Insoluble Invertase Research Articles

Performance of Hevea brasiliensis under drought conditions on osmoregulation and antioxidant activity through evaluation of vacuolar invertase and reducing sugars

Rubber tree cultivation is limited in many regions by abiotic factors such as drought. We investigated the biochemical mechanisms responsible for responses to, and recovery from, drought conditions during the establishment phase of four high latex producing rubber tree clones (RRIM600, IAC40, PR255 and GT1). Five-month-old plants were exposed to 32 days of water restriction, followed by 15 days of soil rehydration. Leaf area, as well as their osmolyte accumulations, saccharolytic enzyme activity, and oxidative stress markers, were accompanied. Although clones IAC40 and PR255 responded more precociously to drought conditions, halting leaf expansion before clones GT1 and RRIM600, they demonstrated slow recuperation after reestablishing irrigation. The greater tolerances of clones RRIM600 and GT1 to drought conditions were related to greater vacuolar invertase (VINV) activity in their leaves, which guaranteed more significant accumulations of vacuolar reducing sugars (RS). Similar to RS, glycine betaine accumulations were related to osmoprotection and to reducing oxidative damage (lipidic peroxidation) caused by water deficit conditions. The observed decreases in cytosol neutral invertase (AINV) and cell wall insoluble invertase (CWINV) activities, which resulted in cytosol hexose decreases, may be related to increases in antioxidant enzyme (superoxide dismutase and ascorbate peroxidase) activities in the leaves in response to water deficit conditions. As such, the introduction of specific sugars (RS) and the modulation of key carbon metabolism enzymes, such as VINV, are promising strategies for promoting drought tolerance in rubber tree clones.

The cytosolic invertase NI6 affects vegetative growth, flowering, fruit set, and yield in tomato.

Sucrose metabolism is important for most plants, both as the main source of carbon and via signaling mechanisms that have been proposed for this molecule. A cleaving enzyme, invertase (INV) channels sucrose into sink metabolism. Although acid soluble and insoluble invertases have been largely investigated, studies on the role of neutral invertases (A/N-INV) have lagged behind. Here, we identified a tomato A/N-INV encoding gene (NI6) co-localizing with a previously reported quantitative trait locus (QTL) largely affecting primary carbon metabolism in tomato. Of the eight A/N-INV genes identified in the tomato genome, NI6 mRNA is present in all organs, but its expression was higher in sink tissues (mainly roots and fruits). A NI6-GFP fusion protein localized to the cytosol of mesophyll cells. Tomato NI6-silenced plants showed impaired growth phenotype, delayed flowering and a dramatic reduction in fruit set. Global gene expression and metabolite profile analyses of these plants revealed that NI6 is not only essential for sugar metabolism, but also plays a signaling role in stress adaptation. We also identified major hubs, whose expression patterns were greatly affected by NI6 silencing; these hubs were within the signaling cascade that coordinates carbohydrate metabolism with growth and development in tomato.

Low source-sink ratio reduces reserve starch in grapevine woody canes and modulates sugar transport and metabolism at transcriptional and enzyme activity levels.

Severe leaf removal decreases storage starch and sucrose in grapevine cv. Cabernet Sauvignon fruiting cuttings and modulates the activity of key enzymes and the expression of sugar transporter genes. Leaf removal is an agricultural practice that has been shown to modify vineyard efficiency and grape and wine composition. In this study, we took advantage of the ability to precisely control the number of leaves to fruits in Cabernet Sauvignon fruiting cuttings to study the effect of source-sink ratios (2 (2L), 6 (6L) and 12 (12) leaves per cluster) on starch metabolism and accumulation. Starch concentration was significantly higher in canes from 6L (42.13±1.44mggDW-1) and 12L (43.50±2.85mggDW-1) than in 2L (22.72±3.10mggDW-1) plants. Moreover, carbon limitation promoted a transcriptional adjustment of genes involved in starch metabolism in grapevine woody tissues, including a decrease in the expression of the plastidic glucose-6-phosphate translocator, VvGPT1. Contrarily, the transcript levels of the gene coding the catalytic subunit VvAGPB1 of the VvAGPase complex were higher in canes from 2L plants than in 6L and 12L, which positively correlated with the biochemical activity of this enzyme. Sucrose concentration increased in canes from 2L to 6L and 12L plants, and the amount of total phenolics followed the same trend. Expression studies showed that VvSusy transcripts decreased in canes from 2L to 6L and 12L plants, which correlated with the biochemical activity of insoluble invertase, while the expression of the sugar transporters VvSUC11 and VvSUC12, together with VvSPS1, which codes an enzyme involved in sucrose synthesis, increased. Thus, sucrose seems to control starch accumulation through the adjustment of the cane sink strength.

