Abstract
In the present study, influence of nitrate concentration on in vitro rooting and its ex vitro hardened tubers were investigated in D. hamiltonii. In vitro multiple shoots established on the MS medium were used as explants for this study and concentration of indole-3-butyric acid (1.23 µM) required for root initiation was determined. The effect of different nitrate concentrations in MS medium for in vitro rooting was investigated and positive (¼ and ½ strength) influence was observed on 14 days inoculation. The in vitro rooted plants (IVP) showed 80% survival upon hardening and exhibited similar growth pattern to seedling plants. The 1-year grew ex vitro hardened plants were evaluated for their tuber quality with reference to their yield and flavor metabolite 2-hydroxy-4-methoxy benzaldehyde (2H4MB) content. The IVP grown from ¼ nitrate strength media produced 155 ± 4.85 g FW of tuber biomass which is observed to be higher than full nitrate strength IVP (112 ± 2.52 g) which are grown under similar conditions in the greenhouse. The flavor metabolite content, total phenolics, total flavonoids, and antioxidant potential of these IVP tubers were evaluated. Upregulation of flavor biosynthetic pathway genes DhPAL, DhC4H and DhCoMT were observed in tubers of potted plants that developed from low nitrate strength culture media. In this study, superiority of tissue cultured plants was evident, wherein in vitro plants developed in low nitrate strength medium and acclimatized could produce a higher yield in tuber biomass and maintain relative content of flavor compounds in this endangered plant.