冷冻条件对植物细胞低温保存的影响
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摘要
为保证食品果蔬冷藏保存的品质,细胞层面的基础研究是必不可少的一部分,细胞内冰的形成(IIF)会导致严重的细胞损伤从而导致低温储存中的诸多问题。预冷过程对胞内冰晶的形成具有重要影响,因此拍摄到细胞内冰晶形成过程为优化冷冻保存方案提供必要的支持。在冷冻洋葱表皮细胞时,采用不同的降温速率,并使用低温显微系统配合高速摄像机,可视化细胞内更清晰的IIF过程。实验结果表明,冷却速率的增加使得在相同温度下细胞内IIF出现的概率降低,但细胞边缘冰晶生长速率增加,使细胞受到的损伤程度变化波动明显。
Basic research at the cellular level is an essential part for ensuring the quality of food and vegetables in cold storage.Intracellular ice formation(IIF) causes severe cellular damage,thus causing many problems in cryopreservation.The pre-cooling process has an important influence on the formation of intracellular ice crystals.Therefore,photographing the intracellular ice crystal formation process provides the necessary support for optimizing the cryopreservation scheme.The onion epidermal cells were frozen at different cooling rates,and a low-temperature microscopy system was used with a high-speed camera to visualize the IIF process in the cells.At the same temperature,the greater the rate of cooling,the lower the probability of intracellular IIF emergence.However,the growth rate of ice crystals at the edge of the cells increased,and the degree of damage to the cells fluctuated significantly.
Basic research at the cellular level is an essential part for ensuring the quality of food and vegetables in cold storage.Intracellular ice formation(IIF) causes severe cellular damage,thus causing many problems in cryopreservation.The pre-cooling process has an important influence on the formation of intracellular ice crystals.Therefore,photographing the intracellular ice crystal formation process provides the necessary support for optimizing the cryopreservation scheme.The onion epidermal cells were frozen at different cooling rates,and a low-temperature microscopy system was used with a high-speed camera to visualize the IIF process in the cells.At the same temperature,the greater the rate of cooling,the lower the probability of intracellular IIF emergence.However,the growth rate of ice crystals at the edge of the cells increased,and the degree of damage to the cells fluctuated significantly.
引文
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[27]LAGERVELD B W.Cryosurgical induced injury of human cancerous tissues-How it works?[J].British Journal of Medical&Surgical Urology,2012,5(5):S24-S27.
[28]MULDREW K,MCGANN L E.The osmotic rupture hypothesis of intracellular freezing injury[J].Biophysical Journal,1994,66(2 Pt 1):532-541.
[29]GORDON-KAMM W J,STEPONKUS P L.The behavior of the plasma membrane following osmotic contraction of isolated protoplasts:implications in freezing injury[J].Protoplasma,1984,123(2):83-94.
[30]FONSECA S C,GIL L,MANSO M C,et al.Modelling the influence of storage temperature and time after cutting on respiration rate of diced red onions(Allium cepa L.cv.Vermelha da Póvoa)[J].Postharvest Biology&Technology,2018,140(C):27-33.
[31]KAWAHARA H.Characterizations of functions of biological materials having controlling-ability against ice crystal growth[J].Zentralblatt Für Arbeitsmedizin Arbeitsschutz Prophylaxe Und Ergonomie,2013,30(11):424-430.
[2]HOFFMANN N E,BISCHOF J C.The cryobiology of cryosurgical injury[J].Urology,2002,60(2):40-49.
[3]LEVITT J.Responses of Plants to Environmental Stress:Volume 1:Chilling,Freezing,and High Temperature Stresses[M].2nd ed.New York:Academic Press,1980.
[4]BROWN M S.Texture of frozen vegetables:effect of freezing rate on green beans[J].Journal of the Science of Food&Agriculture,2010,18(2):77-81.
[5]BARBOSACANOVAS G V,ALTUNAKAR B,MEJIALORIO D.Freezing of fruits and vegetables.An agribusiness alternative for rural and semi-rural areas[J].FAO Agricultural Services Bulletin,2005,36(9):3911-3915.
[6]MANSOORI G A.Kinetics of water loss from cells at subzero centigrade temperatures[J].Cryobiology,1975,12(1):34-45.
[7]BARTELSRAUSCH T,BERGERON V,CARTWRIGHT J H E,et al.Ice structures,patterns,and processes:a view across the ice-fields[J].Review of Modern Physics,2012,84(2):885-944.
[8]ZARAGOTAS D,LIOLIOS N T,ANASTASSOPOULOS E.Supercooling,ice nucleation and crystal growth:a systematic study in plant samples[J].Cryobiology,2016,72(3):239-243.
[9]MATSUMOTO M.Molecular dynamics of ice nucleation and crystal growth[J].Refrigeration(in Japanese),2012,87:554-559.
[10]FUJIKAWA S.Freeze-fracture and etching studies on membrane damage on human erythrocytes caused by formation of intracellular ice[J].Cryobiology,1980,17(4):351-362.
