Cryopreservation and evaluations of vegetative growth, microtuber production and genetic stability in regenerants of purple-fleshed potato

Abstract

Buds of in vitro-grown shoots of two purple-fleshed potato genotypes were successfully cryopreserved by encapsulation-vitrification (Encap-vitri) and droplet-vitrification (Drop-vitri). Optimal time durations of exposure to PVS2 for shoot regrowth of cryopreserved buds were 5–7 h and 6 h for ‘E03-2677’ and for ‘Blue Congo’, respectively, in Encap-vitri, and 30–50 min and 40 min for the former and the latter, respectively, in Drop-vitri. Higher rates of shoot regrowth were obtained in 1.5–2.0 mm-buds than in 1.0–1.4 mm-ones in Encap-vitri for ‘E03-2677’ and ‘Blue Congo’, while opposite results were found in Drop-vitri. In ‘Blue Congo’, only apical shoot tips survived and developed into shoots, with one shoot produced in one cryopreserved bud in Encap-vitri and Drop-vitri. In ‘E03-2677’, survival and shoot regrowth patterns were similar to those of ‘Blue Congo’ in Encap-vitri. However, both apical and axillary shoot tips survived and developed into two shoots in one bud in Drop-vitri. In ‘E03-2677’, histological observations revealed only apical shoot tips survived following Encap-vitri, while both apical and axillary shoot tips survived following Drop-vitri. Vegetative growth in shoots regenerated from Encap-vitri and Drop-vitri after 3 weeks of post-thaw culture was significantly lower than that from control, but markedly increased after 6 months of post-thaw culture. In both ‘E03-2677’ and ‘Blue Congo’, number of microtubers per shoot, per vessel and ≥3 mm in diameter were significantly greater in shoots regenerated from Encap-vitri than in those from the control. Assessments by ISSR and RAPD of genetic stability did not find any polymorphic bands in regenerants recovered from Encap-vitri and Drop-vitri. To the best of our knowledge, this is the first report on cryopreservation of purple-fleshed potato by vitrification-based procedures. Results reported here would provide valuable basic and technical information on cryopreservation of purple-fleshed potato.