Micropropagation of date palm (Phoenix dactylifera L.) and papaya (Carica papaya L.)
McCubbin, Michelle Jacqueline.
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Date palms (Phoenix dactylifera L.) and papayas (carica papaya L.) are two commercially important plantation crops. Their economic potential in South Africa and worldwide is increasing. However, due to disease, pests and socio-economic reasons, planting material is in short supply. Micropropagation provides a method for rapidly propagating selected superior cultivars for commercial and environmental interests. A satisfactory process for the regeneration of elite cultivars should result in individuals phenotypically and genetically identical to the explant from which they were derived. However, due to somaclonal variation generated during in vitro culture, the true-to-typeness is questionable. For this reason a southern African survey for off-types on date palms produced using somatic embryogenesis was conducted. Plant growth variations such as leaf variegation, seedless fruit, broad leaves, compact growth habit and parthenocarpic fruit were recorded and possible explanations for each phenomenon given. Factors influencing the date palm initiation process such as decontaminating agents, plant growth regulators, explant type and nurse cultures were investigated. A double decontamination process with 2.6% and 1.3% sodium hypochlorite was most effective at reducing contamination. Alternative plant growth regulators, TIBA and NAA were ineffective as a substitute to 2,4-D for somatic embryogenesis. The size of the explant and "nurse cultures" played an important role in explant growth and initiating callogenesis. A "nurse culture" reduced the time in culture significantly. The problem areas in the three commercial tissue culture techniques used for date palms were outlined. In the second part of the study, factors influencing initiation, multiplication and rooting of papaya were determined. Presoaking with antibiotic, Rifampicin, and various fungicides had a positive effect on decontaminating papaya explants, while Bronocide™ had little effect. Various methods and materials were used to optimize papaya multiplication and rooting in vitro. The growth and multiplication of papaya was optimal at 50 g l ¯¹ sucrose. Gelling agent, Gelrite, increased multiplication rates significantly but had a negative effect on overall growth causing plants to become vitrified. The addition of activated charcoal reduced vitrification but also reduced multiplication rate. Activated charcoal greatly improved overall growth of papaya and reduced leaf senescence. No vitrification was observed in multiplying papaya cultures where agar and Gelrite combinations were used, but multiplication rate was reduced compared to cultures grown on Gelrite alone. callus removal from the bases of papaya at subculturing reduced multiplication rate and influenced elongation, growth and leaf senescence. Lower concentrations magar and Gelrite improved rooting percentages, but did not provide good support. Damaged roots and lower rooting percentages were observed on plantlets treated with IBA for four weeks compared to those exposed for only two days. A one hour pulse with a higher concentration (5 mg l ¯¹) of IBA greatly improved rooting percentage and further eliminated a second subculture onto an IBA-free medium after two days. Good, strong roots with root hairs were produced on vermiculite medium containing equal volumes of DS salts and vitamins. Modified lids with cotton-wool plugs also reduced leaf abscission. In vitro grafting using stericrepe proved impractical, while grafting in vitro unrooted papaya plants onto ex vitro seedlings was more successful, using wedge and slant grafts. Grafts sealed with pegs and Parafilm™ were less effective.