In-vitro propagation of miracle berry (Synsepalum dulcificum/Richardella dulcifica)

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2001-12-12
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Abstract
An experiment to determine the sterilant, media and types of explants suitable for in-vitro propagation of miracle berry (Synsepalum dulcificurn/Richardella dulcifica) was conducted at the Tissue Culture laboratory of the Ghana Atomic Energy Commission (GAEC), Kwabenya, Ghana. The sterilant used was sodium hypochiorite (NaOCI) at concentrations of 25, 30 and 35% respectively for durations of 10 and 20 minutes in each case. Treating shoot tip explants of miracle berry with 30% v/v NaOCl for 20 minutes was found to be the most effective. Twenty-five percent by volume NaOCl for both 10 and 20 minutes was not effective in decontaminating the shoot tip explants and 35% v/v NaOC1 for both 10 and 20 minutes was found to be toxic to the explants and therefore resulted in their death. Five different explants were used for the experiment namely; shoot tips, cotyledon, embryo, epicotyl and hypocotyl. The shoot tip explants were obtained from Ghana Atomic Energy Commission (GAEC), Nsawam and Plant Genetic Resource Centre (PGRC), Bunso. The immature fruits from which embryo, cotyledon, epicotyl and hypocotyl explants were obtained were harvested from a 26- year-old miracle berry tree at the Horticulture Department of KNUST, Kumasi. The shoot tip explants obtained from Nsawam and PGRC recorded a higher percentage contamination (67 — 100%) compared to those obtained from GAEC that recorded a slightly lower percentage contamination of (27 — 83%). The higher percentage contamination may have caused the failure of the shoot tip explants from Nsawam and PGRC, to regenerate. The shoot tip explants obtained from GAEC resulted in shoot regeneration but the regenerated shoots failed to root. The cotyledon and epicotyl explants, though did not form shoots directly were capable of callus formation. The embryo and hypocotyl explants did not show any form of regeneration. The Murashige and Skoog (MS) medium with various concentrations and combinations of BAP (0.5, 1.0, 1.5, 2.0 and 5.0 mg/l), KIN (0.5, 1.0, 1.5, 2.0 and 5.0 mg/l) and NAA (0.5, 1.0, 1.5 mg/l) supplemented with 30 g/l sucrose and 3.5 g/1 phytagel were used to culture shoot tip explants. The medium with concentrations and combinations of BAP (0.5, 1.0, 1.5, 2.0 mg/l) and 2, 4—D (0.5, 1.0, 1.5, 2.0 mg/l) supplemented with 30g/l sucrose and 3.5g/l phytagel were used to culture the cotyledon, embryo, epicotyl and hypocotyl explants. Shoot tip explants cultured on MS full strength medium supplemented with 5.0 mg/l BAP (MSB5N0) resulted in the highest shoot regeneration whiles those cultured on MS full strength medium supplemented with 1.5 mg/l BAP (MSB3N0) gave the lowest shoot regeneration. Callus formation was highest for the cotyledon and epicotyl explants that were cultured on MS full strength medium supplemented with 2.0 mg/I 2, 4 — D (MSB0D4).
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A thesis submitted to the School of Graduate Studies, Kwame Nkrumah University of Science and Technology in partial fulfilment of the requirements for the award of Master of Science degree in Fruit Crops (Pomology), 2001
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