Effect of abscisic acid (ABA) on sugar accumulation in the flesh tissue of peach fruit at the start of the maturation stage

Experiments were conducted on¹⁴C-sorbitol, fructose, and glucose uptakeinto flesh discs, and sorbitol efflux from thediscs, with and without ABA application toexamine the effect of abscisic acid (ABA) onsugar accumulation in peach fruit flesh at thestart of the maturation stage in relation tomembrane transport. Total uptake of¹⁴C-sorbitol, fructose, and glucose intoflesh discs was effectively promoted by ABA ata concentration of 10^–5 M. PCMBS(p-chloromercuribenzensulfonicacid)-sensitive uptake, which was considered ascarrier-mediated uptake, of sorbitol into thediscs was clearly stimulated by ABA at10^–5 M, compared with glucose andfructose uptake. Sorbitol efflux from the discsacross the tonoplast was restricted by ABA at10^–5 M. ABA application todeveloping fruit increased sugar accumulationin the fruit. Estimated ABA concentration inthis fruit was approximately 10^–5 M. These results indicate that sugar accumulationin peach fruit flesh is stimulated by ABA at aconcentration of 10^–5 M both invitro and in vivo. ABA stimulatesuptake of sugars, especially sorbitol, into theflesh by enhancing carrier-mediated transportpossibly across both tonoplast and plasmamembrane.     

Glyoxylate metabolism and fatty acid oxidation in mango fruit during development and ripening

Throughout the development (maturation) of mango fruit the contents of citric and glyoxylic acids increased steadily. As the fruit matured the levels of isocitrate lyase, malate lyase and alanine: glyoxylate aminotransferase increased and reached maximum values prior to the time of harvesting. At and after harvest the levels of malate lyase and alanine : glyoxylate aminotransferase began to decrease but that of isocitrate lyase remained high until after the harvest when it decreased. The level of glyoxylate reductase was highest in the early developmental stage but declined as the fruit matured and ripened. As the fruit ripened, after harvest, the amounts of citric and glyoxylic acids decreased concomitant with a considerable increase in the levels of isocitrate dehydrogenase, malic dehydrogenase, malic enzyme and glyoxylate dehydrogenase.Fatty acid oxidizing capacity of mitochondria isolated from immature (developing) and postclimacteric fruit pulps was much less than that observed with mitochondria from preclimacteric and climacteric fruit. Glyoxylate stimulated the oxidation of caprylic, lauric, myristic and palmitic acids and inhibited the activity of isocitrate dehydrogenase in vitro.     

Thermal responses in the citrus fruit fly,Dacus tsuneonis: evidence for a pupal diapause

Thermal responses controlling pupariation and adult eclosion in a citrus fruit fly,Dacus tsuneonis (Miyake), were studied to understand the winter biology of this species. When mature larvae were exposed to various temperature conditions, the highest percentage of pupariation was obtained at 15 °C, although the variance at this temperature was greater than at 20 °C or 25 °C. Pupariation occurred most rapidly at 20 °C and an alternating temperature with a mean of 15 °C. At constant 15 °C, pupae failed to emerge as adults. Pupae were characterized by a reduced respiration rate, which is typical of a diapausing pupa. When insects were stored at different temperatures for 45 days after pupariation, and then transferred to 25 °C, adult eclosion occurred earlier when the initial temperature was 10 °C than when it was 5 °C or 15 °C. Adult eclosion occurred most synchronously and pupal mortality was lowest when insects were stored at 15 °C for 90 days before incubation at 25 °C. These results strongly suggest thatD. tsuneonis enters a pupal diapause.     

Properties of proton and sugar transport at the tonoplast of tomato (Lycopersicon esculentum) fruit

Tonoplast vesicles were isolated from tomato (Lycopersicon esculentum Mill.) fruit pericarp and purified on a discontinuous sucrose gradient. ATPase activity was inhibited by nitrate and bafilomycin A₁ but was insensitive to vanadate and azide. PPase hydrolytic activity was inhibited by NaF but was insensitive to nitrate, bafilomycin A₁ vanadate and azide. Kimetic studies of PPase activity gave an apparent K_m, for PP₃ of 18 μM. Identical distributions of bafilomycin- and NO₃-sensitive ATPase activities within continuous sucrose density gradients, confirmed that bafilomycin-sensitive ATPase activity is a suitable marker for the tonoplast. By comparing the distribution of bafilomycin-sensitive ATPase activity with that of PPase activity, it was possible to locate the PPase enzyme exclusively at the tonoplast. The apparent density of the tonoplast did not change during fruit development. Measurements of tonoplast PPase and ATPase activities during fruit development over a 35-day period revealed an 80% reduction in PPase specific activity and a small decrease in ATPase specific activity. ATP- and PP₁-dependent ΔpH generation was measured by the quenching of quinacrine fluorescence in tonoplast vesicles prepared on a discontinuous Dextran gradient. No H⁺ efflux was detected on the addition of sucrose to energized vesicles. Therefore a H⁺/sucrose antiport may not be the mechanism of sucrose uptake at the tomato fruit tonoplast. Similar results were obtained with glucose, fructose and sorbitol. The lack of ATP (or PP₁) stimulation of [¹⁴C]-sucrose uptake also suggested that an antiport was not involved. Initial uptake rates of radiolabelled glucose and fructose were almost double that for sucrose. The inhibition of hexose uptake by p-chloromercuribenzene sulphonate (PCMBS) implicated the involvement of a carrier. Therefore storage of hexose in the tomato fruit vacuole and maintenance of a downhill sucrose concentration gradient into sink cells is likely to be regulated by the activity of sucrose metabolizing enzymes, rather than by energy-requiring uptake mechanisms at the tonoplast.     

