In higher plants, vacuolar invertases play essential roles in sugar metabolism, organ development, and sink strength. In sorghum(Sorghum bicolor), two vacuolar invertase genes,Sb VIN1(Sobic. 004 G004800) and Sb VIN2(S...In higher plants, vacuolar invertases play essential roles in sugar metabolism, organ development, and sink strength. In sorghum(Sorghum bicolor), two vacuolar invertase genes,Sb VIN1(Sobic. 004 G004800) and Sb VIN2(Sobic. 006 G160700) have been reported, but their enzymatic properties and functional differences are largely unknown. We combined molecular, biochemical and genomic approaches to investigate their roles in sorghum stem and grain traits. Sb VIN1 and Sb VIN2 showed different expression levels in internodes,leaves, and panicles, indicating that their importance in each organ was different. In an in vitro sucrose hydrolysis assay, proteins of both genes heterologously expressed in Pichia pastoris displayed similar enzyme properties including the same optimum reaction p H(5)and similar Kmfor sucroe(49 mmol L-1 and 45 mmol L-1 for Sb VIN1 and Sb VIN2,respectively). The optimum reaction temperatures of Sb VIN1 and Sb VIN2 were 45 °C and65 °C, respectively. Sb VIN2 showed higher tolerance to high temperature than Sb VIN1. We characterized the sequence variation of these two vacuolar invertase genes in a panel of 216 diverse inbred lines of sweet and grain sorghum and performed gene-based association analysis. Sb VIN1 showed significant associations with stem traits including stem length,stem diameter, internode number, stem fresh weight, and Brix, as well as grain traits including hundred-grain weight and grain width. Significantly associated variation sites were mainly in 5′ upstream and intron regions. Sb VIN2 only associated with grain width and stem water-soluble carbohydrates(WSCs) content. We conclude that the vacuolar invertase genes Sb VIN1 and Sb VIN2 are differently associated with stem and grain traits in sorghum.展开更多
Abstract Invertase is a key enzyme in sucrose catabolism and crucial for plant assimilate distribution.With the development of molecularbiology,a lot of invertsae genes were cloned recently,and significant progress ha...Abstract Invertase is a key enzyme in sucrose catabolism and crucial for plant assimilate distribution.With the development of molecularbiology,a lot of invertsae genes were cloned recently,and significant progress have been made in regulators on the expression of invertase genes.Thus,this article summarized theresearch progress of invertase in biological characteristics,molecular characteristics and expression regulation.展开更多
A hybrid GMDH neural network model has been developed in order to predict the partition coefficients of invertase from Baker's yeast. ATPS experiments were carried out changing the molar average mass of PEG(1500–...A hybrid GMDH neural network model has been developed in order to predict the partition coefficients of invertase from Baker's yeast. ATPS experiments were carried out changing the molar average mass of PEG(1500–6000 Da), p H(4.0–7.0), percentage of PEG(10.0–20.0 w/w), percentage of MgSO_4(8.0–16.0 w/w), percentage of the cell homogenate(10.0–20.0 w/w) and the percentage of MnSO_4(0–5.0 w/w) added as cosolute. The network evaluation was carried out comparing the partition coefficients obtained from the hybrid GMDH neural network with the experimental data using different statistical metrics. The hybrid GMDH neural network model showed better fitting(AARD = 32.752%) as well as good generalization capacity of the partition coefficients of the ATPS than the original GMDH network approach and a BPANN model. Therefore hybrid GMDH neural network model appears as a powerful tool for predicting partition coefficients during downstream processing of biomolecules.展开更多
