Insoluble Acid Invertase Research Articles

Identification and expression analysis of invertase family genes during grape (Vitis vinifera L.) berry development under CPPU and GA treatment.

Sugar is crucial for grape berry, whether used for fresh food or wine. However, berry enlargement treatment with forchlorfenuron (N-(2-chloro4-pyridyl)-N'-phenylurea) (CPPU, a synthetic cytokinin) and gibberellin (GA) always had adverse effects on sugar accumulation in some grape varieties, especially CPPU. Therefore exploring the molecular mechanisms behind these adverse effects could provide a foundation for improving or developing technology to mitigate the effects of CPPU/GA treatments for grape growers. In the present study, invertase (INV) family, the key gene controlling sugar accumulation, was identified and characterized on the latest annotated grape genome. Their express pattern, as well as invertase activity and sugar content, were analyzed during grape berry development under CPPU and GA3 treatment to explore the potential role of INV members under berry enlargement treatment in grapes. Eighteen INV genes were identified and divided into two sub-families: 10 neutral INV genes (Vv-A/N-INV1-10) and 8 acid INV genes containing 5 CWINV (VvCWINV1-5) and 3 VIN (VvVIN1-3). At the early development stage, both CPPU and GA3 treatment decreased the hexose level in berries of 'Pinot Noir' grape, whereas the activity of three types inverstase (soluble acid INV, insoluble acid INV, and neutral INV) increased. Correspondingly, most of INV members were up-regulated by GA3 /CPPU application at least one sampling time point during early berry development, including VvCWINV1, 2, 3, 4, 5, VvVIN1, 2, 3 and Vv-A/N-INV1, 2, 5, 6, 7, 8, 10. At maturity, the sugar content in CPPU-treated berries is still lower than that in the control. Soluble acid INV and neutral INV, rather than insoluble acid INV, presented lower activity in CPPU-treated berries. Meanwhile, several corresponding genes, such as VvVIN2 and Vv-A/N-INV2, 8, 10 in ripening berries were obviously down-regulated by CPPU treatment. These results suggested that most of INV members could be triggered by berry enlargement treatment during early berry development, whereas VvVINs and Vv-A/N-INVs, but not VvCWINVs, could be the limiting factor resulting in decreased sugar accumulation in CPPU-treated berries at maturity. In conclusion, this study identified the INV family on the latest annotated grape genome and selected several potential members involving in the limit of CPPU on final sugar accumulation in grape berry. These results provide candidate genes for further study of the molecular regulation of CPPU and GA on sugar accumulation in grape.

The effects of arbuscular mycorrhizal fungi and deficit irrigation on the yield and sugar content of watermelons (Citrullus lanatus)

Many studies have demonstrated that arbuscular mycorrhizal fungi (AMF) and deficit irrigation (DI) have positive effects on the fruit yield or quality. This study aims to test whether the watermelon sugar content and yield can be improved by appropriate amounts of AMF and DI methods and to clarify the underlying physiological mechanism. Diploid and triploid watermelon cultivars and their pumpkin rootstock-grafted seedlings were treated with AMF, DI and DI + AMF in a randomised complete block design with five replications. The results showed that DI significantly reduced the relative water content (RWC), PN, alkaline α-galactosidase activity, but increased the insoluble acid invertase (IAI), sucrose synthase (SuSy) and sucrose phosphate synthase (SPS) activities compared with the well-watered (WW) treatment, which led to a decrease in the fruit yield and an increase in the fruit sugar content. Although the AMF improved the RWC, PN and alkaline α-galactosidase, IAI, SuSy and SPS activities in all the watermelon lines under both the DI and WW conditions, the improvement magnitude of these parameters was more pronounced in the pumpkin-root watermelon lines than the corresponding own-root watermelon lines, especially under the DI condition. The integrated application of AMF and DI increased the fruit yield to a level similar to the WW value in the pumpkin-root watermelon lines and sugar content to an optimal level in all the watermelon lines.

