Proceedings of the XLVI Italian
Society of Agricultural Genetics - SIGA Annual Congress
Giardini
Naxos, Italy - 18/21 September, 2002
ISBN 88-900622-3-1
Poster
Abstract - 5.22
STUDY
OF DIFFERENTIALLY EXPRESSED GENES IN APPLE
Toller C., Zini E., Baldi P., Komjanc M.
Istituto Agrario
di San Michele all’Adige- via E. Mach n°1-38010 San Michele
all’Adige (TN)
apple,
PCR-select, differentially expressed genes
For years the Agricultural Institute of San Michele all’Adige
carried out the study of the plant - pathogen interaction in apple and recently
has started a project called “Advanced Biology Applied to Grape, Apple and Salmons” financed
by the Caritro foundation.
In the project the apple research is developed on three principal lines:
MAS (marker assisted selection) for scab and oidium resistance; identification
of molecular markers related to Va scab
resistance gene and isolation and cloning of resistance genes analogous and
differentially expressed sequences related to fungal infection resistance and
salicylic acid treatment.
Our contribution is focused on the isolation and characterisation of
differentially expressed genes and sequences associated to signal transduction
pathways involved in apple defence mechanisms.
In this study the
PCR-Select technique is used (kit “PCR-Select cDNA Subtraction” by
Clontech); method already used by E.Wang et al. (Nature Biotechnology 2001) for
the identification and isolation of rare and differentially expressed genes.
Literature data say that such method is more efficient and effective in
identifying rare genes than the differential display and cDNA AFLP.
The current study is turned to the molecular analysis in order to
identify defence and signal transduction genes during salicylic acid treatment
and SAR (systemical applied resistance) response.
At present 347 clones have been isolated and 121 of these sequenced.
The results revealed 8 most frequent sequences; the two most representative
genes were a proline synthetase and a beta-1,3-glucanase (with a frequency
respectively of 21.5 % and 17.3 %
on the total of the sequenced clones).
It’s interesting to note also the presence of a sequence coding
for a hypothetical protein highly homologue (89% of identity) to the 9
multispanning membrane protein family.
All these genes are known to be induced by salicylic acid treatment
and/or fungal infection, but have never been studied in apple.
Our next step will be to confirm, by Northen analysis, the real
expression of these genes in the leaf
tissue.