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Published on: December 11, 2012
A novel metalloproteinase virulence factor is involved in Bacillus thuringiensis pathogenesis in nematodes and
Donghai Peng1, Jian Lin1, Qiong Huang1
1State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.
Abstract:
The Gram-positive soil bacterium Bacillus thuringiensis has been developed as the leading microbial insecticide for years. The pathogenesis of B. thuringiensis requires common extracellular factors that depend on the PlcR regulon, which regulates a large number of virulence factors; however, the precise role of many of these proteins is not known. In this study, we describe the complete lifecycle of a nematicidal B. thuringiensis strain in the free living nematode Caenorhabditis elegans using in vitro and in vivo molecular techniques to follow host and bacterial effectors during the infection process. We then focus on the metalloproteinase ColB, a collagenase, which was found highly important for destruction of the intestine thereby facilitates the adaptation and colonization of B. thuringiensis in C. elegans. In vivo green fluorescent protein (GFP) reporter-gene studies showed that ColB expression is highly induced and regulated by the global activator PlcR. Finally, we demonstrated that ColB also takes part in B. thuringiensis virulence in an insect model following injection and oral infection. Indeed, addition of purified ColB accelerates the action of Cry toxin proteins in insects, too. These results give novel insights into host adaptation for B. thuringiensis and other B. cereus group bacteria and highlight the role of collagenase metalloproteases to synergize infection process.
Insights
Bacillus thuringiensis uses the collagenase ColB to break down the intestine of Caenorhabditis elegans, aiding its colonization. ColB also enhances the efficacy of insecticidal Cry toxins, improving microbial insecticide performance.
Area of Science:
- Microbiology
- Molecular Biology
- Nematology
Background:
- Bacillus thuringiensis is a key microbial insecticide.
- Virulence factors regulated by the PlcR regulon are crucial for pathogenesis.
- The specific roles of many virulence factors remain unclear.
Purpose of the Study:
- To investigate the lifecycle of a nematicidal B. thuringiensis strain in Caenorhabditis elegans.
- To elucidate the role of the metalloproteinase ColB in B. thuringiensis pathogenesis.
- To explore ColB's contribution to virulence in both nematodes and insects.
Main Methods:
- In vitro and in vivo molecular techniques to track host and bacterial factors.
- Green fluorescent protein (GFP) reporter gene assays to monitor ColB expression.
- Nematode and insect infection models (oral and injection).
Main Results:
- ColB is essential for B. thuringiensis intestinal destruction and colonization in C. elegans.
- ColB expression is induced and regulated by PlcR in vivo.
- ColB enhances B. thuringiensis virulence in insect models and potentiates Cry toxin activity.
Conclusions:
- ColB, a collagenase, plays a significant role in B. thuringiensis virulence against nematodes and insects.
- PlcR-regulated collagenases are important for host adaptation in B. thuringiensis and B. cereus group bacteria.
- ColB synergizes infection by facilitating bacterial colonization and enhancing toxin efficacy.
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