The effects of pumpkin rootstock on photosynthesis, fruit mass, and sucrose content of different ploidy watermelon (Citrullus lanatus)

Pumpkin rootstock affects watermelon scion growth, fruit yield, and quality, but the mechanisms of related key enzymes and photosynthesis remain unclear. In this study, net photosynthetic rate (PN), chlorophyll (Chl) fluorescence parameters, fruit mass and sugar accumulation, and related key enzyme activities were determined during the fruit development stage in diploid and triploid watermelon lines and corresponding pumpkin rootstock-grafted lines. The results showed that pumpkin rootstock increased PN and Chl fluorescence parameters of diploid and triploid watermelon, indicating that pumpkin rootstock could increase photosynthesis, the utilization efficiency of light energy of diploid and triploid watermelon lines. Pumpkin-grafted lines had higher alkaline α-galactosidase activity and lower activities of insoluble acid invertase, sucrose phosphate synthase, and sucrose synthase than corresponding own-root lines in diploid and triploid watermelon. It indicates that pumpkin rootstock could increase the unloading of photoassimilates and reduce the conversion of photoassimilates into sucrose in diploid and triploid watermelon fruits.

Switch from Apoplasmic to Symplasmic Phloem Unloading during Storage Roots Formation and Bulking of Sweetpotato

To clarify the unloading pathway of assimilates in storage roots of sweetpotato (Ipomoea batatas L.), a combination of methods including electron microscopy, movement of the phloem–mobile symplasmic tracer carboxyfluorescein, and assays of invertase and sucrose synthase activities were explored to investigate this pathway from storage roots formation to harvest. The sieve element–companion cell complex was symplasmically connected to surrounding parenchyma cells by plasmodesmata and isolated before storage root bulking (40–140 d after planting). Numerous plasmodesmata appeared among phloem parenchyma throughout storage root bulking. Images of carboxyfluorescein movement indicated that the dye was restricted to phloem and released into surrounding tissues before and after storage root formation, respectively. Sucrose synthase activity increased continually during storage root bulking, and it was much higher than that of insoluble acid invertase. Although this remained low and changed little, that of soluble acid invertase increased and remained high. The starch content in storage roots increased during bulking, but sucrose content decreased. Thus, the predominant unloading pathway switched from apoplasmic to symplasmic during storage root formation and bulking. This switch resulted in enhanced sink potential of storage roots, evidencing opportune sink–source relationships in sweetpotato.

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.

Dynamic characteristics of sugar accumulation and related enzyme activities in sweet and non-sweet watermelon fruits

Sugar content largely determines watermelon fruit quality. We compared changes in sugar accumulation and activities of carbohydrate enzymes in the flesh (central portion) and mesocarp of elite sweet watermelon line 97103 (Citrullus lanatus subsp. vulgaris) and exotic non-sweet line PI296341-FR (C. lanatus subsp. lanatus) to elucidate the physiological and biochemical mechanisms of sugar accumulation in watermelon fruit. The major translocated sugars, raffinose and stachyose, were more unloaded into sweet watermelon fruit than non-sweet fruit. During the fruit development, acid α-galactosidase activity was much higher in flesh of 97103 than in mesocarp of 97103, in flesh and mesocarp of PI296341-FR fruit. Insoluble acid invertase activity was higher in 97103 flesh than in 97103 mesocarp, PI296341-FR flesh or mesocarp from 18 days after pollination (DAP) to 34 DAP. Changes in soluble acid invertase activity in 97103 flesh were similar to those in PI296341-FR flesh and mesocarp from 18 DAP to full ripening. Sucrose synthase and sucrose phosphate synthase activities in 97103 flesh were significantly higher than those in 97103 mesocarp and PI296341-FR fruits from 18 to 34 DAP. Only insoluble acid invertase and sucrose phosphate synthase activities were significantly positively correlated with sucrose content in 97103 flesh. Therefore, phloem loading, distribution and metabolism of major translocated sugars, which are controlled by key sugar metabolism enzymes, determine fruit sugar accumulation in sweet and non-sweet watermelon and reflect the distribution diversity of translocated sugars between subspecies.

Purification and biochemical characterization of insoluble acid invertase (INAC-INV) from pea seedlings.

Invertase (EC 3.2.1.26) catalyzes the hydrolysis of sucrose into D-glucose and D-fructose. Insoluble acid invertase (INAC-INV) was purified from pea (Pisum sativum L.) by sequential procedures entailing ammonium sulfate precipitation, ion exchange chromatography, absorption chromatography, reactive green-19 affinity chromatography, and gel filtration. The purified INAC-INV had a pH optimum of 4.0 and a temperature optimum of 45 °C. The effects of various concentrations of Tris-HCl, HgCl(2), and CuSO(4) on the activities of the purified invertase were examined. INAC-INV was not affected by Tris-HCl and HgCl(2). INAC-INV activity was inhibited by 6.2 mM CuSO(4) up to 50%. The enzymes display typical hyperbolic saturation kinetics for sucrose hydrolysis. The K(m) and V(max) values of INAC-INV were determined to be 4.41 mM and 8.41 U (mg protein)(-1) min(-1), respectively. INAC-INV is a true member of the β-fructofuranosidases, which can react with sucrose and raffinose as substrates. SDS-PAGE and immunoblotting were used to determine the molecular mass of INAC-INV to be 69 kDa. The isoelectric point of INAC-INV was estimated to be about pH 8.0. Taken together, INAC-INV is a pea seedling invertase with a stable and optimum activity at lower acid pH and at higher temperature than other invertases.

