GoldenBraid-compatible infectious clone of apple latent spherical virus (ALSV) and its use for virus-induced gene silencing

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61389030%3A_____%2F25%3A00638827" target="_blank" >RIV/61389030:_____/25:00638827 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00027006:_____/25:10178146

Výsledek na webu

<a href="https://doi.org/10.32615/bp.2025.005" target="_blank" >https://doi.org/10.32615/bp.2025.005</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.32615/bp.2025.005" target="_blank" >10.32615/bp.2025.005</a>

Jazyk výsledku

angličtina

Název v původním jazyce

GoldenBraid-compatible infectious clone of apple latent spherical virus (ALSV) and its use for virus-induced gene silencing

Popis výsledku v původním jazyce

Virus-induced gene silencing (VIGS) using self-replicating viral vectors is a powerful technique for analysing plant genes and proteins. Conversely, analysing gene silencing provides insights into the infectivity, replication, movement and stability of viral vectors. This study investigates the VIGS of phytoene desaturase gene (PDS) in different Nicotiana species using a vector based on apple latent spherical virus (ALSV, species Cheravirus mali, ICTV 2023). The ALSV genome consists of RNA1 and RNA2, which were cloned separately into plasmids using the GoldenBraid 3.0 system and transformed into two Agrobacterium strains. A third plasmid containing p19, a known gene silencing suppressor, was also introduced. These threeAgrobacterium cultures are traditionally grown separately and then inoculated together (as a mixture) in plants. Here, we describe a novel All-in-One system in which RNA1, RNA2 and p19 are co-expressed in plants after inoculation of a single Agrobacterium strain. A RT-qPCR analysis comparing both methods revealed that PDS expression is significantly different in the early phase of inoculation (6 dpi), but becomes statistically similar at later time points. The All-in-One approach therefore enables efficient co-expression of multiple targets, minimises variations in gene expression and reduces handling complexity, space requirements and costs compared to conventional co-inoculation methods.

Název v anglickém jazyce

GoldenBraid-compatible infectious clone of apple latent spherical virus (ALSV) and its use for virus-induced gene silencing

Popis výsledku anglicky

Virus-induced gene silencing (VIGS) using self-replicating viral vectors is a powerful technique for analysing plant genes and proteins. Conversely, analysing gene silencing provides insights into the infectivity, replication, movement and stability of viral vectors. This study investigates the VIGS of phytoene desaturase gene (PDS) in different Nicotiana species using a vector based on apple latent spherical virus (ALSV, species Cheravirus mali, ICTV 2023). The ALSV genome consists of RNA1 and RNA2, which were cloned separately into plasmids using the GoldenBraid 3.0 system and transformed into two Agrobacterium strains. A third plasmid containing p19, a known gene silencing suppressor, was also introduced. These threeAgrobacterium cultures are traditionally grown separately and then inoculated together (as a mixture) in plants. Here, we describe a novel All-in-One system in which RNA1, RNA2 and p19 are co-expressed in plants after inoculation of a single Agrobacterium strain. A RT-qPCR analysis comparing both methods revealed that PDS expression is significantly different in the early phase of inoculation (6 dpi), but becomes statistically similar at later time points. The All-in-One approach therefore enables efficient co-expression of multiple targets, minimises variations in gene expression and reduces handling complexity, space requirements and costs compared to conventional co-inoculation methods.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10607 - Virology

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

Biologia Plantarum

ISSN

0006-3134

e-ISSN

1573-8264

Svazek periodika

69

Číslo periodika v rámci svazku

Jun

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

11

Strana od-do

38-48

Kód UT WoS článku

001537894600001

EID výsledku v databázi Scopus

2-s2.0-105011769560