A Successful Regeneration from Shoot Tips of Chrysanthemum morifolium (Ramat.) following Cryopreservation by Droplet-vitrification

다국어 초록 (Multilingual Abstract)

This study describes an efficient and widely applicable droplet-vitrification following cryopreservation for shoottips of Chrysanthemum morifolium (Ramat.) cvs. ‘Borami’ and ‘Yes morning’. The shoot tips of Chrysanthemum wereprecultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3-0.7 M). Precultured explantswere treated with loading solution (LS, C6) containing glycerol 20% and sucrose 20% for 30 min and exposed todehydration solution (B5) containing 40% of glycerol and 40% of sucrose for 60 min at 25℃, and then transferred ontodroplets containing 2.5 ㎕ PVS3 on sterilized aluminum foils (4 ㎝ × 0.5 ㎝) prior to direct immersion in liquid nitrogen(LN) for 1 h. The highest regeneration rate (%) was obtained when shoot tips were precultured with treatment-2 (exposingof shoot tips to MS + 0.3M sucrose for 30 h and then treated with MS+0.5 M sucrose for 16 h) at 25℃ in both the cultivars.
The viability of cooled samples, followed by culturing on NH4NO3-free MS medium for first 5 days was increased totwo-fold (80.7%) regrowth rate over those cultured on normal MS medium or MS medium containing plant growthregulators. This result shows droplet-vitrification would be a promising method for cryobanking chrysanthemumgermplasm.

참고문헌 (Reference)

1 이윤걸, "작은방울-유리화법에 의한 국화 신초의 초저온동결보존" 농업과학연구소 38 (38): 227-233, 2011

2 Sakai, A, "Vitrification, encapsulationvitrification and droplet-vitrification: a review" 28 : 151-172, 2007

3 Martín, C., "Survival and genetic stability of Dendranthema grandiflora Tzvelev shoot apices after cryopreservation by vitrification and encapsulation-dehydration" 51 : 281-289, 2005

4 Wang, R.R., "Shoot recovery and genetic integrity of Chrysanthemum morifolium shoot tips following cryopreservation by droplet-vitrification" 176 : 330-339, 2014

5 이정윤, "Regeneration of Cryopreserved Pear Shoot Tips Grown in Vitro by Encapsulation-Dehydration" 한국자원식물학회 30 (30): 612-617, 2017

6 Khaing, A.A., "Phylogenetic relationships of chrysanthemums in Korea based on novel SSR markers" 12 : 5335-5347, 2013

7 Teixeira da Silva, J.A., "Ornamental chrysanthemums: improvement by biotechnology" 79 : 1-18, 2004

8 Kuriyama, A., "Inhibitory effect of ammonium ion on recovery of cryopreserved rice cells" 64 : 231-235, 1989

9 Vidal, N., "In Vitro Cell" 41 : 63-68, 2005

10 Lee, Y.G., "Improved cryopreservation of chrysanthemum (Chrysanthemum morifolium) using droplet-vitrification" 32 : 487-497, 2011

11 Yeung, H.C., "Handbook of Chinese Herbs" Institute of Chinese Medicine 1983

12 Martín, C., "Genetic stability analysis of Chrysanthemum (Chrysanthemum × morifolium Ramat.) after different stages of an encapsulation-dehydration cryopreservation protocol" 168 : 158-166, 2011

13 Schwinn, K.E., "Floral flavonoids and their potential for pelargonidin biosynthesis in commercial Chrysanthemum cultivars" 35 : 145-150, 1994

14 이정윤, "Efficient Cryopreservation of Lilium spp. Shoot Tips using Droplet-vitrification" 한국육종학회 1 (1): 131-136, 2013

15 Panis, B., "Droplet-vitrification of apical meristems: a cryopreservation protocol applicable to all Musaceae" 168 : 45-55, 2005

16 Kim, H.H., "Development of cryopreservation protocols through droplet-vitrification and its application to vegetatively propagated crop germplasm" 12-, 2010

17 Kim, H.H., "Development of alternative plant vitrification solutions in droplet-vitrification procedures" 30 : 320-334, 2009

18 Kim, H.H., "Development of alternative loading solutions in droplet-vitrification procedures" 30 : 291-299, 2009

19 Fukai, S., "Cryopreservation of shoot tips of Chrysanthemum morifolium and related species native to Japan" 54 : 201-204, 1991

20 Kim, H.H., "Cryopreservation of potato cultivated varieties and wild species: critical factors in droplet vitrification" 27 : 223-234, 2006

21 Chen, X.L., "Cryopreservation of in vitro-grown apical meristems of Lilium by droplet-vitrification" 77 : 397-403, 2011

22 Fukai, S., "Cryopreservation of chrysanthemum shoot tips" 45 : 167-174, 1990

23 Halmagyi, A., "Cryopreservation of Chrysanthemum morifolium (Dendranthema grandiflora Ramat.) using different approaches" 22 : 371-375, 2004

24 Hitmi, A., "Cryopreservation of Chrysanthemum cinerariaefolium shoot tips" 156 : 408-412, 2000

25 Kulus, D., "Cryopreservation as a tool used in long-term storage of ornamental species – a review" 168 : 88-107, 2014

26 Martín, C., "Cryopreservation and genetic stability of Dendranthema grandiflora Tzvelev in vitro cultures" 18 : 129-135, 2009

27 Kim, H.H., "Cryobanking of Korean Allium germplasm collections: results from 10-yearexperience" 33 : 45-57, 2012

28 Teixeira da Silva, J.A., "Chrysanthemum low-temperature storage and cryopreservation: a review" 120 : 423-440, 2015

29 De Boucaud, M.T., "Biotechnology in Agriculture and Forestry 50" 2002

30 Luyen, B.T., "Anti-inflammatory components of Chrysanthemum indicum flowers" 25 : 266-269, 2015

31 Liang, F., "An arabinogalactan from flowers of Chrysanthemum morifolium: structural and bioactivity studies" 387 : 37-41, 2014

32 Decruse, S.W., "Ammonium nitrate in the culture medium influences regeneration potential of cryopreserved shoot tips of Holostemma annulare" 23 : 55-60, 2002

33 Murashige, T, "A revised medium for rapid growth and bioassay with tobacco tissue cultures" 15 : 473-497, 1962

34 Suzuki, M., "A novel preculture method for induction of desiccation tolerance in gentian axillary buds for cryopreservation" 135 : 69-76, 1998