]中国草原学会.中国草地科学进展(第四届第二次年会暨学术讨论会集)[C].北京:中国农业大学出版社,1998,132-135
[10]娄志义.生物固氮研究的大事记[J].生物学教学.2005.30(4):60-61
[11]韩素芬,周湘泉.我国豆科树种结瘤情况[J].南京林业大学学报,1990,14(3):84-90
[12]唐东阶.大豆品种根瘤菌株最佳组合筛选试验[J].广西农学院学报,1989,8(1):49-55
[13]Kundig C H.Hcnnccke and M.Gottfert.Correlated physical and genetic map of the Bradyrhizobium japonicum 110 genome Bacteriol[J].1993,175:613-622
[14]王逸群,赵仁贵,王玉兰,等.豆科植物结瘤及其结瘤的分子基础[J].吉林农业科学,2001,26(5):20-25
[15]Beringer J E,Beynon J L,Buchanan-wollaston A V,et al.Transfer of the drug-resist anceansposon to Rhizobium.Nature 1978,276:633-634
[16]Kondorosi A,Kondorosi E,John M.The role of nodulation genes in bacterium-plant,communication Genetic Egineering.Plenum Press,New York,1996,13:115-136
[17]Soto M J,Zorzano A,Mercado-Blanco Jet al.Nudeotide sequence and characterization of Rhizobium meliloti nodulation competitiveness genes of nfe[J].Mol Biol,1993,229(2):570-576
[18]Beringer J E,Beynon J L,Buchanan-wollaston A V,et al.Transfer of the Tn5 to Rhizobium[J].Nature.1978,276:633-634
[19]Michiels J,Pelemans H.Identification and characterization of a Rhizobium leguminosarum by.Phaseoli Gene that is important for nodulation competitiveness and shows structural homology to a Rhizobium fredii host-induced gene[M].Plant-Microbe interat.1995,8(3):468-472
[20]Chun J Y,Stacey G.A Bradyrhizobium japonicum gene essential for competitiveness is differentially regulated from two promoters[M].Plant-Microbe Interat.1995,8(3)7(2):248-255
[21]Arcia-Rodriguez F M,Toro N.Sinorhizobium meliloti nfe(nodulation formation etliciency)genes exhibit temporal and spatial expression patterns similar to those of genes involved in symbiotic nitrogen fixation.[M].Plant Microbe Interact,2000,13(6):583-591
[22]Eardly B D,Hannaway D B,Botfemley P J.Characterization of rhizobia from ineffective alfalfa nodules:ability to nodulate bean plants Phaseolus valgaris(L.).Appl.Environ.Microbil,1985,50:1422-1427
[23]周俊初,刘孔鑫,杨润英,等.广谱花生根瘤菌97-1菌株在花生和大豆根瘤中的分化和繁殖能力[J],微生物学报,1992,32(4):278-284
[24]周刚林,武波,唐东阶,等.慢生型大豆根瘤菌nfe基因的分子克隆[J],广西农业生物科学,1999,18(1):29-32
[25]武波,周刚林,龙章德,等.慢生型大豆根瘤菌nfeC基因的定位[J],广西农业生物科学,2001,20(2):89-92
[26]唐咸来,武波,柏学亮,等.慢生型大豆根瘤菌putA基因的功能分析[J],中国农业科学 2002,35(4):399-402
[27]武波,唐咸来,等.慢生大豆根瘤菌与竞争结瘤相关的Irp基因的克隆[J],中国农业科学,2002,35(4):399-402
[28]周俊初等.导入外源DNA片段的花生根瘤菌高效基因=I=程菌株的构建[J],华中农业大学学报1994,03
[29]武波,周刚林,龙章德,等.慢生型大豆根瘤nfeC基因的克隆及序列分析[J],高技术通讯,2001,11(5):6-9
[30]周俊初等.高固氮基因工程大豆根瘤菌的研制及其增氮效应[J],广西农业生物科学,1996,01
[31]周俊初等.外源共生基因对紫云英根瘤菌共生效率的影响[J],中国农业科学,1995,04
