臺灣博碩士論文加值系統

本研究以北蕉(Giant Cavendish. Musa acuminata Colla)組織培養苗為材料，探討玻璃化法超低溫冷凍保存前處理流程對存活率之影響。北蕉以高濃度蔗糖培養基健化與處理冷凍保護劑LS和PVS2，及進行生理分析和切片觀察，並探討流程中各種處理對冷凍保存存活率之影響和貢獻。
北蕉組織培養苗以不同濃度蔗糖培養基進行短期預培養和長期預培養，配合LS處理0~60分鐘，PVS2處理0~3小時，進行超低溫冷凍保存。結果顯示經0.3M蔗糖培養8週後，處理LS 20分鐘、PVS2 2小時，超低溫冷凍保存之存活率可達37.5%，未經液態氮處理可達77.08％。根據生理分析測定，發現北蕉經0.3M蔗糖長期培養後之相對含水量減少、滲透潛勢降低、可溶性醣類含量和可溶性蛋白質含量之也都增加。顯示高糖培養導致北蕉莖頂脫水及累積溶質，有助於適應高滲透逆境及增加冷凍保存時的耐凍性，推論高糖預培養有助於提高超低溫冷凍保存後的存活率。
因此，玻璃化法冷凍保存流程之前處理中，以適當高濃度蔗糖長期預培養與適當時間之冷凍保護劑處理，為北蕉超低溫冷凍保存成功關鍵之兩步驟。

This study investigates effects of cryopreservation pretreatment by vitrification on banana (Giant Cavendish. Musa acuminata Colla). The physiological responses and observed section were analyzed by preculturing plantlets with high concentration of sucrose and cryoprotectant (LS and PVS2) treatment to investigate the effects and contribution of cryopreservation.
The apices from in vitro-grown banana plantlets were precultured for short- and long-time on hormone-free MS medium containing different sucrose concentrations. The precultured apices were then treated with a loading solution (LS) for 0~60 minutes and then dehydrated with a plant vitrification solution (PVS2) for 0~3 hours, and following cryopreservation. Banana plantlet cultured on 0.3M sucrose medium long-time results in best survival rates of 37.5%, and treated without LN survival rates of 77.08%. Based on the physiological measurement，banana apices relative water content reduce, osmotic potential decrease, and both soluble sugar content and soluble protein content increase. These results indicated that banana plantlet culturing with high concentration of sucrose result in dehydration of banana shoot tips and accumulation of soluble sugar which renders high osmotic potential and increased freeze-tolerance for cryopreservation. It suggsets that precultrue in high concentration of sucrose is helpful for increasing survival rates after cryopreservation.
Thus preculture with high concentration sucrose and the duration for pretreatment with cryoprotectant were achieved through investigating two key steps of the vitrification protocol.

目錄………………………………………………………………………Ⅰ
表目錄……………………………………………………………………Ⅱ
圖目錄……………………………………………………………………Ⅲ
縮寫表……………………………………………………………………Ⅴ
摘要…………………………………………………………………………1
Abstract……………………………………………………………………..2
一、前言……………………………………………………………………4
二、前人研究………………………………………………………………8
三、材料與方法…………………………………………………………14
四、結果…………………………………………………………………23
五、討論…………………………………………………………………58
六、結論…………………………………………………………………77
七、參考文獻……………………………………………………………79
附錄………………………………………………………………………99

