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This study examined the relationship between the activity of fruit enzymes involved in metabolizing sucrose and sugar accumulation during fruit development, to clarify the role of these key enzymes in sugar accumulation in papaya fruit. Papaya fruit (Carica papaya L. cv. Sunset) were harvested from 14 to 140 days after anthesis (DAA). Fruit dry matter persent, total soluble solids (TSS), and sugar composition and the activity of enzymes: sucrose phosphate synthetase (SPS), sucrose synthetase (SS), and acid invertase were measured. `Sunset' papaya matured 140 days after anthesis during the Hawaii summer season and in about 180 days in cool season on the same plant. Fruit flesh dry matter persent, TSS, and total sugar did not significantly increase until 30 days before harvest. Sucrose synthetase was very high 2 weeks post-anthesis, then decreased to less than one-third in 42 to 56 DAA, then remained relatively low during the rest of fruit development. Seven to 14 days before fruit maturation, SS increased about 30% at the same time as sucrose accumulation in the fruit. Acid invertase activity was very low in the young fruit and increased more than 10-fold 42 to 14 days before maturation. SPS activity remained very low throughout the fruit development and was about 40% higher in mature-green fruit. The potential roles of invertase and sucrose synthetase in sugar accumulation will be discussed.
Papaya (Carica papaya L.) fruit flesh and seed growth, fruit respiration, sugar accumulation, and the activities of sucrose phosphate synthase (SPS), sucrose synthase (SS), and acid invertase (AI) were determined from anthesis for ≈150 days after anthesis (DAA), the full ripe stage. Sugar began to accumulate in the fruit flesh between 100 and 140 DAA, after seed maturation had occurred. SPS activity remained low throughout fruit development. The activity of SS was high 14 DAA and decreased to less than one-fourth within 56 DAA, then remained constant during the remainder of fruit development. AI activity was low in young fruit and began to increase 90 DAA and reached a peak more than 10-fold higher, 125 DAA, as sugar accumulated in the flesh. Results suggest that SS and AI are two major enzymes that may determine papaya fruit sink strength in the early and late fruit development phases, respectively. AI activity paralleled sugar accumulation and may be involved in phloem sugar unloading.
`Chandler' strawberries (Fragaria ananassa Duck.) were kept in air, 0.25% O2, 21% O2 + 50% CO2, or 0.25 O2 + 50% CO2 (balance N2) at 5C for 1 to 7 days to study the effects of controlled atmospheres (CAs) on volatiles and fermentation enzymes. Concentrations of acetaldehyde, ethanol, ethyl acetate, and ethyl butyrate were greatly increased, while concentrations of isopropyl acetate, propyl acetate, and butyl acetate were reduced by the three CA treatments compared to those of air-control fruit. The CA treatments enhanced activities of pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) but slightly decreased activity of alcohol acetyltransferase (AAT). The results indicate that the enhanced PDC and ADH activities by CA treatments cause ethanol accumulation, which in turn drives the biosynthesis of ethyl esters. The increased ethanol concentration also competes with other alcohols for carboxyl groups for esterification reactions. The reduced AAT activity and limited availability of carboxyl groups due to ethanol competition decrease production of other acetate esters.
`Hass' avocado (Persea americana Mill.) fruit were kept in air, 0.25% O2 (balance N2), 20 % O2 + 80% CO2, or 0.25% O2 + 80% CO2 (balance N2) at 20C for up to 3 days to study the regulation of fermentative metabolism. The 0.25% 02 and 0.25% 02 + 80% CO2 treatments caused accumulations of acetaldehyde and ethanol and increased NADH concentration, but decreased NAD level. The 20% O2 + 80% CO2 treatment slightly increased acetaldehyde and ethanol concentrations without significant effects on NADH and NAD levels. Lactate accumulated in avocadoes kept in 0.25 % 02. The 80% CO, (added to 0.25% O2) did not increase lactate concentration and negated the 0.25% O2-induced lactate accumulation. Activities of PDC and LDH were slightly enhanced and a new isozyme of ADH was induced by 0.25% O2, 20% O2 + 80% CO2, or 0.25 % O2 + 80% CO2; these treatments partly reduced the overall activity of the PDH complex. Fermentative metabolism can be regulated by changes in levels of PDC, ADH, LDH, and PDH enzymes and/or by metabolic control of the functions of these enzymes through changes in pH, ATP, pyruvate, acetaldehyde, NADH, or NAD. Chemical names used: alcohol dehydrogenase (ADH), adenosine triphosphate (ATP), lactate dehydrogenase (LDH), nicotinamide adenine dinucleotide (NAD), reduced NAD (NADH), pyruvate decarboxylase (PDC), pyruvate dehydrogenase (PDH).
