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In Vitro Propagation and Cryopreservation of Golden Paintbrush (Castilleja levisecta Greenm.)

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Title: In Vitro Propagation and Cryopreservation of Golden Paintbrush (Castilleja levisecta Greenm.)
Author: Salama, Ahmed
Department: Department of Plant Agriculture
Program: Plant Agriculture
Advisor: Saxena, Praveen
Abstract: Golden paintbrush (Castilleja levisecta Greenm.) is an herbaceous perennial plant native to the Pacific Northwest and is considered critically endangered with only 11 populations remaining in the wild. The objective of this study was to develop micropropagation and cryopreservation methods for large scale propagation and long-term ex situ germplasm conservation of C. levisecta. Cultures were initiated from nodal explants isolated from seedlings and shoots were successfully multiplied on modified MS medium (MS salt, B5 vitamins, 30 g/L sucrose, and 2.2 g/L phytagel; pH 5.7) supplemented with 2.0 µM 6-benzylaminopurine (BA) and 3.0 µM Kinetin. Micropropagated shoots were rooted ex-vitro with 100% rooting and an average of ~17 roots per shoot. Cryopreservation was achieved using axillary buds from in vitro plants (7, 12, or 21 days old) that were pre-cultured step-wise on media containing 0.3 M and 0.5 M sucrose then treated with vitrification solutions VS A3 (22.5% sucrose, 37.5% glycerol, 15% ethylene glycol and 15% Me2SO; w/v) on ice or with PVS3 (50% glycerol + 50% sucrose; w/v) for 0, 20, 40, 60, 80, and 100 min at room temperature, and processed using droplet-vitrification. Unloading treatments evaluated included 0, 0.8 M, 1.0 M and 1.2 M sucrose solutions for 30 min. There was no significant difference in regrowth of nodal explants of different ages. The highest regrowth after cryopreservation (65%) was observed for axillary buds treated with VS A3 and PVS3 for 60-80 min followed by unloading in 0.8 M sucrose. The recovered shoot tips were successfully rooted and acclimatized in the greenhouse and 268 micropropagated/ cryopreserved plants were re-introduced into their natural habitat in BC.
URI: http://hdl.handle.net/10214/11557
Date: 2017-08-29
Rights: Attribution-NoDerivs 2.5 Canada
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Attribution-NoDerivs 2.5 Canada Except where otherwise noted, this item's license is described as Attribution-NoDerivs 2.5 Canada