Understanding the dissemination of viruses and viroids identified through virome analysis of major grapevine rootstocks and RPA-based detection of prevalent grapevine virus B

被引：3

作者：

Thapa, Pooja ^{[1
]}

Diksha, Damini ^{[1
]}

Sharma, Susheel K. ^{[1
]}

Khan, Zainul A. ^{[1
]}

Gupta, Nitika ^{[1
]}

Shimray, Mailem Yazing ^{[1
]}

Prajapati, Malyaj R. ^{[1
]}

Holkar, Somnath Kadappa ^{[2
]}

Naik, Sharmistha ^{[2
]}

Saha, Sujoy ^{[2
]}

Baranwal, Virendra K. ^{[1
]}

机构：

[1] ICAR Indian Agr Res Inst, Adv Ctr Plant Virol, Div Plant Pathol, New Delhi 110012, India

[2] ICAR Natl Res Ctr Grapes, Pune 412307, Maharashtra, India

来源：

SCIENTIA HORTICULTURAE | 2024年 / 337卷

关键词：

Virome analysis; Grapevine rootstock; Virus and viroid dissemination; Grapevine virus B; Recombinase polymerase amplification; DEEP SEQUENCING ANALYSIS; RT-PCR DETECTION; SCION;

D O I：

10.1016/j.scienta.2024.113538

中图分类号：

S6 [园艺];

学科分类号：

0902 ;

摘要：

Grapevine stocks are generally vegetatively propagated for the commercial cultivation. The propagation is predominantly done on rootstocks with >90 % of plant stocks propagated on Dogridge rootstock alone in India. Three additional rootstocks i.e. Paulsen, SO4 and Ramsey are also used for grapevine propagation on a limited scale. However, no information is available on the health status of these rootstocks being used by nurserymen and growers. Therefore, these rootstocks were collected from nurseries of five different locations and subjected to high-throughput sequencing (HTS) to determine the spectrum of viruses and viroids. The de novo assembly of unaligned reads revealed the presence of genome sequences of nine viruses viz. grapevine rupestris stem pittingassociated virus (GRSPaV), grapevine leafroll-associated virus 3 (GLRaV-3), grapevine leafroll-associated virus 4 (GLRaV-4), grapevine virus A (GVA), grapevine virus B (GVB), grapevine virus D (GVD), grapevine virus F (GVF), grapevine virus L (GVL), grapevine fleck virus (GFkV) and five viroids viz. hopstunt viroid (HSVd), grapevine yellow speckle viroid 1 and 2 (GYSVd-1 and GYSVd-2), Australian grapevine viroid (AGVd) and grapevine latent viroid (GLVd). The identified viruses and viroids were further confirmed through RT-PCR and Sanger sequencing. Indexing for the presence of these graft-transmissible pathogens in 43 rootstocks and 23 scion samples collected through field surveys indicated that infected rootstocks are the probable source for dissemination of these viruses and viroids in grapevine scions. As GVB was the most prevalent virus in all the rootstocks and scions as observed in RT-PCR testing, a simplified crude sap-based recombinase polymerase amplification (RPA) assay was optimized for its rapid detection. The present study highlights that grapevine rootstocks are reservoir of viruses and viroids and source for their dissemination in grapevine vineyards. Findings of the study including the optimized simplified cost-effective isothermal nucleic acid-based detection technique for GVB will be highly useful for production of certified clean rootstocks of grapevine in India.

引用

页数：17