Fungal proteomics

Initial mapping of biological control strain Trichoderma harzianum

Jasmine Grinyer*, Matthew McKay, Helena Nevalainen, Ben R. Herbert

*Corresponding author for this work

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

Trichoderma harzianum is a soil-borne filamentous fungus that exhibits biological control properties. T. harzianum can prevent the growth of pathogenic fungi on many types of plant crops, providing a chemically benign alternative to fungicidal agents currently on the market. A proteomic approach was taken to separate and identify proteins from a strain of T. harzianum with well established biocontrol properties. We developed a method of extracting proteins under acidic conditions that increased the solubilisation of alkaline proteins and eliminated acidic cell wall artefacts from micro-organisms in general. Combined with the use of protease inhibitors, this sample preparation method resulted in hundreds of proteins from T. harzianum being extracted and separated by two-dimensional gel electrophoresis. Proteins were identified by a combination of matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry and liquid chromatography mass spectrometry (LC MS/MS). Manual de novo sequencing was conducted to obtain sequence tags on unidentified proteins. A total of 25 protein spots were positively identified from a whole-cell protein reference map of T. harzianum.

Original languageEnglish
Pages (from-to)163-169
Number of pages7
JournalCurrent Genetics
Volume45
Issue number3
DOIs
Publication statusPublished - Mar 2004

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