[32]Callaham D et al.Isolation and cultivation in vitro of the actinomycete causing root nodulation In Comptonia[J].Science,1978(199):899-902
[33]林稚兰,黄秀梨.现代微生物学与实验技术[M].北京:科学出版社,2000,34-56
[34]尤崇杓,姜涌明,宋鸿遇.生物固氮[M].北京:科学出版社.1987,338
[35]Trinick M J.Symbiotis between Rhizobium and nonlegume-Tremaaspera[J].Nature,1973(244):459-460
[36]F.J.伯杰森.生物固氮研究方法[M].北京:科学出版社,1987,116
[37]黄宝灵,吕成群,唐东阶,等.罗汉松根瘤内生细菌的分离和特性[J].微生物学报,2002,42(5):620-624
[38]黄宝灵,吕成群,于新宁,.等.几种罗汉松根瘤的形态和结构[J].广西植物.2005(3):226-228
[39]王子芳.细菌一植物联合固氮研究进展[J].微生物学通报,1982(9):176-181
[40]林敏,尤崇杓.根际联合固氮作用研究进展[J].植物生理学通讯,1992.28(5):323-329
[41]新楠,连秀芬,樊明寿.非共生生物固氮的重要作用和研究进展[J].内蒙古农业科技,2005,(2):18-19
[42]崔宗均,苏宝林,蔡妙英,等.北方水稻田固氮细菌资源的研究(简报)[J].中国农业大学学报,1996,1(3):63
[43]何恒福,张锦秋,金世芳,等.水稻、玉米、甘蔗根系联合固氮菌的分离和鉴定[J].微生物学通报,1986,13(1):2-6
[44]谢光辉,苏宝林,崔宗均.长江流域水稻根际芽孢杆菌属固氮菌株的分离与鉴定[J].微生物学报,1998,38(6):480-83
[45]Peoples M B,Hetridge D F,Ladha J K Biological nitrogen fixation:an efficient source of nitrogen for sustainable agricultural production[J].Plan and soft,1995(174):3-28
[46]Berge O,Heulin T,Balandreau J.Biology and Fertility of Soft[J].Plant and Soft,1991,11(3):210-215
[47]Seldin L,Van Elsas J D.Bacillus azotofixans sp.nov,a nitrogen fixining species from Brazilian soils and grass roots[J].Bacterial.1984(34);451-456
[48]康丽华.桉树与联合固氮菌相互作用的研究[J].微生物学通报,2002,29(4):14-18
[49]谢莉华,周建,周政贤.马尾松幼树根系固氮细菌的发现及其初步研究[J].林业科学,1997,33(2):152-160
[50]Di Fiore S.Endophytic bacteria and thek possible role in the host plant[A].Istven E Az0spirilum Ⅵ and related microorganisms:genetics,physiology,ecology[c],Heidelberg,Springer,1995
[51]黎万奎,胡之璧.内生菌与天然药物[J].中国天然药物,2005,18:61
[52]Kohayashi D Y.Endophytc bacteria and their effects oft plant and uses in agriculture[A].Charles B W.Microbial endophytes[C].New York,Marcel Dekker,2000
[53]Samish Z,Etinger-Tulezynska R,Bick M.The raicroflora within the tissue of fruits and vegetables[J].Food Sci,1963,28(3):259-266
[54]宋子红,丁立孝,马伯军,等.花生内生菌的种群及动态分析[J].植物保护学报,1999,26(4):309-314
[55]Bell C R,Dickie G A,Harvey W L G.Endophytic Bacteria in Grapevine[J].Can J Microbiol,1995,41:46-53
[56]Gardner J M,Feldman A W,Zablotowicz R M.Identity and behavior of xylem-residing bacteria in rough lemon of Florida citrus trees[J].Appl Environ Microbiol,43:1335-1342
[57]Misaghi I J,Donndelinger C R.Endophytic bacteria in symptom-free cotton plants[J].Phytopathology,1990,80:808-811
[58]Gagn e S,Richard C,Rousseau H,Antoun H.Xylemresiding bacteria in alfalfa roots[J].Can J Microbiol,1987,33:996-1000
