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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front. Agric. China    2010, Vol. 4 Issue (1) : 37-41     DOI: 10.1007/s11703-009-0090-2
Research articles |
Detection of indigenous endophytic bacteria in Eucalyptus urophylla in vitro conditions
Hongmiao SHEN1,Dangyue HAN1,Fenghuan YANG1,Qiuxian HUANG1,Longxian RAN1,Zhengnan LI2,
1.Section Forest Pathology, College of Forestry, Agricultural University of Hebei, Baoding 071001, China; 2.Mountain Area Research Institute of Hebei Province, Agricultural University of Hebei, Baoding 071001, China;
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Abstract  The presence of indigenous endophytic bacteria in aseptically grown seedlings of Eucalyptus urophylla germinated from surface sterilized seeds was investigated using dilution plating, microscopy, and PCR detection. No culturable endophytic bacteria could be detected in suspensions of ground plant tissue incubated on solid or in liquid cultivation media. However, a large number of endophytic bacterial cells, mostly rod-shaped and measured 2–3μm×0.5–0.8μm, were observed in in vitro cultured seedlings of E. urophylla using both light and electron microscopy. Using the universal bacterial 16S rDNA primers, a predicted 190-bp fragment was amplified from total DNA isolated from the seedlings of E. urophylla. We concluded that the endophytic bacteria originated from the seed were present in seedlings of E. urophylla. However, the bacterial cells observed appeared to be nonculturable.
Keywords electron microscopy      endophytic bacteria      Eucalyptus urophylla      PCR      
Issue Date: 05 March 2010
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-009-0090-2     OR     http://academic.hep.com.cn/fag/EN/Y2010/V4/I1/37
Bacilio-Jiménez M, Aguilar-Flores S, del Valle M V, Pérez A, Zepeda A, Zenteno E (2001). Endophytic bacteria in rice seedsinhibits early colonization of roots by Azospirillumbrasilense. Soil Biol Biochem, 33: 167―172

doi: 10.1016/S0038-0717(00)00126-7
Cankar K, Kraigher H, Ravnikar M, Rupnik M (2005). Bacterial endophytes from seeds of Norway spruce. FEMS Microbiol Lett, 244: 341―345

doi: 10.1016/j.femsle.2005.02.008
Cao J D (1982). Investigation of bacterial wilt in Eucalyptus saligna and E. grandis introduced from Brazil. Guangxi ForestrySci Tech, 4: 30―31 (in Chinese)
Chen C, Bauske E M, Musson G, Rodríguez-Kábana R, Kloepper J W (1995). Biologicalcontrol of Fusarium wilt on cotton by use of endophytic bacteria. Biol Control, 5: 83―91

doi: 10.1006/bcon.1995.1009
Coutinho T A, Roux J, Riedel K H, Terblanche J, Wingfield M J (2000). Firstreport of bacterial wilt caused by Ralstoniasolanacearum on eucalypts in South Africa. Forest Pathol, 30: 205―210

doi: 10.1046/j.1439-0329.2000.00205.x
FAO (2001). Global forest resources assessment 2000, main report: forestry paper140, Rome
Ferreira A, Quecine M C, Lacava P T, Oda S, Azevedo J L, Araújo W L (2008). Diversity of endophyticbacteria from Eucalyptus speciesseeds and colonization of seedlings by Pantoeaagglomerans. FEMS MicrobiolLett, 287: 8―14

doi: 10.1111/j.1574-6968.2008.01258.x
Fisher P J, Petrini O, Lappin Scott H M (1992). The distribution ofsome fungal and bacterial endophytes in maize (Zea mays L.). New Phytol, 122: 299―305

doi: 10.1111/j.1469-8137.1992.tb04234.x
Fu Z Q, Xia Z J, Wu A M, Yang Y B, Zheng Q, Gu B K (1999). The mechanism for controlling cotton wilt (Verticillium dahliae) by endophytic bacteria. Jiangsu J Agric Sci, 15: 211―215 (in Chinese)
Han X, Zhang L W (2007). Screeningtest of Bacillus endospores. Food Sci, 28: 347―350 (in Chinese)
He H, Qiu S X, Hu F P, Guan X (2004). Advance in biological effects of endophytic bacteria. J Microbiol, 5: 40―45 (in Chinese)
Keep N H, Ward J M, Robertson G, Cohen-Gonsaud M, Henderson B (2006). Bacterialresuscitation factors: revival of viable but non-culturable bacteria. Cell Mol Life Sci, 63: 2555―2559

