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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bacilio-Jiménez M, Aguilar-Flores S, del Valle M V, Pérez A, Zepeda A, Zenteno E (2001). Endophytic bacteria in rice seeds inhibits early colonization of roots by Azospirillum brasilense. Soil Biol Biochem, 33: 167–172
Cankar K, Kraigher H, Ravnikar M, Rupnik M (2005). Bacterial endophytes from seeds of Norway spruce. FEMS Microbiol Lett, 244: 341–345
Cao J D (1982). Investigation of bacterial wilt in Eucalyptus saligna and E. grandis introduced from Brazil. Guangxi Forestry Sci Tech, 4: 30–31 (in Chinese)
Chen C, Bauske E M, Musson G, Rodríguez-Kábana R, Kloepper J W (1995). Biological control of Fusarium wilt on cotton by use of endophytic bacteria. Biol Control, 5: 83–91
Coutinho TA, Roux J, Riedel K H, Terblanche J, Wingfield M J (2000). First report of bacterial wilt caused by Ralstonia solanacearum on eucalypts in South Africa. Forest Pathol, 30: 205–210
FAO (2001). Global forest resources assessment 2000, main report: forestry paper 140, Rome
Ferreira A, Quecine M C, Lacava P T, Oda S, Azevedo J L, Araújo W L (2008). Diversity of endophytic bacteria from Eucalyptus species seeds and colonization of seedlings by Pantoea agglomerans. FEMS Microbiol Lett, 287: 8–14
Fisher P J, Petrini O, Lappin Scott H M (1992). The distribution of some fungal and bacterial endophytes in maize (Zea mays L.). New Phytol, 122: 299–305
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 LW (2007). Screening test 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). Bacterial resuscitation factors: revival of viable but non-culturable bacteria. Cell Mol Life Sci, 63: 2555–2559
King E O, Ward M K, Raney D E (1954). Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med, 44: 301–307
Kong Q K, Ding AY (2001). Advances of 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 antifungal activities of endophytic fungi isolated from several pharmaceutical plants. Microbiol, 28: 64–68 (in Chinese)
Li Q Q, Luo K, Lin W, Peng H W, Luo X M (2003). Isolation of tomato endophytic 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). Studies on 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 of the maturing seeds of rice plants (Oryza sativa) cultivated in a paddy field. Microbes Environ, 21: 86–100
Mercado-Blanco J, Bakker PA H M (2007). Interactions between plants and beneficial Pseudomonas spp.: exploiting bacterial traits for crop protection. Antonie van Leeuwenhoek. 92: 367–389
Mukhopadhyay K, Garrison N K, Hinton D M, Bacon C W, Khush G S, Peck H D, Datta N (1996). Identification and characterization of bacterial endophytes of rice. Mycopathologia, 134: 151–159
Mundt J O, Hinkle F N (1976). Bacteria within ovules and seeds. Appl Environ Microbiol, 32: 694–698
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant, 15: 473–497
Qiu S X, He H, Ruan H C, Guan X, Hu F P (2004). Selection of endophytic bacteria 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). Induction of systemic resistance against bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp.. Eur J Plant Pathol, 113: 59–70
Ran L X, Liu C Y, Wu G J, van Loon L C, Bakker P A H M (2005a). Suppression of bacterial wilt in Eucalyptus urophylla by fluorescent Pseudomonas spp. in China. Biol Control, 32: 111–120
Reiter B, Pfeifer U, Schwab H, Sessitsch A (2002). Response of endophytic bacterial communities in potato plants to infection with Erwinia carotovora subsp. atroseptica. Appl Environ Microbiol, 68: 2261–2268
Tholozan J L, Cappelier JM, Tissier J P, Delattre G, Federighi M (1999). Physiological characterization of viable-but-nonculturable Campylobacter jejuni cells. Appl Environ Microbiol, 65: 1110–1116
Xiang M L, Ran L X (2004). Research progress 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 activated sludge based on different targeted sequence of 16S rDNA by denaturing gradient 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 research of 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 bacteria from rhododendron and resistance determination. J Neijiang Normal Univ, 23: 267–269 (in Chinese)
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Shen, H., Li, Z., Han, D. et al. Detection of indigenous endophytic bacteria in Eucalyptus urophylla in vitro conditions. Front. Agric. China 4, 37–41 (2010). https://doi.org/10.1007/s11703-009-0090-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11703-009-0090-2