[11]郝保同,刘宝林,林萍,等.冰晶对黏附成骨细胞损伤的实验研究[J].制冷学报,2010,31(1):59-62.HAO B T,LIU B L,LIN P,et al.Experimental study on the damage of ice crystals to adherent osteoblasts[J].Journal of Refrigeration,2010,31(1):59-62.
[12]苏光明,HOSAHALLI S,RAMASWAMY,等.牛肉高压冷冻过程中热变化和冰晶形态研究[J].农业机械学报,2014,45(3):206-214.SU G M,HOSAHALLI S,RAMASWAMY,et al.Thermal changes and ice crystal morphology during high pressure freezing of beef[J].Transactions of the Chinese Society of Agricultural Machinery,2014,45(3):206-214.
[13]华泽钊.人体细胞的低温保存与冷冻干燥[J].制冷技术,2007,(2):17-20.HUA Z Z.Cryopreservation and freeze-drying of human cells[J].Refrigeration Technology,2007,(2):17-20.
[14]张哲,赵静,田津津,等.冷冻-复温过程中葡萄细胞结晶变化研究[J].农业机械学报,2016,47(5):211-217.ZHANG Z,ZHAO J,TIAN J J,et al.Study on the change of grape cell crystallization during freezing-rewarming process[J].Transactions of the Chinese Society of Agricultural Machinery,2016,47(5):211-217.
[15]李维杰,王利红,刘宝林,等.降温速率对聚乙烯吡咯烷酮水溶液结晶的影响[J].制冷技术,2016,36(1):31-34.LI W J,WANG L H,LIU B L,et al.Effect of cooling rate on crystallization of polyvinylpyrrolidone solution[J].Refrigeration Technology,2016,36(1):31-34.
[16]MAZUR P.Freezing of living cells:mechanisms and implications[J].American Journal of Physiology,1984,247(3 Pt 1):C125-C142.
[17]LEEF J L,MAZUR P.Physiological response of neurospora conidia to freezing in the dehydrated,hydrated,or germinated state[J].Applied&Environmental Microbiology,1978,35(1):72-83.
[18]OHNISHI S,FUJII T,MIYAWAKI O.Electrical and rheological analysis of freezing injury of agricultural products[J].International Journal of Food Properties,2002,5(2):317-332.
[19]王益聪,武卫东,张兵.山梨醇水溶液过冷度影响因素研究[J].制冷技术,2018,18(1):26-31.WANG Y C,WU W D,ZHANG B.Study on factors affecting subcooling of sorbitol aqueous solution[J].Refrigeration Technology,2018,18(1):26-31.
[20]MODLIBOWSKA I,ROGERS W S.Freezing of plant tissues under the microscope[J].Journal of Experimental Botany,1955,6(18):384-391.
[21]ASAHINA E.Freezing processes and injury in plant cells[J].Plant Cold Hardiness&Freezing Stress,1978,39(5):17-36.
[22]杨戈尔,张爱丽,徐学敏,等.胞内冰晶形成[J].工程热物理学报,2007,28(s2):55-57.YANG G E,ZHANG A L,XU X M,et al.Formation of intracellular ice crystals[J].Journal of Engineering Thermophysics,2007,28(s2):55-57.
[23]STOTT S L,KARLSSON J O M.Visualization of intracellular ice formation using high-speed video cryomicroscopy[J].Cryobiology,2009,58(1):84-95.
[24]NINAGAWA T,EGUCHI A,KAWAMURA Y,et al.A study on ice crystal formation behavior at intracellular freezing of plant cells using a high-speed camera[J].Cryobiology,2016,73(1):20-29.
[25]SCHNEIDER C A,RASBAND W S,ELICEIRI K W.NIH Image to ImageJ:25 years of Image Analysis[J].Nature Methods,2012,9(7):671-675.
[26]高永毅.植物的细胞力学行为研究及在水果机械损伤研究中的应用[D].北京:中国农业大学,2003.GAO Y Y.Study on the cellular mechanical behavior of plants and its application in fruit machinery damage research[D].Beijing:China Agricultural University,2003.
[27]LAGERVELD B W.Cryosurgical induced injury of human cancerous tissues-How it works?[J].British Journal of Medical&Surgical Urology,2012,5(5):S24-S27.
[28]MULDREW K,MCGANN L E.The osmotic rupture hypothesis of intracellular freezing injury[J].Biophysical Journal,1994,66(2 Pt 1):532-541.
[29]GORDON-KAMM W J,STEPONKUS P L.The behavior of the plasma membrane following osmotic contraction of isolated protoplasts:implications in freezing injury[J].Protoplasma,1984,123(2):83-94.
[30]FONSECA S C,GIL L,MANSO M C,et al.Modelling the influence of storage temperature and time after cutting on respiration rate of diced red onions(Allium cepa L.cv.Vermelha da Póvoa)[J].Postharvest Biology&Technology,2018,140(C):27-33.
[31]KAWAHARA H.Characterizations of functions of biological materials having controlling-ability against ice crystal growth[J].Zentralblatt Für Arbeitsmedizin Arbeitsschutz Prophylaxe Und Ergonomie,2013,30(11):424-430.