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Development and validation of a method
for food quality analysis (MZe âR 1B44068)
The project aims to develop an effective tool for highthroughput
genotyping and identification of plant
material.An approach based on DNA arrays technologies
may allow the identification of numerous genes
of plant species (preferentially on the basis of allelic
discrimination) or detection of several alleles in
thousands of samples. In 2006 we aimed to provide
more information on the variability of GBSSI (granule
bound starch synthase) underlying gene that drives the
type of starch grains in wheat and related species.After
the amplification of GBSSI fragments from different
Triticeae species the respective products were cloned.
In total 120 clones were sequenced and compared.
We detected a new polymorphism in the old wheat
cultivar Stepowa. DNA sequence in the modern
cultivar Nelly was identical to that published for
cv. Chinese Spring. DNA sequences detected in wild
related species were found to be polymorphic. Lr genes
were amplified and prepared as probes for DNA arrays.
Besides, the optimisation of individual steps for the use
of DNA arrays continued. (Ovesná J., Nguyen M.,
Kuãera L., Tomková L., Svejkovská B.)
Development and application of DNA
microarray technology for a large-scale system
of gene and genotype identification
(MZe âR 1G46068)
Within the project we optimise individual steps
necessary in a high-throughput gene detection system.
The optimised procedure should be used to detect
food contaminants effectively. We use DNA arrays
technologies for the detection of specific alleles.
In 2006 we amplified and cloned DNA probes
corresponding to specific transgenes in GMOs
approved in the EU. Subsequently, recombinant
plasmids were transformed into E. coli and preserved
for subsequent use in a deep freezer. They are used as
a source of plasmids and template for amplicon multiplication
by PCR. Several approaches were tested for
the preparation of probes that are spotted on the glass.
Glasses with arrayed probes were hybridised with
assayed DNA. Plasmids, specific PCR products were
labelled with Cy3 and Cy5 fluorescent dye and used
for hybridisation. Different hybridisation conditions
were tested along with the probe type. Sensitivity of
the assay was increased.The project will continue using
another approach to label the analyte. (Ovesná J.,
Hodek J., Kuãera L., Vi‰tejnová L., Pouchová V.,
Demnerová K.)
Selection of garlic clones comprising a high
level of biologically active compound
precursors (MZe âR 1G58084)
Garlic belongs to one of the oldest cultivated plant
species. In the long history of mankind garlic has been
used as a culinary and medicinal species. Garlic (Allium
sativum L.) is a vegetatively propagated species and the
progeny resulting from intended crossing can be
obtained with obstacles. Therefore genetic resources
(GR) are a valuable wealth representing available
biodiversity, i.e. genes/alleles for future breeding programmes.The
gene bank of RICP in Prague maintains
one of the largest collections of short-day garlic. The
main aim of the project is to develop tools for highthroughput
characterisation of garlic (A. sativum)
genetic resources by sequencing and RNA profiling.
In the first year the overall variability of the
collection was analysed and available information on
O-acetylserine(thiol)-lyase EC 2.5.1.47 coding
sequence was gathered. Primers were designed and the
reaction was optimised to amplify sequences from
different sources. (Ovesná J., Mitrová K., Kuãera L.,
Velí‰ek J., Králová J.)
Study of selected factors influencing
the adventitious presence of unwanted GMO
and biodiversity in the context of the
coexistence of genetically modified, conventional
and ecological crops (MZe âR 1B53047)
In 2006 different DNA based marker systems were
investigated for haplotyping of E. repens and E. intermedium.
In total 46 accessions from Moravian and
Bohemian regions were characterised. Acquired input
information on couch-grass biodiversity in monitored
areas will contribute to the optimisation of agricultural
systems.The project is also focused on the monitoring
of changes in farming practices on a set of farms
once GM crops are commercialised. The aim of the
project is to look into technical possibilities of spatial
isolation of genetically modified crops in selected areas
of the Czech Republic. Acquired data can be used
when the Good Agricultural Practice dealing with the
release and monitoring of genetically modified crops is
being put in practice. Outputs of the project will help
to observe the legislative precautions and recommendations.
(Kuãera L., Kuãerová D., Ovesná J.,
Hanu‰ová M., Hodek J.)
The use of polymorphism of cereal genome
repetitive regions for characterisation of gene
resources and breeding materials (MZe âR
1G57064)
The project is focused on the investigation and
application of a new generation of molecular markers
for the characterisation of genetic resources and breeding
materials of wheat and barley.The markers use the
polymorphic potential of transposable element
sequences and they are developed for the scanning of
genome regions abundant in genes, mainly the regions
with R-genes and with genes influencing product
quality.The new sets of primers for haplotyping of the
barley Rht7 locus and primers for the detection of
inter-MITE polymorphism and inverted repeats of DNA
sequences were tested on 101 different genotypes of
H. vulgare. (Kuãera L., Lei‰ová L., Tomková H.
Hanu‰ová M., Ovesná J.)
Establishment of a cryobank for the
conservation of potato and hop apices
(MZe âR QF 3039)
The goal of this project is to store selected potato
cultivars of Czech origin and important hop genotypes
in liquid nitrogen.The keepers of the crop germplasms
– Potato Research Institute in HavlíãkÛv Brod
and Hop Research Institute in Îatec provide the
multiplication of in vitro plants and evaluation of
selected genotypes. The cryopreservation methods
and cryostorage of plant material are performed in
RICP in Prague. In 2006, fifteen potato cultivars
and fifteen hop genotypes were cryopreseved by