Acid invertase in germinating Lactuca sativa seeds: Evidence for de novo synthesis

Acid invertase activity in germinating lettuce seeds is first observed after 15 hr germination, from when it rises steadily at least till 30 hr of germination. The enzyme was purified about 500-fold using ammonium sulphate fractionation followed by isoelectric focussing. Labelling the enzyme with ³⁵SO₄ or leucine-¹⁴C during development of its activity, followed by purification suggests that acid invertase is synthesized de novo during germination. The possible significance of acid invertase in the metabolism of the seed is discussed.     

Invertases in young and mature leaves of Citrus sinensis

The sucrose catabolic enzymes acid invertase (EC 3.2.1.26) and alkaline invertase (EC 3.2.1.27) were studied in young and mature Citrus sinensis leaf tissue. In young, expanding leaves (60 % final length) soluble acid invertase activity predominated, while soluble alkaline invertase activity predominated in mature leaves. The acid and alkaline invertase activities were separated on Sephadex G-200. The acid invertase had an M_r of approximately 60 000, pH maximum of 4.5 and apparent K_m of 3.3 mM sucrose. The alkaline invertase had an M_r of approximately 200 000, pH maxima of 6.8 and an apparent K_m of 20 mM sucrose. Alkaline invertase was strongly inhibited by 10 mM Tris while acid invertase was not. Possible physiological roles for the two invertases are discussed.     

The role of sucrose-metabolizing enzymes in pear fruit that differ in sucrose accumulation

In the paper, the soluble sugar composition and activities of enzymes metabolizing sucrose: invertase (β-fructosidase, EC 3.2.1.26), sucrose synthase (SS; EC 2.4.1.13) and sucrose-phosphate synthase (SPS; EC 2.4.1.14) were investigated during fruit development of two pear species: Pyrus bretschneideri Rehd. cv. ‘Yali’ and P. pyrifolia Nakai cv. ‘Aikansui’, characterized as low and high sucrose types, respectively. It was found that, at the end of fruit development of ‘Aikansui’, the level of sucrose was five times higher than in ‘Yali’ in the same period. It was coincident with the significantly higher activities of SS (synthesis) and SPS and lower activities of invertase (vacuolar and cell wall-bound acid invertase and neutral invertase). The high correlation was found between sucrose level and SS (synthesis) and SPS activities in ‘Aikansui’ pears.     

Properties of Invertases in Sugar Storage Tissues of Citrus Fruit and Changes in Their Activities during Maturation

Both acid and alkaline invertases were present in immature juice sacs of satsuma mandarin (Citrus‘Unshu Marcovitch”) fruit, in which sugar content was low. Maturing and mature juice sacs, in which sugar content increased steadily with time, were characterized by the presence of alkaline invertase and the absence of acid invertase. When the immature juice sacs were homogenized with 0.2 M sodium phosphate-citrate buffer (pH 8.0), almost all of the acid invertase activity was found in the solubilized fraction, whereas almost all of the alkaline invertase activity was present in the insoluble fraction. The distribution of alkaline invertase between the solubilized and insoluble fractions changed with the development of fruit. The acid invertase had a molecular weight of 69,000, optimum pH of 4.8–5.3, and K_m value for sucrose of 7.3 mM. The alkaline invertase had a molecular weight of 200,000, pH optimum of 7.2–7.7, and K_m value of 35.7 mM. The hydrolysing activities of both enzymes for raffinose were considerably less than those for sucrose. The alkaline invertase had lower activity for raffinose than the acid invertase.     

Acid and alkaline invertases in suspension cultures of sugar beet cells

Alkaline invertase was induced during the initiation of suspension cultures of single cells from leaf explants of sugar beets in Murashige-Skoog liquid medium which contained benzyladenine. This activity was barely detectable in the leaves themselves. In suspension cultures, the presence of both acid and alkaline invertases was detected; alkaline invertase was only present in the cytoplasm of the cultured cells, whereas acid invertase was present in the cytoplasm and cell walls, and was also detected in the culture medium. The cell wall contained at least three types of acid invertase; two of these activities were solubilized by saline (saline-released) and EDTA (EDTA-released), respectively, and the third remained tightly associated with the cell wall. Saline-released and EDTA-released invertases from the cell wall showed the significant differences in their properties: the saline-released enzyme had the highest affinity for sucrose among the invertases tested, and was easily bound to cell walls, to DNA, and to a cation exchanger, unlike the EDTA-released enzyme. Sucrose is the source of carbon for plant cells in suspension culture and is probably degraded in the cell wall by the saline-released invertase, which had the highest activity and the highest affinity for sucrose. Hexose products of this degradation would be transported to cytoplasm. Soluble invertase, EDTA-released invertase from the cell wall, and one of two extracellular invertases behaved similarly upon chromatography on DEAE-cellulose. They had similar activity profiles with changing pH, and similar K_m values for sucrose. Thus it appears that they are identical. Two extracellular invertases found in the growth medium of the suspension cultures were probably identical with those in the soluble fraction of callus and seedlings of sugar beets, because they showed similar behaviors during chromatography on DEAE-cellulose, and had similar activity profiles with changing pH and K_m values for sucrose.     