Virus-free sugarcane seedlings have improved biomass and sucrose content compared with ordinary seedlings, and sucrose invertases are key enzymes regulating sugarcane growth and sucrose accumulation. In this study, th...Virus-free sugarcane seedlings have improved biomass and sucrose content compared with ordinary seedlings, and sucrose invertases are key enzymes regulating sugarcane growth and sucrose accumulation. In this study, the differences in the expression levels of 3 invertase genes, CWI, SAI and NI, between virus- free and ordinary sugarcane seedlings were analyzed. Compared with ordinary sugarcane plants, the expression of CWI was mainly up-regulated in immature leaves and stems at elongation stage and leaves and immature internodes at maturation stage, and especially, greatly up-regulated in immature interuedes at maturation stage of virus-free plants. The expression of SAI and NI were mainly up-regnlated in leaves and immature internedes of virus-free plants at maturation stage, which might be beneficial to sugar accumulation and rapid utilization of monosaccharide in the stalks of virus-free plants. It is further indicated that virus-free treatment could significantly improve the expression of sucrose invertases at late growth period, and might facilitate the increase of plant biomass.展开更多
Sugar is an important material basis in fruit development,and strawberry fruit flavour and sweetness largely depend on the sugar content and variety.Invertases(INVs)play an important role in the regulation of sugar ac...Sugar is an important material basis in fruit development,and strawberry fruit flavour and sweetness largely depend on the sugar content and variety.Invertases(INVs)play an important role in the regulation of sugar accumulation because they irreversibly catalyse the hydrolysis of sucrose into the corresponding nucleoside diphosphate-glucose,glucose or fructose in fruit.In this work,we provided a comprehensive analysis of the INV gene family in octoploid strawberry(Fragaria×ananassa),including the gene structure,chromosomal locations,conserved domains,and gene evolution and expression profiles during strawberry fruit development.Our study revealed that polyploid events resulted in the abundant amplification(almost three-or four-fold)of the INV gene in the F.×ananassa genome,and these amplified INV genes showed dominant expression in strawberry fruit.More than half of the FaINVs transcripts with low expression had incomplete coding sequences by alternative splicing.Previous studies have shown that cell wall invertases(CWINV)are involved in the regulation of phloem unloading and sink strength establishment.The expression of FaCWINV1 was markedly upregulated during fruit development and strongly expressed in ripe fruit.Moreover,a significant correlation was observed between the total sugar content and the FaCWINV1 expression level.These findings suggest that FaCWINV1 may be involved in sugar accumulation in strawberry fruit.Taken together,the results of our study will be beneficial for further research into the functions of INVs in the regulation of fruit ripening.展开更多
Invertase(INV),a key enzyme in sucrose metabolism,irreversibly catalyzes the hydrolysis of sucrose to glucose and fructose,thus playing important roles in plant growth,development,and biotic and abiotic stress respons...Invertase(INV),a key enzyme in sucrose metabolism,irreversibly catalyzes the hydrolysis of sucrose to glucose and fructose,thus playing important roles in plant growth,development,and biotic and abiotic stress responses.In this study,we identified 27 members of the BnaINV family in Brassica napus.We constructed a phylogenetic tree of the family and predicted the gene structures,conserved motifs,cis-acting elements in promoters,physicochemical properties of encoded proteins,and chromosomal distribution of the BnaINVs.We also analyzed the expression of the BnaINVs in different tissues and developmental stages in the B.napus cultivar Zhongshuang 11 using qRT-PCR.In addition,we analyzed RNA-sequencing data to explore the expression patterns of the BnaINVs in four cultivars with different harvest indices and in plants inoculated with the pathogenic fungus Sclerotinia sclerotiorum.We used WGCNA(weighted coexpression network analysis)to uncover BnaINV regulatory networks.Finally,we explored the expression patterns of several BnaINV genes in cultivars with long(Zhongshuang 4)and short(Ningyou 12)siliques.Our results suggest that BnaINVs play important roles in the growth and development of rapeseed siliques and the defense response against pathogens.Our findings could facilitate the breeding of high-yielding B.napus cultivars with strong disease resistance.展开更多
