Naslov
GLRaV-3 Genetic Variants in Indigenous Vines of Mediterranean Croatia

Autori
Hančević Katarina, Saldarelli Pasquale, Čarija Mate*, Černi Silvija, Zdunić Goran, Mucalo Ana i Radić Tomislav

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
World Microbe Forum

Mjesto i datum
Online kongres, 20.06.2021. - 24.06.2021

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
GLRaV-3, molecular characterization, phylogenetic analysis

Sažetak
Background Grapevine leafroll-associated virus 3 (GLRaV-3), the main causal agent of leafroll disease, is one of the most important grapevine virus pathogens worldwide, as also proved for the Mediterranean vineyards in Croatia. There, it is continuously detected mostly in mixed infections with other viruses, but always as the most predominant one (Vončina et al., 2019 ; Hančević et al., 2021). Since the first report on GLRaV-3 population structure (Turturo et al., 2005), genetic variability has been brought to the wider attention, so new divergent variants are continuously discovering all around the globe. Objectives and Material In order to identify and characterize GLRaV-3 variants present within each isolate, thirty-three selected GLRaV-3 positive samples from most widespread native grapevine varieties were analyzed on a molecular basis. Samples included sixteen varieties from Croatian Adriatic coast, now present in the germplasm collection vineyard located at the Institute for Adriatic Crops in Split, Croatia. Methods and Results A fragment (545 bp) of the HSP70h gene of GLRAV-3 was amplified using LC1F and LC2R primer pairs (Turturo et al., 2005) to detect GLRaV-3 presence in all grapevine varieties tested. The PCR conditions were as reported by Turturo et al., except for the final extension, which in our case was prolonged to 15 min. To separate different genomic variants presumably present in each isolate, amplicons were cloned in the in E. coli competent cells. Whenever possible, 20 transformed colonies per sample (in total 586 clones) were randomly selected and analyzed by single-stranded conformational polymorphism (SSCP ; Černi et al, 2008) and sequenced. The phylogenetic analysis of all 139 sequenced variants obtained showed clustering into GLRaV-3 phylogenetic groups I and/or II supported by high bootstrap values. Genomic variants from 55% analyzed samples clustered into phylogenetic group II, while those from 27% samples clustered into phylogenetic group I. Approximately one fifth of samples, 18%, were mixed infected with both variants clustering into both phylogenetic groups I and II. This study emphasizes the predominant presence of GLRaV-3 phylogenetic groups I and II, in indigenous vines in Mediterranea Croatia, either individually or in combination.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Poljoprivreda (agronomija)

POVEZANOST RADA