Proteins, non-structural carbohydrates and organic acids in the flowers of sweet orange (Citrus sinensis (L.) osbeck)

The concentration of soluble proteins in open flowers of sweet orange (Citrus sinensis (L.) Osbeck) was about 35% of the protein content found in green leaves, while ovary and stigma, with attached style, contained up to 85%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicates that flowers and leaves contained ribulose bisphosphate carboxylase-oxygenase (Rubisco). At least 12 polypeptides, ranging from 22 to 100 kDa, were more distinct in flowers than in leaves. The polypeptide band at about 100 kDa in the flower extract suggests the presence of phosphoenolpyruvate carboxylase (PEPCase) in citrus flowers. Activity of Rubisco and PEPCase, expressed in μmol g tissue fresh weight⁻¹ hr⁻¹, averaged 14.2 and 64.6 for intact open flowers, and 547.9 and 63.3 for the leaves, respectively. Starch and reducing sugars were 1.2- and 8.4-fold higher, respectively, and dark respiration rates were about five-fold greater for open flowers than fully expanded, sun leaves. Quinic and malic acids made up about 96% of the organic acids found in flowers and leaves. Ascorbic, citric, fumaric, and shikimic acids were in small or trace amounts. The large accumulation of starch and soluble sugars, in addition to the presence and activity of Rubisco and PEPCase, indicates that the flowers of sweet orange would have some capability to perform photosynthetic CO₂ assimilation, the metabolites of which might play some important role in flower development and fruit setting.     

Hydraulic connections of leaves and fruit to the parent plant in Capsicum frutescens (hot pepper) during fruit ripening

Background and Aims

The hydraulic architecture and water relations of fruits and leaves of Capsicum frutescens were measured before and during the fruiting phase in order to estimate the eventual impact of xylem cavitation and embolism on the hydraulic isolation of fruits and leaves before maturation/abscission.

Methods

Measurements were performed at three different growth stages: (1) actively growing plants with some flowers before anthesis (GS1), (2) plants with about 50 % fully expanded leaves and immature fruits (GS2) and (3) plants with mature fruits and senescing basal leaves (GS3). Leaf conductance to water vapour as well as leaf and fruit water potential were measured. Hydraulic measurements were made using both the high-pressure flow meter (HPFM) and the vacuum chamber (VC) technique.

Key Results

The hydraulic architecture of hot pepper plants during the fruiting phase was clearly addressed to favour water supply to growing fruits. Hydraulic measurements revealed that leaves of GS1 plants as well as leaves and fruit peduncles of GS2 plants were free from significant xylem embolism. Substantial increases in leaf petiole and fruit peduncle resistivity were recorded in GS3 plants irrespective of the hydraulic technique used. The higher fraction of resistivity measured using the VC technique compared with the HPFM technique was apparently due to conduit embolism.

Conclusions

The present study is the first to look at the hydraulics of leaves and fruits during growth and maturation through direct, simultaneous measurements of water status and xylem efficiency of both plant regions at different hours of the day.     

The occurrence of crassulacean acid metabolism in Cymbidium (Orchidaceae) and its ecological and evolutionary implications

Crassulacean acid metabolism (CAM) is one of the photosynthetic pathways regarded as adaptations to water stress in land plants. Little is known about correlations among the level of CAM activity, environment of habitat, life form, and phylogenetic relationship of a plant group from an evolutionary perspective. We examined these relationships in 18 species of Cymbidium (Orchidaceae) because the genus shows distinctive diversification of habitats and life forms. The photosynthetic type was classed into three categories, strong CAM, weak CAM, and C₃ on the basis of CAM activity. CAM expression in Cymbidium was confined to the epiphytic and lithophytic species. Especially, all of these species from tropical to subtropical rainforest exhibited CAM activity. On the other hand, the terrestrial species always exhibited C₃ metabolism irrespective of their varied habitats. Regarding the evolution of photosynthetic characters, weak CAM was the ancestral state in Cymbidium and strong CAM and C₃ metabolism occurred subsequently. The evolution of strong CAM likely enabled Cymbidium to extend to exposed sites in tropical lowland where marked water stress exists. Further, different levels of CAM activity characterized each species and such potential plasticity of CAM may realize the radiation of Cymbidium into sites with different environmental conditions.     

