Copper induction of laccases in the plant pathogenic fungus Botrytis cinerea

Research output: Other contribution to conferencePoster

Abstract

Botrytis cinerea, a fungal pathogen of grapevines, produces oxidative enzymes including laccases, during the infection of plant tissues. Laccases are reported to be induced by copper however it is not known if this is the case for B. cinerea. Three laccase genes are known (LAC1, LAC2 and LAC3) but gene specific inducers have not been fully investigated. This study investigates the expression of laccases in response to copper. Liquid cultures of B. cinerea were prepared in potato dextrose broth by inoculation of the media with 100 μL of spore suspension (105/ml). After 5 days mycelia were harvested and placed in a laccase inducing medium with a range of CuSO4 concentrations (0–0.8 mM). Laccase activities in culture filtrates were determined after 2 days. Laccase activity was dependent upon copper concentration up to 0.6 mM and decreased at 0.8 mM indicating 0.6 mM is the optimum concentration of maximum laccase production. Mycelia were harvested for mRNA transcript analysis. LAC2 gene expression was similarly correlated with the copper concentrations in the culture medium with a 5-fold increase at 0.6 mM CuSO4. There were no significant changes in LAC1 and LAC3 gene expression indicative that LAC2 is a copper-inducible laccase gene. Proteins from culture filtrates were concentrated using ultra centrifugal filters (30 KDa) and separated by SDS-PAGE. A single band corresponding to a 75 KDa protein was observed which may correspond to a copper inducible laccase. This is the first time laccase gene expression in response to copper has been investigated in B. cinerea. Copper induction of laccases in the plant pathogenic fungus, Botrytis cinerea. Available from: https://www.researchgate.net/publication/313213225_Copper_induction_of_laccases_in_the_plant_pathogenic_fungus_Botrytis_cinerea [accessed Sep 22, 2017].
Original languageEnglish
Pages122
Number of pages1
Publication statusPublished - 2016
EventComBio2016 - Brisbane Convention Centre, Brisbane, Australia
Duration: 02 Oct 201606 Oct 2016
http://www.asbmb.org.au/combio2016/

Conference

ConferenceComBio2016
CountryAustralia
CityBrisbane
Period02/10/1606/10/16
Internet address

Fingerprint

laccase
plant pathogenic fungi
Botrytis cinerea
copper
copper sulfate
culture filtrates
gene expression
mycelium
genes
plant tissues
proteins
spores
culture media

Cite this

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title = "Copper induction of laccases in the plant pathogenic fungus Botrytis cinerea",
abstract = "Botrytis cinerea, a fungal pathogen of grapevines, produces oxidative enzymes including laccases, during the infection of plant tissues. Laccases are reported to be induced by copper however it is not known if this is the case for B. cinerea. Three laccase genes are known (LAC1, LAC2 and LAC3) but gene specific inducers have not been fully investigated. This study investigates the expression of laccases in response to copper. Liquid cultures of B. cinerea were prepared in potato dextrose broth by inoculation of the media with 100 μL of spore suspension (105/ml). After 5 days mycelia were harvested and placed in a laccase inducing medium with a range of CuSO4 concentrations (0–0.8 mM). Laccase activities in culture filtrates were determined after 2 days. Laccase activity was dependent upon copper concentration up to 0.6 mM and decreased at 0.8 mM indicating 0.6 mM is the optimum concentration of maximum laccase production. Mycelia were harvested for mRNA transcript analysis. LAC2 gene expression was similarly correlated with the copper concentrations in the culture medium with a 5-fold increase at 0.6 mM CuSO4. There were no significant changes in LAC1 and LAC3 gene expression indicative that LAC2 is a copper-inducible laccase gene. Proteins from culture filtrates were concentrated using ultra centrifugal filters (30 KDa) and separated by SDS-PAGE. A single band corresponding to a 75 KDa protein was observed which may correspond to a copper inducible laccase. This is the first time laccase gene expression in response to copper has been investigated in B. cinerea. Copper induction of laccases in the plant pathogenic fungus, Botrytis cinerea. Available from: https://www.researchgate.net/publication/313213225_Copper_induction_of_laccases_in_the_plant_pathogenic_fungus_Botrytis_cinerea [accessed Sep 22, 2017].",
author = "{Udugama Vithanage}, Aruni and Sandra Savocchia and Padraig Strappe and Leigh Schmidtke and Christopher Steel",
year = "2016",
language = "English",
pages = "122",
note = "ComBio2016 ; Conference date: 02-10-2016 Through 06-10-2016",
url = "http://www.asbmb.org.au/combio2016/",

}

Copper induction of laccases in the plant pathogenic fungus Botrytis cinerea. / Udugama Vithanage, Aruni; Savocchia, Sandra; Strappe, Padraig; Schmidtke, Leigh; Steel, Christopher.

