Improved cryopreservation by diluted vitrification solution with supercooling-facilitating flavonol glycoside |
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| Affiliation: | 1. National Agricultural Research Center for Hokkaido Region, Sapporo 062-8555, Japan;2. Research Faculty and Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan;1. Plant Stress Molecular Biology Department, Institute of Plant Physiology and Genetics, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 21, 1113 Sofia, Bulgaria;2. Institute of Plant Sciences and Oeschger Centre for Climate Change Research (OCCR), University of Bern, Altenbergrain 21, CH-3013 Bern, Switzerland;1. Warsaw University of Life Sciences (SGGW), Faculty of Agriculture and Biology, Department of Botany, Nowoursynowska 159, Building 37, 02-776 Warsaw, Poland;2. University of Agriculture, Faculty of Biotechnology and Horticulture, Institute of Plant Biology and Biotechnology, Unit of Botany and Plant Physiology, Al. 29-Listopada 54, 31-425 Krakow, Poland;3. University of Agriculture, Faculty of Agriculture and Economics, Institute of Soil Science and Agrophysics, Department of Soil Science and Soil Protection, Al. Mickiewicza 21, 31-120 Krakow, Poland;4. University of Agriculture, Center for Technology Transfer, Al. Mickiewicza 21, 31-120 Krakow, Poland;1. Department of Science and Engineering of Oxide Materials and Nanomaterials, Faculty of Applied Chemistry and Materials Science, University Politehnica of Bucharest, 1–7 Polizu Street, 011061 Bucharest, Romania;2. AMG Transcend, 1–7 Polizu Street, 011061 Bucharest, Romania;3. Lasers Department, National Institute for Lasers, Plasma & Radiation Physics, P.O. Box MG-36, Magurele, Bucharest, Romania;4. Microbiology Immunology Department, Faculty of Biology, University of Bucharest, 1–3 Portocalelor Lane, Sector 5, 77206 Bucharest, Romania;5. Research Center for Microscopic Morphology and Immunology, University of Medicine and Pharmacy of Craiova, 2 Petru Rareş Street, 200349 Craiova, Romania;6. Department of Pharmacognosy & Phytotherapy, Faculty of Pharmacy, University of Medicine and Pharmacy of Craiova, 2 Petru Rareş Street, 200349 Craiova, Romania;7. National Institute of Materials Physics, 105 bis Atomistilor Street, Bucharest-Magurele, P.O. Box MG-7, 077125, Romania;8. Institute of Cellular Biology and Pathology of Romanian Academy, “Nicolae Simionescu”, Department of Fetal and Adult Stem Cell Therapy, 8, B.P. Hasdeu, Bucharest 050568, Romania;1. Department of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, South Korea;2. Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, RDA, Wan-Ju, 565-852, South Korea |
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| Abstract: | The effect of kaempferol-7-O-glucoside (KF7G), one of the supercooling-facilitating flavonol glycosides which was originally found in deep supercooling xylem parenchyma cells of the katsura tree and was found to exhibit the highest level of supercooling-facilitating activity among reported substances, was examined for successful cryopreservation by vitrification procedures, with the aim of determining the possibility of using diluted vitrification solution (VS) to reduce cryoprotectant toxicity and also to inhibit nucleation at practical cooling and rewarming by the effect of supplemental KF7G. Examination was performed using shoot apices of cranberry and plant vitrification solution 2 (PVS2) with dilution. Vitrification procedures using the original concentration (100%) of PVS2 caused serious injury during treatment with PVS2 and resulted in no regrowth after cooling and rewarming (cryopreservation). Dilution of the concentration of PVS2 to 75% or 50% (with the same proportions of constituents) significantly reduced injury by PVS2 treatment, but regrowth was poor after cryopreservation. It is thought that dilution of PVS2 reduced injury by cryoprotectant toxicity, but such dilution caused nucleation during cooling and/or rewarming, resulting in poor survival. On the other hand, addition of 0.5 mg/ml (0.05% w/v) KF7G to the diluted PVS2 resulted in significantly (p < 0.05) higher regrowth rates after cryopreservation. It is thought that addition of supercooling-facilitating KF7G induced vitrification even in diluted PVS2 probably due to inhibition of ice nucleation during cooling and rewarming and consequently resulted in higher regrowth. The results of the present study indicate the possibility that concentrations of routinely used VSs can be reduced by adding supercooling-facilitating KF7G, by which more successful cryopreservation might be achieved for a wide variety of biological materials. |
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