Construction of Beauveria Bassiana T-DNA Insertion Mutant Collections and Identification of Thermosensitive and Osmosensitive Mutants

Wei Sheng Wu Xue Bao. 2009 Oct;49(10):1301-5.

Abstract

Objective: This study was to identify genes involved in thermotolerance and osmotolerance of Beauveria bassiana.

Methods: A collection of T-DNA random insertion mutagenesis was constructed and thermosensitive and osmosensitive mutants were screened. The DNA fragments flanking to inserted T-DNA sequence in the mutants were isolated by using Y-shaped adaptor dependent extension (YADE) method.

Results: Five mutants impaired in thermotolerance or osmotolerance were identified and characterized. Mutant 212 and 2550 could not grow at 32%, while mutants 139, 2737 and 2812 showed depressed growth under osmotic stress. Conidial viabilities of 212 was decreased by 23%, 52% and 21% after stressed by ultraviolet, heat shock, and H2O2 oxidation, respectively, compared to the wild-type strain. Virulence of mutant 212 and 2737 against aphids Myzus persicae were reduced by 12% and 40%, respectively. Sequencing analysis indicated that two function-unknown genes were interrupted by insertion of T-DNA in mutant 212 and 2737. In other three mutants, the insertions are located in the non-encoding regions.

Conclusion: The results demonstrate the successful use of T-DNA insertion mutagenesis in identification of genes involved in thermotolerance and osmotolerance of B. bassiana.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aphids / microbiology
  • Beauveria / genetics*
  • Beauveria / pathogenicity
  • Beauveria / physiology
  • DNA, Bacterial / genetics*
  • Hot Temperature
  • Mutagenesis, Insertional
  • Mutation*
  • Osmosis
  • Virulence

Substances

  • DNA, Bacterial
  • T-DNA