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Published on: June 1, 2018
Application of representational difference analysis to identify sequence tags expressed by Metarhizium anisopliae
Valéria Dutra1, Luciano Nakazato, Leonardo Broetto
1Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, P.O. Box 15005, 91501-970, Porto Alegre, RS, Brazil.
Abstract:
Metarhizium anisopliae is a well-characterized biocontrol agent of a wide range of plagues, including insects and acari. To identify genes involved in the infection process, representational difference analysis was performed using cDNA generated from germinated conidia of M. anisopliae in the tick Boophilus microplus cuticle, and cDNA generated during fungal growth in glucose-rich medium. Sequence determination of approximately 135 clones and comparison analysis using public databases led to the identification of 34 sequences and 14 expressed sequence tags with known orthologs. As expected, almost all identified sequences showed significant similarity to other fungal genes. The diversity of gene clusters found reflects the participation of several proteins in the early infection process of M. anisopliae in the cattle tick B. microplus.
Insights
Metarhizium anisopliae, a fungal biocontrol agent, has its infection genes identified. Researchers pinpointed genes crucial for its early infection stages in the cattle tick, Boophilus microplus.
Area of Science:
- Mycology
- Entomology
- Biocontrol
Background:
- Metarhizium anisopliae is a known biocontrol agent targeting insects and acari.
- Understanding its infection mechanisms is key to optimizing its use against pests like the cattle tick Boophilus microplus.
Purpose of the Study:
- To identify genes in Metarhizium anisopliae essential for the initial stages of infection in Boophilus microplus.
- To gain insights into the molecular basis of fungal pathogenesis in ticks.
Main Methods:
- Representational difference analysis (RDA) was employed to compare gene expression.
- cDNA libraries were generated from M. anisopliae germinated on tick cuticle versus growth in a glucose-rich medium.
- Sequencing of 135 clones and database comparisons were performed.
Main Results:
- 34 sequences and 14 expressed sequence tags with known fungal orthologs were identified.
- The identified genes showed significant similarity to other fungal genes.
- A diverse array of gene clusters involved in early infection was revealed.
Conclusions:
- The study successfully identified genes involved in the early infection process of Metarhizium anisopliae in Boophilus microplus.
- The findings highlight the complex genetic mechanisms underlying fungal pathogenesis in ticks.
- This research provides a foundation for developing more effective biocontrol strategies against cattle ticks.