doi: 10.1007/s00018-006-6188-2
King E O, Ward M K, Raney D E (1954). Two simple media for the demonstrationof pyocyanin and fluorescin. J Lab ClinMed, 44: 301―307
Kong Q K, Ding A Y (2001). Advancesof study on endophytic bacteria as biological control agents. J Shandong Agric Univ (Nat Sci), 32: 256―260 (in Chinese)
Li G L, Wang J F, Huang Y J, Zheng Z H, Su W J (2001). Study on the antifungalactivities of endophytic fungi isolated from several pharmaceuticalplants. Microbiol, 28: 64―68 (in Chinese)
Li Q Q, Luo K, Lin W, Peng H W, Luo X M (2003). Isolation of tomatoendophytic antagonists against Ralstonia solanacearum. Acta Phytopathologica Sinca, 33: 364―367 (in Chinese)
Li Q Q, Ye Y F, Meng X Y, Peng H W, Lin W, Luo K (2005). Identification of endophytic bacterium strain B47 and its control effects on tomato bacterial wilt. Chin J Biol Control, 21: 178―182 (in Chinese)
Li Z H, Liu Y J (2003). Studieson variety introductions and applicable disseminating of Eucalyptus in Hunan Province. Hunan Forestry Sci Tech, 3: 16―19 (in Chinese)
Mano H, Tanaka F, Watanabe A, Kaga H, Okunishi S, Morisaki H (2006). Culturable surface and endophytic bacterial flora ofthe maturing seeds of rice plants (Oryza sativa) cultivated in a paddy field. MicrobesEnviron, 21: 86―100

doi: 10.1264/jsme2.21.86
Mercado-Blanco J, Bakker P A H M (2007). Interactionsbetween plants and beneficial Pseudomonas spp.: exploiting bacterial traits for crop protection. Antonie van Leeuwenhoek. 92: 367―389

doi: 10.1007/s10482-007-9167-1
Mukhopadhyay K, Garrison N K, Hinton D M, Bacon C W, Khush G S, Peck H D, Datta N (1996). Identificationand characterization of bacterial endophytes of rice. Mycopathologia, 134: 151―159

doi: 10.1007/BF00436723
Mundt J O, Hinkle F N (1976). Bacteriawithin ovules and seeds. Appl Environ Microbiol, 32: 694―698
Murashige T, Skoog F (1962). A revisedmedium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant, 15: 473―497

doi: 10.1111/j.1399-3054.1962.tb08052.x
Qiu S X, He H, Ruan H C, Guan X, Hu F P (2004). Selection of endophyticbacteria with antipathogenic and plant growth promoting functions. Chin J Appl Environ Biol, 10: 655―659 (in Chinese)
Ran L X, Li Z N, Wu G J, van Loon L C, Bakker P A H M (2005b). Inductionof systemic resistance against bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp.. Eur J PlantPathol, 113: 59―70

doi: 10.1007/s10658-005-0623-3
Ran L X, Liu C Y, Wu G J, van Loon L C, Bakker P A H M (2005a). Suppressionof bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp.in China. Biol Control, 32: 111―120

doi: 10.1016/j.biocontrol.2004.08.007
Reiter B, Pfeifer U, Schwab H, Sessitsch A (2002). Response of endophytic bacterial communities in potatoplants to infection with Erwinia carotovora subsp. atroseptica. Appl Environ Microbiol, 68: 2261―2268

doi: 10.1128/AEM.68.5.2261-2268.2002
Tholozan J L, Cappelier J M, Tissier J P, Delattre G, Federighi M (1999). Physiologicalcharacterization of viable-but-nonculturable Campylobacter jejuni cells. Appl Environ Microbiol, 65: 1110―1116
Xiang M L, Ran L X (2004). Researchprogress of bacterial wilt in Eucalyptus. Forest Pest and Disease, 23: 37―40 (in Chinese)
Xing D F, Ren N Q, Song J X, Qu M, Xu X L (2006). Community of activatedsludge based on different targeted sequence of 16S rDNA by denaturinggradient gel electrophoresis. Environ Sci, 27: 1424―1428 (in Chinese)
Yan M H, Cai Z Q, Han J G, Sun L, Song W (2004). The applied researchof endophytic bacteria in biological control of plant disease. Biotechnol Bull, 3: 8―12 (in Chinese)
Yang H L, Sun X L, Song W (1998). Studies on phytoendophytic bacteria. Microbiol, 25: 224―227 (in Chinese)
Zheng H X, Zhou L, Lan X L, Gao Z Q, Gao Y, Li Y (2008). Isolation and identification of endophytic bacteriafrom rhododendron and resistance determination. J Neijiang Normal Univ, 23: 267―269 (in Chinese)
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