[59]Dong Z,Canny M J,McCully M E,et al.A nitrogen-fixing endophyte of sugarcane stems[J].Plant Physiol,1994,105:1 139-1147
[60]Fisher P J,Petrini O,Lappin Scott HM.The distribution of some fungal and bacteria endophytes in maizea(Zea may L.)[J].NewPhytol,1992,122(3):249-305
[61]Misaghi I J,Donndelinger C D.Endophytic bacteria in symptom-free cotton plants[J].Phytopathology,1990,80(9):808-811
[62]Sturz AV,Christie BR,Matheson BG,et al.Biodiversity of endophytic bacteria which colonize red clover nodules,roots,stems and foliage and their influence on host growth[J].Biology and Fertility of Soil,1997,25(1):13-19
[63]杨海莲,孙晓璐,宋未.植物内生细菌的研究[J].微生物学通报,1998,25(4):224-227
[64]Lamb T C,Tonkyn D W,Kluepfel D A.Can J Microbiol,1996,42:1112-1120
[65]Hallmann J,Roddguez R,Kloepper,et al.Soil Biology and Biochemistry,1999,31(5):551-560
[66]孔庆科,丁爱云.内生细菌作为生防因子的研究进展[J].山东农业大学学报(自然科学版),2001,32(2):256-260
[67]Van buren AM,Andre C,Ishimaru CA.Biological control of the bacterial ring rot pathogen by endophytic bacteria isolated from potato[J].Phyropthol,1993,83(2):140-146
[68]Baldani I J,Pot B,Kirchhof G,et al.Emended description of Herbaspirillum,inclusion of Pseudomonas rubrisubalbicans,a milk plant pathogen,as Herbaspirillum rubrisubalbicans comb.Nov and dassitication of a group of clinical isolates(EFgroup 1)as Herbaspirillum species[J].Int J Syst Bacteriol,1996,46:802-810
[69]Rinhold-Hurek B,Httrekt T,Gillis M,et al.Azarcus geno,noc.nitrogen-fixing proteobacteda with roots of Kallar grass and description oftwo species,Azoarcus fp.nov.and Azoarcuscommtmis sp.nov.[J].Int J Syst Bacteriol,1993,43:574-584
[70]Sheath P H A,Batefial classification H,Numerical taxonomy.In:N.R krieg and J.G Holt(cal.),Bergeys Manual of Systematic Bacteriology,Williams and Wilklns,Baltimore[M].MD,USA,1984,1-7
[71]Chen W X,Li G S,Qi Y L,et al.Rhizobiura huakuii sp.nov.isolatd from the root nudules of Astragalus sinicus[J].Int J syst Bacteriol,1991,41:275-280
[72]汪恩涛,陈文新,李季伦,等.SDS一聚丙烯酞胺凝胶电泳在根瘤菌鉴定中的应用[J].微生物学通报.1987.14:86-89
[73]De Lajudi P,Wtllems S A,Bruno P,et al.Poloyphasic taxonomy of Rhizobia:emendation of the genera sinorhizobium and description of sinorhizobium meliloti Comb.nov.Sinorhizobium saheli sp.nov.and sinorhizobium feranga sp.nov.[J].Int.J.cyst.Bacterial,1994,44:715-733
[74]Selander P K,Caugant D A,Ochman H,et al.Methods of multilocus enzyme detroplaresis for bacterial population qentics and systematics[J].Appl.Environ.Microbiol,1986,51:873-884
