"Studium kandid\u00E1tn\u00EDch sekvenc\u00ED z biosyntetick\u00E9 dr\u00E1hy anthokyan\u016F p\u0161enice"@cs . . "Anthokyany, flavonoidn\u00ED l\u00E1tky, jsou zodpov\u011Bdn\u00E9 za rozd\u00EDln\u00E9 zbarven\u00ED obilky p\u0161enice set\u00E9 (Triticum aestivum L.). Pigmenty tvo\u0159en\u00E9 biosyntetickou drahou anthokyan\u016F jsou ulo\u017Eeny v r\u016Fzn\u00FDch \u010D\u00E1stech obilky. Purpurov\u00E9 pigmenty jsou akumulov\u00E1ny v perikarpu a modr\u00E9 pigmenty v aleuronov\u00E9 vrstv\u011B obilky. Z kolekce Agrotestu fyto Krom\u011B\u0159\u00ED\u017E, s.r.o. byly vybr\u00E1ny vzorky jarn\u00ED formy p\u0161enice (Novosibirskaja 67, ANK-28B, Abissinskaja arrasajta, UC66049 a Tschermaks Blauk\u00F6rniger Sommerweizen) s r\u016Fzn\u011B zbarven\u00FDmi obilkami, z kter\u00FDch byla izolov\u00E1na celkov\u00E1 RNA pomoc\u00ED fenol-chloroformov\u00E9 extrakce. Celkov\u00E1 RNA byla p\u0159eps\u00E1na reverzn\u00ED transkripc\u00ED do cDNA. Gen pro GAPDH (glyceraldehyd-3-fosf\u00E1t dehydrogenasu) byl vybr\u00E1n jako provozn\u00ED gen pro ov\u011B\u0159en\u00ED \u00FAsp\u011B\u0161nosti p\u0159episu. N\u00E1sledn\u011B byla na cDNA testov\u00E1na optim\u00E1ln\u00ED teplotn\u00ED profil PCR reakce pro navr\u017Een\u00E9 primery pomoc\u00ED programu Primer3 a aplikace BLAST. Na z\u00E1klad\u011B sekvenace PCR produkt\u016F byly z\u00EDsk\u00E1ny kandid\u00E1tn\u00ED sekvence gen\u016F pro chalkon synthasu (82-100% podobnost), pro chalkon isomerasu (podobnost 98-100%) a pro dihydroflavonol reduktasu (95-100% podobnost). Zji\u0161t\u011Bn\u00E1 variabilita byla zp\u016Fsobena jednonukleotidov\u00FDm polymorfismem a indely. Primery jsou pou\u017E\u00EDv\u00E1ny pro studium genov\u00E9 exprese pomoc\u00ED qPCR." . "Praha" . . . "V\u00DARV Praha-Ruzyn\u011B" . . "Vyhn\u00E1nek, Tom\u00E1\u0161" . "Studium kandid\u00E1tn\u00EDch sekvenc\u00ED z biosyntetick\u00E9 dr\u00E1hy anthokyan\u016F p\u0161enice"@cs . "43210" . "Studium kandid\u00E1tn\u00EDch sekvenc\u00ED z biosyntetick\u00E9 dr\u00E1hy anthokyan\u016F p\u0161enice" . "Ondrou\u0161kov\u00E1, Jana" . "4"^^ . . . . . "Studium kandid\u00E1tn\u00EDch sekvenc\u00ED z biosyntetick\u00E9 dr\u00E1hy anthokyan\u016F p\u0161enice" . . "Anthocyanins, flavonoid substances in nature, are responsible for the different pigmented caryopsis of common wheat (Triticum aestivum L.). Anthocyanin biosynthetic pathway is realized in plants by many enzymes from early (CHS -- chalcon sythase, CHI -- chalcon isomerase, and FH -- flavonone hydroxylase) and late (DFR -- dihydroflavonol reductase, ANS -- anthocyanindin synthase and UFGT -- UDP-glucose flavonoid glycosyltransferase) biosynthetic pathway. Pigments formed in this biosynthetic pathway in grain are stored in different layers. Purple pigments are simultaneously accumulated in pericarp and blue pigments are stored in aleurone layer of caryopsis. The field experiment Agrotest Fyto Krom\u011B\u0159\u00ED\u017E, Ltd. derived samples of ripened caryopsis spring wheat form, selected genotypes (Novosibirskaya 67, ANK-28B, Abyssinskaya Arraseita, UC66049, and Tschermaks Blauk\u00F6rniger Sommerweizen) with