刘菲菲;李赤;刘勇勤;于莉;

• 1:广东海洋大学农学院

摘要(Abstract)：

以健康香蕉植株和感染香蕉枯萎病菌Fusarium oxysporum f.sp.cubense(FOC)后的香蕉植株的根茎叶组织为材料,提取样品中的总DNA,扩增细菌的16S rDNA的V3可变区,对扩增产物进行DGGE电泳分析,并对其中18条优势条带进行切胶回收、克隆测序和系统发育分析。结果表明,香蕉健株与病株各组织中所含内生细菌的种群丰富度为根部>假茎>叶片;感病植株组织内生细菌种类比健康植株丰富;BLAST结果为大部分克隆序列与已知细菌16SrDNA基因序列的同源性较高(94%~100%),分别归属于蓝细菌门(Cyanobacteria)、绿弯菌门(Chloroflexi)、放线菌门(Actinobacteria)、变形杆菌门(Proteobacterium)、拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和绿菌门(Chlorobi)7大类群;其中一个序列同源性较低(91%),可能代表新的分类单元。

关键词(KeyWords)： 香蕉;内生细菌;DGGE;群落结构;多样性

Abstract:

Keywords:

基金项目(Foundation): 湛江市科技局项目:抗枯萎病香蕉品种的引进与应用

作者(Author): 刘菲菲;李赤;刘勇勤;于莉;

Email:

