- Volume 28 Issue 6
DOI QR Code
Morphological and Genetic Stability of Dormant Apple Winter Buds After Cryopreservation
Yi, JungYoon;Lee, GiAn;Chung, JongWook;Lee, YoungYi;Kwak, JaeGyun;Lee, SeokYoung
- 투고 : 2015.05.28
- 심사 : 2015.07.08
- 발행 : 2015.12.31
Twenty apple germplasm accessions from the Korean Genebank were successfully cryopreserved using two-step freezing to back up genetic resources maintained by field collections. This study examined the morphological and genetic stability of cryopreserved dormant apple buds that were stored in liquid nitrogen, and then rewarmed and regrown. Whole plants were regenerated directly from dormant buds through budding without an intermediary callus phase. The cryopreserved buds produced high levels of shoot formation (76.2-100%), similar to those of noncryopreserved buds (91.3-100%), with no observed differences between cryopreserved and noncryopreserved materials. Three of the twenty cryopreserved apple germplasm accessions were used to assess morphological and genetic stability. No differences in morphological characteristics including shoot length, leaf shape, leaf width/length ratio, and root length were observed between controls (fresh control and noncryopreserved) and cryopreserved plantlets. The genetic stability of regenerants (before and after cryopreservation) was investigated using inter simple sequence repeat (ISSR) markers. The ISSR markers produced 253 bands using four primers, ISSR 810, SSR 835, ISSR 864, and ISSR 899. These markers showed monomorphic banding patterns and revealed no polymorphism between the mother plant and regenerants before and after cryopreservation, suggesting that cryopreservation using two-step freezing does not affect the genetic stability of apple germplasm. These results show that two-step freezing cryopreservation is a practical method for long-term storage of apple germplasms.
Apple;Cryopreservation;Dormant bud;Two-step freezing;Genetic stability
- Forsline, P.L., J.R. McFerson, W.F. Lamboy and L.E. Towill. 1999. Development of base and active collections of Malus germplasm with cryopreserved dormant buds. Acta Hort. 484:75-78.
- Dixit, S., B.B. Mandal, S. Ahuja and P.S. Srivastav. 2003. Genetic stability assessment of plants regenerated from cryopreserved embryogenic tissue of Dioscorea bulbifera L. using RAPD, biochemical and morphological analysis. Cryo Lett. 24:77-84.
- De Verno, L.L., Y.S. Park, J.M. Bonga and J.D. Barrett. 1999. Somaclonal variation in cryopreserved embryogenic clones of white spruce. Plant Cell Rep. 18:948-953. https://doi.org/10.1007/s002990050689
- Castillo, N.R.F., N.V. Bassil, S. Wada and B.M. Reed. 2010. Genetic stability of cryopreserved shoot tips of Rubus germplasm. In vitro Cell. Dev. Biol.-Plant. 46:246-256. https://doi.org/10.1007/s11627-009-9265-z
- Benson, E.E., M. Wikinson and A. Todd. 1996. Developmental competence and ploidy stability in plants regenerated from cryopreserved potato shoot tips. Cryo Lett. 17:119-128.
- Sakai, A. and Y. Nishiyama. 1978. Cryopreservation of winter vegetative buds of hardy fruit trees in liquid nitrogen. Hort. Sci. 13:225-227.
- Rostiana, O., M. Niwa and Marubashi. 1999. Efficiency of inter-simple sequence repeat PCR for detecting somaclonal variation among leaf-culture-regenerated plants of horseradish. Br. Sci. 49:245-250. https://doi.org/10.1270/jsbbs.49.245
- Piccioni, F., G. Barcaccia, M. Falcinelli and A. Standardi. 1997. Estimating alfalfa somaclonal variation in axillary branching propagation and indirect somatic embryogenesis by rapid fingerprinting. Int. J. Plant Sci. 158:556-562. https://doi.org/10.1086/297467
- Harding, K. 2004. Genetic integrity of cryopreserved plant cells; A review. Cryo Lett. 25:3-22.
- Harding, K. 1991. Molecular stability of the ribosomal RNA genes in Solanum tuberosum plants recovered from slow growth and cryopreservation. Euphytica 55:141-146. https://doi.org/10.1007/BF00025226
- Harding, K. and E.E. Benson. 2000. Analysis of nuclear and chloroplast DNA in plants regenerated from cryopreserved shoot-tips of potato. Cryo Lett. 21:279-289.
- Haggman, H.M., L.A. Ryynanen, T.S. Aronen and J. Krajnakova. 1998. Cryopreservation of embryogenic cultures of Scots pine. Plant Cell Tiss. Organ Cult. 54:45-53. https://doi.org/10.1023/A:1006104325426
- Gonzalez-Amao, M.T., C. Urra, F. Engelmann, R. Ortiz and C. de la Fe. 1999. Cryopreservation of encapsulated sugarcane apices: effects of storage temperature and storage duration. Cryo Lett. 20:347-352.
