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Related Experiment Videos

Targeted gene mutation in Phytophthora spp.

Kurt H Lamour1, Ledare Finley, Oscar Hurtado-Gonzales

  • 1Department of Entomology and Plant Pathology, The University of Tennessee, Rm 205 Ellington Plant Science, 2431 Joe Johnson Dr., Knoxville 37996, USA. klamour@utk.edu

Molecular Plant-Microbe Interactions : MPMI
|December 13, 2006
PubMed
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Researchers developed a new method to create gene-specific mutants in Phytophthora, a genus of plant pathogens. This technique, targeting induced local lesions in genomes (TILLING), enables functional genomics studies in Phytophthora species.

Area of Science:

  • Plant Pathology
  • Molecular Biology
  • Genetics

Background:

  • Phytophthora species are significant plant pathogens within the oomycetes.
  • Current limitations exist in mutating specific genes within this genus.
  • Availability of whole genome sequences for key Phytophthora species necessitates advanced functional genomics tools.

Purpose of the Study:

  • To adapt and implement the targeting induced local lesions in genomes (TILLING) strategy for generating gene-specific mutants in Phytophthora.
  • To investigate the feasibility of isolating mutants with specific gene disruptions in Phytophthora spp.

Main Methods:

  • Creation of a genomic library of 2,400 ethylnitrosourea (ENU) induced mutants in Phytophthora sojae.
  • Screening the mutant library for point mutations in the PsojNIP and PsPXTM-PLD genes.

Related Experiment Videos

  • Identification and characterization of homozygous mutants, including those with missense and nonsense mutations.

    • Main Results:

    • Successfully detected various point mutations (silent, missense, nonsense) in target genes within individual mutants.
    • Identified homozygous mutants with a missense mutation in PsojNIP and a premature stop codon in PsPXTM-PLD.
    • Observed reduced growth rate and appressed mycelial growth in PsPXTM-PLD homozygous mutants, while PsojNIP mutants showed no apparent phenotypic effect.

    Conclusions:

    • The TILLING approach is feasible for target-selected gene disruption in Phytophthora species.
    • This method provides a valuable new tool for functional genomic investigations in Phytophthora.
    • Gene-specific mutations can be isolated and studied to understand gene function and pathogenicity in these oomycetes.