DOI: 10.13925/j.cnki.gsxb.2012.02.017

参考文献(References)：

• [1]ZINNIEL D K,LAMBRECHT P A,HARRIS B N,FENG Z Y,KUCZMARSKI D,HIGLEY P,ISHIMARU A C,ARUNAKUMARIA,BARLETTA G R,VIDAVER K A.Isolation and characterizationof endophytic colonizing bacteria from agronomic crops and prairieplants[J].Applied and Environmental Microbiology,2002,68(5):2198-2208.
• [2]SURETTE A M,STURZ V A,LADA R R,NOWAK J.Bacterial en-dophytes in processing carrots[J].Plant and Soil,2003(253):381-390.
• [3]TING S A,MEON S,KADIR J,RADU S,SINGH G.Endophytic mi-croorganisms as potential growth promoters of banana[J].Biocontrol,2008,(53):541-553.
• [4]YANG Hai-lian,SUN Xiao-lu,SONG Wei.Study of endophytic bac-teria in plant[J].Microbiology,1998,25(4):224-227.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.
• [5]HALLMANN Q A,HALLMANN J,KLOEPPER W J.Bacterial endo-phytes in cotton location and interaction with other plant-associatedbacteria[J].Can J Microbiol,1997,(43):254-259.
• [6]AMANN R I,LUDWIG W,SCHLEIFER K A.Phylogenetic identifi-cation and in situdetection of individual m icrobial cells without cul-tivation[J].American Society for Microbiology,1995,59(1):143-169.
• [7]LIU Hui-jie,YANG Cai-yun,TIAN Yun,LIN Guang-hui,ZHENGTian-ling.Analysis of microbial community structure in mangrovesediments by PCR-DGGE technique[J].ACTA Microbiologica Sini-ca,2010,50(7):923-930.刘慧杰,杨彩云,田蕴,林光辉,郑天凌.基于PCR-DGGE技术的红树林区微生物群落结构[J].微生物学报,2010,50(7):923-930.
• [8]WU Q,ZHAO X H,ZHAO S Y.Application of PCR-DGGE in re-search of bacterial diversity in drinking water[J].Biomedical andEnvironmental Sciences,2006,(19):371-374.
• [9]SANCHEZ O,GASOL M J,MASSANA R,MAS J,PEDROS-ALIOC.Comparison of different denaturing gradient GEL electrophoresisprimer SETS for the study of Marine bacterioplankton communities[J].Applied and Environmental Microbiology,2007,73(18):5962-5967.
• [10]LIU Lin,LIU Yang,SONG Wei.Application of PCR-DGGE technol-ogy in the research of Micro-Ecology in the plants[J].BiotechnologyBulletin,2009(3):54-56.刘琳,刘洋,宋未.PCR-DGGE技术及其在植物微生态研究中的应用[J].生物技术通报,2009(3):54-56.
• [11]FU Ye-qin,CAI Ji-miao,LIU Xian-bao,HUANG Gui-xiu.Isola-tion,Bioactivity evaluation and quantitydistribution of endophyticbacteria in banana[J].Chinese Journal of Tropical Crops,2007,28(4):78-85.付业勤,蔡吉苗,刘先宝,黄贵修.香蕉内生细菌分离、活性评价及数量分布[J].热带作物学报,2007,28(4):78-85.
• [12]PAN Xian-xin,LIU Xian-bao,SHI Tao,LIN Chun-hua,CAI Ji-miao,HUANG GUI-xiu.A diversity research of endophytic bacterialiving in banana root with16Sr DNA sequences analysis[J].ChineseJournal of Tropical Crops,2010,31(5):772-776.潘羡心,刘先宝,时涛,林春花,蔡吉苗,黄贵修.16SrDNA序列分析法对香蕉根部内生细菌种群多样性初析[J].热带作物学报,2010,31(5):772-776.
• [13]FANG Zhong-da.Research methods of plant disease[M].Beijing:China Agriculture Press,1998.方中达.植病研究方法[M].北京:中国农业出版社,1998.
• [14]REITER B,PFEIFER U,SCHWAB H,SESSITSCH A.Response ofendophytic bacterial communities in potato plants to infection withErwinia carotovora[J].Applied and Environmental Microbiology,2002,68(5):2261-2268.
• [15]SHEN Hong,CHEN Ming-jie,ZHAO Yong-chang,PAN Ying-jie.PCR-DGGE identification of endophytes in morels[J].Acta Agricul-turae Shanghai,2008,24(2):58-60.沈洪,陈明杰,赵永昌,潘迎捷.羊肚菌内生细菌DGGE鉴定[J].上海农业学报,2008,24(2):58-60.
• [16]MUYZER G,WAAL C D,UITIERLINDEN G A.Profiling of complexmicrobial populations by denaturing gradient GEL electrophoresisanalysis of polymerase chain Reaction-Amplified genes coding for16Sr RNA[J].Applied and Environmental Microbiology,1993,59(3):695-700.
• [17]HOU Xiao-jie,CUI Jian-zhou,LI Zheng-nan,YU Zhan-jing,NIUXiao-ke,RAN Long-xian.Diversity analysis of microbial communityfrom the fruit of jujube fruit shrink diseaseby PCR-DGGE[J].Journalof Chinese Institute of Food Science and Technology,2010,10(4):260-266.侯晓杰,崔建州,李正楠,于占晶,牛晓科,冉隆贤.枣缩果病果实内微生物种群多样性的PCR-DGGE分析[J].中国食品学报,2010,10(4):260-266.
• [18]LUO Ming,LU Yun,CHEN Jin-huan,JIANG Yun-qiin,ZHANGXiang-lin.Study on the dynamic change of population density anddistribution ofendophytic bacteria in HAMI melon plants in Xinjiang[J].Arid Zone Research,2007,24(1):28-33.罗明,卢云,陈金焕,姜云琴,张祥林.哈密瓜内生细菌菌群密度及分布动态[J].干旱区研究,2007,24(1):28-33.
• [19]WANG Ai-hua,YAN You-ping,XIONG Gong-li,LI Yan-fang,LIJia,XIAN Jia-xu,WANG Zhong-kang.Endophytic bacterial diver-sity analysis of huanglongbing Pathogen-Infected citrus phloem tis-sue in Guangxi[J].Agricultural Sciences in China,2010,43(23):4823-4833.王爱华,殷幼平,熊红利,李颜方,李佳,贤家旭,王中康.广西柑橘黄龙病植株韧皮部内生细菌多样性分析[J].中国农业科学,2010,43(23):4823-4833.
• [20]MURRAY E A,HOLLIBAUGH T J,ORREGO C.Phylogenetic com-positions of bacterioplankton from TWO California estuaries com-pared by denaturing gradient GEL electrophoresis of 16S RDNAfragments[J].Applied and Environmental Microbiology,1996,62(7):2676-2680.
• [21]CHENG Xiao-yan,LI Wen-jun,WANG Yun,YANG Hong-mei,LOU Kai.Endophytic bacterial diversity in Glycyrrhiza inflata Bat.from Xinjiang by culture independent method[J].Acta Microbio-logica Sinica,1995,49(6):718-725.程晓燕,李文军,王芸,杨红梅,娄恺.新疆野生胀果甘草内生细菌多样性的非培养初步分析[J].微生物学报,1995,49(6):718-725.
• [22]CAO Li-xiang,TIAN Xin-li,ZHOU Shi-ning.Isolation of Endo-phytic Fungi and Actinomycetes from banana(Musa paradisiaca)Plants[J].Acta Scifntiarum Naturalium Universitatis Sunyaatseni,2003,42(2):70-73.曹理想,田新莉,周世宁.香蕉内生真菌、放线菌类群分析[J].中山大学学报:自然科学版,2003,42(2):70-73.
• [23]WOESE C R.Bacterial evolution[J].Microbiol REV,1987,51(2):221-271.
• [24]CHEN Bin.Reconstitution of the Cycad-Cyanobacteria association[D].Fuzhou:Fujian Agrieulture and Forestry University,2004.陈彬.蓝细菌与苏铁重组的研究[D].福州:福建农林大学,2004.
• [25]ZHENG Si-ping.Diversity of endophytic bacteria within Azolla mi-crophylla[D].Fuzhou:Fujian Agriculture and Forestry University,2007.郑斯平.小叶满江红内生细菌多样性研究[D].福建:福建农林大学,2007.