Aspergillus oryzae NRRL 35191 was isolated as an endophyte from coffee leaves and found to produce kojic acid (KA) in culture. When inoculated into cacao seedlings (Theobroma cacao), A. oryzae grew endophytically and synthesized KA in planta. Cacao seedlings inoculated with A. oryzae produced higher levels of caffeine than noninoculated ones. Aspergillus oryzae may be a useful endophyte to introduce to cacao since it grows nonpathogenically and induces the caffeine defense response that may make the plant more tolerant to insects and pathogens.
The causes of poor fruit set of cocoa (Theobroma cacao L.) in the greenhouse were studied by examining factors that may influence pollen germination. Hand pollination of cocoa flowers resulted in 45.8% fruit set when flowers were pollinated within 3 hours of anthesis. Pollen germination did not occur until about 6 hours after pollination. Later pollinations (7 to 9 hours after anthesis) or earlier pollinations (16 to 18 hours before anthesis) did not lead to fruit set. Cocoa pollen did not germinate in vitro unless the excised flowers were incubated for 6 hours at 25C in closed vials. During the incubation period, CO2accumulated to a final concentration of about 85 ml·liter-1 as a result of respiration. Ethylene production was not detectable. Incubation of flowers with a NaOH-saturated wick, to absorb CO2, prevented pollen germination in vitro. Incubation of flowers at 15C also prevented pollen germination in vitro at 25C. Hand pollination of flowers 7 to 9 hours after anthesis or 16 to 18 hours before anthesis using CO2-incubated pollen resulted in about 10% fruit set. Enclosed pollinations in vivo, in which CO2 was allowed to accumulate, resulted in nearly 100% fruit set. The initial failure to set fruit from hand pollinations may result from poor or slow pollen germination. Moreover, CO,-incubated pollen might be used to increase fruit set in cocoa by extending the effective pollination period.
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