[75]Agus J E,Steven D G,Brian N D.Isolation and characterization of 2,3-dichcon-1 propanol-degrading rhizobia[J].Appl.Environ.Microbiol,200,66(7):2882-2887
[76]Willam J G K,Kubelik A R,Livak K J,et al.DNA polymorphisms amplified by arbitrary palmers are useful as genetic markers[J].Nucleic Acids Res,1990,18(22):6531-6535
[77]Welsh J,Muclelland M.Fingerprinting genomes using PCR with arbitrary palmers[J].Nucleic Acids Res,1990,18(24):7231-7281
[78]GaelleTr-baol,Charles Manteau,Yves Ttrilly,et al.LAssessment Ofthe genetic diversity among of xanthomonas eynarae by randomly amplified polymo-hic DNA analysis and development of specific characterized amplified for the rapididentification of X.cynarse[J].Appl Envir Microbiol,2001,67(8):3379-3384
[79]ljn M,et al.Deborah Schwarz intraspecific diversity of vibrio vulnificus in galveston bay water and dysters as determined by andomly amplified Dolymorphic DNA eft[J].Appl Envir Microbiol,2003,69(6):3170-3175
[80]何琳燕,殷永娴,黄为一.一株硅酸盐细菌的鉴定及其系统发育学分析[J].微生物学报,2003,43(2):162-167
[81]叶亚新,黄勇,王金虎.分子生物学技术在环境微生物多相分类中的应用[J].苏州科技学院学报(自然科学版),2004,21(4):46-49
[82]Ursing J B,Lior H,Owen R J.Proposal of minimal st Iiards for describing net species of the family Cpylob elaceae.In syst Bacteriol,1994,44(4):842-845
[83]中国科学院昆明植物研究所编著.云南植物志[M].第四卷,科学出版社,1986
[84]中国科学院微生物研究所.一般细菌常用鉴定方法[M].北京:科学出版社,1978
[85]VinCent JM..A Manual for the Practical Study of Root nodule Bacterial[M].International Biological Programme handbook no.15
[86]中国科学院微生物研究所.一般细菌常用鉴定方法[M].北京:科学出版社,1987
[87]韦革宏,陈春,龚明福,等.刺槐根瘤菌的表型多样性研究[J].西北植物学报,2003,23(12):2094-2098
[88]Jordan DC.Family Ⅲ.Rhizobiaceae,Bergey's Manual of Systematic Bacteriology,Volume Ⅰ,Williams and Wllkins,Baltimore,MD,USA,1984
[89]林德球.旋扭山绿豆快生型根瘤菌高度抗碱及共生固氮[J].微生物学报,1989,29(5):354-359
[90]程丽娟,刘海轮.快生型大豆根瘤菌生物学特性研究[J].西北农业大学学报,1994,22(1):74-71
[91]黄维南等.异叶银欢根瘤菌的分离和特性[J].亚热带植物通讯,1997,26(2):12-17
[92]Guerre G,Allard M R,Rely F,et al.Environ Microbiol.1994,60:56-63
[93]东秀珠.常见细菌鉴定手册[M].北京:科学出版社,1978
[94]屈良鹄.微生物遗传学研究综述集[M].上海:复旦大学出版社,1993
[95]Thompson JD,Gibson TJ,Plewniak F,et al.The CLUSTAL X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools.Nucleic Acids Res,1997,25:4876-4882
[96]冯瑞华.用APLF技术和16S r DNA PCR-RFLP分析毛苜蓿根瘤菌多样性的遗传多样性[J],微生物学报,2000,40(4):339-345
[2]黄群策.生物固氮研究的前景[J].科技导报,1999,26(2)
[3]朱奇,陈彦.生物固氮在我国农业生产中的研究现状及发展对策[J].微生物学杂志,2003,23(5):40-43
[4]陈国安.重视生物固氮在我国农业持续发展中的地位[J].科技导报,1998,2:58-59
[5]窦新田.生物固氮[M].北京:农业出版社,1989,129-133
[6]Kuykcndall L D,Saxcna B,Dcvimc T E,ctal.Genetic diversity in Bradyrhyizobium japonicum Jordan 1982 and a Propcsal for Bradyhizobium clkanii sp.now[J].Can J Microbiol,1992,38:501-505
