Selection of optimal stage for protocorm-like bodies and production of artificial seeds for direct regeneration on diffe
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ORIGINAL PAPER
Selection of optimal stage for protocorm-like bodies and production of artificial seeds for direct regeneration on different media and short term storage of Dendrobium Shavin White Suryanti Bustam • Uma Rani Sinniah • Midhzar Abdul Kadir • Faridah Qamaruz Zaman Sreeramanan Subramaniam
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Received: 25 July 2012 / Accepted: 3 October 2012 / Published online: 13 October 2012 Ó Springer Science+Business Media Dordrecht 2012
Abstract Micropropagation is currently the most popular method for orchid propagation through the production of protocorm-like bodies (PLBs). It is suggested that converting the PLBs into artificial seeds by encapsulation with sodium alginate can be useful for short-term preservation and distribution to the laboratories and commercial nurseries. Prior to the production of artificial seeds, the best developmental stage of PLBs based on sizes for increased conversion to plantlet was determined. PLBs were categorized based on size and presence of shoot namely B2 mm (S1), [2–4 mm (S2), [4–6 mm (S3), [2–4 mm with shoot (S4) and [4–6 mm with shoot (S5). S4 and S5 gave significantly higher conversion percentage (85 and 90 %, respectively) as compared to the PLBs without shoot (S1, S2 and S3). Thus, for uniformity PLBs of 3–5 mm with shoot were used for encapsulation with sodium alginate to form artificial seeds. The feasibility of germinating artificial seeds of Dendrobium Shavin White in different substrates namely; M1 (semi-solid Murashige and Skoog (1962) basal medium), M2 (cotton irrigated with sterilized liquid MS basal medium), M3 (cotton irrigated with sterilized distilled water) and M4 (cotton irrigated with non-sterilized distilled water) was tested. The encapsulated
S. Bustam U. R. Sinniah (&) M. A. Kadir Department of Crop Science, Faculty of Agriculture, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia e-mail: [email protected] F. Q. Zaman Department of Biology, Faculty of Science, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia S. Subramaniam School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden Heights, Penang, Malaysia
PLBs regenerated well in M1 where 96 % of encapsulated PLBs germinated after 12 days of inoculation and 76 % of them converted into plantlet after 37 days of inoculation while PLBs subjected to sterile distilled water gave 56 % germination and 44 % conversion after 42 and 167 days of inoculation respectively. The ability to store encapsulated PLBs would be advantageous for transport of planting materials. Encapsulated PLBs survived longer when stored at 25 ± 2 °C compared to 4 °C, 10 °C and 30 ± 2 °C whereby storage up to 75 days retained 80–92 % survival. Further storage up to 135 days retained 52 % survival. All plantlets survived after acclimatization when transferred to charcoal media under shade. Keywords Acclimatization Conversion Encapsulation Orchid Survival Synthetic seed Abbreviations PLBs Protocorm-like bodies MS Murashige and Skoog (1962) ANOVA Analysis of variance SPSS Statistical
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