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A simple and reproducible protocol for plant regenerationand cryopreservation of Grammatophyllum specinocum BL. |
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| รหัสดีโอไอ | |
| Creator | 1. Nattawadee Wongrasee 2. Sumontip Bunnag |
| Title | A simple and reproducible protocol for plant regenerationand cryopreservation of Grammatophyllum specinocum BL. |
| Publisher | Research and Development Office, Prince of Songkla University |
| Publication Year | 2018 |
| Journal Title | Songklanakarin Journal of Science and Technology |
| Journal Vol. | 40 |
| Journal No. | 1 |
| Page no. | 251 |
| Keyword | cryopreservation, plant regeneration, orchid, RAPD |
| URL Website | http://rdo.psu.ac.th/sjstweb/index.php |
| ISSN | 0125-3395 |
| Abstract | Grammatophyllum specinocum BL. is native to Thailand and is at risk of extinction due to human-causedenvironmental changes and commercial exploitation. Cryopreservation is an efficient approach for conservation of plantgermplasms. The aim of this study was to establish a simple and reproducible protocol for callus induction and plantregeneration, and cryopreservation of Grammatophyllum specinocum BL. The orchid seeds successfully germinated on halfstrengthMurashige and Skoog (? MS) medium. Amendment of 0.5 mg/l 6-benzylaminopurine (6-BA) into MS medium wassuitable for callus induction whilst a combination of 1 mg/l 6-BA and 2 mg/l naphthaleneacetic acid (NAA) was effective inpromoting plantlet regeneration. Protocorms of G. specinocum were successfully cryopreserved using encapsulation-dehydrationmethod with the following protocol: 24-h preculture of protocorms with 0.75 M sucrose solution in the dark followed byencapsulation of the precultured protocorms in Ca-alginate beads, 24-h pretreatment of encapsulated beads with 0.5 M sucrosesolution in the dark, and 8-h dehydration before plunging into liquid nitrogen. Random amplified polymorphic DNA (RAPD)analysis was carried out to detect genetic stability of cryopreserved protocorms and genetic differences not detected between noncryopreservedand cryopreserved protocorms. |