Post-anthesis alternate wetting and moderate soil drying enhances activities of key enzymes in sucrose-to-starch conversion in inferior spikelets of rice

This study tested the hypothesis that a post-anthesis moderate soil drying can improve grain filling through regulating the key enzymes in the sucrose-to-starch pathway in the grains of rice (Oryza sativa L.). Two rice cultivars were field grown and two irrigation regimes, alternate wetting and moderate soil drying (WMD) and conventional irrigation (CI, continuously flooded), were imposed during the grain-filling period. The grain-filling rate and activities of four key enzymes in sucrose-to-starch conversion, sucrose synthase (SuSase), adenosine diphosphate-glucose pyrophosphorylase (AGPase), starch synthase (StSase), and starch branching enzyme (SBE), showed no significant difference between WMD and CI regimes for the earlier flowering superior spikelets. However, they were significantly enhanced by the WMD for the later flowering inferior spikelets. The activities of both soluble and insoluble acid invertase in the grains were little affected by the WMD. The two cultivars showed the same tendencies. The activities of SuSase, AGPase, StSase, and SBE in grains were very significantly correlated with the grain-filling rate. The abscisic acid (ABA) concentration in inferior spikelets was remarkably increased in the WMD and very significantly correlated with activities of SuSase, AGPase, StSase, and SBE. Application of ABA on plants under CI produced similar results to those seen in plants receiving WMD. Applying fluridone, an indirect inhibitor of ABA synthesis, produced the opposite effect. The results suggest that post-anthesis WMD could enhance sink strength by regulating the key enzymes involved, and consequently, increase the grain-filling rate and grain weight of inferior spikelets. ABA plays an important role in this process.

Lyso-phosphatidylethanolamine-enhanced phenylalanine ammonia-lyase and insoluble acid invertase in isolated radish cotyledons

Application of lyso-phosphatidylethanolamine (LPE) is purported to suppress fruit ripening and delay foliar senescence. However, the endogenous LPE response of plants is more typically associated with propagation of wound and stress signals. Experiments were therefore carried out to determine whether exogenous LPE could elicit defense responses in plants by determining the effect of this lyso-phospholipid on activity of two key metabolic enzymes and pathogenesis-related proteins viz phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) and insoluble acid invertase (Ac INV; EC 3.2.1.26) in expanding cotyledons of Raphanus sativus L. cv. Cherry Belle (radish). Activity of both enzymes was increased following exposure of tissue to 18:0-LPE and the response was dose dependent. Soluble Ac INV activity was not enhanced by exogenous 18:0-LPE. Increased PAL activity appeared to coincide with a decline in phenolic acid content and a rise in sinapine and lignin. An increase in insoluble Ac INV by 18:0-LPE was associated with a reduction in sucrose concentration. However, levels of glucose and fructose were unaffected. In view of these findings it is proposed that applied LPE acts to co-ordinate carbohydrate partitioning locally to fulfill anabolic respiratory requirements associated with the propagation of systemic wound and stress responses. Furthermore, the impact of exogenous 18:0-LPE on insoluble Ac INV activity is discussed in relation to the proposed role of this enzyme in cytokinin-mediated senescence delay.

Role of sucrose synthase and invertases during petal senescence in rose (Rosa hybrida L.)

SummaryThe roles of sucrose synthase and invertases were explored in relation to petal senescence in rose (Rosa hybrida L.). A developmental shift in the activities of these enzymes was observed. Higher sucrose synthase activity (0.52 – 0.95 µmol sucrose min–1 mg–1 protein) was observed during the initial stages (S1 and S2) of flower bud development, in contrast to invertases. However, the lower activity (0.56 µmol sucrose min–1 mg–1 protein) of sucrose synthase at a later stage (S6) of senescence could help the mobilisation of vacuolar sucrose. The different isoforms of invertase exhibited variable levels of activity. Insoluble acid invertases (IAI) were the most active (11.01 µmol glucose min–1 mg–1 protein), followed by soluble acid invertases (SAI; 8.02 µmol glucose min–1 mg–1 protein), and soluble neutral invertases (SNI; 0.74 µmol glucose min–1 mg–1 protein) at Stage-4. A significant decline in invertase activities (IAI, 0.98; SAI, 1.25; SNI, 0.32 µmol glucose min–1 mg–1 protein) coincided with higher levels of ethylene production at the later stages (S5 and S6) of flower bud development and senescence. We propose that developmental as well as ethylene-mediated pathways account for petal senescence in rose.