완두콩(Pisum sativum L.) 상처에서 유도되는 불용성 산성 인버타제의 면역조직화

Invertase, that hydrolyzes sucrose into glucose and fructose, plays a great role in carbohydrate reallocation between the photosynthetic source tissue and various sink tissues. Invertase also occurs in a variety of isoforms for various functions in plants. Insoluble invertases were extracted only in buffer solutions containing high concentrations of salt. Within these classes, acid invertase has an optimum activity at acidic pH (pH 4-5). Induction of insoluble acid invertase (INAC-INV) in leaf, stem, and root tissues in response to physical wounding has been investigated. To detect the localization of INAC-INV within the plant, immunolocalization has been performed. In this study, the accumulation of INAC-INV was noticeable to reach maximum levels on 72 hr after mechanical injuries. INAC-INV was induced in wounded leaves 3 times more than control leaves. Immunolocalization results showed that INAC-INV accumulated in wall appositions and intercellular spaces. INAC-INV was also localized at sieve cell walls in phloem tissues close to the site of wounding. Taken together, this study suggested that INAC-INV induction upon wounding injuries can play a role on responses to the high energy demand for wound healing process.

GA 처리 후 급 성장하는 완두콩(Pisum sativum L.) 발아체로부터 분리된 중성 invertase의 특성

Invertase (β-D-fructosfuranosidase, EC 3.2.1.26) catalyzes the hydrolysis of sucrose into D-glucose and D-fructose. Three biochemical subgroups of invertases have been investigated in plants: vacuolar (soluble acid), cytoplasmic (soluble alkaline), and cell wall-bound (insoluble acid) invertases. An isoform of neutral invertase was purified from pea seedlings (Pisum sativum L.) and treated with gibberellic acid (GA) by sequential procedures consisting of ammonium sulfate precipitation, ion-exchange chromatography, absorption chromatography, and reactive green-19 affinity chromatography. The results of the overall insoluble invertase purification were a 430-fold increase. The purified neutral invertase was not glycosylated and had an optimum pH between neutral and alkaline (pH 6.8-7.5). It was inhibited by Tris, as well as by heavy metals, such as Hg 2+ and Cu 2+ . Typical Michaelis?Menten kinetics were observed when the activity of the purified invertase was measured, with sucrose concentrations up to 100 mM. The K m and V max values were 12.95 mM and 2.98 U/min, respectively. The molecular mass was around 20 kDa. The sucrose-cleaving enzyme activity of this enzyme is similar to that of sucrose synthase and fructosyltransferase, but its biochemical characteristics are different from those of sucrose synthase and fructosyltransferase. Based on this biochemical characterization and existing knowledge, neutral INV is an invertase isoform in plants.

Inverted sugar syrup attained from sucrose hydrolysis using a membrane reactor

Invertase, whether adsorbed on styrene-divinylbenzene copolymers or otherwise, was used for continuous sucrose hydrolysis using a cell-type membrane reactor (CTMR), coupled with an ultra (UF-100kDa), or a microfiltration (MF- pore diameter of 5 µm) membrane. In all tests, the pH (5.5), temperature (30 ºC), reaction volume (10 mL) and agitation (100 rpm) were set constant; whereas, variable parameters were: feeding rate (0.4, 0.8 and 1.6 h-1), inlet sucrose concentration (2.5, 6.5, 50 and 100 mM) and enzyme/resin ratio (1.64 mg or 3.28 mg of protein per 25, 50 or 100 mg of resin). The best result (yield of 100%, steady-state duration over 20h and specific reaction rate over 243 x 10-3 mmol/h.mE) was obtained when insoluble invertase (1.64 mg protein/100 mg resin) was used to convert 50 mM or 100 mM of sucrose solution at 0.4 h-1 using a UF-CTMR.

