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Statements

Subject Item
n2:RIV%2F62156489%3A43510%2F09%3A00152163%21RIV14-MSM-43510___
rdf:type
skos:Concept n20:Vysledek
dcterms:description
Many studies describing phenotypic and genetic variation among standard and stressed plants were already published. A possibility to estimate stress-induced changes in genome, epigenome and transcriptome arose by establishing three AFLP-based methods within our laboratory (standard AFLP, methylation-sensitive AFLP and cDNA-AFLP). Three variants of grapevine plants after different stress actions (stress by in vitro cultivation, in vitro thermotherapy and virus infection) were prepared for this study. In case of standard AFLP application, comparison of obtained spectra via 0/1 binary matrix is usually performed. Within our group of variants the results show insignificant changes in its genomes. MS-AFLP spectra evaluation needs more detailed analysis of results to extract more precise information about DNA methylation characters and impact of individual stress factors on epigenetic changes. From the point of view of functional methodology, the cDNA-AFLP appeared as the most difficult to establish. During first experiments only short PCR amplicons (at most 150 bp) were generated. Further, the calculated similarity coefficients showed surprisingly low values. These facts signalised certain unspecific RNA degradation before its conversion onto ds cDNA. The significant improvement of obtained results was recorded by using RNA later reagent, SpectrumTM Plant Total RNA Kit, RNase Out enzyme and MINT cDNA synthesis kit in cDNA-AFLP protocol. In the case of cDNA-AFLP spectra evaluation, it is possible either to prepare 0/1 binary matrix or differences can be estimated on the base of ratio between intensities of individual peaks. This fact should be taken into account if cDNA-AFLP results are compared with other fingerprinting methods as RAPD, SSR or standard AFLP. Many studies describing phenotypic and genetic variation among standard and stressed plants were already published. A possibility to estimate stress-induced changes in genome, epigenome and transcriptome arose by establishing three AFLP-based methods within our laboratory (standard AFLP, methylation-sensitive AFLP and cDNA-AFLP). Three variants of grapevine plants after different stress actions (stress by in vitro cultivation, in vitro thermotherapy and virus infection) were prepared for this study. In case of standard AFLP application, comparison of obtained spectra via 0/1 binary matrix is usually performed. Within our group of variants the results show insignificant changes in its genomes. MS-AFLP spectra evaluation needs more detailed analysis of results to extract more precise information about DNA methylation characters and impact of individual stress factors on epigenetic changes. From the point of view of functional methodology, the cDNA-AFLP appeared as the most difficult to establish. During first experiments only short PCR amplicons (at most 150 bp) were generated. Further, the calculated similarity coefficients showed surprisingly low values. These facts signalised certain unspecific RNA degradation before its conversion onto ds cDNA. The significant improvement of obtained results was recorded by using RNA later reagent, SpectrumTM Plant Total RNA Kit, RNase Out enzyme and MINT cDNA synthesis kit in cDNA-AFLP protocol. In the case of cDNA-AFLP spectra evaluation, it is possible either to prepare 0/1 binary matrix or differences can be estimated on the base of ratio between intensities of individual peaks. This fact should be taken into account if cDNA-AFLP results are compared with other fingerprinting methods as RAPD, SSR or standard AFLP.
dcterms:title
AFLP-derived methods as a tool for study of genomic, epigenomic and transcriptomic changes in stressed grapevine plants AFLP-derived methods as a tool for study of genomic, epigenomic and transcriptomic changes in stressed grapevine plants
skos:prefLabel
AFLP-derived methods as a tool for study of genomic, epigenomic and transcriptomic changes in stressed grapevine plants AFLP-derived methods as a tool for study of genomic, epigenomic and transcriptomic changes in stressed grapevine plants
skos:notation
RIV/62156489:43510/09:00152163!RIV14-MSM-43510___
n4:aktivita
n6:P
n4:aktivity
P(OC 116)
n4:dodaniDat
n9:2014
n4:domaciTvurceVysledku
n12:8526028 n12:2774771 n12:6417752 n12:7910444 n12:9771832
n4:druhVysledku
n13:D
n4:duvernostUdaju
n21:S
n4:entitaPredkladatele
n15:predkladatel
n4:idSjednocenehoVysledku
302109
n4:idVysledku
RIV/62156489:43510/09:00152163
n4:jazykVysledku
n16:eng
n4:klicovaSlova
stress; grapevine; cDNA-AFLP; somaclonal variation; MSAP
n4:klicoveSlovo
n14:somaclonal%20variation n14:MSAP n14:grapevine n14:stress n14:cDNA-AFLP
n4:kontrolniKodProRIV
[F62819E21DA7]
n4:mistoKonaniAkce
Dresden
n4:mistoVydani
Leuven
n4:nazevZdroje
Acta Horticulturae 839
n4:obor
n8:EB
n4:pocetDomacichTvurcuVysledku
5
n4:pocetTvurcuVysledku
5
n4:projekt
n5:OC%20116
n4:rokUplatneniVysledku
n9:2009
n4:tvurceVysledku
Ondrušiková, Eva Baránek, Miroslav Raddová, Jana Křižan, Břetislav Pidra, Miroslav
n4:typAkce
n11:WRD
n4:zahajeniAkce
2008-09-01+02:00
s:numberOfPages
9
n10:hasPublisher
International Society for Horticultural Science
n19:isbn
978-90-6605-612-1
n17:organizacniJednotka
43510