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Limitations in the Use of Cycloheximide in Studies of Apple Ripening
Authors:KNEE  MICHAEL; HATFIELD  STEPHEN G S; RATNAYAKE  MALINIE
Abstract:In order to discover whethor the production of aroma volatilesby apple fruits is dependent on the synthesis of appropriateenzymes during ripening, excised peel, excised cortical tissue,and whole apples were treated with cycloheximide (CH). Volatilerelease, ethylene production, respiration, flesh softening,and peel chlorophyll degradation were measured. The ethylene and volatile compounds produced by excised peelapparently resulted from wounding rather than processes analogousto fruit ripening. Excised cortical tissue was capable of autonomousripening with ethylene production, respiration, and softeningcomparable to that in intact fruits. After infiltration withsucrose solution the same changes occurred, but they were delayedby up to 4 d. Cycloheximide inhibited respiration although theextent of this inhibition decreased after 3 d. Cycloheximideprevented the onset of rapid ethylene production but stimulatedproduction of ethanol, ethyl acetate, and other volatiles. Softeningof CH-treated cortical discs was associated with progressivenecrosis. When whole apples were infiltrated with CH through hypodermicneedles inserted into the core, 14C]valine incorporation wasinhibited from the core to the mid-cortex but not in the peeland outer cortex. Infiltration with sucrose solution delayedmany ripening changes although the time of maximum 14C]valineincorporation was unaffected. Early effects of CH on respirationwere masked by the effects of infiltration, but after 5 d CH-infiltratedfruit contained higher CO2 concentrations and respired morerapidly than controls. Internal ethylene concentrations wereusually lower in CH-treated apples than in controls. CH stimulated release of ethanol and ethyl acetate but inhibitedrelease of higher molecular weight esters such as propyl andbutyl acetates. Cycloheximide-treated fruit softened, but thiswas apparently due to internal necrosis. Peel chlorophyll degradationwas inhibited by CH treatment of whole apples although the tissuehad apparently received no inhibitor.
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