王源龍。2004。番木瓜莖頂之液態氮超低溫保存方法及傷害之探討。國立中興大學生命科學研究所博士論文。
孔慶仁。1979。香蕉之栽培。台灣省青果運銷合作社 印。
李淑英，戴奇協，朱慶國。1988。北蕉不定芽分切代數對變異之影響。中國園藝。34（4）：321。
國際種子檢查規則。2000。行政院農委會種子檢查室 編印。
黃新川。2005。香蕉。台灣農家要覽農作篇（二）。財團法人豐年社。pp.9-18
黃弼臣。1966。台灣栽培之品種。分類與品種。香蕉。中興大學農業教育學系。
張明聰。1992。木瓜水分生理之探討 國立中興大學植物學研究所博士論文。
蔡叔芬。2004。臺農25、臺農64與臺農66品種甘藷玻璃化法超低溫冷凍保存前處理流程之探討。國立中興大學生命科學研究所碩士論文。
蔡疇。1985。台灣外銷香蕉產業之演變。青果研究資料第78期。台灣省青果運銷合作社 編印。
鄧澄欣，劉程江，戴奇協，邱讚秀，黃新川。2000。香蕉園藝性狀改良研究（Ⅰ）中矮性抗黃葉病品系的選育與評估。中國園藝。46 (2) : 173-182
劉淦芝。1974。香蕉的分類與型態。香蕉通論。台灣香蕉研究所。pp.1-13
劉淦芝。1976。世界香蕉栽培品種概況。香蕉栽培法。徐氏基金會。pp.47-50
劉云國，王曉云。2002。蘋果種質資源玻璃化法超低溫保存技術。山東農業大學學報(自然科學版)。33(1)：32-36
賴宏輝。1985。香蕉栽培指導手冊。台灣香蕉研究所。pp.1-71
嚴慶丰，黃純農。1994。植物組織和細胞的玻璃化凍存研究[J]。細胞生物學雜誌。16 (3) : 117-122
Abdelnour-Esquivel A, Mora A, Villalobos V (1992) Cryopreservation of zygotic embryos of Musa acuminata (AA) and M. balbisiana (BB).
CryoLett. 131:59-164
Baek HJ, Kim HH, Cho EG, Chae YA, Engelmann F (2003) Importance of explant size and origin and of preconditioning treatments for cryopreservation of garlic shoot apices by vitrification. CryoLett. 24 : 381-388
Bagniol S, Engelmann F (1992)Effect of thawing and recovery conditions on the regrowth of meristems of date palm (Phoenix dactylifera L.) after cryo-preservation in liquid nitrogen. CryoLett. 13:253-260
Bartolome AP, Ruperez P, Fuster C (1995) Pineapple fruit: morphological characteristics, chemical composition and sensory analysis of Red Spanish and Smooth Cayenne cultivars. Food Chem 53:75-79
Benson EE, Reed BM, Brennam RM, Clacher KA , Ross DA (1996)Use of thermal analysis in the evaluation of cryopreservation protocols for Ribes nigrum L. germplasm. CryoLett. 17:347-362
Bonicel A, Haddad G, Gagnaire J (1987) Seasonal variations of starch and major soluble sugars in the different organs of young poplars. Plant Physiol. Biochem. 25:451-459
Bradford M (1976) A rapid and sensitive method for quantification of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem. 72:248-254
Brison M（1995）Cryopreservation of in vitro grown shoot tips of two interspecific Prunus rootstocks. Plant Sci. 105:235-242
Bryan DM, Ya''acov YL(1994) Stress and Stress Coping In Cultivated Plants. Dordrecht , Boston : Kluwer Academic Publishers, pp:136-139
Bryant G, Koster KL, Wolfe J (2001) Membrane behaviour in seeds and other systems at low water content: the various effects of solutes. Seed Sci Res. 11:17-25
Charoensub R, Phansiri S, Sakai A, Yongmanitchai W(1999) Cryopreservation of cassava in vitro-grown shoot tips cooled to -196℃ by vitrification. CryoLett. 20: 89-94
Chin HF (1996)Germination and storage of banana seeds, In: E.A.
