Hostname: page-component-78c5997874-94fs2 Total loading time: 0 Render date: 2024-11-06T01:01:08.921Z Has data issue: false hasContentIssue false
  • English
  • Français

Diversity analysis of endophytic bacteria within Azolla microphylla using PCR-DGGE and electron microscopy

Published online by Cambridge University Press:  03 March 2009

Zheng Si-Ping ,
Chen Bin ,
Guan Xiong  and
Zheng Wei-Wen
Zheng Si-Ping
Affiliation:
Biotechnology Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Chen Bin
Affiliation:
Biotechnology Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China
Guan Xiong
Affiliation:
Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Zheng Wei-Wen*
Affiliation:
Biotechnology Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China
*
*Corresponding author. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Using 16S rDNA-polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE), electron microscopy and a conventional plating method, the genetic diversity and phenotype polymorphism of the endophytic bacteria within Azolla microphylla were explored. The 16S rDNA-PCR-DGGE profile showed a complex and divergent bacterial community, with Bacillus cereus as the dominant species, within the Azolla–cyanobacteria association. This result was supported by the fact that endobacterial cells exhibited distinct ultrastructural characteristics in vivo and, in vitro, bacteria displayed various colonies with different sizes, shapes and colours. This study demonstrates that the genetic diversity of endophytic bacteria in Azolla can be investigated using the16S rDNA-PCR-DGGE technique.

Keywords

Azollaendophytic bacteriadiversity16S rDNA-PCR-DGGE
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 3 , December 2008 , pp. 269 - 276
Copyright
Copyright © China Agricultural University 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(3): 508–514

References

Bai, KZ, Yu, SL, Chen, WL, Yang, SY and Cui, H (1979) The isolation and culture of Azolla without cyanobacteria and Anabaena azollae. Chinese Science Bulletin 24: 664–666 (in Chinese).Google Scholar
Carrapico, F (1991) Are bacteria the 3rd partner of the Azolla–Anabaena symbiosis? Plant Soil 137: 157–160.CrossRefGoogle Scholar
Carrapico, F and Tavares, R (1989) New data on the Azolla–Anabaena symbiosis. I. Morphological and histochemical aspects. In: Skinner, FA, Boddey, RM and Frederik, I (editors) Nitrogen Fixation with Non-Legumes. Dordrecht, The Netherlands, Kluwer Academic Publishers, pp. 89–94.CrossRefGoogle Scholar
Ekman, M (2005) Mass spectrometric analysis of the nitrogen-fixing cyanobacterium of Azolla symbiosis: Adaptation and NifH modification. Ph.D. thesis, Department of Botany, Stockholm University, Sweden.Google Scholar
Forni, C, Gentili, S, Van Hove, C and Grilli Caiola, MG (1990) Isolation and characterization of the bacteria living in the sporocarps of Azolla filiculoides Lam. Annals of Microbiology 40: 235–243.Google Scholar
Forni, C, Gaegi, A and Del Gallo, M (1998) Polysaccharide composition of the mucilage of the Azolla algal packet. Symbiosis 24: 303–313.Google Scholar
Gates, JE, Fisher, RW and Candle, RA (1980) The occurrence of coryneforme bacteria in the leaf cavity of Azolla. Archives of Microbiology 127: 163–165.CrossRefGoogle Scholar
Hurt, RA, Qiu, X, Wu, L, et al. (2001) Simultaneous recovery of RNA and DNA from soils and sediments. Applied and Environmental Microbiology 67(10): 4495–4503.CrossRefGoogle ScholarPubMed
Lechno, YS and Nierzwicki-Bauer, SA (2002) Azolla–Anabaena symbiosis. In: Rai, AN, Bergman, B and Rasmussen, U (editors) Cyanobacteria in Symbiosis. Dordrecht, The Netherlands: Kluwer Academic Publishers, pp. 153–178.Google Scholar
Liu, CC and Zheng, WW (1989) Azolla in China. Beijing: Agricultural publishing House, pp. 111–117, 196–198 (in Chinese).Google Scholar
Murry, AE, Hollibaugh, JT and Orrego, C (1996) Phylogenetic compositions of acterioplankton from two California estuaries compared by denaturing gradient gel electrophoresis of 16S rDNA fragments. Applied and Environmental Microbiology 62: 2676–2680.CrossRefGoogle Scholar
Muyzer, G, de Waal, EC and Uitterlinden, AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Applied and Environmental Microbiology 59: 695–700.CrossRefGoogle Scholar
Nickell, LG (1958) Physiological studies with Azolla under aseptic conditions. I. Isolation and preliminary growth studies. American Fern Journal 48: 103–108.CrossRefGoogle Scholar
Nierzwicki-Bauer, SA and Aulfinger, H (1990) Ultrastructural characterization of eubacteria residing within leaf cavities of symbiotic and cyanobiont-free Azolla mexicana. Current. Microbiology 21: 123–129.CrossRefGoogle Scholar
Nierzwicki-Bauer, SA and Aulfinger, H (1991) Occurrence and ultrastructural characterization of bacteria in association with Azolla. Applied and Environmental Microbiology 57: 3629–3636.CrossRefGoogle ScholarPubMed
Peters, GA and Meeks, JC (1989) The Azolla–Anabaena symbiosis: Basic biology. Annual Review of Plant Physiology and Plant Molecular Biology 40: 193–210.CrossRefGoogle Scholar
Peters, GA, Calvert, HE, Kaplan, D, Ito, O and Toia, RE Jr (1982) The Azolla–Anabaena symbiosis: Morphology, physiology and use. Israel Journal of Botany 31: 305–323.Google Scholar
Plazinski, J, Taylor, R, Shaw, W, Croft, L, Rolfe, BG and Gunning, BES (1990) Isolation of Agrobacterium sp. strain from the Azolla leaf cavity. FEMS Microbiology Letters 70: 55–59.CrossRefGoogle Scholar
Ren, YH, Xu, XY and Tang, Q (2005) Molecular characterization of microbial populations in anaerobic PCP-degrading bioreactor. Acta Scientiae Circumstantiae 25(2): 214–219 (in Chinese with English abstract).Google Scholar
Serrano, R, Carrapico, F and Vidal, R (1999) The presence of lectins in bacteria associated with Azolla–Anabaena symbiosis. Symbiosis 15: 169–178.Google Scholar
Zheng, WW (1991) Occurrence of bacteria in Azolla–Anabaena association and their interaction. Journal of Electron Microscopy of China 15: 54–56 (in Chinese with English abstract).Google Scholar
Zheng, WW, Lin, YH, Lu, PJ and Liu, CC (1987) Scanning electron microscopic observation of symbiotic relationship of Azolla–Anabaena azollae during the vegetative growth. Acta Botanica Sinica 29: 588–593 (in Chinese with English abstract).Google Scholar