Cryopreservation of shoot primordia cultures of melon using a slow prefreezing procedure

• Published in: Plant cell, tissue and organ culture Vol. 49; no. 3; pp. 171 - 177
• Main Authors: Ogawa, R, Ishikawa, M, Niwata, E, Oosawa, K
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 1997
• Subjects: benzyladenine; Biological and medical sciences; Biotechnology; cell growth; cryopreservation; cryoprotectants; cryoprotective solution; Cucumis melo; culture media; dose response; Establishment of new cell lines, improvement of cultural methods, mass culture; Eukaryotic cell cultures; fluorescein diacetate; freezing; Fundamental and applied biological sciences. Psychology; germplasm; liquid nitrogen; methodology; Methods. Procedures. Technologies; micropropagation; Plant cells and fungal cells; regenerative ability; shelf life; shoots; temperature; tissue culture; viability; Culture medium; Vegetals; Cucurbitaceae; Tissue culture; Cryogenic storage; Environmental factor; Survival; Optimization; Regeneration; Cryopreservation; Fruit crop; Dicotyledones; Angiospermae; Slow cooling; Spermatophyta; Cryoprotective agent; Cucumis melo; Pretreatment
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1023/A:1005856617676

Tissue-cultured shoot primordia of melon (Cucumis melo L. cv. prince melon) were successfully cryopreserved in liquid nitrogen (LN) using a slow prefreezing method. The highest survival and recovery were obtained with the following procedure. Three week-old shoot primordia clumps were dissected into pieces of 2-3 mm of diameter and precultured in standard medium for 3 days. They were directly soaked in CSP1 cryoprotective solution (10%w/v sucrose, 10%w/v dimethylsulfoxide and 5%w/v glycerol) and incubated at room temperature for 30 min. Samples were ice-inoculated at -8 degrees C and cooled at a rate of between 0.3 and 1 degrees C min-1 with a programmable freezer to -30 degrees C for prefreezing. They were then plunged into LN for storage. After rapid thawing in 40 degrees C water, the cryoprotective solution was slowly diluted 5 fold in a dropwise manner with 3% sucrose and the shoot primordia were transferred onto regeneration medium. Under optimal conditions, more than 80% of cryopreserved shoot primordia were viable and 50 to 80% regenerated shoots after one month of reculture. Cryopreserved shoot primordia could be used both for reproducing a shoot primordia culture and for regenerating plants.