Micropropagation of raspberry (Rubus idaeus L.) in liquid immersion system
| Autor: | Jaredić, Ivana |
|---|---|
| Přispěvatelé: | Stanisavljević, Aleksandar, Popović, Brigita, Agić, Dejan |
| Jazyk: | chorvatština |
| Rok vydání: | 2018 |
| Předmět: | |
| Popis: | Osjetljivost maline (Rubus idaeus L.) na virusna oboljenja naglašava potrebu njenog razmnožavanja u kulturi tkiva in vitro – bezvirusni sadni materijal. Konvencionalni in vitro model mikropropagacije limitiran je uporabom polučvrstih hranjivih medija bez mogućnosti ventilacije i kontrole vitrifikacije. Cilj ovog diplomskog rada usmjeren je na ispitivanje mogućnost optimizacije i standardizacije protokola mikropropagacije maline uporabom imerznih bioreaktora (TIB) nove generacije uz modifikaciju mineralnih komponenti medija. Istraživanje je provedeno u laboratoriju za kulturu biljnog tkiva pri Katedri za voćarstvo, vinogradarstvo i vinarstvo na Fakultetu agrobiotehničkih znanosti Osijek. U istraživanju su korištene hranjive podloge MS i DKW s različitim kelatnim oblicima željeza. Tretman s modificiranom MS podlogom (MS(m)) sadržavao je kelatni oblik željeza FeEDDHA. Tretman DKW FeNaEDTA sadržavao je standardnu formulaciju DKW podloge s kelatnim oblikom željeza FeNaEDTA, te tretman DKW FeEDDHA standardnu formulaciju DKW podloge s kelatnim oblikom željeza FeEDDHA. Uspoređivan je i sustav imerznih bioreaktora koji koristi tekući medij s konvencionalnim in vitro modelom proizvodnje na polučvrstom mediju (agar). Praćeni su sljedeći parametri: morfološke karakteristike biljaka, stopa multiplikacija i pH tekućeg medija. Tretman s modificiranim MS medijem (MSm) i kelatnim oblikom željeza EDDHA rezultirao je značajno većim listovima i biljkama te brojem listova, odnosno ukupna vegetativna masa na oba sustava (TIB i konvencionalni) pri tretmanu MS(m) FeEDDHA bila je bolja u odnosu na ostale tretmane s DKW medijem. Stopa multiplikacije na polučvrstom mediju pri istom tretmanu MS(m) bila je najveća (5.7). Svi promatrani parametri na biljkama (visina biljaka, broj listova, broj stabljika, dužina i širina lista) bili su značajno veći na tekućem mediju. Imerzni sustav opravdao je recentne navode o svojoj učinkovitosti u skraćenju proizvodnog ciklusa (15 dana) i značajno boljim morfološkim parametrima (broj i veličina listova, broj izdanaka) ali multiplikacija je izostala. Jedini uočeni nedostatak kod sustava s tekućim medijem je stabilizacija pH. Uporaba MS medija s modifikacijom mezo komponenti makro hraniva (KH2PO4, MgSO4 i CaCl2) uz upotrebu željeznog kelata EDDHA sa sigurnošću možemo reći da je vrlo učinkovita u mikropropagaciji maline. The sensitivity of raspberry (Rubus idaeus L.) to viral diseases emphasizes need for her propagation in tissue culture (virus-free plant material). The conventional in vitro model of micropropagation is limited by the use of semi-solid media without possibility of ventilation and hyperhydricity control. The aim of this graduate thesis was to investigate the possibility of optimization and standardization of the micropropagation protocol for raspberry by using a new generation of immersion bioreactors (TIB) with the modification of mineral components in certain media. The research was carried out at the Tissue Culture Laboratory within the Chair of Pomology, Viticulture and Enology at the Faculty of Agrobiotechnical Sciences in Osijek. Growing media MS and DKW was used in the study with different chelating forms of iron. The modified MS media treatment (MS (m)) contained the chelating form of iron FeEDDHA. The DKW FeNaEDTA treatment contained a standard formulation of the DKW salts with the chelating form of iron FeNaEDTA and the DKW FeEDDHA treatment also contained DKW base salts with the chelating form of iron FeEDDHA. The system of immersion bioreactors that utilizes liquid media with conventional in vitro production model on solid medium (agar) has also been compared. The following parameters were observed: morphological characteristics of the plants, multiplication rate and the pH of the liquid medium. The treatment with modified MS medium (MSm) and chelating form of EDDHA iron resulted in significantly bigger leaves and plants, as well as the number of leaves; respectively the total vegetative mass of both systems (TIB and conventional) in the treatment of MS (m) FeEDDHA was better compared to other treatments with the DKW media. Multiplication rate on semi solid medium in the same treatment of MS (m) was the biggest (5.7). All the observed plant parameters (plant height, number of leaves, number of shoots, length and width of leaves) were significantly bigger on the liquid medium system. The immersion system justified the recent findings regarding its efficiency in shortening the production cycle (15 days) as well as regarding the significantly better morphological parameters (number and leaf size, number of outputs), but multiplication didn't occur. The only observed deficiency in the liquid media system is the pH stabilization. The use of MS medium with the modification of the macro-nutrient meso components (KH2PO4, MgSO4 and CaCl2) with the use of the EDDHA chelating iron can safely be said to be very effective in raspberry micropropagation. |
| Databáze: | OpenAIRE |
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