Pregled bibliografske jedinice broj: 25259
Transformacija stanica somatskih embrija Pančićeve smreke (Picea omorika) posredstvom agrobakterija
Transformacija stanica somatskih embrija Pančićeve smreke (Picea omorika) posredstvom agrobakterija // Šesti kongres biologa Hrvatske s međunarodnim sudjelovanjem / Huber, Đuro (ur.).
Zagreb: Hrvatsko biološko društvo, 1997. str. 82-83 (poster, domaća recenzija, sažetak, znanstveni)
CROSBI ID: 25259 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Transformacija stanica somatskih embrija Pančićeve smreke (Picea omorika) posredstvom agrobakterija
(Agrobacterium-mediated cell transformation of Picea omorika somatic embryos)
Autori
Mihaljević, Snježana ; Tramišak-Milaković, Tamara ; Jelaska, Sibila
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Izvornik
Šesti kongres biologa Hrvatske s međunarodnim sudjelovanjem
/ Huber, Đuro - Zagreb : Hrvatsko biološko društvo, 1997, 82-83
Skup
Šesti kongres biologa Hrvatske s međunarodnim sudjelovanjem
Mjesto i datum
Opatija, Hrvatska, 22.09.1997. - 26.09.1997
Vrsta sudjelovanja
Poster
Vrsta recenzije
Domaća recenzija
Ključne riječi
somatska embriogeneza; Picea omorika; transformacija; A. tumefaciens; strain LBA4404
(somatic embryogenesis; Picea omorika; transformation; A. tumefaciens; strain LBA4404)
Sažetak
Agrobacterium-mediated gene transfer system that targets cells of Picea omorika somatic embryos was developed. Somatic embryos derived from the highly embryogenic line BB19 were infected with A. tumefaciens strain LBA4404 containing a binary vector with genes for kanamycin resistance (NPTII) and beta-glucuronidase (GUS). Transient GUS expression events were quantified histochemically by determing the frequency of explants exhibiting blue spots indicative of GUS at four to eight days after cocultivation with Agrobacterium. Critical factors influencing an optimal transformation were the embryo stage, and a short preculture treatment. A. tumefaciens strain LBA4404 that did not contain the GUS gene (control) did not elicit any blue staining characteristic of GUS activity. The effect of carbenicillin and cefotaxime (antibiotics used to eliminate Agrobacterium from cultures) on somatic embyogenesis of Picea omorika was determined. Carbenicillin strongly reduced embryogenic tissue proliferation (60% reduction) and ought to be omitted in transformation procedure of Picea omorika. Kanamycin also inhibited embryogenic tissue initiation in somatic embryos of this species. The lethal kanamycin concentration for non-transformed embryogenic tissue cells was 10 mg/L. Up to 60% of embryos remained GUS positive after one month in culture. These somatic embryos were used to re-induce embryogenic tissue. The putative transformed embryogenic tissu were subjected to PCR and Southern blot analysis.
Izvorni jezik
Hrvatski
Znanstvena područja
Biologija