Frison, J.-P. Horry, D. De Waele (Eds.), in: Proceedings of a
Workshop on New Frontiers in Resistance Breeding for Nematode,
Fusarium and Sigatoka, Kuala Lumpur, Malaysia, pp. 218-227
Chu C, Dai Z, Maurice SB, Edward GE (1990) Induction of crassulacean acid metabolism in the facultative halophyte Mesembryanthemum Crystallium by abscisic acid. Plant Physiol. 93:1253-1260
Crowe JH, Crowe LM, Carpenter JF, Wistrom CA (1987) Stabilization
of dry phospholipid bilayers and proteins by sugars. Biochem J 242:1-10
Crowe JH, Crowe LM, Carpenter JF, Rudolph AS, Wistrom CA,
Spargo BJ, Anchordoguy TJ (1988) Interactions of sugars with membranes, Biochim. Biophys. Acta 947:367-384
de Boucaud MT, Brison M, Helliot B, Herve´-Paulus V (2002) Cryopreservation of prunus, in: L.E. Towill, Y.P.S. Bajaj (Eds.), Biotechnology in Agriculture and Forestry. Cryopreservation of Plant
Germplasm II, vol. 50, Springer-Verlag , Berlin, Heidelberg. pp. 288-311
Dereuddre J, Blandin S, Hassen N (1991) Resistance of alginate-coated
somatic embryos of carrot (Daucus carota L.) to desiccation and freezing in liquid nitrogen. 1. Effect of preculture. CryoLett. 12:125-134
Dereuddre J, Fabre J, Bassaglia C (1988) Resistance to freezing in liquid nitrogen of carnation (Dianthus caryophyllus L. var Eolo) apical and axillary shoot tips excised from different aged in vitro plantlets Plant Cell Rep. 7: 170-173
Dumet D, Engelmann F, Chabrillange N, Duval Y, Dereuddre J (1993)
Importance of sucrose for the acquisition of tolerance to desiccation and cryopreservation of oil palm somatic embryos. CryoLett. 14: 243-250
Engelmann F (1997) Importance of desiccation for the cryopreservation of recalcitrant seed and vegetatively propagated species. Plant Genetic Resources Newsletter 112: 9-18
Engelmann F, Dereuddre J (1988) Cryopreservation of oil palm somatic embryos: importance of the freezing process. CryoLett. 9: 220-235
Escobar RH, Mafla G, Roca WM (1997) A methodology for recovering
cassava plants from shoot tips maintained in liquid nitrogen. Plant Cell
Rep. 16:474-478
Fahy GM, MacFarlane DR, Angell CA, Meryman HT (1984) Vitrification as an approach to cryopreservation. Cryobiol. 21(4) :407-426
Forsline PL, Towill LE, Waddell JW, Stushnoff C, Lamboy WF, Mcferson JR (1998) Recovery and longevity of cryopreserved dormant apple buds. J. Am. Soc. Hort. Sci. 123:365-370
Frison EA, Sharrock S (1999) The economic, social and nutritional importance of banana in the world. In: Picq C, Foure E,Frison EA (eds) Banana and food security. INIBAP, Montpellier,France, pp 21-35
Goldstein G, Nobel PS (1994)Water relations and low-temperature acclimation for cactus species varying in freezing tolerance. Plant Physiol. 104:675-681
Gonzalez-Arnao MT, Moreira T, Urra C (1996) Importance of
pregrowth with sucrose and vitrification for the cryopreservation
of sugarcane apices using encapsulation-dehydration. CryoLett. 17: 141-148
Gonzalez-Arnao MT, Ravelo MM, Villavicencio CU, Montero MM,
Engelmann F (1998)Cryopreservation of pineapple (Ananas Comosus)
apices. CryoLett. 19 : 375-382
Guy CL, Haskell D (1987) Induction of freezing tolerance in spinach is associated with the synthe-sis of cold acclimation-induced proteins. Plant Physiol. 84：872-878
Guy CL, Niemi K J, Brambl R (1985) Altered gene expression during cold acclimation of spinach. Proc. Natl. Acad. Sci. USA 82：3673-3677
Haskins RH, Kartha KK (1980) Freeze preservation of pea meristems: cell survival. Canadian Journal of Botany. 58:833-840
Helliot B, Swennen R, Poumay Y, Frison E, Lepoivre P, Panis B (2003)
Ultrastructural changes associated with cryopreservation of banana (Musa spp.) highly proliferating meristems. Plant Cell Rep. 21:690-698
Hincha DK (1989) Low concentrations of trehalose protect isolated thylakoids against mechanical freeze– thaw damage. Biochem. Biophys. Acta 987:231- 234
Hirai D , Sakai A (1999a) Cryopreservation of in vitro-grown
axillary shoot tips of mint (Mentha spicata L.) by encapsulation vitrification. Plant Cell Rep. 19: 150-155
Hirai D, Sakai A (1999b) Cryopreservation of in vitro-grown
meristems of potato (Solanum tuberosum L.) by encapsulation vitrification. Potato Res 42: 153-160.
