Abstract

Yukon Draba (Draba yukonensis) is a small, short-lived perennial mustard species that is endemic to southwestern Yukon in Canada. This plant has been categorized as a species of Special Concern. It faces the threat of habitat loss due to natural and man-made causes and a population that is unevenly distributed to a few large and several small subpopulations in the area. It will therefore be judicious to undertake investigations on the conservation of this species to save it from further deterioration which may lead to its extinction. In this study, a protocol was developed for in vitro propagation and cryopreservation of Yukon Draba. The micropropagation protocol was optimized using shoot tips which enabled clonal propagation and in vitro storage of the species. Shoots grew best in the medium containing MS basal salts and had the highest multiplication with the addition of 2 µM 6-benzylaminopurine or 5 µM Kinetin with 3% sucrose. The addition of 10 µM Indole Butyric Acid (IBA) produced the highest number of adventitious roots on the shoots and the longest root length was observed at 2 µM IBA. The rooted plantlets were transferred to greenhouse and the highest survival (87.5%) was observed for the plantlets treated with a lower concentration of IBA (2 µM). Cryopreservation protocol was developed using the droplet-vitrification method for in vitro shoot tips. Two-week-old shoots had the highest survival and regrowth following exposure to plant vitrification solution 3 (PVS3) for 30 min, prior to direct immersion of the droplets into the liquid nitrogen. The optimized protocols for the micropropagation and cryopreservation may be useful for the long-term germplasm conservation and reintroduction of this species in its natural habitat.

Highlights

  • Our results showed that the use of 2-week-old in vitro stock cultures resulted in higher recovery after cryopreservation as compared to 1, 3 and 4-week-old plants

  • The protocol for micropropagation and cryopreservation of Yukon Draba was successfully developed from the seedling which served as the source for establishing shoot cultures

  • The shoot tip explants were multiplied in vitro with optimization of basal medium, sugar, and plant growth regulators and rooted plantlets were successfully transferred to the greenhouse

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Summary

Introduction

Yukon Draba (Draba yukonensis A.E. Porsild) is a small, short-lived, perennial species of mustard (family Brassicaceae) with untoothed leaves covered with distinctive stiff and unforked hairs. Porsild) is a small, short-lived, perennial species of mustard (family Brassicaceae) with untoothed leaves covered with distinctive stiff and unforked hairs This species is endemic to southwestern Yukon, Canada. The Yukon Draba population is estimated between 160,200 to 333,000 individuals [1]. The plant survives extreme cold and dry weather conditions and requires full sun and well-drained soils. The physiology of the life cycle of this plant is understudied and the mechanisms of cold tolerance remain unknown. Yukon Draba can disperse via seeds, but this ability is apparently limited, with no known method of long-distance dispersal, and an inability to colonize nearby apparently suitable habitat

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