© Benaki Phytopathological Institute
Abstracts - 16th Hellenic Phytopathological Congress
81
Characterization and distribution of fungicide resistant phenotypes in
Botrytis cinerea
originating from lettuce crops in Greece
M. C
HATZIDIMOPOULOS
and A. C. P
APPAS
University of Thessaly, Department of Agriculture Crop Production and Rural
Environment, GR-384 46 N. Ionia Volos, Greece
The minimum inhibitory concentration (MIC)
for spore germination and mycelial growth of
447
B. cinerea
isolates to nine fungicides from
different chemical groups was determined
in
vitro
, by the point inoculation method. Three
phenotypes with multiple resistance to sto-
bilurins (QoI), carboximides (Bos), hydroxya-
nilides (Hyd), anilinopyrimidines (Ani), phe-
nylpyrroles (Phen), dicarboximides (Dic) and
benzimidazoles(Βen),[QoI
ΗR
Bos
ΗR
Hyd
HR
Ani
ΗR
P
hen
MR
Dic
MR
Ben
HR
, QoI
ΗR
Bos
ΗR
Ani
ΗR
Dic
MR
Ben
HR
,
QoI
ΗR
Bos
ΗR
Dic
MR
Ben
HR
] were detected at fre-
quency 7%, 12% and 6%, respectively. These
phenotypes were mainly isolated from a
glasshouse lettuce crop located in Thessaly.
Isolates with single high resistance to benz-
imidazoles counted 11% in total, and mainly
originated from open field lettuce grown in
the Macedonia or Peloponnese regions. Phe-
notypes with various resistance combina-
tions to 1-5 chemical groups were found less
frequently (<5%). No resistant phenotypes to
fluazinam (dinitroanilides) and chlorothalonil
(phthalonitriles) were detected.
B. cinerea
phenotypes
exhibiting multiple resistance
to 7 different groups of fungicides, high re-
sistance to fenhexamid (Hyd) and moderate
resistance to fludioxonil (Phen), were record-
ed for the first time in Greece. To reduce grey
mould losses the application of anti-resis-
tance strategies is considered necessary.
Molecular characterization, methodology of
sdh
B mutation identification,
and cross-resistance in SDHI-resistant
Botrytis cinerea
isolates
T. V
ELOUKAS
1
, M. L
EROCH
2
, M. H
AHN
2
and G.S. K
ARAOGLANIDIS
1
Plant Pathology Laboratory, Faculty of Agriculture, Aristotles University of
Thessaloniki, GR-541 24 Thessaloniki, Greece
Succinate dehydrogenase inhibitors (SDHIs)
have been used against grey mould since
the end of the previous decade. Howev-
er, shortly after their introduction into the
spray programmes resistance emerged
both in Greece and worldwide. The current
study was conducted to: i) investigate the
molecular mechanism of resistance in 25
B.
cinerea
isolates showing different levels of
resistance to the SDHI fungicide boscalid,
ii) develop molecular methods for the rap-
id identification of resistance mutations
and iii) investigate the cross-resistance re-
lationships among members of the SDHI
group. Sequence analysis of the
sdh
B
, sdh
C
and
sdh
D
subunits of the gene
revealed five
mutations leading to amino acid substitu-
tion in the
Sdh
B subunit (P225F, N230I και
H272L/R/Y). To facilitate rapid detection of
these mutations associated with resistance
to boscalid, a primer introduced restriction
analysis PCR (PIRA PCR) was developed. The
method was successfully applied to the en-
tire resistant subpopulation. To study the
cross-resistance relationships, 30 isolates (5
per genotype) were characterized for their
sensitivities to 8 SDHI fungicides. The results
showed different sensitivities and cross re-
sistance patterns between structurally dif-
ferent SDHIs. P225F mutants were resistant
in vitro
to all SDHIs tested. Similarly, isolates
possessing the H272L mutation were high-
ly resistant to boscalid, but showed low to
moderate levels of resistance to other SDHIs.
The N230I mutants were moderately resis-
tant to boscalid, fluopyram and fluxapyrox-
ad and showed low levels of resistance to
isopyrazam, bixafen, fenfuram, benodanil
and carboxin. The H272R mutants showed
moderate levels of resistance to boscalid
