Effects of culture conditions and nutrient medium composition on the physiological properties of Ficus carica L. plant in vitro cultures / In vitro kültürlerde Ficus carica L. bitkisinin fizyolojik özellikleri üzerine kültür koşulları ve besin ortamı kompozisyonun etkileri
Creators: name: Abacı, Altın Kardelen
Publisher: name: Zenodo
Year: 2024

description: This thesis study aims to establish an effective protocol for determining the most suitable nutrient medium composition and light-based culture conditions in terms of shoot and root regeneration in order to ensure a successful clonal micropropagation procedure in in vitro conditions on the physiological properties of Sarılop cultivar, which belongs to the Ficus carica L. (fig) plant species. Node explants of Sarılop cultivar of fig plantlets is taken from in vitro donor stocks in Ege University Engineering Faculty Bioengineering Department Plant Cell, Tissue and Organ Culture Laboratory, were cultured in 4 different hormone-free nutrient medium (WPM, DKW, MS and B5) in order to determine the best nutrient medium for shoot regeneration. Shoot regeneration percentage (%) in Sarılop cultivar was achieved with 100% success from all nutrient media and newly developed shoots emerged from the bottom parts of the explants. The highest average number of shoots per explant (1,23) in Sarılop cultivar was obtained on DKW nutrient medium. In terms of average values obtained per explant, the highest shoot length (1,60 cm) obtained per explant in WPM nutrient medium; number of nodes (1,62), number of leaves (2,70) and leaf length (1,27 cm) in DKW nutrient medium; root number (0,57) in WPM and MS nutrient mediums, root length (0,37 cm) in MS nutrient medium and the highest root regeneration percentage (50%) was obtained in WPM and MS nutrient mediums. In order to determine the best auxin and cytokine for shoot regeneration, node explants of Sarılop cultivar were cultured in DKW nutrient medium including different concentrations of BAP, KIN, IBA and IAA plant growth regulators. The highest average number of shoots per explant (2,67) was obtained in DKW nutrient medium containing 2 mg/L BAP. In terms of average values per explant, the highest average shoot length (1,48 cm) and number of nodes (2,56) containing 2 mg/L KIN; leaf number (8,20) including 2 mg/L BAP; leaf length (1,07 cm) containing 1 mg/L KIN; root number (2,17) and root length (0,63 cm) were achieved in DKW nutrient medium supplemented with 0,5 mg/L IBA. Node explants of Sarılop cultivar were cultured in DKW nutrient medium containing combinations of BAP and IBA at different concentrations in order to ensure better shoot regeneration. In terms of average values per explant, the highest number of shoots per explant (3,2) was achieved in DKW nutrient medium containing the combination of 2 mg/L BAP and 0,5 mg/L IBA. In the study, it was observed that in the cultured node explants that shoot formation generally occurred from the bottom and mostly 1 or 2 shoots were formed. The formation of multiple shoots (3-5 numbers) was achieved in DKW nutrient medium containing combination of 2 mg/L BAP and 0,5 mg/L IBA. The highest average shoot length (1,43 cm) and number of nodes (2,53) in the combination of 0,5 mg/L BAP and 0,5 mg/L IBA; number of leaves (6,10) in the combination of 1 mg/L BAP and 0,5 mg/L IBA; leaf length (0,91 cm) in combination of 0,5 mg/L BAP and 0,5 mg/L IBA; root number (0,53) in combination of 2 mg/L BAP and 0,5 mg/L IBA; root length (0,17 cm) was obtained in DKW nutrient medium containing growth regulators in the combination of 0,5 mg/L BAP and 0,5 mg/L IBA. Node explants of Sarılop cultivar were cultured in DKW basal nutrient medium containing a combination of 2 mg/L BAP and 0,5 mg/L IBA in six different culture vessels (55 mL glass tube, 170 mL, 300 mL and 265 mL glass jars, Vitrovent, Magenta GA-7) and under two different LEDs (4500 lux white and 600 lux, 2:1 red-blue) conditions in order to ensure shoot regeneration. The highest average number of shoots per explant (2,33) was obtained in the interaction with a 265 mL glass jar culture vessel under white LED light condition, in DKW nutrient medium including 2 mg/L BAP and 0,5 mg/L IBA. The highest average shoot length (1,57 cm) and number of nodes (2,68) per explant were observed under red-blue LED light conditions with a 55 mL glass tube culture vessels. The highest average number of leaves (4,96) with 265 mL glass jar culture vessel under white LED light conditions, the highest average leaf length (0,89 cm) with Magenta culture vessel under white LED light conditions, the highest average root number (0,90) and root length (0,34 cm) with 265 mL glass jar culture vessel under red-blue LED light condition achieved in DKW nutrient medium containing 2 mg/L BAP and 0,5 mg/L IBA. Explants obtained from shoot regeneration experiments were subcultured 4 times (in DKW nutrient medium containing 2 mg/L BAP and 0,5 mg/L IBA) at 4-week intervals with 55 mL glass tubes in order to evaluate the effect of subculture number on multiplication rate, average shoot length, node number, leaf number and length, root number and length in Sarılop cultivar. The highest multiplication rate (3,27) was obtained in the 3rd subculture. The highest average shoot length per explant (1,62 cm) was in the 1st subculture, the highest average node number (2,39) and leaf number (6,06) in the 2nd subculture. The highest average leaf length (1,30 cm), number of roots (0,57) and root length (0,28 cm) per explant were obtained in 1st subculture. In all experiments, physiological problems such as hyperhydricity, browning, chlorosis and necrosis were not observed in the produced plantlets. However, in B5 nutrient medium the leaves of some explants chlorosis was determined. The rooted plantlets are acclimatized with 70% success percentage.