00 ZLOM VURV 06 v roce 07

25
cial seed of wheat, barley and potatoes. In 2006 a set of
comparative electrophoretic analyses of potato tuber
proteins was realized. The experiments were done in
RICP and in Potato Research Institute in HavlíãkÛv
Brod. Both laboratories achieved consistent results.
Analyses of wheat and barley samples were done in
2006 in order to check the variety declaration using
storage protein (gliadins and hordeins) electrophoresis.
Several paid expertises were realized by request.
(S˘korová S., Bradová J.)
Department of Molecular Biology
Study and utilisation of biodiversity, genetic
mechanisms and new methods with the aim
to improve the biological potential of cultivars
and sustainable development of agriculture
(MZe âR 0002700602)
Phase 3: Methods for improving the storability
of seeds and plant parts of vegetatively
propagated crops
Development and utilisation of effective methods of
preservation of plant germplasm resources – seed and
explants.
Eleven cultivars of Vitis sp. were introduced into
conditions in vitro. Differential scanning calorimetric
measurements of dormant grapevine shoots were
carried out to determine the freezing of water in
dormant buds. It was concluded that the freezing of
buds consisted of three exothermic events although
only one endotherm was found during heating. The
obtained results were used to propose the optimal
temperature limits for the first step of dormant bud
cryopreservation by the slow cooling method. A new
method for the regeneration of apple dormant buds by
chip budding was evaluated. The influence of the size
and desiccation procedure of shoot tips from in vitro
potato cultures on water content and time necessary to
achieve the optimal dehydration of shoot tips was
studied. 42 Allium genotypes were introduced into conditions
in vitro.The dependence of the dehydration level
of Allium shoot tips on the glass transition was defined.
Seeds of selected crop plants harvested in previous seasons
were analysed. After the application of natural compounds
the seeds were evaluated on the basis of germination
and laboratory defined germination in order to
quantify the influence of environment and treatment on
their storability. Calorimetric determination of seed
energy, chemical analysis, and measurement of the
amount of phytohormones (auxins, cytokinins and
abscissic acid) were performed. (Zámeãník J., Bilavãík
A., Faltus M., Bláha L., Jandurová O.)
Phase 4: Characterisation and development
of new genotypes of cultivated species by
molecular biology approaches
The development, optimisation and validation of
methods that are suitable for effective characterisation
of the structure and function of plant genome are main
goals of the project. New approaches may allow more
effective characterisation of genotype and genetic
resources. They can further be used for the improvement
of cultivated species.
In-house developed cDNA based DNA arrays were
fully optimised in 2006. In total 386 probes corresponding
to the genes expressed after cold stress in barley
plants cloned into the plasmids were amplified, purified
and set up to appropriate concentrations. Probes were
spotted on a glass support and hybridised with
Cy3/Cy5 labelled RNA isolated from leaves and
crowns of cold stressed barley cultivars. After washing,
scanning, appropriate scaling and normalisation up and
down regulated genes were identified. Differences
between cultivars and tissues were identified.
Wild Allium species were analysed by AFLP. MseI x
EcoRI restriction enzymes were used prior to preselective
and selective amplifications. In total 12 selective
fluorescently labelled primer pairs were used to amplify
236 polymorphic signals, i.e. 19 polymorphic signals per
primer combination. Similarity matrices were calculated
and data were processed in STATISTICA program to
perform cluster analysis.The respective dendrogram was
drawn. PCA and dendrogram divided the genotypes
according to the species and place of origin.
PC software that allows easy identification of
reliable markers was developed, validated and the
program is ready for users. (Ovesná J., Svejkovská B.,
Mal˘ M., Lei‰ová L., Kuãera L., Stavûlíková H.,
Udavská H., Pouchová V.)
Example of cold stressed barley RNA analysis by in-house
developed DNA array (a) raw scan of the array (b) data
processing resulting in cold regulated genes identification
(author M. Malý)