Papaya (Carica papaya L.) source size and sink strength were modified by a single defoliation or continual defoliation and fruit thinning. Fruit set, development, weight, total sugar (sum of sucrose, fructose, and glucose), sucrose phosphate synthase (SPS), sucrose synthase (SS), and acid invertase (AI) enzyme activities in response to defoliation and fruit thinning were determined. The effects of defoliation and fruit thinning varied with weather conditions, plant growth conditions, and cultivar. Removal of 75% of the leaves significantly reduced new flower production and fruit set, and decreased ripe fruit total soluble solids (TSS), while 50% defoliation did not reduce new fruit set or ripe fruit TSS. When every third leaf from the oldest leaf was not removed, the number of new flowers was reduced by 47% more than when the same number of leaves was removed from the oldest to younger leaves. Continual removal of old leaves reduced new fruit set, fruit weight, and TSS in the 168 day experimental period. Fruit thinning increased new fruit set and ripe fruit TSS. Larger fruit size, faster fruit development, lower respiration rate, and higher sugar contents and AI activity were observed in immature (young) fruit when old fruit were removed. AI activity was reduced during early fruit development and increased again in mature fruit in plants subjected to defoliation, and suggested a role for AI in mature fruit sugar accumulation, while SS activity declined significantly in fruit 154 and 175 days after anthesis and in mature fruit when plants were subjected to continual defoliation. SPS activity was not affected significantly by defoliation or fruit thinning. Source-sink balance was critical for papaya fruit set, development, and sugar accumulation and each mature leaf was able to provide photoassimilate for about three fruit.
Amplified fragment length polymorphisms (AFLPs) were used to analyze the relationships between sweet cherry (Prunus avium L.) cultivars and selections from the breeding program at the Pacific Agri-Food Research Centre in Summerland, Canada. Six pairs of preselected primers were used for the analysis of a total of 67 cultivars and selections. Scoring the absence and presence of 118 polymorphic DNA fragments produced a unique binary code for each cultivar and selection. Two phylogenetic trees were constructed using these 118 polymorphic fragments, one tree for 55 related cultivars and selections from the Summerland breeding program and the other for 23 self-incompatible cultivars of differing origins. The reliability of AFLP DNA fingerprints was confirmed by correlating relationships revealed by AFLP profiles with known genetic relationships of some sweet cherry cultivars and by a blind test for cultivar identification. Results indicate that AFLP analysis is a good technique to evaluate genetic distance and relationships in a sweet cherry breeding population.
Papaya (Carica papaya L.) leaves are large, up to 70 cm wide, and frequently deeply lobed, with seven to 13 major veins. The scan width of current handheld digital leaf area instruments is generally less than 15 cm. A rapid method is needed to estimate the total leaf area of a plant in the field with leaves at different stages of growth from the apex. The length of the main and side veins of papaya leaves can be used to estimate the area of a single leaf and the total leaf area of the plant. The relationship between main vein lengths and total leaf area was determined for mature leaves from the cultivars Sunset, Line-8, and Kapoho. A simple relationship exists between leaf area and the length of the two main side midribs (L3 and L4): Leaf area (cm2) = −2280 + 87.7*L3 + 55.6*L4 (P > F = 0.0001; r 2 = 0.969), explaining ≈94% of the variation between estimated leaf area and leaf area. The most recently matured leaf is the third or fourth discernable leaf from the apex, with a positive net photosynthetic CO2 assimilation rate and an average area of 2331 cm2 that could fix up to 1.6 g carbon per 10-hour day under full sun. The rate of photosynthesis declined with leaf age, and the overall net photosynthetic CO2 assimilation rate of the plant can be predicted. Following 80% leaf defoliation of the plant, the net photosynthetic CO2 assimilation rate of the most recently matured leaf increased 30% to 50% on days 11 and 19 after treatment when the photosynthetic active radiation (PAR) was approximately half of that on day 15 under full sun when no difference in net photosynthetic CO2 assimilation rate was seen. Fruit removal did not affect the net photosynthetic CO2 assimilation rate. Papaya adjusts its single-leaf net photosynthetic CO2 assimilation rate under lower light levels to meet plant growth and fruit sink demand.
An invertase gene was isolated and its mRNA activity and protein levels were determined during papaya (Carica papaya L.) fruit development. A complete invertase cDNA (AF420223) and a partial sucrose synthase cDNA (AF420224) were isolated from papaya fruit cDNA libraries. The invertase cDNA encoded a predicted polypeptide of 582 residues (MW 65,537 Da), and was 68% and 45% identical with carrot apoplastic and vacuolar invertases, respectively. Key amino acids indicative of an apoplastic invertase were conserved. A full-length gene corresponding to the putative apoplastic invertase cDNA was isolated and was organized into seven exons and six introns. Exon 2 (9 bp long) encoded part of a highly conserved region (NDPNG/A). Invertase mRNA and activity levels increased during fruit maturation and sugar accumulation just before ripening. In contrast, sucrose synthase mRNA levels were high during early fruit growth and low during the fruit sugar accumulation stage. A 73-kDa cell wall extractable protein that cross-reacted with carrot apoplastic invertase antisera substantially increased during papaya fruit maturation and declined in full ripe fruit. The increase in invertase protein levels occurred 2 to 4 weeks before maturity and was markedly higher than the overall increase in enzyme activity at this stage. Subsequently, the increase in enzyme activity was higher than the increase in protein levels between 2 weeks before maturity and fully ripe. The results suggested that mRNA level and invertase activity were related to maturity. The data suggested that the invertase was apoplastic, and that post-translational control of enzyme activity occurred, in which a significant accumulation of invertase occurred before the peak of enzymes activity.