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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.
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Keywords
electron microscopy
endophytic bacteria
Eucalyptus urophylla
PCR
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Issue Date: 05 March 2010
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