2016. 122 Poster session presented at ComBio2016, Brisbane, Australia.

Research output: Other contribution to conferencePoster

TY - CONF

T1 - Copper induction of laccases in the plant pathogenic fungus Botrytis cinerea

AU - Udugama Vithanage, Aruni

AU - Savocchia, Sandra

AU - Strappe, Padraig

AU - Schmidtke, Leigh

AU - Steel, Christopher

PY - 2016

Y1 - 2016

N2 - Botrytis cinerea, a fungal pathogen of grapevines, produces oxidative enzymes including laccases, during the infection of plant tissues. Laccases are reported to be induced by copper however it is not known if this is the case for B. cinerea. Three laccase genes are known (LAC1, LAC2 and LAC3) but gene specific inducers have not been fully investigated. This study investigates the expression of laccases in response to copper. Liquid cultures of B. cinerea were prepared in potato dextrose broth by inoculation of the media with 100 μL of spore suspension (105/ml). After 5 days mycelia were harvested and placed in a laccase inducing medium with a range of CuSO4 concentrations (0–0.8 mM). Laccase activities in culture filtrates were determined after 2 days. Laccase activity was dependent upon copper concentration up to 0.6 mM and decreased at 0.8 mM indicating 0.6 mM is the optimum concentration of maximum laccase production. Mycelia were harvested for mRNA transcript analysis. LAC2 gene expression was similarly correlated with the copper concentrations in the culture medium with a 5-fold increase at 0.6 mM CuSO4. There were no significant changes in LAC1 and LAC3 gene expression indicative that LAC2 is a copper-inducible laccase gene. Proteins from culture filtrates were concentrated using ultra centrifugal filters (30 KDa) and separated by SDS-PAGE. A single band corresponding to a 75 KDa protein was observed which may correspond to a copper inducible laccase. This is the first time laccase gene expression in response to copper has been investigated in B. cinerea. Copper induction of laccases in the plant pathogenic fungus, Botrytis cinerea. Available from: https://www.researchgate.net/publication/313213225_Copper_induction_of_laccases_in_the_plant_pathogenic_fungus_Botrytis_cinerea [accessed Sep 22, 2017].

AB - Botrytis cinerea, a fungal pathogen of grapevines, produces oxidative enzymes including laccases, during the infection of plant tissues. Laccases are reported to be induced by copper however it is not known if this is the case for B. cinerea. Three laccase genes are known (LAC1, LAC2 and LAC3) but gene specific inducers have not been fully investigated. This study investigates the expression of laccases in response to copper. Liquid cultures of B. cinerea were prepared in potato dextrose broth by inoculation of the media with 100 μL of spore suspension (105/ml). After 5 days mycelia were harvested and placed in a laccase inducing medium with a range of CuSO4 concentrations (0–0.8 mM). Laccase activities in culture filtrates were determined after 2 days. Laccase activity was dependent upon copper concentration up to 0.6 mM and decreased at 0.8 mM indicating 0.6 mM is the optimum concentration of maximum laccase production. Mycelia were harvested for mRNA transcript analysis. LAC2 gene expression was similarly correlated with the copper concentrations in the culture medium with a 5-fold increase at 0.6 mM CuSO4. There were no significant changes in LAC1 and LAC3 gene expression indicative that LAC2 is a copper-inducible laccase gene. Proteins from culture filtrates were concentrated using ultra centrifugal filters (30 KDa) and separated by SDS-PAGE. A single band corresponding to a 75 KDa protein was observed which may correspond to a copper inducible laccase. This is the first time laccase gene expression in response to copper has been investigated in B. cinerea. Copper induction of laccases in the plant pathogenic fungus, Botrytis cinerea. Available from: https://www.researchgate.net/publication/313213225_Copper_induction_of_laccases_in_the_plant_pathogenic_fungus_Botrytis_cinerea [accessed Sep 22, 2017].

UR - http://www.asbmb.org.au/combio2016/docs/POSTER%20PRESENTATIONS%20-%20FINAL%20FOR%20PROGRAM%2023.08.16.pdf

M3 - Poster

SP - 122

ER -