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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2010, Vol. 4 Issue (4) : 443-448     DOI: 10.1007/s11703-010-1038-2
RESEARCH ARTICLE |
Presence of indigenous endophytic bacteria in jujube seedlings germinated from seeds in vitro
Xiaojie HOU1, Zhengnan LI2, Dangyue HAN1, Qiuxian HUANG1, Longxian RAN1()
1. Section Forest Pathology, Forestry College, Agricultural University of Hebei, Baoding 071001, China; 2. National Engineering Research Center for Agriculture in Northern Mountainous Areas, Baoding 071000, China
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Abstract  

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.

Keywords DGGE      optical microscope      scanning electron microscopy (SEM)      unculturable bacteria      Zizyphus jujuba     
Corresponding Authors: RAN Longxian,Email:longxianran@hebau.edu.cn   
Issue Date: 05 December 2010
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-010-1038-2     OR     http://academic.hep.com.cn/fag/EN/Y2010/V4/I4/443
Fig.1  Presence of endophytic bacteria in seedlings germinated from seeds of var. Fupingdazao observed by optical microscope
Fig.2  Observation of endophytic bacteria inside the phloem cell of seedling germinated from seeds of var. Fupingdazao by SEM
Fig.3  Amplification of 16S rDNA fragments of endophytic bacteria in jujube seedlings germinated from seeds
Note: M is Marker DL3000; Lanes 1 and 2 represent two seedlings germinated from seeds of var. Fupingdazao.
Fig.4  DGGE profiles of 16S rDNA of endophytic bacteria in jujube seedlings germinated from seeds
Note: Lanes 1 and 2 represent two seedlings germinated from var. Fupingdazao seeds.
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