中国十三省市土壤中非共生固氮微生物菌种资源研究

:2008-10-06:2008-12-29:948(2007-Z1);863(2006AA10Z417);(2005DKA21201-14-1):(1963),,,,,Tel:[phone],E2mail:[e-mail],,,,,,(;100081):137018116SrDNA,2466,,,65,36%;52,29%;19,11%;376%,:;;;:S154138:A:1008-505X(2009)06-1450-16Collectionandinvestigationonasymbioticnitrogen2fixingmicrobialresourcesfrom13provincesoverChinaSUNJian2guang,XUJing,HUHai2yan,ZHANGYan2chun,LIUJun,WANGWen2bo,SUNYan2hua(MinistryofAgricultureKeyLaboratoryofCropNutritionandFertilization/InstituteofAgriculturalResourcesandRegionalPlanning,ChineseAcademyofAgriculturalSciences,Beijing100081,China)Abstract:Onehundredandeightyoneasymbioticnitrogen2fixingbacterialstrainswereisolatedfrom70soilsamplescollectedfrom13provincesoverChina.Resultsbasedonthedeterminationofmorphology,physiologyandchemicalproper2ties,togetherwith16SrDNAsequencing,indicatedthatthesestrainsbelongto66speciesof24genera,whichaccountforaboutahalfofcurrentlyreportedgeneraofasymbioticnitrogen2fixingbacteria.Taxonomicdistributionofthesestrainswasrelativeconverging,65strainsbelongstoPaenibacillus,52strainsbelongstoBacillusand19strainsbelongstoArthrobac2ter,whichaccountsfor36%,29%and11%ofgrossresource,respectively.Strainsofthese3groupsmakeup76%ofgrossresource.StrainsofPaenibacillusandBacillusshowedsignificantdiversityonthedistributionregardingtogeographyandcrops.Namelystrainsofthese2groupscouldbeisolatedfromalmosteverysamplecollectedfromrootareaofcropsalloverChina.Keywords:asymbiotic;biologicalnitrogenfixation;microbialresources;phylogeny,,,[1],,;,,,,,2009,15(6):1450-1465PlantNutritionandFertilizerScience,,,,,,,,,[2],,,,111170,13,,,1811121121110g90mL20min,5mL30mLACCC55[3](10gK2HPO43H2O015gNaCl012gCaCO31gMgSO47H2O012g1000mLpH710712),28100r/min72h,,31121210-210-310-410-5,011mL(ACCC55115%210%),2923d,ACCC55(10gK2HPO43H2O015gNaCl012gCaCO31gMgSO47H2O012g015g1000mL115%210%,pH710712),,411311311[4],15mm150mm5mL,2872h,10%,72h,100L:[nmol/(mgh)]=C2H4(nmol)/[(mg)(h)]:C2H4nmol=1000C2H4(L)273P/[2214(273+t)760][5];P(mm),t()113125mL,3mL015mol/LNaOH5min,3mL015mol/LHCl,,110mL5mL,3min,595nmA595nm114,[6][7]11516SrDNA,DNA27f:52AGAGTTTGATCCTGGCTCAG23,1492r:52TACGGTTACCTTGTTACGACTT23PCR:9530s551min722min,30DNA,DNAStar,NCBI()2211,300,C2H4121nmol/(mgproh),212181,,,,,0175112m21515416,:515m,,,,,460,6%10%NaCl16SrDNA,NCBIGenBank97%,:1,72;266;34321335,Achromobacterxylosoxidans;ArthrobacterarilaitensiA.globiformisA.oryzaeA.oxydansA.pascens;BacilluscirculansB.flexusB.funiculusB.licheniformisB.megateriumB.nealsoniiB.pichinotyiB.racemilacticus;BrevibacilluschoshinensisB.parabrevis;Caulobacterhenricii;Chitinophagaginsengisegetis;DyadobacterfermentansEnterobacteraerogenes;Flavobacteriumanhuiense;Inquilinusginsengisoli;Lysobacterantibioticus;PaenibacillusalginolyticusP.alkaliterraeP.amylolyticusP.barengoltziiP.glycanilyticusP.sabina;Phyllobacteriumifriqiyense;Pseudomonasfluorescens;Pseudoxanthomonasmexicana;Sphingobacteriumsiyangensis;Stenotrophomonasmaltophilia;Variovoraxparadoxus16SrDNA(1),7:1Paeni2bacillus,6;2Bre2vibacillus,2;3Bacil2lus,8;6Arthrobacter,5;44;5;7457214181,2466,65Paenibacillus,36%;52Bacillus,29%;19Arthrobacter,11%376%(4)215,BacillusPaenibacillus,(5),1%14%,1371%,,76%,,,Arthrobacter,,,3,,,,,2007Azospirarestricta[8],Azospirillumcanadense[9],Azospirillumzeae[10],Burkholderiano2dosa[11],Gluconacetobacterkombuchae[12],Novosphingo2biumnitrogenifigens[13],Pseudacidovoraxintermedius[14],Paenibacillusdonghaensis[15],Paenibacillusforsythi2ae[16],Paenibacillussabinae[17],Paenibacillustaiwa2nensis[18],Paenibacilluszanthoxyli[19],Phytobacterdia2zotrophicus[20],Pseudoxanthobactersoli[21],Rhizobiumoryzae[22],Vibrioporteresiae[23],16,25411535416,:45411555416,:65411575416,:85411595416,:0641151Fig.1Phylogenetictreeofcollectednitrogen2fixingbacteria16416,:4Table4Taxonomicdistributionofasymbioticnitrogen2fixingmicrobialresources()Genus()SpeciesAmmount01Achromobacterxylosoxidans(1)1(015%)02Arthrobactersarilaitensis(1)globiformis(3)oryzae(1)oxidant(3)pascens(7)polychromogenes(2)ramosus(2)19(1015%)03Bacilluscereus(1)circulans(9)edaphicus(1)firmus(1)flexus(3)funiculus(13)licheniformis(1)megaterium(9)mucilaginosus(1)nealsonii(1)niacini(1)pichinotyi(4)racemilacticus(3)subtilis(2)vireti(1)yunchengensis(1)52(2817%)04Brevibacillusagri(2)choshinensis(3)formosus(2)parabrevis(1)8(414%)05Burkholderiavietnamiensis(1)1(015%)06Caulobacterhenricii(1)1(015%)07Chitinophagaginsengisegetis(2)2(111%)08Dyadobacterfermentans(2)2(111%)09Enterobacteraerogenes(1)1(015%)()Genus()SpeciesAmmount10Flavobacteriumanhuiense(2)2(111%)11Inquilinusginsengisoli(2)2(111%)12Lysobactergummosus(1)1(015%)13Micrococcusluteus(2)2(111%)14Paenibacillusagarexedens(6)alginolyticus(1)alkaliterrae(2)amylolyticus(1)barengoltzii(1)caespitis(6)daejeonensis(2)forsythia(1)glycanilyticus(4)graminis(2)humicus(11)kobensis(4)panacisoli(2)polymyxa(2)sabina(13)stellifer(2)thailandensis(2)validus(3)65(3519%)15Phyllobacteriumbrassicacearum(3)ifriqiyense(5)8(414%)16Pocheoniasoli(1)1(015%)17Pseudomonasfluorescens(4)4(212%)18Pseudoxanthomonasmexicana(1)1(015%