- Forsline, P.L., L.E. Towill, J.W. Waddell, C. Stushnoff, W.F. Lamboy and J.R. McFerson. 1998. Recovery and longevity of cryopreserved apple buds. J. Amer. Soc. Hort. Sci. 123:365-370.
- Withers, L.A. and F. Engelmann. 1997. In vitro conservation of plant genetic resources: In Altman, A. (ed.), Biotechnology in Agriculture, Marcel Dekker, New York, USA. pp. 57-88.
- Ward, A.C.W., E.E. Benson, N.W. Blackhall, S. Cooper-Bland, W. Powell, J.B. Power and M.R. Davey. 1993. Flow-cytometric assessments of ploidy stability in cryopreserved dihaploid Solanum tuberosum and wild Solanum species. Cryo Lett. 14:145-152.
- Vendrame, W.A., G. Kochert and H.Y. Wetzstein. 1999. AFLP analysis of variation in pecan somatic embryos. Plant Cell Rep. 18:853-857. https://doi.org/10.1007/s002990050673
- Tyler, N.J., C. Stushnoff and L.V. Gusta. 1988. Freezing of water in dormant vegetative apple buds in relation to cryopreservation. Plant Physiol. 87:201-205. https://doi.org/10.1104/pp.87.1.201
- Tyler, N.J. and C. Stushnoff. 1988. Dehydration of dormant apple buds at different stages of cold acclimation to induce cryopreservability in different cultivars. Can. J. Plant Sci. 68:1169-1176. https://doi.org/10.4141/cjps88-145
- Towill, L.E., P.L. Forsline, C. Walters, J.W. Waddell and J. Laufman. 2004. Cryopreservation of Malus germplasm using a winter vegetative bud method: results from 1915 accessions. Cryo Lett. 25:323-334.
- Towill, L.E. and D.D. Ellis. 2008. Cryopreservation of dormant buds: In Reed, B.M. (ed.), Plant Cryopreservation―A Practical Guide, Springer, New York, USA. pp. 421-439.
- Stushnoff, C. 1987. Cryopreservation of apple genetic resources. Can. J. Plant Sci. 67:1151-1154. https://doi.org/10.4141/cjps87-154
- Scocchi, A., M. Faloci, R. Medina, S. Oimos and L. Mroginski. 2004. Plant recovery of cryopreserved apical meristem-tips of Melia azedarach L. using encapsulation-dehydration and assessment of their genetic stability. Euphytica 135:29-38. https://doi.org/10.1023/B:EUPH.0000009538.01279.d3
- Zhang, Z.H. and Z.L. Hu. 2000. Regenerating plants from cryopreserved adventitious buds of haploids and their genetic stability in rice. L. Wuhan Bot. Res. 18:169-173.
- Zhang, Y.J., X.G. Zhang and J. Pang. 2001. Cryopreservation of in vitro-grown shoot tips of Amorphophallus by vitrification. Acta Agronom. Sin. 27:97-102.
- Zhai, Z., Y. Wu, F. Engelmann, R. Chen and Y. Zhao. 2003. Genetic stability assessment of plantlets regenerated from cryopreserved in vitro cultured grape and kiwi shoot tips using RAPD. Cryo Lett. 24:315-322.
- Yi, J.Y., G.A. Lee, S.Y. Lee, J.W. Chung and I.S. Shin. 2013. Cryopreservation of dormant apple winter buds using ‘two-step freezing.’ Plant Breed. Biotech. 1:283-289.
- Wu, Y., F. Engelmann and M. Zhao. 1999. Cryopreservation of apple shoot tips: importance of cryopreservation technique and of conditioning of donor plants. Cryo Lett. 20:121-130.
- Morphological Characteristics and Genetic Diversity Analysis of Cultivated Sancho (Zanthoxylum schinifolium) and Chopi (Zanthoxylum piperitum) in Korea vol.29, pp.5, 2016, https://doi.org/10.7732/kjpr.2015.28.6.697
- Cryobiotechnology of apple (Malus spp.): development, progress and future prospects 2018, https://doi.org/10.7732/kjpr.2015.28.6.697
- Response of Germination Rate to Variable Drying Conditions and Moisture Contents for Storage of Dehisced Korean Ginseng Seeds vol.29, pp.6, 2016, https://doi.org/10.7732/kjpr.2015.28.6.697
- Cryopreservation of in Vitro Grown Shoot Tips of Sweet Potato (Ipomoea batatas L.) by the Encapsulation-Vitrification Method vol.29, pp.6, 2016, https://doi.org/10.7732/kjpr.2015.28.6.697
연구 과제번호 : "사과, 배 유전자원의 초저온동결보존 기술개발 및 실용화"