Imaging and Quantifying Carbohydrate Transport to the Developing Ovaries of Maize

Shade or inadequate water can inhibit photosynthesis and limit the development of maize (Zea mays) ovaries around the time of pollination, potentially reducing the number of kernels at harvest. This study investigated whether the decreased photosynthesis diminished only the sugar supply or also altered the transport path to the ovaries. Photosynthesis and water potentials (Psiw) were measured in the leaves while dry matter delivery was monitored in the ovaries. Ovary glucose, starch and acid invertase activities were measured in situ. Stems were fed xylem-mobile safranin or phloem-mobile carboxyfluorescein (CF), and the dye transport to the ovaries was determined. Under normal conditions, the ovaries gained in dry mass, and starch accumulated in the pedicel and ovary wall. Glucose accumulated in the pedicel, apparently in the apoplast where insoluble (cell-wall-bound) acid invertase acted on the arriving sucrose. A glucose gradient developed from pedicel to nucellus. Safranin moved in the xylem and did not reach the ovary, but CF moved in the phloem and arrived at the ovary. CF also spread into the pedicel but unlike glucose it did not enter the nucellus. Low Psiw or shade decreased leaf photosynthesis, ovary dry mass accumulation, invertase activities, pedicel glucose, starch accumulation and CF delivery. Removal of these treatments reversed the effects. The success of CF in tracing the general path and rate of carbohydrate transport gave visual evidence that phloem transport to the ovary decreased at low Psiw or in the shade but otherwise remained functional. The decreases indicated that losses in carbohydrate delivery are central features of failed ovary development under these conditions. The selectivity of transport into the nucellus resembled the situation later when embryo and endosperm are present and selective uptake occurs from the apoplast.

Expression of several genes involved in sucrose/starch metabolism as affected by different strategies to induce phosphate deficiency in Arabidopsis

The effects of inorganic phosphate (Pi) deficiency on expression of genes encoding ADP-glucose pyrophosphorylase small and large subunits (ApS and ApL1, ApL2, ApL3 genes), UDP-glucose pyrophosphorylase (Ugp gene), sucrose synthase (Sus1), soluble and insoluble acid invertases (Inv and Invcw) and hexokinase (Hxk1 gene), all involved in carbohydrate metabolism, were investigated in Arabidopsis thaliana (L.) Heynh. We used soil-grown pho mutants affected in Pi status, as well as wild-type (wt) plants grown under Pi deficiency conditions in liquid medium, and leaves of wt plants fed with D-mannose. Generally, ApS, ApL1, Ugp and Inv genes were upregulated, although to a varied degree, under conditions of Pi-stress. The applied conditions had differential effects on expression of other genes studied. For instance, Sus1 was downregulated in pho1 (Pi-deficient) mutant, but was unaffected in wt plants grown in liquid medium under P-deficiency. Mannose had distinct concentration-dependent effects on expression of genes under study, possibly reflecting a dual role of mannose as a sink for Pi and as glucose analog. Feeding Pi (at up to 200 mM) to the detached leaves of wt plants strongly affected the expression of ApL1, ApL2, Sus1 and Inv genes, possibly due to an osmotic effect exerted by Pi. The data suggest that ADP-glucose and UDP-glucose pyrophosphorylases (enzymes indirectly involved in Pi recycling) as well as invertases (sucrose hydrolysis) are transcriptionally regulated by Pi-deficiency, which may play a role in homeostatic mechanisms that acclimate the plant to the Pi-stress conditions.

