Micropropagation and Cryopreservation of Yukon Draba (Draba yukonensis), a Special Concern Plant Species Endemic to Yukon Territory, Canada

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.

Temperature Dependence of Membrane Permeability Parameters for Five Cell Types Using Nonideal Thermodynamic Assumptions to Mathematically Model Cryopreservation Protocols

“…). The cell volume changes before reaching the equilibrium result from water and CPA transport across the cell membrane which can be generally modeled by the following equations: (14) where V w in and V p in are the volume of water and volume of cryoprotectant inside the cell, respectively, L p * is the hydraulic conductivity, P s * is the membrane permeability to permeating solutes or cryoprotectants, T is the absolute temperature, A cell is the surface area of the cell, and a p in and a p ex are the intra-and extracellular activity of the permeating CPA, respectively. The extracellular solution is assumed to be an infinite solution with constant properties.…”

“…Cryopreservationthe process of preserving biological matter at low subzero temperatures for long-term storagehas become the prevalent way of preserving cells used in medical treatments − and genes of rare and endangered species of animals − or plants − over the past few decades. Cells and tissues at low temperatures are far from their physiological environment, making them susceptible to various injuries due to intracellular ice formation, increasing concentrations of solutes inside and outside the cells, and dehydration. , A successful cryopreservation procedure relies on maximizing cell viability by minimizing the injuries incurred during cryoprotective agent (CPA) addition, cooling, storage, thawing, and CPA removal.…”

In Vitro Propagation of Peumus boldus Mol, a Woody Medicinal Plant Endemic to the Sclerophyllous Forest of Central Chile

“…In this context, the need to establish comprehensive large-scale propagation methods has become crucial to increasing the population size and contributing to the conservation of this medicinal plant. The use of in vitro mass propagation for habitat restoration has emerged as a viable strategy, given that conventional methods involving cuttings or seeds fall short in guaranteeing substantial production and the creation of pathogen-free germplasm [23][24][25]. Therefore, the aim of this study was to develop and implement a protocol for mass in vitro regeneration of P. boldus.…”

In Vitro Technologies for American Chestnut (Castanea dentata (Marshall) Borkh) Conservation

“…Mass propagation and reintroduction in natural habitats is an effective strategy for increasing the declining populations of endangered species [ 5 , 6 , 7 ]. Traditional propagation systems of the American chestnut are not efficient for large-scale propagation of plants for conservation or breeding programs to develop disease- resistant germplasm [ 8 , 9 ].…”