Wall-bound enzymes in callus of Convolvulus arvensis

Isolated cell walls of Convolvulus callus contain α- and β-galactosidase, α- and β-glucosidase, α- and β-mannosidase, acid invertase and acid phosphatase activities. No neutral invertase or alkaline phosphatase activities could be detected. Acid invertase activity per mg cell wall increased considerably during incubation of callus fragments in nutrient solution, as opposed to the activities of the other enzymes mentioned.     

Genome-Wide Identification of the Invertase Gene Family in Populus

Invertase plays a crucial role in carbohydrate partitioning and plant development as it catalyses the irreversible hydrolysis of sucrose into glucose and fructose. The invertase family in plants is composed of two sub-families: acid invertases, which are targeted to the cell wall and vacuole; and neutral/alkaline invertases, which function in the cytosol. In this study, 5 cell wall invertase genes (PtCWINV1-5), 3 vacuolar invertase genes (PtVINV1-3) and 16 neutral/alkaline invertase genes (PtNINV1-16) were identified in the Populus genome and found to be distributed on 14 chromosomes. A comprehensive analysis of poplar invertase genes was performed, including structures, chromosome location, phylogeny, evolutionary pattern and expression profiles. Phylogenetic analysis indicated that the two sub-families were both divided into two clades. Segmental duplication is contributed to neutral/alkaline sub-family expansion. Furthermore, the Populus invertase genes displayed differential expression in roots, stems, leaves, leaf buds and in response to salt/cold stress and pathogen infection. In addition, the analysis of enzyme activity and sugar content revealed that invertase genes play key roles in the sucrose metabolism of various tissues and organs in poplar. This work lays the foundation for future functional analysis of the invertase genes in Populus and other woody perennials.     

Effects of gibberellic Acid and gold light on germination, enzyme activities, and amino Acid pool size in a dwarf strain of watermelon

Gibberellic acid (GA₃) promotes and continuous gold light inhibits germination of seeds of a dwarf strain (WB-2) of watermelon [Citrullus lanatus (Thunb.) Matsu. and Nakai]. Osmotic inhibition of germination with mannitol in light-grown seeds of WB-2 was only slightly reversed by GA₃ at the concentrations used, whereas, GA₃ substantially relieved osmotic inhibition in dark-grown seeds.

The effects of GA₃ and gold light on development of catalase and invertase activities and on levels of free amino acids in germinating seeds of WB-2 were examined. Light depressed development of catalase and invertase activity. Levels of free amino acids increased more slowly in embryonic axes of light- than dark-incubated seeds, but in cotyledons higher levels of amino acids were maintained in light-grown seeds. GA₃ accelerated the development of catalase activity in whole embryos and invertase activity in embryonic axes, but did not significantly affect invertase activity in cotyledons during germination. GA₃ had little effect on amino acid pools in cotyledons and embryonic axes.

     

Sucrose metabolism in lima bean seeds

Developing and germinating lima bean (Phaseolus lunatus var Cangreen) seeds were used for testing the sucrose synthase pathway, to examine the competition for uridine diphosphate (UDP) and pyrophosphate (PPi), and to identify adaptive and maintenance-type enzymes in glycolysis and gluconeogenesis. In developing seeds, sucrose breakdown was dominated by the sucrose synthase pathway; but in the seedling embryos, both the sucrose synthase pathway and acid invertase were active. UDPase activity was low and seemingly insufficient to compete for UDP during sucrose metabolism in seed development or germination. In contrast, both an acid and alkaline pyrophosphatase were active in seed development and germination. The set of adaptive enzymes identified in developing seeds were sucrose synthase, PPi-dependent phosphofructokinase, plus acid and alkaline pyrophosphatase; and, the adaptive enzymes identified in germinating seeds included the same set of enzymes plus acid invertase. The set of maintenance enzymes identified during development, in the dry seed, and during germination were UDP-glucopyrophosphorylase, neutral invertase, ATP and UTP-dependent fructokinase, glucokinase, phosphoglucomutase, ATP and UTP-dependent phosphofructokinase and sucrose-P synthase.     