[7]陈丹明,曾昭海,隋新华,等.紫花首稽高效共生根瘤菌的筛选[J].草业科学,2002,19(6):27-31
[8]Denarie J,Roche P.R hizbitun nodulaton signals[M].Vcnma DPS.Molecular signals in plant-mirobe communications.London,CRC Press,1991:296-324
[9]中国草原学会.中国草地科学进展(第四届第二次年会暨学术讨论会集)[C].北京:中国农业大学出版社,1998,132-135
[10]娄志义.生物固氮研究的大事记[J].生物学教学.2005.30(4):60-61
[11]韩素芬,周湘泉.我国豆科树种结瘤情况[J].南京林业大学学报,1990,14(3):84-90
[12]唐东阶.大豆品种根瘤菌株最佳组合筛选试验[J].广西农学院学报,1989,8(1):49-55
[13]Kundig C H.Hcnnccke and M.Gottfert.Correlated physical and genetic map of the Bradyrhizobium japonicum 110 genome Bacteriol[J].1993,175:613-622
[14]王逸群,赵仁贵,王玉兰,等.豆科植物结瘤及其结瘤的分子基础[J].吉林农业科学,2001,26(5):20-25
[15]Beringer J E,Beynon J L,Buchanan-wollaston A V,et al.Transfer of the drug-resist anceansposon to Rhizobium.Nature 1978,276:633-634
[16]Kondorosi A,Kondorosi E,John M.The role of nodulation genes in bacterium-plant,communication Genetic Egineering.Plenum Press,New York,1996,13:115-136
[17]Soto M J,Zorzano A,Mercado-Blanco Jet al.Nudeotide sequence and characterization of Rhizobium meliloti nodulation competitiveness genes of nfe[J].Mol Biol,1993,229(2):570-576
[18]Beringer J E,Beynon J L,Buchanan-wollaston A V,et al.Transfer of the Tn5 to Rhizobium[J].Nature.1978,276:633-634
[19]Michiels J,Pelemans H.Identification and characterization of a Rhizobium leguminosarum by.Phaseoli Gene that is important for nodulation competitiveness and shows structural homology to a Rhizobium fredii host-induced gene[M].Plant-Microbe interat.1995,8(3):468-472
[20]Chun J Y,Stacey G.A Bradyrhizobium japonicum gene essential for competitiveness is differentially regulated from two promoters[M].Plant-Microbe Interat.1995,8(3)7(2):248-255
[21]Arcia-Rodriguez F M,Toro N.Sinorhizobium meliloti nfe(nodulation formation etliciency)genes exhibit temporal and spatial expression patterns similar to those of genes involved in symbiotic nitrogen fixation.[M].Plant Microbe Interact,2000,13(6):583-591
[22]Eardly B D,Hannaway D B,Botfemley P J.Characterization of rhizobia from ineffective alfalfa nodules:ability to nodulate bean plants Phaseolus valgaris(L.).Appl.Environ.Microbil,1985,50:1422-1427
[23]周俊初,刘孔鑫,杨润英,等.广谱花生根瘤菌97-1菌株在花生和大豆根瘤中的分化和繁殖能力[J],微生物学报,1992,32(4):278-284
[24]周刚林,武波,唐东阶,等.慢生型大豆根瘤菌nfe基因的分子克隆[J],广西农业生物科学,1999,18(1):29-32
[25]武波,周刚林,龙章德,等.慢生型大豆根瘤菌nfeC基因的定位[J],广西农业生物科学,2001,20(2):89-92
[26]唐咸来,武波,柏学亮,等.慢生型大豆根瘤菌putA基因的功能分析[J],中国农业科学 2002,35(4):399-402
