Development of a Reverse-Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Brassica Yellows Virus in China

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027006%3A_____%2F25%3A10179518" target="_blank" >RIV/00027006:_____/25:10179518 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.mdpi.com/2073-4395/15/12/2727/pdf" target="_blank" >https://www.mdpi.com/2073-4395/15/12/2727/pdf</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/agronomy15122727" target="_blank" >10.3390/agronomy15122727</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Development of a Reverse-Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Brassica Yellows Virus in China

Popis výsledku v původním jazyce

Brassica yellows virus (BrYV) mainly infects cruciferous crops and has been widely prevalent across China. To develop a rapid and highly sensitive method for detecting BrYV in oilseed rape, a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was established. Four specific primers were designed to target the conserved gene of BrYV, with total RNA extracted from BrYV-infected oilseed rape leaves used as the template for the RT-LAMP assay. The optimal reaction conditions were determined, including a primer concentration ratio of 1:8, MgSO4 concentration of 4 mM, reaction temperature of 64 ◦C, and a suitable reaction time of 60 min. Sensitivity analysis demonstrated that the RT-LAMP assay could detect total RNA at a concentration of 0.091 × 10−3 μg/μL, which was 100-fold more sensitive than conventional RT-PCR for BrYV detection. In addition to visualizing results by electrophoresis, the RT-LAMP assay could also be easily visualized using calcein-MnCl2. These results indicate the potential of the established RT-LAMP assay for rapid BrYV detection in oilseed rape plants, which can provide better technical support for field diagnosis, disease forecasting, and the implementation of effective control strategies against the virus.

Název v anglickém jazyce

Development of a Reverse-Transcription Loop-Mediated Isothermal Amplification Assay for Detecting Brassica Yellows Virus in China

Popis výsledku anglicky

Brassica yellows virus (BrYV) mainly infects cruciferous crops and has been widely prevalent across China. To develop a rapid and highly sensitive method for detecting BrYV in oilseed rape, a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was established. Four specific primers were designed to target the conserved gene of BrYV, with total RNA extracted from BrYV-infected oilseed rape leaves used as the template for the RT-LAMP assay. The optimal reaction conditions were determined, including a primer concentration ratio of 1:8, MgSO4 concentration of 4 mM, reaction temperature of 64 ◦C, and a suitable reaction time of 60 min. Sensitivity analysis demonstrated that the RT-LAMP assay could detect total RNA at a concentration of 0.091 × 10−3 μg/μL, which was 100-fold more sensitive than conventional RT-PCR for BrYV detection. In addition to visualizing results by electrophoresis, the RT-LAMP assay could also be easily visualized using calcein-MnCl2. These results indicate the potential of the established RT-LAMP assay for rapid BrYV detection in oilseed rape plants, which can provide better technical support for field diagnosis, disease forecasting, and the implementation of effective control strategies against the virus.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

40106 - Agronomy, plant breeding and plant protection; (Agricultural biotechnology to be 4.4)

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

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

Název periodika

Agronomy-Basel

ISSN

2073-4395

e-ISSN

2073-4395

Svazek periodika

15

Číslo periodika v rámci svazku

12

Stát vydavatele periodika

CH - Švýcarská konfederace

Počet stran výsledku

8

Strana od-do

2727

Kód UT WoS článku

001646088500001

EID výsledku v databázi Scopus

2-s2.0-105027531155