Impact of temperature, length of storage and postharvest disease on sucrose catabolism in sugarbeet

Abstract Sucrose catabolism during postharvest storage of sugarbeet (Beta vulgaris L.) root has been the subject of several studies; yet, no consensus exists about the contribution of the major sucrolytic activities to postharvest sucrose loss. Because differences in storage temperature, length of storage, and the presence of storage pathogens may have contributed to the discrepant results from earlier studies, the impact of these three factors on sugarbeet root postharvest sucrose catabolism was determined. Sucrolytic activities and soluble carbohydrate concentrations were measured in roots exhibiting no pathological symptoms during storage at 6, 12, and 21 °C and in roots exhibiting severe rotting symptoms due to infection by Penicillium spp. and Botrytis cinerea during storage at 6 °C. Sucrose synthase was the predominant sucrolytic activity throughout storage, regardless of storage temperature, length of storage, or pathogenesis, and accounted for more than 90% of the total soluble sucrolytic activity present in roots. In disease-free roots, no significant change in sucrose synthase activity, soluble acid invertase activity, or insoluble acid invertase activity occurred in roots stored at 6 or 12 °C, although an increase in sucrose synthase activity was observed in roots stored at 21 °C. Alkaline invertase activity was impacted by the length of storage and exhibited a transient decline in activity at all storage temperatures. Glucose and fructose concentrations generally increased as a function of time in storage at 6, 12, and 21 °C. In roots with severe rot, insoluble acid invertase activity declined, sucrose synthase and alkaline invertase activities were unchanged, and soluble acid invertase increased seven-fold. The increase in soluble acid invertase activity was primarily due to the presence of fungal acid invertase isoforms. These results indicate that sugarbeet sucrolytic activities change little during storage, regardless of storage temperature, length of storage, and pathogenesis, and suggest that sucrose synthase, as the predominant sucrolytic activity in stored roots, is central to postharvest sucrose catabolism in sugarbeet roots.

Activities of key enzymes in sucrose-to-starch conversion in wheat grains subjected to water deficit during grain filling.

This study tested the hypothesis that a controlled water deficit during grain filling of wheat (Triticum aestivum) could accelerate grain-filling rate through regulating the key enzymes involved in Suc-to-starch pathway in the grains. Two high lodging-resistant wheat cultivars were field grown. Well-watered and water-deficit (WD) treatments were imposed from 9 DPA until maturity. The WD promoted the reallocation of prefixed 14C from the stems to grains, shortened the grain-filling period, and increased grain-filling rate or starch accumulation rate (SAR) in the grains. Activities of Suc synthase (SuSase), soluble starch synthase (SSS), and starch branching enzyme (SBE) in the grains were substantially enhanced by WD and positively correlated with the SAR. ADP Glc pyrophosphorylase activity was also enhanced in WD grains initially and correlated with SAR with a smaller coefficient. Activities of granule-bound starch synthase and soluble and insoluble acid invertase in the grains were less affected by WD. Abscisic acid (ABA) content in the grains was remarkably enhanced by WD and very significantly correlated with activities of SuSase, SSS, and SBE. Application of ABA on well-watered plants showed similar results as those by WD. Spraying with fluridone, an ABA synthesis inhibitor, had the opposite effect. The results suggest that increased grain-filling rate is mainly attributed to the enhanced sink activity by regulating key enzymes involved in Suc-to-starch conversion, especially SuSase, SSS, and SBE, in wheat grains when subjected to a mild water deficit during grain filling, and ABA plays a vital role in the regulation of this process.

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.

Sucrolytic activities during fruit development of Lycopersicon genotypes differing in tolerance to salinity.

The different growth responses under control and moderate salinity (70 mM NaCl) in relation to the carbon partitioning and sucrose metabolism in developing tomato fruits [20 days after anthesis (DAA), start of ripening and ripe stages] were studied in the cultivated tomato Lycopersicon esculentum Mill (cv. H-324-1), in the wild relative species L. cheesmanii (ac. LA-530) (hexose-accumulators), L. chmielewskii (ac. LA-1028) (sucrose-accumulator) and in two interspecific F1 hybrids (hexose-accumulators) (F1-530: H-324-1 x A-530, F1-1028: H-324-1 x A-1028). The higher salt-tolerance of the wild species and hybrids with respect to the domestic tomatoes was also observed at the fruit level because these genotypes were less affected in the assimilation of dry weight (DW) under salinity. With the exception of the wild tomatoes, the sink strength, evaluated as the dry matter accumulation rate (mg DW day-1) and the sink activity, evaluated as a relative growth rate (mg DW mg-1 day-1), were reduced during the early fruit growing period (20 DAA-start ripening). However, a total recovery of growth was registered in the salinized hybrid fruits during the late growing period (start of ripening-ripe fruits). The early reduction in sink activity in the hybrid and domestic fruits was related to a sucrose accumulation and a decrease in the total sucrolytic activity at 20 DAA, especially the cytoplasmic sucrolytic activities sucrose synthase (EC 2.4.1.13) and neutral invertase (EC 3.2.1.26). The further recovery in sink strength of the hybrid fruits was related to the maintenance of the insoluble acid invertase (EC 3.2.1.25) and the induction of the cytoplasmic sucrolytic activities, namely at the start of ripening stage, demonstrating the existence of an inverse relationship between these activities, which suggests a regulatory mechanism in order to maintain the sink capacity. The roles of different enzymes in the control of assimilate import under salinity in relation to the sucrose transport and possible regulatory mechanisms are discussed.