[27]武波,唐咸来,等.慢生大豆根瘤菌与竞争结瘤相关的Irp基因的克隆[J],中国农业科学,2002,35(4):399-402
[28]周俊初等.导入外源DNA片段的花生根瘤菌高效基因=I=程菌株的构建[J],华中农业大学学报1994,03
[29]武波,周刚林,龙章德,等.慢生型大豆根瘤nfeC基因的克隆及序列分析[J],高技术通讯,2001,11(5):6-9
[30]周俊初等.高固氮基因工程大豆根瘤菌的研制及其增氮效应[J],广西农业生物科学,1996,01
[31]周俊初等.外源共生基因对紫云英根瘤菌共生效率的影响[J],中国农业科学,1995,04
[32]Callaham D et al.Isolation and cultivation in vitro of the actinomycete causing root nodulation In Comptonia[J].Science,1978(199):899-902
[33]林稚兰,黄秀梨.现代微生物学与实验技术[M].北京:科学出版社,2000,34-56
[34]尤崇杓,姜涌明,宋鸿遇.生物固氮[M].北京:科学出版社.1987,338
[35]Trinick M J.Symbiotis between Rhizobium and nonlegume-Tremaaspera[J].Nature,1973(244):459-460
[36]F.J.伯杰森.生物固氮研究方法[M].北京:科学出版社,1987,116
[37]黄宝灵,吕成群,唐东阶,等.罗汉松根瘤内生细菌的分离和特性[J].微生物学报,2002,42(5):620-624
[38]黄宝灵,吕成群,于新宁,.等.几种罗汉松根瘤的形态和结构[J].广西植物.2005(3):226-228
[39]王子芳.细菌一植物联合固氮研究进展[J].微生物学通报,1982(9):176-181
[40]林敏,尤崇杓.根际联合固氮作用研究进展[J].植物生理学通讯,1992.28(5):323-329
[41]新楠,连秀芬,樊明寿.非共生生物固氮的重要作用和研究进展[J].内蒙古农业科技,2005,(2):18-19
[42]崔宗均,苏宝林,蔡妙英,等.北方水稻田固氮细菌资源的研究(简报)[J].中国农业大学学报,1996,1(3):63
[43]何恒福,张锦秋,金世芳,等.水稻、玉米、甘蔗根系联合固氮菌的分离和鉴定[J].微生物学通报,1986,13(1):2-6
[44]谢光辉,苏宝林,崔宗均.长江流域水稻根际芽孢杆菌属固氮菌株的分离与鉴定[J].微生物学报,1998,38(6):480-83
[45]Peoples M B,Hetridge D F,Ladha J K Biological nitrogen fixation:an efficient source of nitrogen for sustainable agricultural production[J].Plan and soft,1995(174):3-28
[46]Berge O,Heulin T,Balandreau J.Biology and Fertility of Soft[J].Plant and Soft,1991,11(3):210-215
[47]Seldin L,Van Elsas J D.Bacillus azotofixans sp.nov,a nitrogen fixining species from Brazilian soils and grass roots[J].Bacterial.1984(34);451-456
[48]康丽华.桉树与联合固氮菌相互作用的研究[J].微生物学通报,2002,29(4):14-18
[49]谢莉华,周建,周政贤.马尾松幼树根系固氮细菌的发现及其初步研究[J].林业科学,1997,33(2):152-160
[50]Di Fiore S.Endophytic bacteria and thek possible role in the host plant[A].Istven E Az0spirilum Ⅵ and related microorganisms:genetics,physiology,ecology[c],Heidelberg,Springer,1995
[51]黎万奎,胡之璧.内生菌与天然药物[J].中国天然药物,2005,18:61
[52]Kohayashi D Y.Endophytc bacteria and their effects oft plant and uses in agriculture[A].Charles B W.Microbial endophytes[C].New York,Marcel Dekker,2000
[53]Samish Z,Etinger-Tulezynska R,Bick M.The raicroflora within the tissue of fruits and vegetables[J].Food Sci,1963,28(3):259-266
[54]宋子红,丁立孝,马伯军,等.花生内生菌的种群及动态分析[J].植物保护学报,1999,26(4):309-314
[55]Bell C R,Dickie G A,Harvey W L G.Endophytic Bacteria in Grapevine[J].Can J Microbiol,1995,41:46-53
[56]Gardner J M,Feldman A W,Zablotowicz R M.Identity and behavior of xylem-residing bacteria in rough lemon of Florida citrus trees[J].Appl Environ Microbiol,43:1335-1342