Dealt with in this paper is the relationship between Cu contents of soil and activity parameters for soil invertase and urease from the Changtai loquat orchards in Fujian Province. The soil invertase activity and maxi...Dealt with in this paper is the relationship between Cu contents of soil and activity parameters for soil invertase and urease from the Changtai loquat orchards in Fujian Province. The soil invertase activity and maximum reaction velocity (enzyme kinetic parameters: Vmax and Vmax/Km) have no remarkable negative correlation with Cu concentrations, but Cu concentrations have an obvious positive effect on invertase Km, indicating a reversible competitive restraint on the reaction between Cu and invertase. The soil urease activity and maximum reaction velocity (Vmax,Vmax/Km) have a remarkable negative correlation with Cu concentrations, and in this aspect, they can be used as ecological indices of the soil polluted by Cu. A slightly variable Km indicates the non-reversible competitive restraint between Cu and soil urease. Cu concentration= 2.00 mg·L-1 in soil is an indicative of the moderate Cu pollution level.展开更多
The production of invertases by anamorph (A. nidulans) and teleomorph (E. nidulans) was investigated. The best level of extracellular enzymatic production for anomorph was obtained in Khanna medium containing sucrose ...The production of invertases by anamorph (A. nidulans) and teleomorph (E. nidulans) was investigated. The best level of extracellular enzymatic production for anomorph was obtained in Khanna medium containing sucrose as carbon source, whereas for teleomorph the best production was archived using M5 medium containing inulin as carbon source. Despite this, rye flour was selected as carbon source. The extracellular enzyme production was higher for teleomorph than that observed for anomorph for all carbon sources used. The enzyme production was inhibited by the addition of fructose and glucose in the medium containing rye flour as carbon source. The best conditions to recover the higher enzymatic activity were temperature of 54℃ - 62℃ and pH of 4.8 5.6 for both enzymes determined by experimental design (CCRD). The stability of the temperatures at 40℃ and 50℃were similar for both enzymes. The invertases from the anomorph and teleomorph were activated by Mn2+, but the response of each one towards the presence of this cation was different with best activation observed for the anomorph enzyme (+80%). The extracellular enzymes were able to hydrolyze inulin, sucrose and raffinose. However, the affinity was higher for sucrose than inulin. In conclusion, the carbon source assimilation and the invertase production, as well as the enzymes properties, were different for the anomorph and teleomorph mycelia.展开更多
基金the National Natural Science Foundation of China(No.31471570,No.31461143023)Ministry of Science and Technology of the People's Republic of China(2015BAD15B03).
文摘In higher plants, vacuolar invertases play essential roles in sugar metabolism, organ development, and sink strength. In sorghum(Sorghum bicolor), two vacuolar invertase genes,Sb VIN1(Sobic. 004 G004800) and Sb VIN2(Sobic. 006 G160700) have been reported, but their enzymatic properties and functional differences are largely unknown. We combined molecular, biochemical and genomic approaches to investigate their roles in sorghum stem and grain traits. Sb VIN1 and Sb VIN2 showed different expression levels in internodes,leaves, and panicles, indicating that their importance in each organ was different. In an in vitro sucrose hydrolysis assay, proteins of both genes heterologously expressed in Pichia pastoris displayed similar enzyme properties including the same optimum reaction p H(5)and similar Kmfor sucroe(49 mmol L-1 and 45 mmol L-1 for Sb VIN1 and Sb VIN2,respectively). The optimum reaction temperatures of Sb VIN1 and Sb VIN2 were 45 °C and65 °C, respectively. Sb VIN2 showed higher tolerance to high temperature than Sb VIN1. We characterized the sequence variation of these two vacuolar invertase genes in a panel of 216 diverse inbred lines of sweet and grain sorghum and performed gene-based association analysis. Sb VIN1 showed significant associations with stem traits including stem length,stem diameter, internode number, stem fresh weight, and Brix, as well as grain traits including hundred-grain weight and grain width. Significantly associated variation sites were mainly in 5′ upstream and intron regions. Sb VIN2 only associated with grain width and stem water-soluble carbohydrates(WSCs) content. We conclude that the vacuolar invertase genes Sb VIN1 and Sb VIN2 are differently associated with stem and grain traits in sorghum.
