Laccase gene expression during the infection of grape berries by Botrytis cinerea

Research output: Other contribution to conferenceAbstract

Abstract

Laccases produced by Botrytis cinerea during infection of grape bunches can lead to wineoxidation and subsequent detrimental effects on wine quality. The role of laccases in thedevelopment of Botrytis bunch rot has not been fully elucidated even though the researchsuggests the involvement of three genes (LAC1, LAC2, LAC3) in laccase production by B. cinerea.The expression of these genes during infection by B. cinerea on grapes was examined by selectingone botrytis isolate (TN080) following an initial screening of virulence. Sterilized wine grapes(Chardonnay) were inoculated with a 10µl spore suspension (107spores∙ml-1) and incubated at24°C. Berries were collected at different time intervals post-inoculation, frozen in liquid nitrogenand total RNA extracted, followed by cDNA synthesis, RT-qPCR with primers designed for the threeLAC genes. Relative gene expression was normalized to the internal control, actin. Laccase enzymeactivity was also assessed in berry extracts collected at the same time and expressed as U∙ml-1. Allexperiments were done using three biological replicates. The expression of LAC3 was greatest (15fold) at one day post-inoculation and reduced gradually up to day 3. No expression of LAC2 andLAC1 genes were observed. Despite this gene expression, laccase enzyme activity in infected berryextracts was not observed until 3 days post-inoculation. The expression pattern of laccasetranscripts implies the involvement of the LAC3 gene in disease development. LAC3 may also be adeterminant of virulence during the infection of B. cinerea on grapes. This study implies thatlaccases may be important during the early stages of B. cinerea infection on grape berries.
Original languageEnglish
Pages61
Number of pages1
Publication statusPublished - 2016
EventXVII International Botrytis symposium - Santa Cruz Plaza Hotel, Santa Cruz, Chile
Duration: 22 Oct 201627 Oct 2016
http://www.xvii.botrytisymposium.agronomia.uchile.cl/

Conference

ConferenceXVII International Botrytis symposium
CountryChile
CitySanta Cruz
Period22/10/1627/10/16
Internet address

Fingerprint

laccase
Botrytis cinerea
grapes
gene expression
infection
Botrytis
small fruits
virulence
genes
wine grapes
wine quality
actin
spores
RNA
enzyme activity
screening
synthesis
liquids
extracts

Cite this

Udugama Vithanage, A., Steel, C., Savocchia, S., Strappe, P., & Schmidtke, L. (2016). Laccase gene expression during the infection of grape berries by Botrytis cinerea. 61. Abstract from XVII International Botrytis symposium, Santa Cruz, Chile.
@conference{1efbd6ef498048ae8f7240c5b449e012,
title = "Laccase gene expression during the infection of grape berries by Botrytis cinerea",
abstract = "Laccases produced by Botrytis cinerea during infection of grape bunches can lead to wineoxidation and subsequent detrimental effects on wine quality. The role of laccases in thedevelopment of Botrytis bunch rot has not been fully elucidated even though the researchsuggests the involvement of three genes (LAC1, LAC2, LAC3) in laccase production by B. cinerea.The expression of these genes during infection by B. cinerea on grapes was examined by selectingone botrytis isolate (TN080) following an initial screening of virulence. Sterilized wine grapes(Chardonnay) were inoculated with a 10µl spore suspension (107spores∙ml-1) and incubated at24°C. Berries were collected at different time intervals post-inoculation, frozen in liquid nitrogenand total RNA extracted, followed by cDNA synthesis, RT-qPCR with primers designed for the threeLAC genes. Relative gene expression was normalized to the internal control, actin. Laccase enzymeactivity was also assessed in berry extracts collected at the same time and expressed as U∙ml-1. Allexperiments were done using three biological replicates. The expression of LAC3 was greatest (15fold) at one day post-inoculation and reduced gradually up to day 3. No expression of LAC2 andLAC1 genes were observed. Despite this gene expression, laccase enzyme activity in infected berryextracts was not observed until 3 days post-inoculation. The expression pattern of laccasetranscripts implies the involvement of the LAC3 gene in disease development. LAC3 may also be adeterminant of virulence during the infection of B. cinerea on grapes. This study implies thatlaccases may be important during the early stages of B. cinerea infection on grape berries.",
author = "{Udugama Vithanage}, Aruni and Christopher Steel and Sandra Savocchia and Padraig Strappe and Leigh Schmidtke",
year = "2016",
language = "English",
pages = "61",
note = "XVII International Botrytis symposium ; Conference date: 22-10-2016 Through 27-10-2016",
url = "http://www.xvii.botrytisymposium.agronomia.uchile.cl/",

}

Udugama Vithanage, A, Steel, C, Savocchia, S, Strappe, P & Schmidtke, L 2016, 'Laccase gene expression during the infection of grape berries by Botrytis cinerea' XVII International Botrytis symposium, Santa Cruz, Chile, 22/10/16 - 27/10/16, pp. 61.

