Microtuberization of Potato (Solanum tuberosum L.) as Influenced by Supplementary Nutrients, Plant Growth Regulators, and In Vitro Culture Conditions

Five experiments with in vitro-grown plantlets of potato cvs ‘Superior,’ ‘Early Valley,’ ‘Golden Valley,’ and ‘Winter Valley’ were carried out to investigate in a stepwise approach the potential for enhancing microtuber production of (1) adding supplementary nutrients (SN) including KNO3, NH4H2PO4, Ca(NO3)2.4H2O, and MgSO4 in full- or half-strength Murashige and Skoog (MS) medium, (2) adding a pretested combination of plant growth regulators (PGRs), i.e., 10 mg l−1 6-benzyl aminopurine plus 200 mg l−1 succinic acid 2,2-dimethylhydrazide, (3) varying the timing of the supplemental applications, and (4) varying the replenishment of the medium. Plantlets of ‘Superior’ for microtuberization were grown from two-node explants for 75 days under environmental conditions consisting of 22 ± 2 °C and a 16-h photoperiod on a solid basal medium consisting of half-strength MS (1/2 MS) medium containing 6 % sucrose and a moderate concentration of supplementary nutrients, SN-2 (KNO3, 1.22 g l−1; NH4H2PO4, 1.41 g l−1; Ca(NO3)24H2O, 0.23 g l−1; and MgSO4, 0.74 g l−1). Thereafter, plantlets were grown for 60 days under dark conditions. Liquid media containing different combinations of MS, SN, and PGR levels plus 8 % sucrose were added as a supplement and were evaluated for their ability to increase tuberization compared to the control composition of the supplement (MS with 8 % sucrose). In the first experiment, addition of 1/2 MS liquid medium containing 8 % sucrose and SN-2 before dark incubation induced the highest total number of microtubers (on average 1.4 per plantlet), a significantly higher weight and size than the control, and the highest percentage (50 %) of large-sized (>500 mg) microtubers. In the second experiment, addition of 1/2 MS liquid medium containing 8 % sucrose, SN-2, and PGRs produced the highest number (on average 1.5 per plantlet) and total yield (on average 971 mg per plantlet) of microtubers, and significantly the highest percentage (66 %) of large-sized (>500 mg) microtubers. The size of the microtubers was higher than for the treatment without growth regulators. The optimum time for addition of supplemental medium containing 8 % sucrose, SN-2, and PGRs was shown in experiment 3 to be latest 1 week before dark incubation. This timing resulted in both the highest microtuber number (on average 1.6 per plantlet) and yield (on average 1208 mg per plantlet), and the highest percentage (64 %) of large-sized microtubers. Finally, it was found that a second supplemental treatment consisting of 50 % replacement of the liquid medium (by volume) 2 weeks after transfer to dark conditions gave rise to the highest number (on average 1.9 per plantlet), yield (on average 1346 mg per plant), and also significantly the highest percentage (25 %) of large-sized microtubers (>1000 mg) as compared to replacement of a greater or smaller proportion of the medium. Four potato cultivars, ‘Superior,’ ‘Early Valley,’ ‘Golden Valley,’ and ‘Winter Valley’ produced acceptable numbers and yield of microtubers of relatively large-sized tubers using a process that included supplemental liquid medium with 1/2 MS, 8 % sucrose, moderate concentration (SN-2) of SNs and PGRs, followed by a 50 % replacement of the same medium.