In vitro regeneration and effects of methyl jasmonate elicitor on biomass production in cell suspension cultures in vanilla (Vanilla planifolia L.) plant

Vanilla is a high demanding product sold at high prices in world markets. Natural vanilla has high economic value and is the world's second most expensive spices after saffron. Despite its economic value, it has been one of the least studied species of vanilla plants among other orchid species (Arenas and Dressler, 2010). Vanilla's synthetic form (vanillin) can be obtained from various chemical methods by using starting materials (coniferin, guaiacol, eugenol and lignin), but the vanilla flavor can naturally be obtained only from vanilla fruit (Rao and Ravishankar, 2000). Although some regions of Turkey have the appropriate climate to grow vanilla plant, however this plant is not cultivated in Turkey. For this reason, in the thesis study, regeneration of the vanilla plant in vitro conditions and experiments to increase the vanillin compound which is the active substance of the plant were carried out. The highest callus regeneration (80%) from node explants was obtained in VK4 (MS+1 mg/L 2,4-D + 1 mg/L BAP) nutrient medium, while 100% direct shoot regeneration was provided in VK3 (MS+1 mg/L 2,4-D + 0,5 mg/L BAP) and VK6 (MS+1 mg/L BAP + 0,5 mg/L NAA) nutrient media. In addition, 50% rooting percentage was obtained in VK6 medium. Indirect shoot regeneration rate was determined as 33,3% in VK2 (MS + 1 mg / L 2,4-D + 0,5 mg / L KIN) and 71,4% of indirect embryo formation was detected in VK4 medium. All embryos stayed in globules stages. 0,5 g wet weight calli obtained from seeds and other explants were cultured in the liquid nutrient medium provided with the most successful callus regeneration (stirring speed was approximately 150 rpm and incubation temperature 24 ± 2 °C). The cultures are kept in the dark. Growth parameters were determined in established cell suspension cultures. İn the result of investigated after MeJa application at 0, 7, 14, 21 and 42 days old cultures, the highest wet and dry weight values (0,0519 g/ml, 0,0104 g/ml) with 100 μM of methyl jasmonate was obtained at day 21. At the 6th week of culture, globular embryo formation was observed in cell suspension cultures and the highest dry biomass ratio (30,8%) was taken with applying 100 μM MeJa. Key words: Vanilla, Vanilla planifolia L., In Vitro, Suspension Culture, Methyl Jasmonate