Alkaline β-fructofuranosidases of tuberous roots: Possible physiological function

Summary Alkaline invertase of roots of carrot (Daucus carota L.) did not hydrolyze raffinose while the acid invertase from the same tissue showed with this sugar ca. 60% of the activity found with sucrose. The activity of the two invertases was inhibited by fructose to a different extent, the K _i value being ca. 4×10^–2 M and 3×10^–1M, respectively, for the alkaline and the acid invertases from the roots of both carrot and turnip (Brassica rapa L.). It is proposed that fructose inhibition of acid invertase is of no physiological significance but that, in contrast, hexoses might regulate the activity of alkaline invertase.Comparing several species and cultivars, it was found that the content of reducing sugars and the activity of alkaline invertase of mature tuberous roots showed a positive correlation. This indicates that alkaline invertase may participate in the regulation of the hexose level of the cell, as was previously suggested for sugar-cane. A scheme is presented which proposes a way of participation of alkaline invertase in such a regulation, assuming that this enzyme is located in the cytoplasm and acid invertase is membrane-bound and mainly located at the cell surface.     

Differences in invertase activity in embryogenic and non-embryogenic calli from Medicago arborea

The different invertase activities in embryogenic and non-embryogenic calli induced from explants (cotyledons, petioles, hypocotyls and leaves) obtained from Medicago arborea L. subsp. arborea seedlings were evaluated. Total invertase activity was lower in the calli with the greatest embryogenic capacity. The greatest fraction of this activity corresponded to soluble invertase. Wall-bound invertase showed maximum activity during the first two months of culture and the highest activities of this type were found in non-embryogenic calli. Extracellular invertase formed the smallest fraction of the total invertase activity evaluated. Acid and alkaline invertase activities were found in all calli but differences were detected between the embryogenic and non-embryogenic calli. In the former, the activity of both types of invertase exhibited a similar type of behaviour but different from that observed in the non-embryogenic calli. The calli with the greatest embryogenic capacity had very low levels of acid invertase and very high levels of the alkaline form. Soluble invertase – both acid and alkaline – accounted for the highest fraction after the first two months of culture and was present in lower amounts in the embryogenic than in the non-embryogenic calli. Regarding bound invertase, the highest production was seen to correspond to acid invertase. The extracellular invertase evaluated corresponded to the acid form since the alkaline extracellular invertase did not show any physiologically significant activity.     

套袋对梨果实发育过程中糖组分及其相关酶活性的影响

以翠冠和黄金梨为试材,测定套袋和未套袋(对照)梨果实发育时期果实中蔗糖、葡萄糖、果糖和山梨醇含量以及蔗糖代谢相关酶酸性转化酶(AI)、中性转化酶(NI)、蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)的活性,并对果实中糖组分积累与酶活性的关系进行了分析.结果表明:(1)两梨品种套袋果实在发育过程中蔗糖、葡萄糖、果糖、山梨醇和糖代谢相关酶活性变化趋势与对照基本一致,套袋果实糖含量均低于对照但差异不显著,而各相关酶活性在两类果实间差异表现各异.(2)在梨果实发育早期,果实中以分解酶类为主,糖分积累低;发育后期以合成酶类为主,糖分积累多.(3)两品种套袋和对照果实AI活性与葡萄糖含量均呈显著或极显著正相关,SS合成方向活性与蔗糖含量均为极显著正相关,且翠冠对照果SPS活性与蔗糖含量呈极显著正相关.可见,套袋通过提高果实发育早期转化酶(Inv)活性,降低果实后期蔗糖磷酸合成酶(SPS)、蔗糖合成酶(SS)的活性来影响糖分积累,从而影响梨果品质.     

