Small RNA sequencing and multiplex RT-PCR for diagnostics of grapevine viruses and virus-like organisms

Miljanić, Vanja (avtor), Jakše, Jernej (avtor), Rusjan, Denis (avtor), Škvarč, Andreja (avtor), Štajner, Nataša (avtor)

Metagenomic approaches used for virus diagnostics allow for rapid and accurate detection of all viral pathogens in the plants. In order to investigate the occurrence of viruses and virus-like organisms infecting grapevine from the Ampelographic collection Kromberk in Slovenia, we used Ion Torrent small RNA sequencing (sRNA-seq) and the VirusDetect pipeline to analyze the sRNA-seq data. The used method revealed the presence of: Grapevine leafroll-associated virus 1 (GLRaV-1), Grapevine leafroll-associated virus 2 (GLRaV-2), Grapevine leafroll-associated virus 3 (GLRaV-3), Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine fanleaf virus (GFLV) and its satellite RNA (satGFLV), Grapevine fleck virus (GFkV), Grapevine rupestris vein feathering virus (GRVFV), Grapevine Pinot gris virus (GPGV), Grapevine satellite virus (GV-Sat), Hop stunt viroid (HSVd), and Grapevine yellow speckle viroid 1 (GYSVd-1). Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for validation of sRNA-seq predicted infections, including various combinations of viruses or viroids and satellite RNA. mRT-PCR could further be used for rapid and cost-effective routine molecular diagnosis, including widespread, emerging, and seemingly rare viruses, as well as viroids which testing is usually overlooked.

• Jezik: Angleški jezik
• Ključne besede: Vitis vinifera L., grapevine, virome, metagenomic analysis, viruses, sRNA-seq, molecular diagnostics, mRT-PCR
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: BF - Biotehniška fakulteta
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2022
• Št. strani: 11 str.
• Številčenje: Vol. 14, iss. 5, art. 921
• PID: 20.500.12556/RUL-136449
• UDK: 577.2
• ISSN pri članku: 1999-4915
• DOI: 10.3390/v14050921
• COBISS.SI-ID: 106521859
• Datum objave v RUL: 04.05.2022

Gradivo je del revije

• Naslov: Viruses
• Skrajšan naslov: Viruses
• Založnik: MDPI
• ISSN: 1999-4915
• COBISS.SI-ID: 517597977

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: vinska trta, virusne bolezni, virusi, viroidi, molekularna diagnostika, metagenomska analiza

Projekti

• Financer: ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
• Številka projekta: P4-0077
• Naslov: Kmetijske rastline - genetika in sodobne tehnologije

• Financer: Drugi - Drug financer ali več financerjev
• Program financ.: Public Scholarship, Development, Disability and Maintenance Fund of the Republic of Slovenia, Scholarship for nationals of the Western Balkans for postgraduate study in Slovenia