PMID- 25455904
OWN - NLM
STAT- MEDLINE
DCOM- 20160113
LR  - 20191210
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 211
DP  - 2015 Jan
TI  - Quantitation and localization of pospiviroids in aphids.
PG  - 51-4
LID - S0166-0934(14)00393-0 [pii]
LID - 10.1016/j.jviromet.2014.10.003 [doi]
AB  - In this paper, the potential role of aphids in viroid transmission was explored. 
      Apterous aphids were fed on pospiviroid-infected plants and viroid targets in the 
      aphids were consequently quantified through RT-qPCR and localized within the 
      aphid body using fluorescence in situ hybridization (FISH). Based on the 
      analytical sensitivity test, the limit of detection (LOD) was estimated at 
      1.69x10(6) viroid copies per individual aphid body. To localize the viroids in 
      the aphids, a pospiviroid-generic Cy5-labelled probe was used and the fluorescent 
      signal was determined by confocal microscopy. Viroids were clearly observed in 
      the aphid's stylet and stomach, but not in the embryos. Viroids were detected in 
      29% of the aphids after a 24h feeding period, which suggests only a partial and 
      low concentration viroid uptake by the aphid population including viroid 
      concentrations under the LOD. However, these results show that viroids can be 
      ingested by aphids while feeding on infected plants, thus potentially increasing 
      the transmission risk. The combination of FISH and RT-qPCR provides reliable and 
      fast localization and quantitation of viroid targets in individual aphids and 
      thus constitutes a valuable tool in future epidemiological research.
CI  - Copyright (c) 2014 Elsevier B.V. All rights reserved.
FAU - Van Bogaert, N
AU  - Van Bogaert N
AD  - Department of Crop Protection, Faculty of Bioscience Engineering, Ghent 
      University, Coupure Links 653, 9000 Ghent, Belgium; Plant Sciences Unit - Crop 
      Protection, Institute for Agricultural and Fisheries Research (ILVO), 
      Burgemeester Van Gansberghelaan 96, 9820 Merelbeke, Belgium.
FAU - De Jonghe, K
AU  - De Jonghe K
AD  - Plant Sciences Unit - Crop Protection, Institute for Agricultural and Fisheries 
      Research (ILVO), Burgemeester Van Gansberghelaan 96, 9820 Merelbeke, Belgium.
FAU - Van Damme, E J M
AU  - Van Damme EJ
AD  - Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Ghent 
      University, Coupure Links 653, 9000 Ghent, Belgium.
FAU - Maes, M
AU  - Maes M
AD  - Plant Sciences Unit - Crop Protection, Institute for Agricultural and Fisheries 
      Research (ILVO), Burgemeester Van Gansberghelaan 96, 9820 Merelbeke, Belgium.
FAU - Smagghe, G
AU  - Smagghe G
AD  - Department of Crop Protection, Faculty of Bioscience Engineering, Ghent 
      University, Coupure Links 653, 9000 Ghent, Belgium. Electronic address: 
      guy.smagghe@ugent.be.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20141018
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
SB  - IM
MH  - Animal Structures/virology
MH  - Animals
MH  - Aphids/*virology
MH  - Entomology/methods
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Insect Vectors/*virology
MH  - Microscopy, Confocal
MH  - Real-Time Polymerase Chain Reaction/*methods
MH  - Sensitivity and Specificity
MH  - Viroids/genetics/*isolation & purification
MH  - Virology/methods
OTO - NOTNLM
OT  - Fluorescent in situ hybridization
OT  - Myzus persicae
OT  - Potato spindle tuber viroid
OT  - Tomato apical stunt viroid
OT  - Transmission
EDAT- 2014/12/03 06:00
MHDA- 2016/01/14 06:00
CRDT- 2014/12/03 06:00
PHST- 2014/06/10 00:00 [received]
PHST- 2014/10/06 00:00 [revised]
PHST- 2014/10/10 00:00 [accepted]
PHST- 2014/12/03 06:00 [entrez]
PHST- 2014/12/03 06:00 [pubmed]
PHST- 2016/01/14 06:00 [medline]
AID - S0166-0934(14)00393-0 [pii]
AID - 10.1016/j.jviromet.2014.10.003 [doi]
PST - ppublish
SO  - J Virol Methods. 2015 Jan;211:51-4. doi: 10.1016/j.jviromet.2014.10.003. Epub 
      2014 Oct 18.