Effects of water gradients on soil enzyme activity and active organic carbon composition under Carex lasiocarpa marsh

The responses of soil enzyme activity of freshwater marsh, microbial biomass carbon (MBC), dissolved organic carbon (DOC) and aboveground biomass to water gradients were studied with Carex lasiocarpa pot culture experiment. The relationships between soil enzyme activity and MBC, DOC and aboveground biomass were discussed. The water gradients were W1, 15 cm; W2, ?5 cm; W3, ?5–5 cm; W4, submerged. The results indicated that acid phosphatase, invertase and urease activities were decreased with the increase of water level, while catalase activity was increased with moisture content increasing. Drying-wetting alternation (W3) increased soil enzyme activities if compared with W1. MBC content followed the order of W3 > W1 > W2 > W4, and the activities of invertase, urease and catalase were significantly positively correlated with MBC (p < 0.05). DOC content presented the order of W4 > W1 > W3 > W2, and the activities of urease and acid phosphatase were most significantly negatively correlated with DOC (p < 0.01). In addition, drying-wetting alternation promoted the growth of Carex lasiocarpa. When water submerged plants, the growth of Carex lasiocarpa was significantly inhibited. The aboveground biomass was positively related to soil enzyme activities. There were close relationships between the activities of invertase, urease and catalase and the growth situation of Carex lasiocarpa.     

Influence of yeast-derived invertase gene expression in potato plants on membrane lipid peroxidation at low temperature

Tolerance to chilling was compared in potato plants (Solanum tuberosum L., cv. Désirée) transformed with a yeast-derived invertase gene under the control of the B33 class 1 tuber-specific promoter and with the proteinase inhibitor II leader peptide sequence providing for the apoplastic enzyme localization (the B33-inv plants) and potato plants transformed only with a reporter gene (the control plants) under in vitro conditions. The total activities of acid and alkaline invertases and contents of sugars in the B33-inv plants exceeded those in the control plants. The exposures at low temperature produced a significant less lipid peroxidation activity in the B33-inv plants, as compared to the control. The authors presume that the B33-inv plants acquire a higher tolerance to low temperatures apparently due to the changes in sugar ratio produced by the yeast invertase.     

Invertases of pollen of Haemanthus albiflos

Soluble and insoluble invertase occurs in dormant pollen of Haemanthus albiflos, with pH optima of 5·7 and 5·5 respectively. At their pH optima the activity of the soluble enzyme is 3·5-fold higher. After 2 hr germination the pH optimum of the insoluble invertase is increased to 6·0 and the activity is increased 2-fold while the activity of the soluble invertase is decreased by 26%.     

Enzymatic studies during spontaneous cell rupture of Saccharomyces cerevisiae

Summary Some of the extract and intracellular enzyme activities in K2nB strain of Saccharomyces cerevisiae that growing in the condition which induce spontaneous cell rupture, were measured. B-1-3-glucanase, invertase, acid phosphatase and active chitin synthetase zymogen showed a reduced activity in ruptured cell while alkaline phosphatase shows no differences in its activity.     

The resting metabolism of dodder seeds

Extracts of mature seeds of Cuscuta reflexa were examined for any deficiency in key enzymes. The activities of malate dehydrogenase, β-amylase and fructose 1,6-diphosphate aldolase exceeded 5.0 μmol substrate/min/g, while those of starch phosphorylase, α-amylase, acid phosphatase, phosphogluconate dehydrogenase (decarboxylating), aspartate aminotransferase, glucose 6-phosphate dehydrogenase, fructose 1,6-diphosphatase and alanine aminotransferase fell within the range 1 to 5 μmol/min/g and hexokinase, isocitrate dehydrogenase and alkaline phosphatase were below 1 μmol substrate/min/g seed powder. No activity of the following were found: acid invertase, alkaline invertase, phytase and glutamate dehydrogenase. Some of these observations were made also for seeds of Cuscuta campestris and Cuscuta indicora.     

Differential invertase activity and root growth between Cu-tolerant and non-tolerant populations in Kummerowia stipulacea under Cu stress and nutrient deficiency

