Investigation on possibility of cryopreservation of Eucalyptus microtheca seeds

Document Type : Research article

Authors

1 M.Sc. Student, Faculty of Natural Resources, Tarbiat Modares University

2 Assistant Prof., Research Institute of Forests and Rangelands

3 Senior research expert, Research Institute of Forests and Rangelands

Abstract

The aim of this study was to investigate the possibility of cryopreservation of Eucalyptus microtheca seeds at -196 ºC, using Liquid Nitrogen (LN). Before transferring the seeds into LN, three pre-treatments including Plant Vitrification Solution 2 (PVS2), Desiccation and Glycerol 30% was applied. The treated seeds transferred into LN for period of one week. The seeds removed from the LN, subjected to heat shock (+42 ºC) for 2 minutes, blotted and transferred both onto moist paper in petri dishes and pots, filled with soil and pit moss. The petri dishes and pots, transferred into germinator (+24 ºC) or greenhouse (+20ºC), respectively. Subsequent of cryopreservation period, seed germination and recovery was high, there were no significant differences between cryopreserved and control seeds in germinator and greenhouse conditions. Further more, there were no adverse effect or abnormality observed in seedlings developed from cryopreserved seeds. Results showed that, long-term preservation of E. microtheca seeds under LN (cryopreservation) is promising.                     
 