Hirai D, Shirai K, Shirai S , Sakai A (1998) Cryopreservation of in vitro-grown meristems of strawberry (Fragaria x ananassa Duch.) by encapsulation vitrification. Euphytica. 101: 109-115
Hirsh AG, Williams RJ, Meryman HT (1985) A novel method of natural cryopreservation: Intra-cellular glass formation in deeply frozen Populus. Plant Physiol. 79:41-56
Jensen WA (1973) Botanical histochemistry W.H. Freeman and Company, San Francisco London. pp. 55-99
Jitsuyama Y, Suzuki T, Harada T , Fujikawa S (1997) Ultrastructural study on mechanism of increased freezing tolerance due to extracellular glucose in cabbage leaf cells. CryoLett. 18:33-44
Kartha KK (ed.) (1985) Cryopreservation of Plant Cell and Organs. CRC Press, Boca Raton, Florida. pp.276
Kartha KK , Engelmann F (1994) Cryopreservation and germplasm
storage. Plant Cell and Tissue Culture. ed. Vasil IK,Thorpe TA. pp:95-230
Kartha K, Leung KL, Pahl K (1980) Cryopreservation of strawberry meristems and mass propagation of plantlets. J. Amer. Soc. Hort. Sci. 105: 481-484
King GA, Henderson KG, O’Donoghue EM, Martin W, Lill RE (1988) Flavour and metabolic changes in asparagus during storage. Hort. Sci. 36: 183-190
Kobayashi S, Sakai A, Ohgawara T, Nakamura Y (1994) Stable maintenance of an integrated gene in nucellar cells of navel orange (Citrus sinensis Osb.) under storage in LN2, J. Jpn. Soc. Hort. Sci. 63(3)553-558
Koster KL, Lei YP, Anderson M, Martin S, Bryant G (2000) Effects of vitrified and nonvitrified sugars on phosphatidylcholine fluid-to-gel phase transitions. Biophysical J 78 : 1932-1946
Kumu Y, Harada T, Yakuwa T (1983)Development of a whole plant from a shoot tip of Asparagus officinalis L. frozen down to -196℃. J. Fac. Agr. Hokkaido Univ. 61:285-294
Lambardi M, Fabbri A, Caccavale A (2000) Cryopreservation of white poplar (Populus alba L.) by vitrification of in vitro-grown shoot tips. Plant Cell Rep. 19:213-218
Langis R, Schnable B, Earle ED, Steponkus PL (1989) Cryopreservation
of Brassica campestris L. cell suspension by vitrification. CryoLett. 10: 421-428
Leborgne N, Teulieres C, Travert S, Rols MP, Teissie J, Boudet AM (1995) Introduction of specific carbohydrates into Eucalyptus gunnii cells increases their freezing tolerance. Eur. J. Biochem. 229：710-717
Leslie SB, Israeli E, Lighthart B, Crowe JH, Crowe LM (1995) Trehalose and sucrose protect both membranes and proteins in intact bacteria during drying. Appl. Environ. Microbiol. 61: 3592-3597
Lockhart BEL, Jones DR (2000) Banana mosaic and banana streak,
diseases caused by viruses. In: Jones DR (ed) Diseases of banana,
Abaca and Enset. CABI Publishing, Wallingford, UK, pp 241–293
Mari S, Engelmann F, Chabrillange N, Huet C, Michaux-Ferriere N (1995) Histocytological study of apices of coffee (Coffea racemosa and C. sessiliflora) in vitro plantlets during their cryopreservation using the encapsulation–dehydration technique. CryoLett. 16:289-298
Marthe B; de Boucaud, Marie-Therese; Dosba, Francoise (1995) Cryopreservation of in vitro grown shoot tips of two interspecific Prunus rootstocks. Plant Sci. 105: 235-242
Matsumoto T, Nako Y, Takahashi C, Sakai A (1997) Induction of in vitro cultured masses of shoot primordia of hybrid statice and its cryopreservation by vitrification. Hort Science 32: 309-311
Matsumoto T, Sakai A (1995) An approach to enhance dehydration
tolerance of alginate-coated dried meristems cooled to -196 °C. CryoLett. 16: 299-306
Matsumoto T , Sakai A (2003) Cryopreservation of axillary shoot tips of in vitro-grown grape (Vitis) by a two-step vitrification protocol. Euphytica 131: 299-304
Matsumoto T, Sakai A , Nako Y (1998) A novel preculturing for enhancing the survival of in vitro-grown meristems of wasabi (Wasabia japonica) cooled to –196°C by vitrification. CryoLett. 19:27-36
Matsumoto T, Sakai A, Yamada K (1994) Cryopreservation of in vitro-
grown apical meristems of wasabi (Wasabia japonica) by vitrification
and subsequent high plant regeneration. Plant Cell Rep. 13:442-446
Matsumoto T, Sakai A ,Yamada K (1995) Cryopreservation of in vitro-grown apical meristems of lily (Lilium japonicum) by vitrification. Plant Cell Tiss Org Cult. 41: 231-241.
