Submitted to: CryoLetters
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 3/1/2004
Publication Date: 6/1/2004
Citation: Volk G.M., N. Maness, and K. Rotindo. 2004. Crypreservation of garlic (Allium sativum L.) using plant vitrification solution 2. CryoLetters 25:219-226. Interpretive Summary: Garlic (Allium sativum L.) shoot tips can be successfully cryopreserved using a vitrification technique. Shoot tips are excised from bulbs, sterilized, treated with dehydration solutions, a cryoprotectant solution containing sucrose, glycerol, ethylene glycol, and DMSO, and plunged into liquid nitrogen. Shoot tips are thawed in a sucrose solution and then placed onto media for regrowth. We found that 11 of 18 diverse garlic accessions could be successfully cryopreserved using this method. This is the first report of the cryopreservation of garlic accessions known to be genetically diverse.
Technical Abstract: Most cryopreservation procedures are optimized for a selected portion of germplasm collections. We used a phylogenetic approach to select a subset of garlic (Allium sativum L.) accessions for cryopreservation. We were able to regenerate shoots from at least 30% of the explants in 11 of 18 diverse garlic cultivars using a vitrification protocol with PVS2 as the cryoprotectant. Garlic shoot tips were excised from cloves, surface sterilized, and placed on media at 5C for 2 days prior to cryopreservation. Shoot tips were then treated with sucrose-glycerol for 20 minutes, PVS2 on ice for 15 or 30 minutes, and then plunged on foils into super-cooled liquid nitrogen. Explants were recovered in 1.2 M sucrose for 20 minutes and then plated onto B5 media containing NAA and 2iP. This medium was shown to be superior for shoot regrowth when compared with an MS-based medium containing Gibberelic Acid. Our results demonstrate that phenotypically diverse accessions of garlic can be successfully cryopreserved.