Functional Analysis of an Arabidopsis thaliana Abiotic Stress-inducible Facilitated Diffusion Transporter for Monosaccharides

Sugars play indispensable roles in biological reactions and are distributed into various tissues or organelles via transporters in plants. Under abiotic stress conditions, plants accumulate sugars as a means to increase stress tolerance. Here, we report an abiotic stress-inducible transporter for monosaccharides from Arabidopsis thaliana that is termed ESL1 (ERD six-like 1). Expression of ESL1 was induced under drought and high salinity conditions and with exogenous application of abscisic acid. Promoter analyses using beta-glucuronidase and green fluorescent protein reporters revealed that ESL1 is mainly expressed in pericycle and xylem parenchyma cells. The fluorescence of ESL1-green fluorescent protein-fused protein was detected at tonoplast in transgenic Arabidopsis plants and tobacco BY-2 cells. Furthermore, alanine-scanning mutagenesis revealed that an N-terminal LXXXLL motif in ESL1 was essential for its localization at the tonoplast. Transgenic BY-2 cells expressing mutated ESL1, which was localized at the plasma membrane, showed an uptake ability for monosaccharides. Moreover, the value of K(m) for glucose uptake activity of mutated ESL1 in the transgenic BY-2 cells was extraordinarily high, and the transport activity was independent from a proton gradient. These results indicate that ESL1 is a low affinity facilitated diffusion transporter. Finally, we detected that vacuolar invertase activity was increased under abiotic stress conditions, and the expression patterns of vacuolar invertase genes were similar to that of ESL1. Under abiotic stress conditions, ESL1 might function coordinately with the vacuolar invertase to regulate osmotic pressure by affecting the accumulation of sugar in plant cells.

Sucrose accumulation in watermelon fruits: Genetic variation and biochemical analysis

Sugar accumulation, the key process determining fruit quality, is controlled by both the translocation of sugars and their metabolism in developing fruits. Sugar composition in watermelon, as in all cucurbit fruits, includes sucrose, fructose and glucose. The proportions of these three sugars are determined primarily by three enzyme families: invertases, sucrose synthases (SuSys) and sucrose phosphate synthases (SPSs). The goal of the present research was to explore the process of sugar metabolism in watermelon fruits. Crosses between the domestic watermelon ( Citrullus lanatus) and three wild species provided a wide germplasm to explore genetic variability in sugar composition and metabolism. This survey demonstrated great genetic variability in sugar content and in the proportions of sucrose, glucose and fructose in mature fruits. Genotypes accumulating high and low percentage of sucrose provided an experimental system to study sugar metabolism in developing fruits. Insoluble invertase activity was high and constant throughout fruit development in control lines and in genotypes accumulating low levels of sucrose, while in genotypes accumulating high levels of sucrose, activity declined sharply 4 weeks after pollination. Soluble acid invertase activity was significantly lower in genotypes accumulating high levels of sucrose than in low-sucrose-accumulating genotypes. Conversely, activities of SuSy and SPS were higher in the high-sucrose-accumulating genotypes. The present results establish that, within the genus Citrullus, there are genotypes that accumulate a high percentage of sucrose in the fruit, while others accumulate high percentages of glucose and fructose. The significant negative correlation between insoluble invertase activity and fruit sucrose level suggests that sucrose accumulation is affected by both phloem unloading and sugar metabolism.

Glucose localization in maize ovaries when kernel number decreases at low water potential and sucrose is fed to the stems.