文摘Abstract Invertase is a key enzyme in sucrose catabolism and crucial for plant assimilate distribution.With the development of molecularbiology,a lot of invertsae genes were cloned recently,and significant progress have been made in regulators on the expression of invertase genes.Thus,this article summarized theresearch progress of invertase in biological characteristics,molecular characteristics and expression regulation.
基金CAPES and Brazilian National Council of Research (CNPq) (Grant 407684/2013-1) for the financial support
文摘A hybrid GMDH neural network model has been developed in order to predict the partition coefficients of invertase from Baker's yeast. ATPS experiments were carried out changing the molar average mass of PEG(1500–6000 Da), p H(4.0–7.0), percentage of PEG(10.0–20.0 w/w), percentage of MgSO_4(8.0–16.0 w/w), percentage of the cell homogenate(10.0–20.0 w/w) and the percentage of MnSO_4(0–5.0 w/w) added as cosolute. The network evaluation was carried out comparing the partition coefficients obtained from the hybrid GMDH neural network with the experimental data using different statistical metrics. The hybrid GMDH neural network model showed better fitting(AARD = 32.752%) as well as good generalization capacity of the partition coefficients of the ATPS than the original GMDH network approach and a BPANN model. Therefore hybrid GMDH neural network model appears as a powerful tool for predicting partition coefficients during downstream processing of biomolecules.
基金Supported by"863"Program(2013AA102604-1)Natural Science Foundation of Hainan Province(20163124)+1 种基金Basal Research Fund for Central Public-interest Scientific Institute(ITBB140503)Earmarked Fund for China Agriculture Research System(CARS-20-2-5)
文摘Virus-free sugarcane seedlings have improved biomass and sucrose content compared with ordinary seedlings, and sucrose invertases are key enzymes regulating sugarcane growth and sucrose accumulation. In this study, the differences in the expression levels of 3 invertase genes, CWI, SAI and NI, between virus- free and ordinary sugarcane seedlings were analyzed. Compared with ordinary sugarcane plants, the expression of CWI was mainly up-regulated in immature leaves and stems at elongation stage and leaves and immature internodes at maturation stage, and especially, greatly up-regulated in immature interuedes at maturation stage of virus-free plants. The expression of SAI and NI were mainly up-regnlated in leaves and immature internedes of virus-free plants at maturation stage, which might be beneficial to sugar accumulation and rapid utilization of monosaccharide in the stalks of virus-free plants. It is further indicated that virus-free treatment could significantly improve the expression of sucrose invertases at late growth period, and might facilitate the increase of plant biomass.
基金This work was funded by the Major Project for Breeding New Varieties of Jiangsu Province,China(PZCZ201721)the National Horticulture Germplasm Resources Center,China(NHGRC2020-NH16)the National Crop Germplasm Resources Protection of the Ministry of Agriculture and Rural Affairs of China(19200361).
文摘Sugar is an important material basis in fruit development,and strawberry fruit flavour and sweetness largely depend on the sugar content and variety.Invertases(INVs)play an important role in the regulation of sugar accumulation because they irreversibly catalyse the hydrolysis of sucrose into the corresponding nucleoside diphosphate-glucose,glucose or fructose in fruit.In this work,we provided a comprehensive analysis of the INV gene family in octoploid strawberry(Fragaria×ananassa),including the gene structure,chromosomal locations,conserved domains,and gene evolution and expression profiles during strawberry fruit development.Our study revealed that polyploid events resulted in the abundant amplification(almost three-or four-fold)of the INV gene in the F.×ananassa genome,and these amplified INV genes showed dominant expression in strawberry fruit.More than half of the FaINVs transcripts with low expression had incomplete coding sequences by alternative splicing.Previous studies have shown that cell wall invertases(CWINV)are involved in the regulation of phloem unloading and sink strength establishment.The expression of FaCWINV1 was markedly upregulated during fruit development and strongly expressed in ripe fruit.Moreover,a significant correlation was observed between the total sugar content and the FaCWINV1 expression level.These findings suggest that FaCWINV1 may be involved in sugar accumulation in strawberry fruit.Taken together,the results of our study will be beneficial for further research into the functions of INVs in the regulation of fruit ripening.