Matsumoto T, Mochida K, Itamura H, Sakai A (2001)Cryopreservation of persimmon (Diospyros kaki Thunb.) by vitrification of dormant shoot
tips, Plant Cell Rep. 20:398-402
Mazur P(1966) Theoretical and experimental effects of cooling and warming velocity on the survival of frozen and thawed cells. Cryobiol. 2:181-192
Mazur P (1977)The role of intracellular freezing in the death of cells cooled at supraoptimal rates. Cryobiol. 14:251-272
Meryman HT (1971) Osmotic stress as a mechanism of freezing injury. Cryobiol. 8:489-500
Morton J (1987) Banana. In: Fruits of warm climates. Morton JF, Miami FL. pp. 29–46
Niino T, Sakai A (1992) Cryopreservation of alginate-coated in vitro-
grown shoot tips of apple, pear and mulberry. Plant Sci. 87:199–206
Niino T, Sakai A, Yukuwa H, Nojiri K (1992) Cryopreservation of in vitro-grown shoot tips of apple and pear by vitrification. Plant Cell Tissue Organ Cult. 28: 261-266
Nishizawa S, Sakai A, Amano Y, Matuzawa T (1993) Cryopreservation
of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by vitrification. Plant Sci. 88: 67-73
Palta JP, Weiss LS (1993) Ice formation and freez-ing injury: An overview on the survival mecha-nisms and molecular aspects of injury and cold acclimation in herbaceous plants. In: Advances in Plant Cold Hardiness, Li PH and Chris-tersson L, Ed. CRC Press, Boca Ra-ton/Ann Arbor/London/Tokyo pp. 143–176
Panis B, Piette B, Swennen R (2005) Droplet vitrification of apical meristems: a cryopreservation protocol applicable to all Musaceae.
Plant Sci. 168:45–55
Panis B, Strosse H, Hende SVD, R Swennen (2002)Sucrose preculture to simplify cryopreservation of banana meristem cultures, CryoLett. 23: 375-384
Panis B, Totte N, Vannimmen K, Withers LA, Swennen R (1996)
Cryopreservation of banana (Musa spp.) meristem cultures after preculture on sucrose. Plant Sci. 121: 95-106
Panis B, Withers LA, Langhe ED (1990) Cryopreservation of Musa
suspension cultures and subsequent regeneration of plants. CryoLett.
11:337-350
Pennycooke JC, Towill LE (2000) Cryopreservation of shoot tips from in
vitro plants of sweet potato [Ipomoea batatas (L.) Lam.] by vitrification.
Plant Cell Rep. 19:733-737
Peschke VM, Phillips RL (1992) Genetic implication of somaclonal in plants. Adv.Genet. 30:41-75
Plessis P, Steponkus PL (1996) Effect of preculture in a sucrose-containing medium on the sugar content of potato shoot-tips. Cryobiol. 33: 654-655.
