Applied Entomology and Zoology (2013) 48, 477-487

Zhangxun Wang, Huimin Meng, Zonglan Zhuang, Mingjun Chen, Ling Xie and Bo Huang (2013)
Molecular cloning of a novel subtilisin-like protease (Pr1A) gene from the biocontrol fungus Isaria farinosa
Applied Entomology and Zoology 48 (4), 477-487
Abstract: Fungal proteases, such as subtilisin-like serine protease (Pr1) and trypsin-like serine protease (Pr2), play an important role in penetration through the host cuticle in entomopathogenic fungi, including Isaria farinosa. In the present study, one gene of I. farinosa subtilisin-like protease (Ifa-pr1A), showing high homology to a Beauveria bassiana cuticle-degrading protease, was amplified by rapid amplification of cDNA ends PCR. The resulting full-length cDNA displayed an open reading frame (ORF) of 960 bp, encoding a protein of 319 amino acids. The Ifa-Pr1A protein was expressed in Escherichia coli to verify its protease activity. The recombinant Ifa-Pr1A protein exhibited high enzymatic activity according to the enzyme assay using a synthetic substrate. The expression profiles of Ifa-pr1A and Ifa-pr1H (another subtilisin-like protease gene from I. farinosa) were analyzed at different induction times by adding cuticle material to the media. Quantitative reverse transcriptase PCR (RT-PCR) analysis revealed that Ifa-pr1A transcripts increased 58,800-fold at 12 h post-induction, whereas Ifa-pr1H transcripts peaked at 12 h with only an 11-fold increase, indicating that Ifa-pr1A may be a major gene of cuticle-degrading proteases in I. farinosa. Furthermore, Ifa-pr1A gene expression under in vivo conditions showed a clear upward trend during the early stages of the infection process. These results suggest that Pr1A cloned from I. farinosa is a potential virulence factor for the development of engineered biopesticides.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website

Research topic(s) for pests/diseases/weeds:
biocontrol - natural enemies
Research topic(s) for beneficials or antagonists:
molecular biology - genes

---

Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
Cordyceps farinosa (entomopathogen)			China (south)