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Proteomic analysis of the biological control fungus Trichoderma

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thesis
posted on 28.03.2022, 22:43 by Jasmine Grinyer
Trichoderma harzianum and T. atroviride are filamentous fungi commonly found in soil. Both display biocontrol capabilities against a range of phytopathogenic fungi including Rhizoctonia solani and Botrytis cinerea which are known pests of hundreds of commercially important crops including tomatoes, potatoes, beans, cucumber, strawberries, cotton and grapes. These Trichoderma species secrete a combination of enzymes degrading cell walls and antibiotics to overgrow and kill fungal phytopathogens. They are seen as an environmentally friendly alternative to chemical fungicides currengly used on crops. A proteomic approach was taken to separate and identify proteins from a strain of T. harzianum with well established biocontrol properties. Several methods were developed in this thesis to display the whole proteome content and several subcellular proteome fractions from T. harzianum. Proteins were separated by two-dimensional electrophoresis and identified by mass spectrometric methods. The resulting proteomic maps represent the first extensive array of cellular and sub-cellular proteomes for T. harzianum. Cellular protein patterns of T. atroviride (T. harzianum P1) grown on media containing either glucose or R. solani cell walls were compared by differential gel electrophoresis to identify a suite of new proteins involved in the biological control response. Twenty four T. atroviride protein spots up-regulated in the presence of the R. solani cell walls were identified by mass spectrometry and N-terminal sequencing. Proteins identified from this study included previously implicated enzymes degrading cell walls and three novel proteases, vacuolar serine protease, vacuolar protease A and trypsin-like protease. The genes encoding two of these proteases, vacuolar protease A and vacuolar serine protease have been cloned by degenerate primer PCR and genomic walking PCR and sequenced. The gene sequences and protein sequences derived from these genes have been partially characterised.

History

Table of Contents

1. Introduction -- 1.1. Proteomics and two-dimensional electrophoresis -- 1.2. A proteomic approach to study the filamentous fungus Trichoderma -- 1.3. Aims of the thesis -- 2. Materials and methods -- 3. Results and discussion -- 3.1. Method development for the display and identification of fungal proteins by 2DE and mass spectrometry -- 3.2. Discovery of novel determinants in the biological control of phytopathogens by Trichoderma atroviride -- 3.3. Summary and concluding remarks.

Notes

Bibliography: leaves 157-183 "August 2006" Thesis by publication.

Awarding Institution

Macquarie University

Degree Type

Thesis PhD

Degree

Thesis (PhD), Macquarie University, Division of Environmental & Life Sciences, Dept. of Biological Sciences & Dept. of Chemistry & Biomolecular Sciences)

Department, Centre or School

Dept. of Biological Sciences | Dept. of Chemistry and Biomolecular Sciences

Year of Award

2007

Principal Supervisor

Helena Nevalainen

Additional Supervisor 1

Ben Herbert

Rights

Copyright disclaimer: http://www.copyright.mq.edu.au Copyright Jasmine Grinyer 2006.

Language

English

Extent

194 leaves ill

Former Identifiers

mq:1375 http://hdl.handle.net/1959.14/12407 1115567