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Research articles

ScienceAsia 38 (2012): 244-249 |doi: 10.2306/scienceasia1513-1874.2012.38.244


Effect of cryopreservation on seed germination and protocorm development of Vanda tricolor


Nipawan Jitsopakula, Kanchit Thammasirib,*, Tomohisa Yukawac, Keiko Ishikawad

 
ABSTRACT:     Germination and subsequent development was assessed after seeds of Vanda tricolor were stored in liquid nitrogen. Mature seeds, harvested 7 months after self-pollination, were directly plunged into liquid nitrogen. Germination of cryopreserved seeds on solid New Dogashima (ND) medium supplemented with 1 mg/l 6-benzyladenine, 0.5 mg/l 1-naphthaleneacetic acid and 2% sucrose was faster than non-cryopreserved seeds (28 days versus 60 days after sowing). Immature seeds, harvested 6 months after self-pollination, were treated with or without loading solution (LS) of 2 M glycerol and 0.4 M sucrose in liquid ND medium, pH 5.4 at 25 °C for 15 min and dehydrated with PVS2 solution for 0–210 min on ice and then cryopreserved by vitrification. The results showed that the germination percentage of cryopreserved seeds treated with LS was higher than without LS. After 90 days of sowing, the highest germination percentage of cryopreserved seeds was 13.6% which was higher than non-cryopreserved seeds (10.5%) when seeds were treated with LS for 15 min and then dehydrated with PVS2 solution for 180 min. After 150 days of sowing, protocorms of non-cryopreserved and cryopreserved seeds were able to form new protocorms (budding protocorms) and developed into shoots after 180 days of sowing. There were no significant differences between growth and development of protocorms derived from non-cryopreserved and cryopreserved seeds.

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a Department of Agro-Industry, Faculty of Agriculture and Technology, Rajamangala University of Technology Isan, Surin Campus, Surin 32000, Thailand
b Department of Plant Science, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
c Tsukuba Botanical Garden, National Science Museum, Tsukuba 305-0005, Japan
d Department of Research and Development, Japan Horticultural Production and Research Institute, Chiba 270-2221, Japan

* Corresponding author, E-mail: kanchitthammasiri@gmail.com

Received 18 May 2012, Accepted 3 Sep 2012