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加拿大育空地区特有的、受特别关注的植物物种育空鼠耳芥的微繁殖与超低温保存

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

作者信息

Saxena Akansha, Bi Wen-Lu, Shukla Mukund R, Cannings Syd, Bennett Bruce, Saxena Praveen K

机构信息

Gosling Research Institute for Plant Preservation, Department of Plant Agriculture, University of Guelph, Guelph, ON N1G 2W1, Canada.

Environment and Climate Change Canada, 91780 Alaska Highway, 1st Floor, Whitehorse, YK Y1A 5X7, Canada.

出版信息

Plants (Basel). 2021 Oct 3;10(10):2093. doi: 10.3390/plants10102093.

Abstract

Yukon Draba () 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.

摘要

育空葶苈(Yukon Draba)是一种小型、一年生的多年生芥菜物种,原产于加拿大育空地区西南部。这种植物已被列为特别关注物种。由于自然和人为原因,它面临着栖息地丧失的威胁,且种群分布不均,集中在该地区的几个大的和几个小的亚种群中。因此,对该物种进行保护调查,以防止其进一步恶化并可能导致灭绝是明智的。在本研究中,开发了一种用于育空葶苈离体繁殖和冷冻保存的方案。使用茎尖优化了微繁殖方案,实现了该物种的克隆繁殖和离体保存。茎尖在含有MS基本盐类的培养基中生长最佳,添加2 μM 6-苄基腺嘌呤或5 μM激动素和3%蔗糖时增殖率最高。添加10 μM吲哚丁酸(IBA)时,茎尖上不定根数量最多,在2 μM IBA时观察到最长的根长。生根的小植株被转移到温室中,较低浓度IBA(2 μM)处理的小植株存活率最高(87.5%)。采用玻璃化液滴法对离体茎尖进行冷冻保存方案的开发。两周大的茎尖在暴露于植物玻璃化溶液3(PVS3)30分钟后,直接将液滴浸入液氮中,其存活率和再生率最高。微繁殖和冷冻保存的优化方案可能有助于该物种的长期种质保存及其在自然栖息地的重新引入。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c9/8540741/fdc6a5a91a7c/plants-10-02093-g001.jpg

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