There has been no study on key enzymes in sucrose cleavage in metallophyte plants so far, which may be crucial for the plants’ root growth and heavy-metal tolerance maintenance. Here, we tested the hypothesis that the roots of copper tolerant plants should manifest a higher activity of acid invertases that are rate-limiting in sucrose catabolism than non-tolerant plants both for supporting growth and for their maintaining tolerance under Cu stress. Two populations of Kummerowia stipulacea, one from an ancient waste heap at a Cu mine, and the other from a non-contaminated site, were used in the experiments. The plants were grown in 1/2-fold (control) or 1/20-fold (nutrient deficiency) Hoagland’ solution, with (Cu stress) or without (control) 10 μmol/L Cu²⁺. Plants from the mine proved to be of Cu tolerance. Cu exposure had a stronger inhibition on root growth and thus resulting in a lower root/shoot ratio in the plants of non-mine population compared to the mine population. Cu exposure showed a stronger inhibition of acid invertase activity of Cu non-tolerant plants than Cu-tolerant plants, while neutral/alkaline invertase was insensitive to Cu. A positive correlation between the activity of acid invertases and the root growth and root/shoot ratio was observed. The results indicated an important role of acid invertases in governing root growth and root/shoot biomass allocation in the plants of mine population. The results also suggested that the higher activities in acid invertases of mine population plants might at least partly associate with the plants’ Cu tolerance, and their higher activities in acid invertases in turn played an role in maintenance of the Cu tolerance by supplying carbon and energy for tolerance mechanisms. In addition, the results showed evidence that neutral/alkaline invertase might play a role in compensating for the depression in sucrose catabolism due to Cu-induced inhibition in acid invertases.     

Effect of GA3, kinetin and indole acetic acid on carbohydrate metabolism in chickpea seedlings germinating under water stress

Enhanced amylase activity was observed during a 7-day-growth period in the cotyledons of PEG imposed water stressed chickpea seedlings grown in the presence of GA₃ and kinetin, when compared with stressed seedlings. During the first 5 days of seedling growth, the seedlings growing under water deficit conditions as well as those growing in the presence of PGRs had a higher amylase activity in shoots than that of control seedlings. Neither GA₃ nor kinetin increased the amylase activity of roots whereas IAA reduced root amylase activity. Activity of acid and alkaline invertases was maximum in shoots and at a minimum in cotyledons. Compared with alkaline invertase, acid invertase activity was higher in all the tissues. The reduced acid and alkaline invertase activities in shoots of stressed seedlings were enhanced by GA₃ and kinetin. Roots of stressed seedlings had higher alkaline invertase activity and GA₃ and IAA helped in bringing the level near to those in the controls. GA₃ and kinetin increased the sucrose synthase (SS) and sucrose phosphate synthase (SPS) activities in cotyledons of stressed seedlings, whereas they brought the elevated level of SPS of stressed roots to near normal level. The higher level of reducing sugars in the shoots of GA₃ and kinetin treated stressed seedlings could be due to the high acid invertase activity observed in the shoots, and the high level of bound fructose in the cotyledons of stressed seedlings could be due to the high activity of SPS in this tissue.     

Seasonal Dynamics of Soil Labile Organic Carbon and Enzyme Activities in Relation to Vegetation Types in Hangzhou Bay Tidal Flat Wetland

Soil labile organic carbon and soil enzymes play important roles in the carbon cycle of coastal wetlands that have high organic carbon accumulation rates. Soils under three vegetations (Phragmites australis, Spartina alterniflora, and Scirpusm mariqueter) as well as bare mudflat in Hangzhou Bay wetland of China were collected seasonally. Seasonal dynamics and correlations of soil labile organic carbon fractions and soil enzyme activities were analyzed. The results showed that there were significant differences among vegetation types in the contents of soil organic carbon (SOC) and dissolved organic carbon (DOC), excepting for that of microbial biomass carbon (MBC). The P. australis soil was with the highest content of both SOC (7.86 g kg^-1) and DOC (306 mg kg^-1), while the S. mariqueter soil was with the lowest content of SOC (6.83 g kg^-1), and the bare mudflat was with the lowest content of DOC (270 mg kg^-1). Soil enzyme activities were significantly different among vegetation types except for urease. The P. australis had the highest annual average activity of alkaline phosphomonoesterase (21.4 mg kg^-1 h^-1), and the S. alterniflora had the highest annual average activities of β-glycosidase (4.10 mg kg^-1 h^-1) and invertase (9.81mg g^-1 24h^-1); however, the bare mudflat had the lowest activities of alkaline phosphomonoesterase (16.2 mg kg^-1 h^-1), β-glycosidase (2.87 mg kg^-1 h^-1), and invertase (8.02 mg g^-1 24h^-1). Analysis also showed that the soil labile organic carbon fractions and soil enzyme activities had distinct seasonal dynamics. In addition, the soil MBC content was significantly correlated with the activities of urease and β-glucosidase. The DOC content was significantly correlated with the activities of urease, alkaline phosphomonoesterase, and invertase. The results indicated that vegetation type is an important factor influencing the spatial-temporal variation of soil enzyme activities and labile organic carbon in coastal wetlands.