A real-time PCR quantitative analysis of the Venturia inaequalis cytb gene G143A mutation and its prevalence in the Czech Republic

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F25271121%3A_____%2F20%3AN0000114" target="_blank" >RIV/25271121:_____/20:N0000114 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.pubhort.org/ejhs/85/3/4/85_3_4.pdf" target="_blank" >https://www.pubhort.org/ejhs/85/3/4/85_3_4.pdf</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.17660/eJHS.2020/85.3.4" target="_blank" >10.17660/eJHS.2020/85.3.4</a>

Jazyk výsledku

angličtina

Název v původním jazyce

A real-time PCR quantitative analysis of the Venturia inaequalis cytb gene G143A mutation and its prevalence in the Czech Republic

Popis výsledku v původním jazyce

Venturia inaequalis (Cooke) G. Winter (1875) is a fungal pathogen that causes a disease in apples known as apple scab, which is a major source of economic losses for apple growers. To contain the pathogen, fungicides have been usually applied, often from the strobilurin family. Unfortunately, only two years after the introduction of strobilurin fungicides, resistant V. inaequalis populations appeared, and a substitution of glycine at position 143 with alanine (G143A) in the mitochondrial cytb gene was found to be the mutation responsible. Many methods exist to detect this mutation, but they are generally not suitable for simple routine use, do not provide quantitative data, or are not sensitive enough. Moreover, the quality and quantity of isolated DNA from fungi may also be an issue due to the frequent presence of PCR inhibitors. A new real-time PCR-based assay was therefore designed that can detect 0.1% of the G143A mutation in a wild type background and vice versa; in parallel, DNA isolation was optimized, using 10 million conidia in a suspension prepared from leaves harvested in orchards instead of the commonly used scab mycelia. The assay was then used for screening of the resistant V. inaequalis in orchards. The G143A mutation was present in all conventional and organic orchards tested, and in the vast majority it was present at nearly or at 100%, indicating that the mutation does not likely affect the fitness of V. inaequalis and the mutation persists over time. Furthermore, the G143A mutation (average 2.3%) was also observed in V. inaequalis isolates harvested from alley trees along roads, implying that the mutation also occurs naturally at low levels.

Název v anglickém jazyce

A real-time PCR quantitative analysis of the Venturia inaequalis cytb gene G143A mutation and its prevalence in the Czech Republic

Popis výsledku anglicky

Venturia inaequalis (Cooke) G. Winter (1875) is a fungal pathogen that causes a disease in apples known as apple scab, which is a major source of economic losses for apple growers. To contain the pathogen, fungicides have been usually applied, often from the strobilurin family. Unfortunately, only two years after the introduction of strobilurin fungicides, resistant V. inaequalis populations appeared, and a substitution of glycine at position 143 with alanine (G143A) in the mitochondrial cytb gene was found to be the mutation responsible. Many methods exist to detect this mutation, but they are generally not suitable for simple routine use, do not provide quantitative data, or are not sensitive enough. Moreover, the quality and quantity of isolated DNA from fungi may also be an issue due to the frequent presence of PCR inhibitors. A new real-time PCR-based assay was therefore designed that can detect 0.1% of the G143A mutation in a wild type background and vice versa; in parallel, DNA isolation was optimized, using 10 million conidia in a suspension prepared from leaves harvested in orchards instead of the commonly used scab mycelia. The assay was then used for screening of the resistant V. inaequalis in orchards. The G143A mutation was present in all conventional and organic orchards tested, and in the vast majority it was present at nearly or at 100%, indicating that the mutation does not likely affect the fitness of V. inaequalis and the mutation persists over time. Furthermore, the G143A mutation (average 2.3%) was also observed in V. inaequalis isolates harvested from alley trees along roads, implying that the mutation also occurs naturally at low levels.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

40100 - Agriculture, Forestry, and Fisheries

Projekt

<a href="/cs/project/QJ1510353" target="_blank" >QJ1510353: Zvýšení efektivity postupů ochrany jabloní proti strupovitosti.</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

European Journal of Horticultural Science

ISSN

1611-4426

e-ISSN

—

Svazek periodika

85

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

DE - Spolková republika Německo

Počet stran výsledku

7

Strana od-do

169-175

Kód UT WoS článku

000543446600004

EID výsledku v databázi Scopus

2-s2.0-85086832193