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1、应用与环境生物学报2010，16(6):840~8442010-12-25ChinJApplEnvironBiol=ISSN1006-687XDOI:10.3724/SP.J.1145.2010.00840泸州古酿酒窖池中两株产甲烷杆菌比较研究王俪鲆1,4张良2刘来雁3张宿义2许德富2刘光烨1*1（中国科学院成都生物研究所成都610041）2（泸州老窖股份有限公司泸州646000）3（农业部沼气科学研究所，农业部能源微生物与利用重点开放实验室成都610041）4（中国科学院研究生院北京100049）摘要采用厌氧操作技术，从泸州老窖古
2、酿酒窖池窖泥中分离到两株产甲烷杆菌0372-D1和0072-D2.0372-D1菌体形态为长杆状，略弯，两端整齐，不运动，可由多个菌体形成长链；在固体培养基中难以长出菌落，只利用H+CO产生甲22烷.0072-D2菌体形态为弯曲杆状，淡黄色圆形菌落，利用H+CO或甲酸盐作为唯一碳源生长.两株菌最适生长温度22均为35℃、菌株0372-D1最适生长pH为6.5~7.0，生长pH范围5.0~8.0；菌株0072-D2最适生长pH则为7.5.在各自最适条件下培养，两株菌的最短增代时间分别为19h和8h.通过形态、生理生化特征和16SrDN
3、A序列的同源性分析，表明菌株0372-D1为产甲烷杆菌属的一个新种.0072-D2则为甲酸甲烷杆菌（Methanobacteriumformicicum）的新菌株，相似性为99%.图6表1参16关键词产甲烷菌；比较研究；新种分离；窖泥；16SrDNA序列；系统发育分析；泸州老窖CLCQ939.97:TQ92ComparativeStudyofTwoMethanobacteriumStrainsintheAncientFermentationPitsofLuzhouWANGLiping1,4,ZHANGLiang2,LIULaiyan
4、3,ZHANGSuyi2,XUDefu2&LIUGuangye1*(1ChengduInstituteofBiology,ChineseAcademyofSciences,Chengdu610041,China)(2LuzhoulaojiaoCo.,Ltd.,Luzhou646000,Sichuan,China)(3BiogasInstituteofMinistryofAgricultural,KeyLaboratoryofEnergyMicrobiologyandApplication,MinistryofAgriculture,
5、Chengdu610041,China)(4GraduateUniversityofChineseAcademyofSciences,Beijing100049,China)AbstractTwostrainsofMethanobacterium,namedas0372-D1and0072-D2,wereisolatedfromtheancientfermentationpitsofLuzhoulaojiaobyusingtheHungateanaerobictechniques.Thecellsof0372-D1arelongro
6、d,slightlycurved,bothendsneatlyandnonmotile,whichmayformlongchainsbysomecells.However,theyaredifficulttoformcolonyinthesolidmedium.ThisstrainutilizesonlyH+COtoproducemethane.Strain0072-D2iscurvedinrod-shape,lightyellowandforms22roundcolonies,anditutilizesformateorH+COas
7、itssolecarbonsourceforgrowth.Theoptimalgrowthtemperatureoftwo22strainswasfoundat35℃.TheoptimalgrowthpHofstrain0372-D1was6.5~7.0anditsgrowthpHrangedfrom5.0to8.0,andtheoptimalgrowthpHofstrain0072-D2occurredatpH7.5.Thedoubletimewas19hfor0372-D1and8hfor0072-D2whenculturedu
8、ndertheoptimalconditions.Theresultfromtheanalysisoftheirmorphologicalandphysicobiochemicalcharacteristicsandhomologyo