Pritchard HW, Grout BWW, Short KC (1986) Osmotic stress as a
pregrowth procedure for cryopreservation 1. Growth and ultrastructure
of sycamore and soybean cell suspensions. Ann. Bot 57: 41-48
Reed BM (1989) The effect of cold hardening and cooling rate on the
survival of apical meristems of Vaccinium species frozen in liquid
nitrogen. CryoLett. 10: 315-322
Reed BM (1990)Survival of in vitro-grown apical meri-stems of Pyrus following cryopreservation. Hort Science 25: 111-113
Reinhoud PJ(1996) Cryopreservation of Tobacco Suspension
Cells by Vitrification. Doctoral Thesis, Leiden U, Institute of Molecular Plant Sci., Leiden. pp. 1-95
Reinhoud PJ, Schrijnemakers EWM, Iren F , Kijne JW (1995) Vitrification and a heat-shock treatment improve cryopreservation of tobacco cell suspension compared to two-step freezing. Plant Cell Tiss Org Cult. 42: 261-267
Sakai A (1962) Studies on the frost-hardiness of woody plants. I. The causal relation between sugar content and frost-hardiness. Cont. Inst. Low Temp. Sci. Hokkaido Univ. Ser. B 11: 1-40
Sakai A (1995) Cryopreservation for germplasm collection in woody plants. Somatic embryogenesis in woody plants, Vol. 1. History, Molecular and Biochemical Aspects, and Applications. ed. Jain SM et al., pp.293-315
Sakai A (2000) Development of cryopreservation techniques, In: F. Engelmann, H. Takagi (Eds.), Cryopreservation of Tropical Plant Germplasm. Current Research Progress and Application, IPGRI, Rome, Italy. pp. 1-7
Sakai A, Kobayashi S, Oiyama I (1990) Cryopreservation of nucellar
cells of navel orange (Citrus sinensis Osb. var. brasiliensis
Tanaka) by vitrification. Plant Cell Rep. 9:30-33
Sakai A, Kobayashi S, Oiyama I (1991) Cryopreservation of nucellar
cells of navel orange (Citrus sinensis Osb.) by a simple freezing
method. Plant Sci. 74:243-248
Sakai A, Nishiyama N (1978) Cryopreservation of winter vegetative buds of hardy fruit trees in liquid nitrogen. Hort Science. 13:225-227
Sakai A, Yoshida S (1967) Survival of plant tissue at super-low temperatures. I. Effects of cooling and rewarming rates on survival.
Plant Physiol. 42:1965-1701
Sakai A, Yoshida S (1968) The role of sugar and related compounds in variations of freezing resistance. Cryobiol. 5：160-174
Santarius KA (1973) The protective effect of sugars on chloroplast membranes during temperature and water stress and its relationship to frost desiccation and heat resistance. Planta. 113:105-114
Sarkar D, Naik PS (1998) Cryopreservation of shoot tips of tetraploid potato (Solanum tuberosum L.) clones by vitrification. Ann. Bot 82: 455-461
Scottez C, Chevreau E, Godard N, Arnaud Y, Duron M, Dereuddre J (1992)Cryopreservation of cold-acclimated shoot tips of pear in vitro cultures after encapsulation–dehydration. Cryobiol. 29:691-700
Shiomi N (1989)Properties of fructosyltransferases in-volved in the synthesis of fructan in Liliaceous plants. J. Plant Physiol. 134: 151–155
Simmonds NW (1966) Bananas.2nd ed, Longman, London. pp.512
Slavik B (1974) water in cells and tissues. In: B Slavik ed. Methods of Studying plant water Relations. Academid Publishing House of the Czevhoslovak Academy of Science Prague, New York .pp.1-20
Steponkus PL (1984) Role of the plasma membrane in freezing injury and cold acclimation. Annu. Rev. Plant Physiol. 35:543-584
Steponkus PL, Dowgert MF, Gordon-Kamm WJ (1983) Destabilization of plasma membrane of isolated plant protoplasts during freeze-thaw cycle: The influence of cold acclimation. Cryobiol. 20:448-465
Steponkus PL, Garber MP, Myers SP, Lineberger RD (1977) Effects of cold acclimation and freezing on structure and function of chloroplast thylakoids. Cryobiol. 14:303-321
Steponkus PL, Langis R, Fujikawa S (1992) Cryopreservation of plant tissues by vitrification. In: Advances in Low-Temperature Biology, Vol. 1. P.L. Steponkus (ed.). JAI Press Ltd., Hampton Mill, UK. pp. 1–61
Su HJ, Hwang SC, Ko WH (1986) Fusarial wilt of Cavendish banana in Taiwan .Plant Dis.70:814-818
Suzuki T, Kaneko M, Harada T (1997) Increase in Freezing Resistance of Excised Shoot Tips of Asparagus officinalis L. by Preculture on Sugar-Rich Media. Cryobiol. 34: 264-275
Takagi H (2000) Recent developments in cryopreservation of shoot apices of tropical species, in: Engelmann F, Takagi H (Eds.), Cryopreservation of Tropical Plant Germplasm. Current Research Progress and Application,IPGRI, Rome, Italy. pp. 178-193
Takagi H, Thinh NT, Islam OM, Senboku T, Sakai A (1997) Cryopreservation of in vitro-grown shoot tips of taro(Colocasia esculenta (L.) Schott) by vitrification. 1. Investigation of basic conditions of the vitrification procedure. Plant Cell Rep. 16:594-599
Takahashi T, Hirsh A, Erbe EF, Bross JB, Steere RL, Williams RJ (1986) Vitrification of human monocytes. Cryobiol. 23:103-115
Tao D, Li PH, Carter V (1983) Role of cell wall in freezing tolerance of cultured potato cells and their protoplasts. Plant Physiol. 58: 527-532
Thinh NT (1997) Cryopreservation of germplasm of vegetatively propagated tropical monocots by vitrification. Doctoral Papers of Kobe University, Department of Agronomy, Japan.
