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Extractable activities of α-amylase, β-amylase, and starch phosphorylase were investigated in order to understand the mechanism of starch degradation in buttercup squash (Cucurbita maxima Duchesne ex Lam. `Delica') with the ultimate goal of improving the conversion of starch into sweet sugars. During rapid starch synthesis (0 to 30 days after flowering), extractable activities of α-amylase and β-amylase were low, but those of starch phosphorylase increased. After harvest, during ripening at 12 °C, or in fruit left in the field, activities of α-amylase and β-amylase increased. Starch contained 20% to 25% amylose soon after starch synthesis was initiated and until 49 days after harvest irrespective of whether the crop remained in the field or in storage at 12 °C. Maltose concentrations were low prior to harvest, but levels increased during fruit ripening. Data suggest starch breakdown is hydrolytic in buttercup squash, with α-amylase being the primary enzyme responsible for initiating starch breakdown.
During this study, we divided the developmental growth pattern of buttercup squash into three phases: 1) early growth, from flowering up to 30 days after flowering; 2) maturation, from 30 days until 60 days after flowering (or harvest); and 3) ripening, from 60 days (or harvest) until ≈100 days after flowering. Harvest occurred at 48 days after flowering. Fruit growth (expansion), starch, and dry matter accumulation were largely completed during early growth, and there was a progressive decline in the respiration rate. Extractable activities of acid and alkaline invertases, sucrose synthase, alkaline α-galactosidase, and sucrose phosphate synthase (assayed with saturating substrates) were high initially but declined markedly during this phase. Glucose, fructose, and low concentrations of raffinose saccharides were present, but no sucrose was detected. During maturation, starch and dry matter remained nearly constant and sucrose began to accumulate. During ripening, starch was degraded, sucrose synthase activity was significant but relatively constant, sucrose phosphate synthase activity increased, and sucrose continued to accumulate.
The pathogenic fungus Colletotrichum musae infects developing green bananas (Musa spp. AAA group), but remains latent until the fruit ripens. The aim of this research was to determine whether the appearance of disease symptoms is regulated by chitinase gene expression following treatment of fruit with benzothiadiazole (BTH) and methyl jasmonate (MeJA), and with physical (heat) and chemical (H2O2 and Ca2+-related) treatments. In bananas inoculated with C. musae, BTH and MeJA lowered disease severity and stimulated higher gene expression compared with the untreated controls during ripening. However, in naturally infected bananas, BTH and MeJA treatments slightly reduced transcription of the chitinase gene in green bananas, but they prolonged gene expression in ripe bananas and significantly reduced disease severity. The combination of H2O2 and the NADPH oxidase inhibitor, diphenylene iodonium, down-regulated chitinase gene expression and compromised disease resistance compared with H2O2 alone. Heat treatment (HT) or the combination of HT followed by CaCl2 reduced disease, but only the latter significantly upregulated chitinase gene expression. The combination of HT and a calcium ionophore (A23187) resulted in different disease indicies and different levels of gene expression depending upon the order of application: HT followed by A23187 induced higher gene expression and lower disease. The results suggest that disease resistance of green bananas could be related to high and prolonged levels of chitinase gene expression, and chitinase could be involved in harvested banana's anthracnose resistance activated by different defense pathway signals, such as BTH, MeJA, H2O2, and Ca2+.