基金This work was supported by grants from the National Natural Science Foundation of China(31830067)National Key Research and Development Program of China(2018YFD0100504-05)the“111”Project(B12006).
文摘Invertase(INV),a key enzyme in sucrose metabolism,irreversibly catalyzes the hydrolysis of sucrose to glucose and fructose,thus playing important roles in plant growth,development,and biotic and abiotic stress responses.In this study,we identified 27 members of the BnaINV family in Brassica napus.We constructed a phylogenetic tree of the family and predicted the gene structures,conserved motifs,cis-acting elements in promoters,physicochemical properties of encoded proteins,and chromosomal distribution of the BnaINVs.We also analyzed the expression of the BnaINVs in different tissues and developmental stages in the B.napus cultivar Zhongshuang 11 using qRT-PCR.In addition,we analyzed RNA-sequencing data to explore the expression patterns of the BnaINVs in four cultivars with different harvest indices and in plants inoculated with the pathogenic fungus Sclerotinia sclerotiorum.We used WGCNA(weighted coexpression network analysis)to uncover BnaINV regulatory networks.Finally,we explored the expression patterns of several BnaINV genes in cultivars with long(Zhongshuang 4)and short(Ningyou 12)siliques.Our results suggest that BnaINVs play important roles in the growth and development of rapeseed siliques and the defense response against pathogens.Our findings could facilitate the breeding of high-yielding B.napus cultivars with strong disease resistance.
基金project is funded jointly by Fujian Provincial Natural Science Foundation (Z0516048)Fuujian Educational Department Fundation (JA06038) and Putian Key Science and Technology Project (2005SO2)
文摘Dealt with in this paper is the relationship between Cu contents of soil and activity parameters for soil invertase and urease from the Changtai loquat orchards in Fujian Province. The soil invertase activity and maximum reaction velocity (enzyme kinetic parameters: Vmax and Vmax/Km) have no remarkable negative correlation with Cu concentrations, but Cu concentrations have an obvious positive effect on invertase Km, indicating a reversible competitive restraint on the reaction between Cu and invertase. The soil urease activity and maximum reaction velocity (Vmax,Vmax/Km) have a remarkable negative correlation with Cu concentrations, and in this aspect, they can be used as ecological indices of the soil polluted by Cu. A slightly variable Km indicates the non-reversible competitive restraint between Cu and soil urease. Cu concentration= 2.00 mg·L-1 in soil is an indicative of the moderate Cu pollution level.
基金supported by grants from the Fundacao de Amparo à Pesquisa do Estado de Sao Paulo(FAPESP)and Conselho de Desenvolvimento Científico e Tecnol-ógico(CNPq).
文摘The production of invertases by anamorph (A. nidulans) and teleomorph (E. nidulans) was investigated. The best level of extracellular enzymatic production for anomorph was obtained in Khanna medium containing sucrose as carbon source, whereas for teleomorph the best production was archived using M5 medium containing inulin as carbon source. Despite this, rye flour was selected as carbon source. The extracellular enzyme production was higher for teleomorph than that observed for anomorph for all carbon sources used. The enzyme production was inhibited by the addition of fructose and glucose in the medium containing rye flour as carbon source. The best conditions to recover the higher enzymatic activity were temperature of 54℃ - 62℃ and pH of 4.8 5.6 for both enzymes determined by experimental design (CCRD). The stability of the temperatures at 40℃ and 50℃were similar for both enzymes. The invertases from the anomorph and teleomorph were activated by Mn2+, but the response of each one towards the presence of this cation was different with best activation observed for the anomorph enzyme (+80%). The extracellular enzymes were able to hydrolyze inulin, sucrose and raffinose. However, the affinity was higher for sucrose than inulin. In conclusion, the carbon source assimilation and the invertase production, as well as the enzymes properties, were different for the anomorph and teleomorph mycelia.