Around the time of anthesis, young ovary development in maize (Zea mays) is vulnerable to 2 or 3 d of water deficits that inhibit photosynthesis. Abortion can result, and fewer kernels are produced. A breakdown of stored ovary starch is associated with the abortion and was investigated in the present study by localizing the breakdown product glucose in the ovaries. Ovary glucose was localized with fluorescent Resorufin. Insoluble invertase was localized in vivo and soluble invertase in situ. Sucrose was infused into the stems to vary the sugar flux to the ovaries. At high water potential (high Psi(w)), photosynthesis was rapid in the parent. The upper pedicel of the ovaries had a high activity of insoluble acid invertase and a large amount of glucose and starch. Because the invertase was wall-bound, sucrose hydrolysis appeared to occur in the pedicel apoplast. Soluble invertase was undetected inside the pedicel cells but was present in the nucellus cells where low concentrations of glucose occurred. This created a glucose gradient between pedicel and nucellus that favoured glucose uptake by the developing ovary. At low Psi(w), photosynthesis was inhibited, pedicel glucose and starch were depleted, the glucose gradient became negligible, and abortion occurred. When sucrose was fed, glucose, starch and the glucose gradient were maintained somewhat and were normally distributed in the ovaries. Abortion was diminished. The apoplast hydrolysis of sucrose unloaded from phloem is similar to that described by others during later development when embryo and endosperm are present and separated from the parent by an apoplast. The disappearance of the glucose gradient at low Psi(w) may have inhibited glucose movement into the ovary. The low glucose in the ovaries may have a role in the abortion response.

Activities of enzymes involved in sucrose-to-starch metabolism in rice grains subjected to water stress during filling

Slow grain filling of rice ( Oryza sativa L.) is a problem due to the heavy use of nitrogen fertilizer or the high lodging resistance of some cultivars. This study investigated if controlled water deficit during grain filling could enhance sink strength by regulating key enzymes involved and lead to faster grain filling. Two rice cultivars with high lodging resistance and slow grain filling were grown in the field and treated with either normal nitrogen (NN) or high nitrogen (HN) at heading. Well-watered (WW) and water-deficit stressed (WS) treatments were imposed from 9 days after anthesis (DAA) until maturity. Leaf water potentials of both cultivars markedly decreased during the day as a result of WS treatments, but completely recovered by early morning. WS promoted the reallocation of prefixed 14 C from stems to grains, facilitated starch accumulation in grains and increased grain-filling rate although it shortened grain-filling period. In contrast, HN behaved in the opposite way. Sucrose synthase (EC 2.4.1.13) activity was substantially enhanced by water stress, and was positively correlated with starch accumulation rate (SAR) in the grains. Both soluble and insoluble invertase (EC 3.2.1.26) activities were less enhanced by WS and showed no significant correlation with SAR. Starch branching enzyme (BE) (EC 2.4.1.18) and soluble starch synthase (EC 2.4.1.18) activities were also enhanced by the WS, with the former enhanced more than the latter, and were significantly correlated with SAR. Adenine diphosphoglucose pyrophosphorylase (EC 2.7.7.27) activity was little affected by WS. The results suggest that WS-increased remobilization and grain-filling rate were attributed to enhanced sink strength by regulating sucrose synthase and starch BE activities in rice grains when subjected to water stress during the grain-filling period.

Soluble invertase expression is an early target of drought stress during the critical, abortion-sensitive phase of young ovary development in maize.

To distinguish their roles in early kernel development and stress, expression of soluble (Ivr2) and insoluble (Incw2) acid invertases was analyzed in young ovaries of maize (Zea mays) from 6 d before (-6 d) to 7 d after pollination (+7 d) and in response to perturbation by drought stress treatments. The Ivr2 soluble invertase mRNA was more abundant than the Incw2 mRNA throughout pre- and early post-pollination development (peaking at +3 d). In contrast, Incw2 mRNAs increased only after pollination. Drought repression of the Ivr2 soluble invertase also preceded changes in Incw2, with soluble activity responding before pollination (-4 d). Distinct profiles of Ivr2 and Incw2 mRNAs correlated with respective enzyme activities and indicated separate roles for these invertases during ovary development and stress. In addition, the drought-induced decrease and developmental changes of ovary hexose to sucrose ratio correlated with activity of soluble but not insoluble invertase. Ovary abscisic acid levels were increased by severe drought only at -6 d and did not appear to directly affect Ivr2 expression. In situ analysis showed localized activity and Ivr2 mRNA for soluble invertase at sites of phloem-unloading and expanding maternal tissues (greatest in terminal vascular zones and nearby cells of pericarp, pedicel, and basal nucellus). This early pattern of maternal invertase localization is clearly distinct from the well-characterized association of insoluble invertase with the basal endosperm later in development. This localization, the shifts in endogenous hexose to sucrose environment, and the distinct timing of soluble and insoluble invertase expression during development and stress collectively indicate a key role and critical sensitivity of the Ivr2 soluble invertase gene during the early, abortion-susceptible phase of development.

