• Korean Journal of Environmental Biology
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• v.27 no.4
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• pp.406-413
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• 2009

The enzyme invertase contributes to sugar unloading, pathogen defense, differentiation and development in plants. We cloned the complete cDNA of a soluble acid invertase from pea seedlings (Pisum sativum) via RT-PCR and the rapid amplification of the cDNA end (RACE) technique. The full-length cDNA of the soluble pea invertase comprised 2237 bp and contained a complete open reading frame encoding 647 amino acids. The deduced amino acid sequence showed high homology to soluble acid invertases from various plants. Northern blot analysis demonstrated the soluble acid invertase gene of P. sativum was strongly expressed in sink organs such as shoot tips and root tips, and induced by abscisic acid, gibberellic acid and jasmonic acid in shoots. Especially, gibberellic acid enhanced the gene expression of the soluble acid invertase in a time-dependent manner. This study presents that the gene expression patterns of a soluble acid invertase from pea are strongly consistent with the suggestion that individual invertase gene product has different functions in the growing plant.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• v.2 no.1
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• pp.37-48
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• 1998

Effect of plant hormones on the leaf senescence of mung bean (Phaseolus radiatus) was investigated by measuring the changes of reducing sugar contents and invertase isozyme activities in detached leaves treated with NAA, $GA_3$ or BA. During dark-induced senescence, reducing sugar contents in the detached leaves increased temporarily at 4 d, thereafter decreased rapidly and reached minimum values within 7-14 d. The pattern of soluble acid invertase activity in the senescing leaves kept in the dark was similar to that of reducing sugar accumulation, whereas the activities of alkaline and extracellular invertases were not significantly changed during leaf senescence. Therefore, these results suggest that soluble acid invertase, but not alkaline and extracellular invertases, induces the accumulation of reducing sugar during leaf senescence of mung bean plants. Exogenous NAA application had little or no effect in the increase of soluble acid invertase activity during dark-induced senescence compared to the control. However, exogenous applications of $GA_3$ and BA led to the increase of soluble acid invertase activity in the senescing leaves. Particularly, BA application was very effective in enhancing the activity of soluble acid invertase as well as in delaying chlorophyll breakdown during dark-induced senescence. These results suggest, therefore, that BA regulates the activity of soluble acid invertase, which leads to the accumulation of reducing sugar, and the stability of photosynthetic apparatus to delay leaf senescence.

• Journal of Environmental Science International
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• v.2 no.1
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• pp.37-48
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• 1993

Effect of plant hormones on the leaf senescence of mung bean (Phseoln radiatus) was investigated by measuring the changes of reducing sugar contents and invertase isozyme activities in detached leaves treated with NAA, $GA_3$ or BA. During dark-induced senescence, reducing sugar contents in the detached leaves increased temporarily at 4 6, thereafter decreased rapidly and reached minimum values within 7-14 6. The pattern of soluble acid invertase activity in the senescing leaves kept in the dark was similar to that of reducing sugar accumulation, whereas the activities of alkaline and extracellular invertases were not significantly changed during leaf senescence. Therefore, these results suggest that soluble acid invertase, but not alkaline and extracellular invertases, induces the accumulation of reducing sugar during leaf senescence of Rung bean plants. Exogenous NAA application had little or no effect In the increase of soluble acid invertase activity during dark-induced senescence compared to the control. However, exogenous applications of $GA_3$ and BA led to the increase of soluble acid invertase activity in the senescing leaves. Particularly, BA application was very effective In enhancing the activity of soluble acid invertase as well as in delaying chlorophyll breakdown during dark-induced senescence. These results suggest, therefore, that BA regulates the activity of soluble acid invertase, which leads to the accumulation of reducing sugar, and the stability of photosynthetic apparatus to delay leaf senescence.

• Journal of Plant Biology
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• v.38 no.3
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• pp.251-258
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• 1995

The soluble acid invertase ($\beta$-D-fructofuranoside fructohydrolase, EC 3.2.1.26) was isolated and characterized from the hypocotyls of mung bean (Phaseolus radiatus L.). The enzyme was purified to apparent homogeneity by consecutive step using diethylaminoethyl (DEAE)-cellulose anion exchange, Concanavalin (Con) A affinity and Sephacryl S-300 chromatography. The overall purification was about 148-fold with a yield of about 15%. The finally purified enzyme exhibited a specific activity of about 139 $\mu$mol of glucose produced mg-1 protein min-1 at pH 5.0 and appeared to be a single protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and nondenaturing PAGE. The enzyme had the native molecular weight of 70 kD and subunit molecular weight of 70 kD as estimated by Sephadex G-200 chromatography and SDS-PAGE, respectively, suggesting that the enzyme was composed of a monomeric protein. On the other hand, the enzyme appeared to be a glycoprotein containing N-linked high mannose oligosaccharide chain on the basis of its ability to bind to the immobilized C on A. The enzyme had a Km for sucrose of 1.8 mM at pH 5.0 and maximum activity around pH 5.0. The enzyme showed highest enzyme activity with sucrose as substrate, but the activity was slightly measured with raffinose and cellobise. No activity was measured with maltose and lactose. These results indicate the soluble acid invertase is a $\beta$-fructofuranosidase.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.499-504
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• 2005

