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 μmÂ0.5-0.8 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.
The presence of indigenous endophytic bacteria in tissue-cultured seedlings germinated from seeds of Zizyphus jujuba var. Fupingdazao was investigated using cultivation, light microscopy and scanning electronic microscopy (SEM). In addition, bacteria specific 16S rDNA PCR followed by denaturing gradient gel electrophoresis (DGGE) was used. No cultivable bacteria could be detected on plates or in liquid cultures. However, large quantities of endophytic bacteria in jujube seedlings were observed under the light microscope. The bacterial cells were round (measuring 1.5 μm -1.8 μm), short rods (2.2 μm -2.9 μm  1.1 μm -1.9 μm) and rod shaped (3.7 μm -4.4 μm  1.6 μm -1.9 μm). Rod-shaped bacterial cells (measuring 3.5 m -4.0 m  1.5 m -2.0 m) were observed by SEM as well. A 16S rDNA fragment of 1.5 kb could be amplified from the total DNA of seedlings of jujube using the bacterial primer pair 27F/1525R. The V3 fragment of 16S rDNA was separated by DGGE, and the 16S rDNA-PCR-DGGE showed that there were at least six dominant bands within the seedlings of Z. jujuba var. Fupingdazao. It can be concluded that endophytic bacteria that cannot be detected by cultivation are present with high densities in jujube seeds.
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