The Decreased Cold-Resistance of Chilling-Sensitive Plants Is Related to Suppressed CO2 Assimilation in Leaves and Sugar Accumulation in Roots

Tomato (Lycopersicon esculentum L., cv. Sibirskii skorospelyi) and cucumber (Cucumis sativus L., cv. Konkurent) plants were grown in a soil culture in a greenhouse at an average daily temperature of 20°C and ambient illumination until the development of five and eight true leaves, respectively. During the subsequent three days, some plants were kept in a climatic chamber at 6°C in the light, whereas other plants remained in a greenhouse (control). The cold-resistance of cucumber leaves and roots, as assayed from the electrolyte leakage, was reduced after cold exposure stronger than cold-resistance of tomato organs. The ratio photosynthesis/dark respiration was lower in cucumber than in tomato leaves at all measurement temperatures. The concentrations of sugars (sucrose + glucose + fructose) increased in chilled tomato roots but decreased in cucumber roots. Cold exposure changed the activities of various invertase forms (soluble and insoluble acidic and alkaline invertases). The total invertase activity and the ratio of mono- to disaccharides increased. The lower cucumber cold-resistance is related to the higher sensitivity of its photosynthetic apparatus to chilling and, as a consequence, insufficient root supply with sugars.

Sucrose Metabolism in Leaves and Roots of Bean ( Phaseolus vulgaris L.) during Phosphate Deficiency

Summary The influence of phosphate deficiency on the accumulation and metabolism of sugars in the source and sink leaves and roots of bean ( Phaseolus vulgaris L.) was studied after 16 days of plant culture. Glucose, sucrose and starch contents increased in all tissues of phosphate-deficient plants as compared with control plants. Phosphate deficiency increased activities of enzymes involved in sucrose synthesis in the leaves and root; sucrose phosphate synthase (EC 2.4.1.14) activity increased about twice, while sucrose synthase (EC 2.4.1.13) activity increased about 3-fold in the mature leaves and 50% and 90% in the young leaves and roots, respectively. Phosphate deficiency also increased the activities of enzymes hydrolyzing sucrose in the leaves and roots; neutral invertase (EC 3.2.1.26) activity increased twice in the source leaves and about 50% in sink leaves and roots. Acid invertases (EC 3.2.1.26) were divided into soluble and insoluble forms; insoluble invertases activities were several times lower than those of soluble invertases, both in the leaves and roots. Pi starvation increased mainly the activities of insoluble forms of acid invertases in all tissues in bean plants.

Characterization of Invertase Entrapped into Calcium Alginate Beads

A solution of 10 g/L of sodium alginate (Satialgine types used [Sanofi trademark]: SG800 and S1100 with manuronic/guluronic ratio of 0.5 and 1.2, respectively) containing invertase (0.08 g of protein/L) was dropped into 0.1 M CaCl2 solution buffered at pH 4.0, 7.0, or 8.0. The beads were left to harden in CaCl2 solution for 24 h. The high immobilization yield of 60% occurred with SG800 at pH 8.0. The activity of soluble and insoluble invertase was measured against pH (2.8-8.0), sucrose concentration (4.5-45 mM), and temperature (30-60 degrees C). Both forms presented an optimum pH of 4.6. However, the soluble invertase was stable at the overall pH interval studied, whereas insoluble invertase lost 30% of its original activity at pH > 5.0. At temperatures above 40 degrees C, the insoluble form was more stable than the soluble one. The kinetic constants and activation energies (Ea) for free invertase were KM = 41.2 mM, Vmax = 0.10 mg of TRS/(min.mL), and Ea 28 kJ/mol for entrapped invertase they were (KM)ap = 7.2 mM, (Vmax)ap = 0.060 mg of TRS/(min.mL), and (Ea)ap = 24 kJ/mol.