different caryopsis pigment, which from total RNA was isolated using a phenol-chloroform extraction. Total RNA was reverse transcribed into cDNA. House keeping GAPDH gene (the gene for glyceraldehyde-3-phosphate dehydrogenase) was chosen as a control for PCR to verify the successful transcription of cDNA. Proven cDNA was subsequently used as a template for gradient PCR to test the optimum temperature annealing for designed primers. There were we searched sequences with a high level of similarity in application BLAST, which served for primer design by Primer3. We detected sequences for CHS genes (82-100% similarity), for CHI genes (98-100% similarity) and for DFR genes (95-100% similarity). The variability was due to single nucleotide polymorphism and indels. The primers will be used for gene expression study by qPCR."@en . . "T. aestivum L.; anthocyanins; wheat; chalcon isomerase; dihydroflavonol reductase; chalcone synthase"@en . "172066" . "P\u0161enice 2012 - od genomu po chleba" . "2012-01-01+01:00"^^ . . "Trojan, V\u00E1clav" . "Studies of candidate sequences from wheat anthocyanin bisynthetic pathway"@en . "RIV/62156489:43210/12:00192817" . . . . "[97BDA223D35A]" . "Musilov\u00E1, Milena" . . "RIV/62156489:43210/12:00192817!RIV13-GA0-43210___" . . . . . . . "978-80-7427-122-9" . "V\u00FDzkumn\u00FD \u00FAstav rostlinn\u00E9 v\u00FDroby, v. v. i." . "5"^^ . . "5"^^ . "P(GD204/09/H002)" . "Studies of candidate sequences from wheat anthocyanin bisynthetic pathway"@en . "Anthokyany, flavonoidn\u00ED l\u00E1tky, jsou zodpov\u011Bdn\u00E9 za rozd\u00EDln\u00E9 zbarven\u00ED obilky p\u0161enice set\u00E9 (Triticum aestivum L.). Pigmenty tvo\u0159en\u00E9 biosyntetickou drahou anthokyan\u016F jsou ulo\u017Eeny v r\u016Fzn\u00FDch \u010D\u00E1stech obilky. Purpurov\u00E9 pigmenty jsou akumulov\u00E1ny v perikarpu a modr\u00E9 pigmenty v aleuronov\u00E9 vrstv\u011B obilky. Z kolekce Agrotestu fyto Krom\u011B\u0159\u00ED\u017E, s.r.o. byly vybr\u00E1ny vzorky jarn\u00ED formy p\u0161enice (Novosibirskaja 67, ANK-28B, Abissinskaja arrasajta, UC66049 a Tschermaks Blauk\u00F6rniger Sommerweizen) s r\u016Fzn\u011B zbarven\u00FDmi obilkami, z kter\u00FDch byla izolov\u00E1na celkov\u00E1 RNA pomoc\u00ED fenol-chloroformov\u00E9 extrakce. Celkov\u00E1 RNA byla p\u0159eps\u00E1na reverzn\u00ED transkripc\u00ED do cDNA. Gen pro GAPDH (glyceraldehyd-3-fosf\u00E1t dehydrogenasu) byl vybr\u00E1n jako provozn\u00ED gen pro ov\u011B\u0159en\u00ED \u00FAsp\u011B\u0161nosti p\u0159episu. N\u00E1sledn\u011B byla na cDNA testov\u00E1na optim\u00E1ln\u00ED teplotn\u00ED profil PCR reakce pro navr\u017Een\u00E9 primery pomoc\u00ED programu Primer3 a aplikace BLAST. Na z\u00E1klad\u011B sekvenace PCR produkt\u016F byly z\u00EDsk\u00E1ny kandid\u00E1tn\u00ED sekvence gen\u016F pro chalkon synthasu (82-100% podobnost), pro chalkon isomerasu (podobnost 98-100%) a pro dihydroflavonol reduktasu (95-100% podobnost). Zji\u0161t\u011Bn\u00E1 variabilita byla zp\u016Fsobena jednonukleotidov\u00FDm polymorfismem a indely. Primery jsou pou\u017E\u00EDv\u00E1ny pro studium genov\u00E9 exprese pomoc\u00ED qPCR."@cs . "Havel, Ladislav" . . .