Suppressive effect of H2O2 on increase in activities of acid invertases in rye roots

Although various effects of H2O2 on plant cellular functions have been reported, the effects of H2O2 on glycolytic enzymes remain unknown. It was shown that H2O2 has a suppressive effect on increase in activities of soluble and insoluble acid invertases among a number of glycolytic enzymes during a growth stage of rye roots.

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.

Acid invertase is predominantly localized to cell walls of both the practically symplasmically isolated sieve element/companion cell complex and parenchyma cells in developing apple fruits

AbstractThe acid invertase (β‐fructosidase, EC 3·2·1·26) was localized at subcellular level via immunogold electron microscopy in the phloem‐unloading zone of developing apple fruit. The enzyme (immunogold particles) was found to reside predominantly in the cell walls of the sieve element/companion cell (SE/CC) complex, phloem parenchyma cells and other parenchyma cells. There was almost no gold particle found in cytoplasm and vacuole. This distribution pattern remained unchanged throughout the growing season, but the enzyme numbers varied. The density of immunogold particles increased during fruit development. The immunoblotting of soluble and insoluble acid invertases provided a supporting proof for the assays of immunolocalization. The biochemical analysis showed a predominantly cell‐wall‐distributed activity of acid invertase that corresponds essentially with its amount distribution. The ultrastructural observations showed that there were numerous plasmodesmata between the parenchyma cells, but almost no plasmodesmium between the SE/CC complex and its surrounding parenchyma cells, practically resulting in the symplasmic isolation of the SE/CC complex. It is therefore suggested that the unloading pathway of sucrose from the SE/CC complex may be predominantly apoplasmic in the developing apple fruit, and that the unloaded sucrose may be hydrolysed by the functional acid invertase localized in the cell wall before it is loaded in sink cells.

Factors affecting ‘Hass’ avocado fruit size: Carbohydrate, abscisic acid and isoprenoid metabolism in normal and phenotypically small fruit

Carbohydrate and abscisic acid (ABA) metabolism were investigated in normal and phenotypically small ‘Hass’ avocado (Persea americana Mill.) fruit in an attempt to link alterations in sugar and ABA content with changes in 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase (HMGR, EC 1.1.1.34) activity and fruit size. The small‐fruit phenotype was characterized by reduced seed HMGR activity, increased seed insoluble acid invertase (β‐ d‐fructofuranosidase, EC 3.2.1.26), decreased sucrose synthase (SS; UDP‐ d‐glucose: d‐fructose‐2‐α‐glucosyl‐transferase, EC 2.4.1.13) activity, decreased sucrose content, and increased glucose as a proportion of the total soluble sugar. Sucrose phosphate synthase (SPS; UDP‐ d‐glucose: d‐fructose 6‐phosphate 2‐α‐ d‐glucosyltransferase, EC 2.4.1.14) activity was unaffected in seed but reduced in mesocarp of the small fruit. In addition, the small‐fruit variant displayed enhanced respiration and both seed and mesocarp tissue showed increased ABA metabolism. Applied ABA caused an increase in insoluble acid invertase activity in seed tissue of normal fruit while mevastatin reduced HMGR activity in this tissue, caused sucrose depletion and increased the proportion of glucose from 5 to 57% of total soluble sugars. Exogenous glucose suppressed HMGR activity in seed tissue whereas in mesocarp tissue, HMGR activity was reduced to 38% of the control after 6 h but enhanced by 46% by 48 h. Glucose increased ABA biosynthesis and turnover in competent tissues. These results suggest that ABA turnover is mediated, in part, by carbohydrate content and composition which also affects HMGR activity. It is proposed that sugar and ABA signals act in concert to modulate expression and/or activity of HMGR in the control of ‘Hass’ avocado fruit growth and final fruit size.