© Benaki Phytopathological Institute
Abstracts - 16th Hellenic Phytopathological Congress
49
mitted persistently by the aphid species
Aphis gossypii
and
Myzus persicae
. During ex-
tensive surveys conducted in 2011-2012, leaf
samples were collected from cucurbits ex-
hibiting yellowing symptoms in different ar-
eas of Greece. From the affected crops, we
also collected arable weeds and adult white-
flies. RT-PCR was used for the determina-
tion and discrimination of viruses and Taq-
Man PCR was used for typing of whitefly
species. In total, 128 samples from 4 cucur-
bit crops (cucumber, zucchini, melon, water-
melon), 34 samples from 6 weed species and
241 adult whiteflies were collected. Results
showed that the etiology of CYD is correlat-
ed with the presence of the whitely-vector
species that exists in the geographic area.
Moreover, CYSDV is the predominant virus
(35.64%) in greenhouse crops, followed by
CABYV (23.76%) and BPYV (23.76%). In con-
trast, CABYV prevails (74.1%) in open-field
crops, followed by CYSDV (66.6%), where-
as BPYV was not detected in open-field cu-
curbit crops. Infected weeds seem to play an
important role in the epidemiology of these
viruses, as neither of CYSDV, BPYV or CABYV
is seed-transmitted. It is worth mentioning
that for the first time, CYSDV and CABYVwere
detected in watermelon plants that showed
mild yellowing symptoms in Greece.
Detection and characterization of
Citrus tristeza virus
(CTV) in Cyprus
using molecular techniques
L.C. P
APAYIANNIS
, A. K
YRIAKOU
and T. K
APARI
-I
SAIA
Agricultural Research Institute, 1516, Nicosia, Cyprus
Citrus tristeza virus
(CTV) was first reported
in Cyprus in 1968 and until recently virus de-
tection has been mainly based on Mexican
lime (
Citrus aurantifolia
) indexing and ELI-
SA tests. In view of a national project for dis-
ease management and characterization, a
new diagnostic protocol based on real-time
reverse transcription and the polymerase
chain reaction (Real-Time TaqMan RT-PCR)
was developed and optimized. The proto-
col is suitable for the generic detection of
all virus isolates associated with severe or
mild symptoms in the Mediterranean ba-
sin. In addition, to discriminate between
virus strains identified in Cyprus, six prim-
er pairs were designed suitable for applica-
tion in conventional or real-time PCR assays.
Primer specificity was based on short prim-
er lengths (12-15 nucleotides), and on the
incorporation of modified bases known as
locked nucleic acids (LNAs), which increase
hybridization range and melting tempera-
ture. Evaluation of these primers in isolates
from Cyprus, Greece, and from other geo-
graphical regions showed that they were
able to discriminate different CTV strains ef-
ficiently and rapidly.
Development of a quantitative one-tube Real Time Reverse Transcription
PCR (qRT-PCR) for the detection and quantification of EMDV isolates in
different species
P.G. P
APPI
1
, C.I. D
OVAS
2
, K.E. E
FTHIMIOU
1
and N.I. K
ATIS
1
1
Aristotle University of Thessaloniki, Faculty of Agriculture, Forestry and Natural
Environment, School of Agriculture, Laboratory of Plant Pathology, GR-541 24
Thessaloniki, Greece.
2
Aristotle University of Thessaloniki, School of Veterinary
Medicine, Laboratory of Microbiology and Infectious Diseases, GR-541 24
Thessaloniki, Greece
Eggplant mottled dwarf virus
(EMDV) is en-
demic in countries surrounding the Medi-
terranean basin since 1969. Recently, the en-
tire genome of a Greek isolate originating