Laccase gene expression during the infection of grape berries by Botrytis cinerea. / Udugama Vithanage, Aruni; Steel, Christopher; Savocchia, Sandra; Strappe, Padraig; Schmidtke, Leigh.

2016. 61 Abstract from XVII International Botrytis symposium, Santa Cruz, Chile.

Research output: Other contribution to conferenceAbstract

TY - CONF

T1 - Laccase gene expression during the infection of grape berries by Botrytis cinerea

AU - Udugama Vithanage, Aruni

AU - Steel, Christopher

AU - Savocchia, Sandra

AU - Strappe, Padraig

AU - Schmidtke, Leigh

PY - 2016

Y1 - 2016

N2 - Laccases produced by Botrytis cinerea during infection of grape bunches can lead to wineoxidation and subsequent detrimental effects on wine quality. The role of laccases in thedevelopment of Botrytis bunch rot has not been fully elucidated even though the researchsuggests the involvement of three genes (LAC1, LAC2, LAC3) in laccase production by B. cinerea.The expression of these genes during infection by B. cinerea on grapes was examined by selectingone botrytis isolate (TN080) following an initial screening of virulence. Sterilized wine grapes(Chardonnay) were inoculated with a 10µl spore suspension (107spores∙ml-1) and incubated at24°C. Berries were collected at different time intervals post-inoculation, frozen in liquid nitrogenand total RNA extracted, followed by cDNA synthesis, RT-qPCR with primers designed for the threeLAC genes. Relative gene expression was normalized to the internal control, actin. Laccase enzymeactivity was also assessed in berry extracts collected at the same time and expressed as U∙ml-1. Allexperiments were done using three biological replicates. The expression of LAC3 was greatest (15fold) at one day post-inoculation and reduced gradually up to day 3. No expression of LAC2 andLAC1 genes were observed. Despite this gene expression, laccase enzyme activity in infected berryextracts was not observed until 3 days post-inoculation. The expression pattern of laccasetranscripts implies the involvement of the LAC3 gene in disease development. LAC3 may also be adeterminant of virulence during the infection of B. cinerea on grapes. This study implies thatlaccases may be important during the early stages of B. cinerea infection on grape berries.

AB - Laccases produced by Botrytis cinerea during infection of grape bunches can lead to wineoxidation and subsequent detrimental effects on wine quality. The role of laccases in thedevelopment of Botrytis bunch rot has not been fully elucidated even though the researchsuggests the involvement of three genes (LAC1, LAC2, LAC3) in laccase production by B. cinerea.The expression of these genes during infection by B. cinerea on grapes was examined by selectingone botrytis isolate (TN080) following an initial screening of virulence. Sterilized wine grapes(Chardonnay) were inoculated with a 10µl spore suspension (107spores∙ml-1) and incubated at24°C. Berries were collected at different time intervals post-inoculation, frozen in liquid nitrogenand total RNA extracted, followed by cDNA synthesis, RT-qPCR with primers designed for the threeLAC genes. Relative gene expression was normalized to the internal control, actin. Laccase enzymeactivity was also assessed in berry extracts collected at the same time and expressed as U∙ml-1. Allexperiments were done using three biological replicates. The expression of LAC3 was greatest (15fold) at one day post-inoculation and reduced gradually up to day 3. No expression of LAC2 andLAC1 genes were observed. Despite this gene expression, laccase enzyme activity in infected berryextracts was not observed until 3 days post-inoculation. The expression pattern of laccasetranscripts implies the involvement of the LAC3 gene in disease development. LAC3 may also be adeterminant of virulence during the infection of B. cinerea on grapes. This study implies thatlaccases may be important during the early stages of B. cinerea infection on grape berries.

M3 - Abstract

SP - 61

ER -

Udugama Vithanage A, Steel C, Savocchia S, Strappe P, Schmidtke L. Laccase gene expression during the infection of grape berries by Botrytis cinerea. 2016. Abstract from XVII International Botrytis symposium, Santa Cruz, Chile.