izpis_h1_title_alt

Možnosti genske transformacije navadne konoplje (Cannabis sativa L.) z uporabo bakterije Agrobacterium tumefaciens
ID Bordon, Aleks (Author), ID Murovec, Jana (Mentor) More about this mentor... This link opens in a new window

.pdfPDF - Presentation file, Download (1,54 MB)
MD5: E7CD23185787B05CFEF3D47FBACC0105

Abstract
Razvili smo uspešno metodo za odpravo okužb semen navadne konoplje (Cannabis sativa L.) in hkrati uspešno transformacijo hipokotilov, s katero smo v celice vnesli selekcijski gen hptII in reporterski gen ZsGreen, ki preko proteina izraža zeleno fluorescenco. Genske transformacije so se izvajale s tri dnevno kokultivacijo hipokotilov s suspenzijo bakterije Agrobacterium tumefaciens. Adventivno regeneracijo hipokotilov smo dosegli z gojenjem izsečkov na MS gojišču z vitamini (Duchefa, M0222), 30 g/l saharoze, 8 g/l agarja, 0,2 mg/l hormona TDZ, 0,5 mg/l hormona NAA, 200µM acetosiringona in pH 5,8 (t.i. gojišče KON3). Uspešnost transformacije smo potrdili z uporabo epifluorescentne lupe, saj so regenerirani hipokotili svetili fluorescentno zeleno. V drugem poskusu, kjer smo narezane hipokotile inokulirali na KON3 gojišče, smo preverjali ali oddaljenost izsečka od koreninskega vratu hipkotilov vpliva na adventivno regeneracijo tkiva. Ugotovljeno je bilo, da razlik v regeneraciji ni bilo in, da je za uspeh poskusa ključna izbira nepoškodovanih semen, ki imajo značilno marmoriran vzorec in nerazpokano semensko ovojnico. V nasprotnem primeru pride do večjega števila bakterijskih in glivičnih okužb predvsem endogenega izvora, saj le-teh ni mogoče odpraviti s klasičnim površinskim razkuževanjem.

Language:Slovenian
Keywords:Cannabis sativa, industrijska konoplja, medicinska konoplja, genska transformacija, Agrobacterium tumefaciens
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[A. Bordon]
Year:2020
PID:20.500.12556/RUL-120652 This link opens in a new window
UDC:602.6:582.630.1(043.2)
COBISS.SI-ID:30091523 This link opens in a new window
Publication date in RUL:24.09.2020
Views:2360
Downloads:293
Metadata:XML DC-XML DC-RDF
:
Copy citation
Share:Bookmark and Share

Secondary language

Language:English
Title:Gene transformation experiments on cannabis (Cannabis sativa L.) using Agrobacterium tumefaciens
Abstract:
We have developed a successful method for the elimination of cannabis seed infections (Cannabis sativa L.) and at the same time the successful transformation of hypocotyls, with which we introduced the hptII selection gene and the ZsGreen reporter gene into plant cells, which expresses green fluorescence via protein. Gene transformations were performed by three-day cocultivation of hypocotyls with Agrobacterium tumefaciens. Adventitious regeneration of callus was achieved by growing explants on MS medium supplemented with vitamins (Duchefa, M0222), 30 g/l sucrose, 8 g/l agar, 0.2 mg/l TDZ, 0.5 mg/l NAA, 200µM acetosiringone and pH 5.8 (the KON3 medium). The success of the transformation was confirmed using an epifluorescent macroscope, as the regenerated hypocotyls glowed fluorescent green. In another experiment, where sliced hypocotyls were placed on KON3 medium, we examined whether the position of explants on the hypocotyl influenced adventitious regeneration. It was found that there were no differences in regeneration. For the success of the experiment, we found that the key is to choose undamaged seeds with a characteristically marbled seed coat pattern. Otherwise, a larger number of bacterial and fungal infections, mainly of endogenous origin, occur, as these cannot be eliminated by classical sterilization.

Keywords:Cannabis sativa, industrial hemp, medical hemp, gene transformation, Agrobacterium tumefaciens

Similar documents

Similar works from RUL:
Similar works from other Slovenian collections:

Back