[57]Misaghi I J,Donndelinger C R.Endophytic bacteria in symptom-free cotton plants[J].Phytopathology,1990,80:808-811
[58]Gagn e S,Richard C,Rousseau H,Antoun H.Xylemresiding bacteria in alfalfa roots[J].Can J Microbiol,1987,33:996-1000
[59]Dong Z,Canny M J,McCully M E,et al.A nitrogen-fixing endophyte of sugarcane stems[J].Plant Physiol,1994,105:1 139-1147
[60]Fisher P J,Petrini O,Lappin Scott HM.The distribution of some fungal and bacteria endophytes in maizea(Zea may L.)[J].NewPhytol,1992,122(3):249-305
[61]Misaghi I J,Donndelinger C D.Endophytic bacteria in symptom-free cotton plants[J].Phytopathology,1990,80(9):808-811
[62]Sturz AV,Christie BR,Matheson BG,et al.Biodiversity of endophytic bacteria which colonize red clover nodules,roots,stems and foliage and their influence on host growth[J].Biology and Fertility of Soil,1997,25(1):13-19
[63]杨海莲,孙晓璐,宋未.植物内生细菌的研究[J].微生物学通报,1998,25(4):224-227
[64]Lamb T C,Tonkyn D W,Kluepfel D A.Can J Microbiol,1996,42:1112-1120
[65]Hallmann J,Roddguez R,Kloepper,et al.Soil Biology and Biochemistry,1999,31(5):551-560
[66]孔庆科,丁爱云.内生细菌作为生防因子的研究进展[J].山东农业大学学报(自然科学版),2001,32(2):256-260
[67]Van buren AM,Andre C,Ishimaru CA.Biological control of the bacterial ring rot pathogen by endophytic bacteria isolated from potato[J].Phyropthol,1993,83(2):140-146
[68]Baldani I J,Pot B,Kirchhof G,et al.Emended description of Herbaspirillum,inclusion of Pseudomonas rubrisubalbicans,a milk plant pathogen,as Herbaspirillum rubrisubalbicans comb.Nov and dassitication of a group of clinical isolates(EFgroup 1)as Herbaspirillum species[J].Int J Syst Bacteriol,1996,46:802-810
[69]Rinhold-Hurek B,Httrekt T,Gillis M,et al.Azarcus geno,noc.nitrogen-fixing proteobacteda with roots of Kallar grass and description oftwo species,Azoarcus fp.nov.and Azoarcuscommtmis sp.nov.[J].Int J Syst Bacteriol,1993,43:574-584
[70]Sheath P H A,Batefial classification H,Numerical taxonomy.In:N.R krieg and J.G Holt(cal.),Bergeys Manual of Systematic Bacteriology,Williams and Wilklns,Baltimore[M].MD,USA,1984,1-7
[71]Chen W X,Li G S,Qi Y L,et al.Rhizobiura huakuii sp.nov.isolatd from the root nudules of Astragalus sinicus[J].Int J syst Bacteriol,1991,41:275-280
[72]汪恩涛,陈文新,李季伦,等.SDS一聚丙烯酞胺凝胶电泳在根瘤菌鉴定中的应用[J].微生物学通报.1987.14:86-89
[73]De Lajudi P,Wtllems S A,Bruno P,et al.Poloyphasic taxonomy of Rhizobia:emendation of the genera sinorhizobium and description of sinorhizobium meliloti Comb.nov.Sinorhizobium saheli sp.nov.and sinorhizobium feranga sp.nov.[J].Int.J.cyst.Bacterial,1994,44:715-733