Properties of invertases in mycelium of Hymenoscyphus ericae and in endomycorrhizal association with cranberry seedlings

The properties of extracellular, soluble and insoluble invertases in mycelium of Hymenoscyphus ericae an ericoid mycorrhizal fungus, were investigated. Soluble and insoluble acid invertases were more active in sucrose- compared with glucose-grown mycelium, whereas extracellular invertase activities were similar in both glucose- and sucrose-grown cultures. Acid (pH 5·5) and alkaline (pH 9·0) invertases were detected in soluble and insoluble fractions but extracellular activity was predominantly acid (pH 5·5). Insoluble alkaline invertase was released completely by freeze-thawing. Freeze-thawing and Triton X-100 released similar proportions of acid invertase from the wall (42 and 31% respectively). The K m values varied from 9·1 m m (extracellular acid invertase) to 139·3 m m sucrose (soluble alkaline invertase). Fructose inhibited soluble alkaline invertase completely but it competitively inhibited soluble acid invertase. Increased soluble acid invertase activity was demonstrated in endomycorrhizas compared with uninoculated roots of cranberry.

Heat stress effects on sink activity of developing maize kernels grown in vitro

The objective of this study was to determine the effect of short‐term (4 days) and long‐term (8 days) heat stress (35°C) on sink activity of maize (Zea mays L.) kernels. Beginning at 3 days after pollination (DAP) kernels were grown in vitro at 25°C and 24 h later were transferred to 35°C for either 4 or 8 days. Each treatment had a control that was maintained continously at 25°C. Two experiments were designed to examine the uptake and distribution of 14C among hexoses, sucrose and starch in the pedicel placento‐chalazal (pedicel/p‐c). endosperm, and pericarp tissues of kernels exposed to heat stress for 4 or 8 days. Kernels cultured in vitro were placed in 14C‐sucrose medium either during the period of heat stress (experiment 1; 5 to 13 DAP) or immediately following heat‐stress treatments (experiment 2; 10 to 22 DAP). In both experiments no significant effect of heat stress was observed on the total radioactivity accumulated in the kernels until about 17 DAP, after which heat‐stressed kernels accumulated less 14C than the control. During the linear fill period, the endosperm of kernels exposed to heat stress accumulated more radioactivity associated with hexoses and sucrose and less radioactivity incorporated into starch, as compared to the control. Kernels heat stressed for 4 days showed a partial recovery in starch synthesis by 21 DAP, but to levels of only 65% of that of the control. Kernels heat stressed for 8 days did not recover. When 14C‐sucrose was supplied during the heat stress period (5–13 DAP). kernels from all treatments accumulated more hexoses that sucrose in the pedicel/p‐c. However, during the period following heat stress (10–22 DAP), pedicel/p‐c accumulated sucrose, but only in kernels exposed to long‐term heat stress. Soluble invertase activity was inhibited by both short‐term and long‐term heat stress, whereas the activity of insoluble invertase was affected only by long‐term heat stress. These results support the hypothesis that the disruption of kernel growth and more particularly endosperm starch biosynthesis, in response to heat stress, is mainly associated with changes in carbon utilization and partitioning between the different nonstructural carbohydrates within the endosperm rather than with a limitation in carbon supply to the kernel. Therefore, the effect on sink activity does not seem to be attributable to a thermal disruption of kernel uptake of sugars, but rather it is a consequence of heat perturbation of other physiological processes such as endosperm sugar metabolism and starch biosynthesis.

Soluble and insoluble invertase activity in elongating Tulipa gesneriana flower stalks

Previously ‘frozen’Tulipa gesneriana L. bulbs cv. Apeldoorn, were planted and grown at higher temperatures to study the role of invertase (EC 3.2.1.26) in the cold‐induced elongation of the flower stalk internodes. After planting, flower stalks were left intact, or, the leaves and flower bud were both removed to inhibit internode elongation. In intact flower stalks, elongation of the internodes was accompanied by an accumulation of glucose and an initial decrease in the sucrose content g,−1 dry weight. Insoluble invertase activity g,−1 dry weight hardly changed, but soluble invertase activity showed a peak pattern, that was related, at least for the greater part, to the changes in the sugar contents. Peak activities of soluble invertase were found during (lower‐ and uppermost internodes) or around the onset of the rapid phase of internode elongation (middle internodes). Internode elongation and glucose accumulation immediately ceased when the leaves and flower bud were removed. Insoluble invertase activity g,−1 dry weight remained at its initial level (lowermost internode) or increased more towards the upper internodes. Soluble invertase activity did not further increase (uppermost internode) or decreased abruptly to a low level. It is concluded that soluble invertase may be one of the factors contributing to glucose accumulation and internode elongation in the tulip flower stalk.