This experiment was conducted to identify the effect of several plant growth retardants on growth of Sedirea japonica seedlings cultured in vitro and their changes of invertase activities. When seedlings of Sedirea japonica were treated with ancymidol and paclobutrazol, as the concentrations were increased, leaf length was gradually shortened and leaf width became wider than that of control. On the other hand, root length was shorter, but the number of root and the root's diameters were greatly increased, compared with control. In 0.05mg/L uniconazole, growth of leaf and root were enhanced, compared with the control and higher concentrations of uniconazole. As concentration of each growth retardants was increased, leaf shape became round and smaller. Both soluble acid invertase activity and soluble alkaline invertase activity in leaf were decreased in higher concentrations of each growth retardant, but those of the root were contrary to those of the leaf. To confirm the estimated invertase activities, starch content of leaf was higher in low concentration treatments in each growth retardant, but in the root was contrary to content that of the leaf.

• Journal of Life Science
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• v.25 no.9
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• pp.1021-1026
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• 2015

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 Hg²⁺ and Cu²⁺. 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.

• Journal of Life Science
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• v.11 no.1
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• pp.14-21
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• 2001

To investigate the changes of carbohydrate metabolism in the senescing leaves of Zea mays during dark-induced senescence, the changes in the contents of reducing sugar, sucrose and starch as well as the activities of sucrose synthase, three isozymes of invertase, and ${\alpha}$-amylase were measured. In the senescing leaves, the content of reducing sugars temporarily increased at 4 d and rapidly decreased thereafter, whereas sucrose contents gradually decreased thereafter, whereas sucrose contents gradually decreased until 3 d of senscence and significantly decreased thereafter. The activities of intracellular invertases such as soluble acid and alkaline formed gradually enhanced until 4 d of leaf senescence and significantly declined thereafter. The extracellular invertase activity showed no significant changes during leaf senescence. The deactivation of sucrose synthase was observed within 3 d of leaf senscence. On the other hand, the starch contents gradually declined during 2 d of leaf senescence, and showed a temporary increase at 3 d, which is similar to the pattern of sucrose synthase activity., These results imply that sucrose in the senescing leaves. The major enzymes which correlated to the breakdown of sucrose during dark-induced senescence were soluble acid and alkaline invertases, not sucrose and ABA accelerated leaf senescence by inducing the accumulation of reducing sugar. These result, therefore, that leaf senescence may be mediated by the temporary quantitative changes of reducing sugar induced by the activation of intracellualr inveertases.

• Horticultural Science & Technology
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• v.34 no.6
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• pp.898-911
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• 2016

Sugars play many important roles in plant metabolism and directly influence fruit quality. The effects of two edible coatings, 2% calcium chloride and 1% pullulan, on sugar metabolism in postharvest Huangguan pear fruit were investigated during cold storage. The respiration rate, sugar content and composition, sucrose metabolism enzyme activities [acid invertase (AIV), neutral invertase (NI), sucrose synthase (SS), and sucrose phosphate synthase (SPS)] and expression of the AIV gene were analyzed during storage at $0^{\circ}C$ for 210 days. Coating treatments slowed the decrease of sucrose and hexose, the respiration rate, and the activities of AIV, NI, SS and SPS, thus maintaining high total soluble solids (TSS) and titratable acid (TA) contents in the fruit. There were no significant differences in AIV expression or activity between the treated and control groups of fruits. Both of the coatings could inhibit the activities of sucrose-cleaving enzymes, thus slowing the decrease of sugar content and maintaining high fruit quality during cold storage.

• Journal of Plant Biology
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• v.35 no.4
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• pp.333-338
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• 1992

Ginkgo (Ginkgo biloba L.) seeds were analyzed to determine the level of soluble sugars and insoluble starch during germination. Also the activities of the hydrolytic enzymes such as amylase, invertase and phosphatase were compared. As amylase activity was sharply increased, significant decline of starch was observed in the female gametophyte and increase of soluble sugars occurred concurrently. Invertase activity was gradually increased in cotyledon and radicle, while it was very low in dry seeds. In addition, phosphatase activity was variable only in radicle, and acid phosphatase showed higher activity than alkaline phosphatase.hatase.

• Journal of Plant Biology
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• v.34 no.2
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• pp.113-119
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• 1991

The contents of reducing sugar, sucrose, starch and fructose-2,6-bisphosphate (F-Z,$6-P_2$) in relation to the activities of amylase, invertase and fructose-1,6-bisphosphatase (FBPase) were investigated from the leaves of rice (Oryza sativa L. cv. Samjin) seedlings grown at $4^{\circ}C$ for 3 days_ In the seedlings, the contents of reducing sugar and sucrose were increased, but soluble and insoluble starch were declined. Under this condition, amylase activity was increased. but acid invertase activity was declined and alkaline invertase activity was not changed. Cytosolic and stromal FBPase activities were increased. But F-2,$6-P_2$ content was declined. It seemed that the increase of reducing sugar content might be due to the increased activity of amylase and the increase of sucrose content might be related to the increased activity of cytosolic FBPase, reduced content of F-Z,$6-P_2$ and reduced rate of hydrolysis of sucrose during the cold treatment. These results suggested that the changes in carbohydrate rnetabolim of rice seedlings under low temperature reflect one of the protection mechanism to the low temperature during the cold treatment.atment.