CRYOPRESERVATION OF PEAR MERISTEMATIC TISSUES
Main Article Content
Authors
Т.Т. Turdiyev
Institute of plant biology and biotechnology, Almaty, Kazakhstan, Timiryazev str., 45, 050040
S.N. Frolov
Institute of plant biology and biotechnology, Almaty, Kazakhstan, Timiryazev str., 45, 050040
G.А. Madiyeva
Institute of plant biology and biotechnology, Almaty, Kazakhstan, Timiryazev str., 45, 050040
N.K. Rymkhanova
Al-Farabi Kazakh National University, Almaty, Kazakhstan, 71, Al- Farabi ave., 050040
I.Yu. Kovalchuk
Institute of plant biology and biotechnology, Almaty, Kazakhstan, Timiryazev str., 45, 050040
Abstract
The southern and south-eastern regions of Kazakhstan have exceptionally favorable conditions for the cultivation of pear. However, in recent years, the areas occupied with this culture are gradually decreasing. Many foreign and domestic cultivars, valuable hybrids and local traditional cultivars of folk breeding carrying genes of high productivity and adaptability have been lost in the field genebanks (orchards and nurseries). It is necessary to study, collect and preserve valuable genotypes that have important economic and biological characteristics to restore and prevent further loss of the gene pool.
Conservation of plant genetic resources in the field is associated with significant financial costs for the care of plantations, as well as the risk of loss of cultivars due to disease, pests and adverse environmental factors. The most promising and effective approach to solve this problem is the cryopreservation of plant germplasm in liquid nitrogen.
Studies on optimization of cryopreservation protocols for pear meristematic tissues were carried out based on vitrification method with 0.3 M sucrose. It has been established that the viability of meristematic tissues after cryopreservation depends on the duration of hardening, the method of thawing, the type of cryoprotectant and genotype. Optimum duration of hardening by variable temperatures (8 h, 22°C, light 5.9 lx, then 16 h, –1°C, dark) is 3-4 weeks, and the best cryoprotectant in preparing apical meristems for cryopreservation is PVS2. An effective way for viability recovery is thawing in water at a temperature of +45°C, and then at +25°C followed by planting on a nutrient medium.
Keywords
pear, biotechnology, gene pool, genetic resources, meristematic tissues, cryopreservation, cryobank
Article Details
References
Verzhuk V.G. Kriokonservatsiya – effektivnyy metod sokhraneniya geneticheskikh resursov plodovykh i yagodnykh kul'tur. Trudy po prikladnoy botanike, genetike i selektsii, 2012, vol. 169, ss. 270-279.
Benson E.E., Harding K. Cryopreservation of shoot tips and meristems: an overview of contemporary methodologies.Methods in molecular biology (Clifton, NJ), 2012, vol. 877, pp. 191-226.doi: 10.1007/978-1-61779-818-4_16.
Angelo E., Iverson V.E. Studies on Some Factors Relating to Hardiness in the Strawberry.Univ. Minn. Agric. Sta., Tech., 1939,vol.101, pp. 473-497.
Koval'chukI.Yu., Turdiyev T.T. Optimizatsiya metodov kriokonservatsii germoplazmy chornoy smorodiny (Ribesnigrum l.).Biotekhnologiya.Teoriya i praktika, 2010, no. 2, ss. 54-60.
Zhao Y., Wu Y., Engelmann F., Zhon M., Chen S. Cryopreservation of apple in vitro shoot tips by the droplet freezing methods.CryoLetters, 1999,vol. 20, no. 2, pp. 109-112.
Reed B.M. Responses to ABA and cold acclimation are genotyre development for cryopreserved blackberry and raspberry meristems. Criobiology, 1993, vol. 30, pp. 179-184.
Kovalchuk I., Turdiev T., Kushnarenko S., Rakhimbaev I., Reed B. Cryopreservation of Raspberry Cultivars: Testing Techniques for Long-Term Storage of Kazakhstan’s Plant Germplasm.The Asian and Australian Journal of Plant Science and Biotechnology: Global Science Books, 2010,no. 4, pp. 1-4.
Uragami A., Sakai A., Nagai M. Cryopreservation of dried axillary buds from plantlets of Asparagus officinalis L grown in vitro.Plant Cell Report, 1990,no. 9, pp. 328-331.
Hirai D., Sakai A. Cryopreservation of in vitro grown meristems of potato by encapsulation-vitrification // Cryopreservation of tropical plant germplasm. Current research progress and application. Eds. Engelmann F& Takagi H., JIRCAS, Tsukuba & IPGRI, Rome, 2000, pp. 205-211.
March G.,Boucaud M., Chmielarz P. Cryopreservation of Prunusavium L. embryogenic tissues. CryoLetters, c/o Royal Veterinary Co, 2005, no. 26,pp. 341-348.
Caswell K.L., Kartha K.K. Recovery of plants from pea and strawberry meristems cryopreserved for 28 years.CryoLetters, c/o University of Bedfordshire, Luton LU2 8DL, 2009,no. 30,pp. 41-46.
Niino T., Sakai A., Enomoto S., Magosi J., Kato S. Cryopreservation of in vitro-grown shoot tips of mulberry by vitrification. Cryo-Letters, 1992, vol.13, pp. 303-312.
Reed B.M. США The Basics of in vitro storage and cryopreservation National Clonal Gemplasm Repository Corvallis.O.R. USA, 2000, 34 p.
Matsumoto T., Sakai A., Nako Y. A novel precutting for enhansing the survival of in vitro grown meristems of wasabi (Wasabia japonica) cooled to –196ºC by vitrification. CryoLetters, 1998, no. 19, pp. 27-36.
Reed B.M. Cryopreservation of Temperate Berry Crops / Reed B.M. (Ed) // Plant Cryopreservation: A Practical Guide. – New York: Springer Science + Business Media LLC, 2008, pp. 333-364.
Höfer M. Cryopreservation of in Vitro Shoot Tips of Strawberry by the Vitrification Method–Establishment of a Duplicate Collection of Fragaria Germplasm. CryoLetters., 2016,vol. 37,no. 3, pp. 163-172.
Paul H., Daigny G., Sangwan-Norreel B.S. Cryopreservation of apple (Malus x domestica Borkh.) shoot tips following encapsulation-dehydration or encapsulation-vitrification. Plant Cell Reports, 2000,vol.19, pp. 768-774.
Chang Y., Reed B.M. Extended Alternating-Temperature Cold Acclimation and Culture Duration Improve Pear Shoot Cryopreservation.Cryobiology, 2000,no. 40, pp. 311-322.
Dali T., Li Paul H. Classification of plant cell cryoprotectants. Journal Theoretical Biology, 1986, no. 123, pp. 305-310.
Chang Y., Reed B.M. Preculture conditions influence cold hardiness and regrowth of Pyrus cordata shoot tips after cryopreservation. HortScience, 2001,no. 36, pp. 1329-1333.
Chang Y., Reed B.M. Effects of photoperiod and alternating temperature on the cryopreservation and cold hardiness of in vitro-grown Pyrus meristems // In Vitro Cellular and Developmental Biology, 1998, pp. 34-61.
Senula A., Keller J., Sanduijav T., Yohannes T. Cryopreservation of cold acclimated mint explants using a simple vitrification protocol. CryoLetters, 2007, no. 28, pp. 1-12.