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Research Project: Innovative Strategies and Methods for Improving the Management, Availability, and Utility of Plant Genetic Resource Collections

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Title: Cryopreservation of jerusalem artichoke cultivars

item Volk, Gayle
item Wang, Ling

Submitted to: Cryobiology
Publication Type: Abstract Only
Publication Acceptance Date: 7/10/2018
Publication Date: 12/1/2018
Citation: Volk, G.M., Wang, L. 2018. Cryopreservation of jerusalem artichoke cultivars. Cryobiology.

Interpretive Summary: Jerusalem artichoke (Helianthus tuberosusL.) cultivars are conserved in genebanks for use in breeding and horticultural research programs. As vegetatively propagated collections, Jerusalem artichoke collections are vulnerable to environmental and biological threats. We developed an improved cryopreservation method using four Jersualem artichoke cultivars (‘Shudi’,‘M6’,‘Stampede’, and‘Relikt’), that will enable Jerusalem artichoke field collections to be securely conserved in genebanks. Four steps were optimized, and the resulting procedure is as follows: preculture excised shoot tips (2~3 mm) in liquid MS medium supplemented with 0.4M sucrose for 3 days, osmoprotect shoot tips in loading solution for 30 min,dehydrate with plant vitrification solution 2 for 15 min before rapid cooling in liquid nitrogen, store in liquid nitrogen, rapidly rewarm in MS liquid medium containing 1.2 M sucrose, and recover on MS mediumsupplemented with 0.1 mg L-1GA3for 3 to 5 days in the dark and then onthe same medium for 4 to 6 weeks in the light (14h light/ 10h dark). Jerusalem artichoke cultivar‘Shudi’had the highest survival (93%) and regrowth (83%) percentages. After cryopreservation, cultivars ‘M6’,‘Stampede’, and‘Relikt’achieved regrowth percentages ranging from 37%to 53%. No genetic changes, as assessed by microsatellites, were detected in plants regenerated after LN exposure in Jerusalem artichoke cultivar ‘Shudi’. Differential scanning calorimetry analyses were used to investigate the thermal activities of the tissues during the cryopreservation process and determined that loading with 2.0 M sucrose and 0.4 M sucrose dehydrated the shoot tips prior to treatment with PVS2. Histological observations revealed that the optimized droplet vitrification protocol caused minimal cellular damage within the meristem cells of the shoot tips.

Technical Abstract: N/A