[74]Selander P K,Caugant D A,Ochman H,et al.Methods of multilocus enzyme detroplaresis for bacterial population qentics and systematics[J].Appl.Environ.Microbiol,1986,51:873-884
[75]Agus J E,Steven D G,Brian N D.Isolation and characterization of 2,3-dichcon-1 propanol-degrading rhizobia[J].Appl.Environ.Microbiol,200,66(7):2882-2887
[76]Willam J G K,Kubelik A R,Livak K J,et al.DNA polymorphisms amplified by arbitrary palmers are useful as genetic markers[J].Nucleic Acids Res,1990,18(22):6531-6535
[77]Welsh J,Muclelland M.Fingerprinting genomes using PCR with arbitrary palmers[J].Nucleic Acids Res,1990,18(24):7231-7281
[78]GaelleTr-baol,Charles Manteau,Yves Ttrilly,et al.LAssessment Ofthe genetic diversity among of xanthomonas eynarae by randomly amplified polymo-hic DNA analysis and development of specific characterized amplified for the rapididentification of X.cynarse[J].Appl Envir Microbiol,2001,67(8):3379-3384
[79]ljn M,et al.Deborah Schwarz intraspecific diversity of vibrio vulnificus in galveston bay water and dysters as determined by andomly amplified Dolymorphic DNA eft[J].Appl Envir Microbiol,2003,69(6):3170-3175
[80]何琳燕,殷永娴,黄为一.一株硅酸盐细菌的鉴定及其系统发育学分析[J].微生物学报,2003,43(2):162-167
[81]叶亚新,黄勇,王金虎.分子生物学技术在环境微生物多相分类中的应用[J].苏州科技学院学报(自然科学版),2004,21(4):46-49
[82]Ursing J B,Lior H,Owen R J.Proposal of minimal st Iiards for describing net species of the family Cpylob elaceae.In syst Bacteriol,1994,44(4):842-845
[83]中国科学院昆明植物研究所编著.云南植物志[M].第四卷,科学出版社,1986
[84]中国科学院微生物研究所.一般细菌常用鉴定方法[M].北京:科学出版社,1978
[85]VinCent JM..A Manual for the Practical Study of Root nodule Bacterial[M].International Biological Programme handbook no.15
[86]中国科学院微生物研究所.一般细菌常用鉴定方法[M].北京:科学出版社,1987
[87]韦革宏,陈春,龚明福,等.刺槐根瘤菌的表型多样性研究[J].西北植物学报,2003,23(12):2094-2098
[88]Jordan DC.Family Ⅲ.Rhizobiaceae,Bergey's Manual of Systematic Bacteriology,Volume Ⅰ,Williams and Wllkins,Baltimore,MD,USA,1984
[89]林德球.旋扭山绿豆快生型根瘤菌高度抗碱及共生固氮[J].微生物学报,1989,29(5):354-359
[90]程丽娟,刘海轮.快生型大豆根瘤菌生物学特性研究[J].西北农业大学学报,1994,22(1):74-71
[91]黄维南等.异叶银欢根瘤菌的分离和特性[J].亚热带植物通讯,1997,26(2):12-17
[92]Guerre G,Allard M R,Rely F,et al.Environ Microbiol.1994,60:56-63
[93]东秀珠.常见细菌鉴定手册[M].北京:科学出版社,1978
[94]屈良鹄.微生物遗传学研究综述集[M].上海:复旦大学出版社,1993
[95]Thompson JD,Gibson TJ,Plewniak F,et al.The CLUSTAL X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools.Nucleic Acids Res,1997,25:4876-4882
[96]冯瑞华.用APLF技术和16S r DNA PCR-RFLP分析毛苜蓿根瘤菌多样性的遗传多样性[J],微生物学报,2000,40(4):339-345