Altered fungal sensitivity to a plant antimicrobial peptide through over-expression of yeast cDNAs

Stephens, C, Harrison, SJ, Kazan, K, Smith, FWN, Goulter, KC, Maclean, DJ and Manners, JM (2005) Altered fungal sensitivity to a plant antimicrobial peptide through over-expression of yeast cDNAs. Current Genetics, 47 3: 194-201.


Author Stephens, C
Harrison, SJ
Kazan, K
Smith, FWN
Goulter, KC
Maclean, DJ
Manners, JM
Title Altered fungal sensitivity to a plant antimicrobial peptide through over-expression of yeast cDNAs
Journal name Current Genetics   Check publisher's open access policy
ISSN 0172-8083
1432-0983
Publication date 2005
Sub-type Article (original research)
DOI 10.1007/s00294-005-0562-8
Volume 47
Issue 3
Start page 194
End page 201
Total pages 8
Place of publication New York
Publisher Springer
Collection year 2005
Language eng
Subject C1
270201 Gene Expression
770804 Control of pests and exotic species
Abstract A yeast cDNA expression library was screened to identify genes and cellular processes that influence fungal sensitivity to a plant antimicrobial peptide. A plasmid-based, GAL1 promoter-driven yeast cDNA expression library was introduced into a yeast genotype susceptible to the antimicrobial peptide MiAMP1 purified from Macadamia integrifolia. Following a screen of 20,000 cDNAs, three yeast cDNAs were identified that reproducibly provided transformants with galactose-dependent resistance to MiAMP1. These cDNAs encoded a protein of unknown function, a component (VMA11) of the vacuolar H+-ATPase and a component (cytochrome c oxidase subunit VIa) of the mitochondrial electron transport chain, respectively. To identify genes that increased sensitivity to MiAMP1, the yeast cDNA expression library was introduced into a yeast mutant with increased resistance to MiAMP1. From 11,000 cDNAs screened, two cDNA clones corresponding to a ser/thr kinase and a ser/thr phosphatase reproducibly increased MiAMP1 susceptibility in the mutant in a galactose-dependent manner. Deletion mutants were available for three of the five genes identified but showed no change in their sensitivity to MiAMP1, indicating that these genes could not be detected by screening of yeast deletion mutant libraries. Yeast cDNA expression library screening therefore provides an alternative approach to gene deletion libraries to identify genes that can influence the sensitivity of fungi to plant antimicrobial peptides.
Keyword Genetics & Heredity
Antifungal Peptide
Cytochrome C Oxidase
H+-atpase
Signal Transduction
Fungal Pathogenesis
Plant Defence Protein
Fungal Adaptation
Saccharomyces-cerevisiae Genome
Dahlia Dahlia-merckii
Killer Toxin
Macadamia-integrifolia
Antifungal Protein
Budding Yeast
Nmr Structure
Gene
Resistance
Membrane
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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