Keywords


- عصاره، م.ح. و سردابی، ح، 1386. اکالیپتوس. شناخت، معرفی و ازدیاد با استفاده از فناوریهای نوین (جلد اول). انتشارات مؤسسه تحقیقات جنگلها و مراتع کشور. 672 صفحه.
- Abdul-Baki, A.A. and Anderson, J.D., 1973. Vigor determination in soybean seed by multiplication. Crop Science, 3: 630-633.
- Bhat, S.R., Bhat, K.H. and Chandel, K.P.S., 1994. Studies on germination and cryopreservation of Musa balbisiana seed. Seed Science and Technology, 22(3): 637-640.
- Bhat, S.N., Sharma, A. and Bhat, S.V., 2005. Vitrification and glass transition of water: insights from spin probe ESR. Phys, Rev. Lett., 95 (23):     4 p.
- Blakesley, D. and Kiernan, R.J., 2001. Cryopreservation of axillary buds of a Eucalyptus grandis×Eucalyptus camaldulensis hybrid. CryoLetters, 22(1): 13-8.
- Dussert, S., Chabrillange, N., Engelmann, F., Anthony, F., Louarn, J. and Hamon, S., 1998. Cryopreservation of seeds of four coffee species (Coffea arabica, C. costatifructa, C. racemosa and C. sessiliflora): importance of water content and cooling rate. Seed Science Research, 8: 9-15.
- Engelmann, F., 1990. Use of cryopreservation for plant germplasm long-term conservation casehistory: Oil Palm somatic embryos. International Journal of Refrigeration, 13 (1):     26-30.
- Fahy, G.M., Levy, D.I. and Ali, S.E., 1987. Some emerging principles underlying the physical properties, biological actions and utility of vitrification solutions. Cryobiology, 24: 196–213.
- Ishikawa, K., Harata, K., Mii, M. and Sakai, A., 1997. Cryopreservation of zygotic embryos of a Japanese terrestrial orchid (Bletilla striata) by vitrification. Plant Cell Reports, 16: 754–757.
- Jeyendran, R.S., Van der Ven, H.H., Perez-Pelaez, M. and Zaneveld, L.J., 1985. Effect of glycerol and cryopreservation on oocyte penetration by human spermatozoa. Andrologia, 17(3): 241-8.
- Lambardi, M. and De Carlo, A., 2003. Application of tissue culture to the germplasm conservation of temperate broad-leaf trees. In: Jain, S.M. and Ishii, K., (Eds.), Micropropagation of Woody Trees and Fruits. Kluwer Ac. Pub., Dordrecht: 815-840.
- Leborgne, N., Teulieres, C., Travert, S., Rols, M-P., Teissie, J. and Boudet, A.M., 2004. Introduction of specific carbohydrates into Eucalyptus gunnii cells increases their freezing tolerance. European Journal of Biochemistry, 229 (3): 710-717.
- Little, E.L., 1983. Common fuelwood crops: a handbook for their identification. Communi-Tech Associates. 354 p.
- Liu, H., Yu, H., Dai, J., Gong, Q., Yang, K. and Lu, X., 2003. Cryopreservation of protoplasts of the alga Porphyra yezoensis by vitrification. Plant Science, 166 (1): 97-102.
- Matsumoto, T., Mochida, K., Itamura, H. and Sakai, A., 2001. Cryopreservation of persimmon (Diospyros kaki Thunb) by vitrifiation of dormant shoot tips. Plant Cell Report, 20: 398-402.
- Nishizawa, S., Sakai, A., Amano, Y. and Matuzawa, T., 1993. Cryopreservation of asparagus (Asparagus officinalis L.) embryogenic suspension cells and subsequent plant regeneration by vitrification. Plant Science, 88: 67–73.
- Normah, M.N. and Siti Dewi Serimala, M.N., 1997. Cryopreservation of seeds and embryonic axes of several citrus species. In: Ellis, R.H., Black, M., Murdoch, A.J. and Hong, T.D., (Eds.). Basic and Applied Aspects of Seed Bioligy. Dordrecht: Kluwer Academic Published, 817-823.
- Ozkavukcu, S. and Erdemli, E., 2002. Cryopreservation: basic knowledge and biophysical effects. Journal of AnkaraMedicalSchool, 24(4): 187-196.
- Panis, B., and Lambardi, M., 2005. Status of cryopreservation technologies in plants (crops and forest trees). The role of biotechnology. Villa Gualino, Turin, Italy: 5-7.
- Phunchindawan, M., Hirata, K., Sakai, A., 1997. Cryopreservation of encapsulated shoot primordia induced in horseradish (Armoracia rusticana) hairy root cultures. Plant Cell Reports, 16: 469–473.
- Purohit, S.S., 2004. Plant Tissue Culture. Student Edition, 366 p.
- Rao, N.K., 2004. Plant Genetic Resources: Advancing conservation and use through biotechnology. African Journal of Biotechnology, 3: 136-145.
- Roberts, E.H. and Ellis, R.H., 1989. Water and seed survival. Annals of Botany, 63: 39-52.
- Sakai, A., Kobayashi, S. and Oiyama, I., 1990. Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) by vitrification. Plant Cell Report, 9: 30–33.
- Sakai, A., Kobayashi, S. and Oiyama, I., 1991. Survival by vitrification of nucellar cells of navel orange (Citrus sinensis var. brasiliensis Tanaka) cooled to –196°C. Plant Physiology, 137: 465–470.
- Sakai, A. and Engelmann, F., 2007. Vitrification, Encapsulation and Droplet-Vitrification: a review. CryoLetters, 28 (3): 141-172.
- Scocchi, F.D., Vila, S., Mroginski, L. and Engelmann, F., 2007. Cryopreservation of somatic embryos of Paradise tree (Melia azedarach L.). CryoLetters,  28 (4): 281-290.
- Schoenweiss, K., Meier-Dinkel, A. and Grotha, F., 2005. Comparison of cryopreservation techniques for long-term storage of Ash (Fraxinus excelsior L.). CryoLetters, 26 (3): 201-212.  
- Stanwood, P.C., 1985. Cryopreservation of seed germplasm for genetic conservation. In: Kartha, K.K., (Ed.) Cryopreservation of Plant Cells and Tissues. Boca Raton, FL: CRC: 199-226.
- Susiluoto, S. and Berninger, F., 2007. Interactions between morphological and physiological drought responses in Eucalyptus microtheca. Silva Fennica, 41(2): 221-233.
- Tuomela, K., 1997. Physiological and morphological responses of Eucalyptus microtheca provenances to water availability in tropical drylands. Ph.D. Thesis, Univ. Helsinki, Tropic. For. Rep. 13, 60 p.
- Turner, S., Seranatna, T., Touchell, D., Bunn, E., Dixon, K. and Tan, B., 2001. Stereochemical arrangement of hydroxyl groups in sugar and polyalcohol molecules as an important factor in effective cryopreservation. Plant Science, 160: 489-497.
- Volk, G.M., Harris, J.L. and Rotindo, K.E., 2006. Survival of mint shoot tips after exposure to cryoprotectant solution components. Cryobiology, 52: 305-308.
- Wang, Y.L., Fan, M.J. and Liaw, S.I., 2005. Cryopreservation of in vitro-grown shoot tips of papaya (Carica papaya L.) by vitrification. Bot. Bull. Acad. Sin., 46: 29-34.
- Webb, D.E., Wood, P.J., Smith, J. and Heman, G.S., 1984. Guide to species selection for tropical and sub-tropical plantations. Unit of Tropical Silviculture, Commonwealth Forestry Institute, University of Oxford, Oxford, GB. Tropical forestry papers; No. 15, Second Edition, 256 p.
- Wood, C.B., Pritchard, H.W. and Lindegard, K., 2003. Seed cryopreservation and longevity of two Salix hybrids. CryoLetters, 24 (1): 17-26.