CARBOHYDRATE METABOLISM IN GA3-TREATED AND POLLINATED RABBITEYE BLUEBERRY FRUITS.

Carbohydrate metabolizing enzymes and carbohydrate accumulation were examined in pollinated, non-pollinated and GA3-treated blueberry fruit. Our hypothesis was that GA3 induces fruit set in blueberry by increasing the carbohydrate metabolizing enzyme activity in the ovary with a subsequent increase in carbohydrate mobilization. FW of non-pollinated fruits increased little between 0-45 days, when abscission occurred. FW of GA3-treated and pollinated fruits increased from 20 mg at anthesis to 1 and 2 g, respectively, at ripening. Insoluble invertase activity decreased from 15 μmol gFW-1 h-1 at anthesis to 3 μmol gFW-1 h-1 at 45 DAA for GA3-treated and pollinated fruits, and 9 μmol gFW-1 h-1 in non-pollinated fruits. Soluble invertase was undetectable in any treatment from 0 to 45 days, but rose sharply, averaging 57 μmol gFW-1 h-1 at ripening for both GA3-treated and pollinated fruits. No consistent correlation was found between ovary carbohydrate concentration and fruit growth among treatments. Differences in fruit set and size among treatments were not due to differences in enzyme activity and subsequent differences in carbohydrate accumulation.

Enzyme Activities of Starch and Sucrose Pathways and Growth of Apical and Basal Maize Kernels

Apical kernels of maize (Zea mays L.) ears have smaller size and lower growth rates than basal kernels. To improve our understanding of this difference, the developmental patterns of starch-synthesis-pathway enzyme activities and accumulation of sugars and starch was determined in apical- and basal-kernel endosperm of greenhouse-grown maize (cultivar Cornell 175) plants. Plants were synchronously pollinated, kernels were sampled from apical and basal ear positions throughout kernel development, and enzyme activities were measured in crude preparations. Several factors were correlated with the higher dry matter accumulation rate and larger mature kernel size of basal-kernel endosperm. During the period of cell expansion (7 to 19 days after pollination), the activity of insoluble (acid) invertase and sucose concentration in endosperm of basal kernels exceeded that in apical kernels. Soluble (alkaline) invertase was also high during this stage but was the same in endosperm of basal and apical kernels, while glucose concentration was higher in apical-kernel endosperm. During the period of maximal starch synthesis, the activities of sucrose synthase, ADP-Glc-pyrophosphorylase, and insoluble (granule-bound) ADP-Glc-starch synthase were higher in endosperm of basal than apical kernels. Soluble ADP-Glc-starch synthase, which was maximal during the early stage before starch accumulated, was the same in endosperm from apical and basal kernels. It appeared that differences in metabolic potential between apical and basal kernels were established at an early stage in kernel development.

An Association Between Acid Invertase Activity and Cell Growth during Leaf Expansion inPhaseolus vulgarisL.

Changes in lamina area, dimensions of epidermal and palisade cells, acid invertase activity and content of sucrose and hexose in the primary leaves of Phaseolus vulgaris L. were determined between emergence of the hypocotyl hook and the completion of leaf expansion. Growth in area and thickness of the primary leaf after emergence was attributable to the expansion of cells already present in the lamina at emergence. The major invertase in the expanding leaf was a readily soluble acid invertase; little insoluble invertase activity was detected. Soluble and insoluble fractions of leaf homogenates contained little neutral invertase activity. The specific activity of the soluble acid invertase increased rapidly during the early stages of leaf expansion, reaching a peak at the time of most rapid cell enlargement (5 d after emergence) and then declining as the leaf matured. Highly significant positive correlations were found between enzyme specific activity and the rates of cell and leaf enlargement. The early, rapid phase of lamina expansion was characterized by high concentrations of hexose sugar and low concentrations of sucrose. As the rates of leaf cell enlargement declined the concentration of hexose fell and that of sucrose increased. Between 5 d and 11 d after hypocotyl emergence, the hexose/sucrose ratio in the primary leaf decreased approximately 10-fold as the specific activity of acid invertase decreased. The results are discussed with reference to sources of carbon substrates for cell growth and to the sink/source transition during leaf development.