Thinh NT, Takagi H, Sakai A (2000) Cryopreservation of in vitro grown
apical meristems of some vegetatively propagated tropical monocots by vitrification. In: F. Engelmann, H. Takagi (Eds.),Cryopreservation of Tropical Plant Germplasm. Current Research Progress and Application, IPGRI, Rome, Italy. pp. 227-232
Thinh NN, Takagi H, Yashima S (1999) Cryopreservation of in vitro-grown shoot tips of banana (Musa spp.) by vitrification method. CryoLett. 20: 163-174
Tkagi H, Thinh NT, Kyesmu PM (1998) Cryopreservation of vegetatively propagated tropical crops by vitrificayion. Acta Hort. 461:485-490
Towill LE (1984) Survival of ultra-low temperatures of shoot-tips from Solanum tuberosum groups andigena, phureja, stenotomum and other tuber-bearing Solanum species. CryoLett. 5: 319-326
Towill LE, Jarret RL (1992) Cryopreservation of sweet potato (Ipomea
batatas (L.) Lam.) shoot tips by vitrification. Plant Cell Rep.11: 175-178
Turner SR, Touchell DH, Dixon K, Tan B (2000) Cryopreservation of Anigozanthos viridis spp. viridis and related taxa from the south west of Western Australia. Aust. J. Bot. 48:739-744
Uemura M, Joseph RA, Steponkus PL (1995) Cold acclimation of Arabidopsis thaliana. Plant Physiol. 109: 15-30
Uragami A, Sakai A, Nakai M (1990) Cryopreservation of dried axillary buds from plantlets of Asparagus officinalis L. grown in vitro. Plant Cell Rep. 9: 328-331
Uragami A, Sakai A, Nagai N (1993) Cryopreservation of asparagus (Asparagus officinalis L.) cultured in vitro. Japan Agri Res Quart. 27(2):112-115
Van den Houwe I, De Smet K, Tezenas de Montcel H, Swennen R (1995) Variability in storage potential of banana shoot cultures under medium term storage conditions. Plant Cell Tissue Org. 42:269-274.
Webb MS, Steponkus PL (1990) Dehydration-induced hexagonal phase
formation in phospholipid bilayers. Cryobiol. 27: 666-667
Withers LA, Street HE (1977) Freeze-preservation of cultured plant
cells. III. The pregrowth phase. Plant Physiol. 39: 171-178
Yamada T, Sakai A, Matsumura T, Higuchi S (1991) Cryopreservation
of apical meristems of white clover (Trifolium repens L.) by vitrification. Plant Sci. 78: 81-87
Yoshimatsu K, Yamaguchi H, Shimomura K (1996) Traits of Panax
ginseng hairy roots after cold storage and cryopreservation. Plant Cell Rep. 15:555-560
Yoshida S, Forno JH, Cook H, Gornez KA (1976) Determination of chlorophyll in plant tissue. Laboratory mannal of physiological studies for rice. IRRI 26:43-49