Next Article in Journal
RNASeq Analysis of Aedes albopictus Mosquito Midguts after Chikungunya Virus Infection
Next Article in Special Issue
High-Throughput Sequencing Analysis of Small RNAs Derived from Coleus Blumei Viroids
Previous Article in Journal
Cell Density Effects in Different Cell Culture Media and Their Impact on the Propagation of Foot-And-Mouth Disease Virus
Previous Article in Special Issue
The Avocado Sunblotch Viroid: An Invisible Foe of Avocado
Open AccessArticle

A Highly Sensitive Method to Detect Avocado Sunblotch Viroid for the Maintenance of Infection-Free Avocado Germplasm Collections

1
Subtropical Horticultural Research Station, Agricultural Research Service, United States Department of Agriculture, Miami, FL 33158, USA
2
Centre for Horticulture Science, Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, QLD 4072, Australia
3
Tropical Research and Education Center, University of Florida, Homestead, FL 33031, USA
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Viruses 2019, 11(6), 512; https://doi.org/10.3390/v11060512
Received: 29 March 2019 / Revised: 22 May 2019 / Accepted: 28 May 2019 / Published: 4 June 2019
(This article belongs to the Special Issue Viroid-2018: International Conference on Viroids and Viroid-Like RNAs)
The United States Department of Agriculture (USDA) Agricultural Research Service (ARS) Subtropical Horticulture Research Station (SHRS) in Miami, FL holds a large germplasm collection of avocado (Persea americana). The recent threat of infection by laurel wilt has encouraged the creation of a backup collection at a disease-free site. Creating the backup collection is complicated by infection of some trees in the germplasm collection with avocado sunblotch viroid (ASBVd). Infected trees are frequently asymptomatic, necessitating the use of a molecular diagnostic assay. Although a reverse-transcription based assay already exists and has been used to assay all germplasm at the station, some trees showed inconsistent results. We have developed a more sensitive and specific assay involving pre-amplification of the entire viroid cDNA followed by detection using real-time PCR and a TaqMan assay. A second screening of all germplasm identified additional ASBVd -infected trees and allowed us to confidently remove these trees from the station. This method enables avocado germplasm curators to proceed with the creation of a viroid-free backup collection. View Full-Text
Keywords: ASBVd; RT-PCR; germplasm curation; viroid ASBVd; RT-PCR; germplasm curation; viroid
Show Figures

Figure 1

MDPI and ACS Style

Kuhn, D.N.; Freeman, B.; Geering, A.; Chambers, A.H. A Highly Sensitive Method to Detect Avocado Sunblotch Viroid for the Maintenance of Infection-Free Avocado Germplasm Collections. Viruses 2019, 11, 512.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop