Abstract
The genotypic diversity of insoluble macromolecules degraded myxobacteria, provided an opportunity to discover new bacterial resources and find new ecological functions. In this study, we developed a semi-nested-PCR-denaturing gradient gel electrophoresis (DGGE) strategy to determine the presence and genotypic diversity of myxobacteria in soil. After two rounds of PCR with myxobacteria-specific primers, an 194 bp fragment of mglA, a key gene involved in gliding motility, suitable for DGGE was obtained. A large number of bands were observed in DGGE patterns, indicating diverse myxobacteria inhabiting in soils. Furthermore, sequencing and BLAST revealed that most of the bands belonged to the myxobacteria-group, and only three of the twenty-eight bands belonged to other group, i.e., Deinococcus maricopensis. The results verified that myxobacterial strains with discrepant sequence compositions of gene mglA could be discriminated by DGGE with myxobacteria-specific primers. Collectively, the developed semi-nested-PCR-DGGE strategy is a useful tool for studying the diversity of myxobacteria.
Highlights
Myxobacteria are Gram-negative bacteria that are capable of multicellular, social behaviour [1,2]
Primer specificity A comparison of thirteen mglA gene sequences representing all identified myxobacteria from GenBank were aligned using Clustal W program version 1.8 [16], and results suggested that three regions were potentially taxon-specific for myxobacteria
Except for the quality of genomic DNA (Fig. 1A), no amplification was observed for seventeen non-myxobacterial strains (Table 1) using the same set of primers (Fig. 1B and 1C)
Summary
Myxobacteria are Gram-negative bacteria that are capable of multicellular, social behaviour [1,2]. The distinctive feature of myxobacteria is that the vegetative cells aggregate into large mounds and a matured fruiting body formed, by means of a peculiar gliding motility under starvation conditions [3,4]. Myxobacteria play important roles in establishing ecological function, as result of gliding motility, complex life cycle, fruiting bodies formation, degrade insoluble macromolecules and the production of bioactive compounds. Isolation technologies for myxobacteria usually involve the incubation of natural samples on a selective solid medium, to allow for the formation of fruiting body [3,4]. Many myxobacteria species could not formed its fruiting body under current culture conditions and resulted in missing numerous strains. The works about the myxobacteria were very limited mainly due to insufficiency incubation strategy and limited strain information
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