Oleic acid improves poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production by Ralstonia eutropha in inverted sugar and propionic acid

With the objective of verifying the influence of oleic acid as a nutritional supplement in the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Ralstonia eutropha, cultures were established with 0.3 g oleic acid l^–1 and without this supplement, in 30 g inverted sugar l^–1 and 1 g propionic acid l^–1. The use of this supplement increased the accumulation of polymer from 18.3% to 28.3% (w/w) although the mass of 3-hydroxyvalerate in the polymer remained constant for both cultures.     

Extracellular invertase of Rhizobium japonicum and its role in free sugar metabolism in the developing root nodules of Sesbania grandiflora

Extracellular invertase of Rhizobium japonicum and its role in free sugar metabolism in the developing root nodules of Sesbania grandiflora L. was studied. The enzyme hydrolysed sucrose extracellularly, and its release was substrate inducible. 0.1 Mβ-mercaptoethanol released the cell-bound form of this enzyme. The production of invertase was low when glucose, galactose, mannose, fructose and raffinose were used as carbon sources in the growth medium. In the developing nodules sucrose was the major sugar. The content of fructose was low in comparison with that of glucose – suggesting that in the nodules, fructose is converted to glucose prior to its entry into the bacterial cell. The content of glucose synchronised with the pattern of change in the activity of invertase in the nodules.     

Methyl de-esterification as a major factor regulating the extent of pectin depolymerization during fruit ripening: a comparison of the action of avocado (Persea americana) and tomato (Lycopersicon esculentum) polygalacturonases

Pectinmethylesterase (PME, EC 3.2.1.11) and polygalacturonase (PG, EC 3.2.1.15) are known to operate in tandem to degrade methylesterified polyuronides. In this study, PGs purified from tomato and avocado fruit were compared in terms of their capacity to hydrolyze water-soluble polyuronides from avocado before and following enzymic or chemical de-esterification. When assayed using polygalacturonic acid or polyuronides from avocado fruit, the activity of PG from tomato fruit was 3-4 times higher than that from avocado fruit. High molecular mass, low methylesterified (33%) water-soluble polyuronides (WSP) from pre-ripe avocado fruit (day 0) were partially depolymerized upon incubation with purified avocado and tomato PGs. In contrast, middle molecular mass, highly methylesterified (74%) WSP from day 2 fruit were largely resistant to the action of both PGs. PME or weak alkali treatment of highly methylesterified WSP decreased the methylesterification values to 11 and 4.5%, respectively. Treatment of de-esterified WSP with either avocado or tomato PGs caused extensive molecular mass downshifts, paralleling those observed during avocado fruit ripening. Although PME and PG are found in many fruits, the pattern of depolymerization of native polyuronides indicates that the degree of cooperativity between these enzymes in vivo differs dramatically among fruits. The contribution of PME to patterns of polyuronide depolymerization observed during ripening compared with physically compromised fruit tissues is discussed.     

Effect of ethrel treatment on the postharvest changes in the turnover of ascorbic acid and reducing sugar inAchras sapota fruits

There was a definite relationship between growth and ascorbic acid content inAchras sapota. Increase in ethrel concentration from 250 ppm to 500 ppm hastened early ripening and increased the amount of reducing sugars but depleted the ascorbic acid content. Other aspects of ascorbic acid turnover viz. ascorbigen, bound form of ascorbic acid, ascorbic acid utilization, net ascorbic acid bound and ascorbic acid oxidase were also studied.     

Depolymerisation of mucilage isolated from ruredzo (Dicerocaryum zanguebarium) by ascorbic acid in the presence of catalysts

No change was observed in the viscosity of samples of mucilage from Dicerocaryum zanguebarium when treated with 5 mM ascorbic acid. Mucilage was treated with ascorbic acid in the presence of copper and iron, and a mixture of the two ions. The effect of treatment was followed by gel filtration of products on a Sepharose 6B column and by measuring the amount of reducing sugars generated by the dinitrosalicylic acid (DNSA) method. The molecular weight of treated samples was lower than that of untreated controls. The amount of reducing groups was found to increase. When the mucilage was treated in a system containing hydrogen peroxide and copper ions, the polymer was also degraded, yielding fragments of lower molecular weight as reflected by the broadened gel filtration peak.