Cloning and targeted disruption, via Agrobacterium tumefaciens-mediated transformation, of a trypsin protease gene from the vascular wilt fungus Verticillium dahliae

Katherine F. Dobinson, Sandra J. Grant, Seogchan Kang

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

A gene encoding a trypsin protease was isolated from a tomato isolate of Verticillium dahliae. The gene, designated VTP1, contains two introns and is predicted to encode a protein of 256 amino acids. The gene is present in V. dahliae isolates from different host plants and in V. albo-atrum; weakly hybridizing sequences are present in V. tricorpus. VTP1 cDNA sequences were identified in a sequence tag analysis of genes expressed under growth conditions that promote microsclerotia development. Replacement of the gene, by Agrobacterium tumefaciens-mediated transformation (ATMT), with a mutant allele construct did not noticeably alter either pathogenicity or growth in culture. Searches of expressed sequence tag databases showed that, in addition to the VTP1 gene, V. dahliae contains two genes encoding subtilisin-like proteases similar to those produced by pathogenic Aspergillus spp. This is the first description of the application of ATMT to the molecular analysis of phytopathogenic Verticillium spp.

Original languageEnglish (US)
Pages (from-to)104-110
Number of pages7
JournalCurrent Genetics
Volume45
Issue number2
DOIs
StatePublished - Feb 1 2004

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Verticillium
Agrobacterium tumefaciens
Trypsin
Blood Vessels
Organism Cloning
Peptide Hydrolases
Fungi
Genes
Subtilisin
Expressed Sequence Tags
Aspergillus
Lycopersicon esculentum
Growth
Introns
Sequence Analysis
Virulence
Complementary DNA
Alleles
Databases
Amino Acids

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

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title = "Cloning and targeted disruption, via Agrobacterium tumefaciens-mediated transformation, of a trypsin protease gene from the vascular wilt fungus Verticillium dahliae",
abstract = "A gene encoding a trypsin protease was isolated from a tomato isolate of Verticillium dahliae. The gene, designated VTP1, contains two introns and is predicted to encode a protein of 256 amino acids. The gene is present in V. dahliae isolates from different host plants and in V. albo-atrum; weakly hybridizing sequences are present in V. tricorpus. VTP1 cDNA sequences were identified in a sequence tag analysis of genes expressed under growth conditions that promote microsclerotia development. Replacement of the gene, by Agrobacterium tumefaciens-mediated transformation (ATMT), with a mutant allele construct did not noticeably alter either pathogenicity or growth in culture. Searches of expressed sequence tag databases showed that, in addition to the VTP1 gene, V. dahliae contains two genes encoding subtilisin-like proteases similar to those produced by pathogenic Aspergillus spp. This is the first description of the application of ATMT to the molecular analysis of phytopathogenic Verticillium spp.",
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Cloning and targeted disruption, via Agrobacterium tumefaciens-mediated transformation, of a trypsin protease gene from the vascular wilt fungus Verticillium dahliae. / Dobinson, Katherine F.; Grant, Sandra J.; Kang, Seogchan.

In: Current Genetics, Vol. 45, No. 2, 01.02.2004, p. 104-110.

Research output: Contribution to journalArticle

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