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Post-Translational Derepression of invertase Activity in Source Leaves via Down-Regulation of invertase Inhibitor Expression Is Part of the Plant Defense Response

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Molecular Plant 2010年第6期3卷 1037-1048页

作者： Katharina B. Bonfig Andrea Gabler Uwe K. Simon Nora Luschin-Ebengreuth Martina Hatz Susanne Berger Naseem Muhammad Jurgen Zeier Alok K. Sinha Thomas Roitsch Julius-von-Sachs-Institut fuer Biowissenschaften Lehrstuhl for Pharmazeutische Biologie Universitaet Wuerzburg Julius-von-Sachs-Platz 2 97082 Wuerzburg Germany Institute of Plant Sciences Department of Plant Physiology University of Graz 8010 Graz Austria Zentrum for rnedizinische Grundlagenforschung (ZMF) Medizinische Universitat Graz Stiftingtalstraβe 24 A-8010 Graz Department of Biology Plant Biology Section University of Fribourg Route Albert Gockel 3 CH-1700 Fribourg Switzerland National Institute for Plant Genome Research Aruna Asaf Ali Road New Delhi 110067 India

There is increasing evidence that pathogens do not only elicit direct defense responses, but also cause pronounced changes in primary carbohydrate metabolism. Cell-wall-bound invertases belong to the key regulators of carbohydrate partitioning and source-sink relations. Whereas studies have focused so far only on the transcriptional induction of invertase genes in response to pathogen infection, the role of post-translational regulation of invertase activity has been neglected and was the focus of the present study. Expression analyses revealed that the high mRNA level of one out of three proteinaceous invertase inhibitors in source leaves of Arabidopsis thaliana is strongly repressed upon infection by a virulent strain of Pseudomonas syringae pv. tomato DC3000. This repression is paralleled by a decrease in invertase inhibitor activity. The physiological role of this regulatory mechanism is revealed by the finding that in situ invertase activity was detectable only upon infection by P. syringae. In contrast, a high invertase activity could be measured in vitro in crude and cell wall extracts prepared from both infected and non-infected leaves. The discrepancy between the in situ and in vitro invertase activity of control leaves and the high in situ invertase activity in infected leaves can be explained by the pathogen-dependent repression of invertase inhibitor expression and a concomitant reduction in invertase inhibitor activity. The functional importance of the release of invertase from post-translational inhibition for the defense response was substantiated by the application of the competitive chemical invertase inhibitor acarbose. Posttranslational inhibition of extracellular invertase activity by infiltration of acarbose in leaves was shown to increase the susceptibility to P. syringae. The impact of invertase inhibition on spatial and temporal dynamics of the repression of photosynthesis and promotion of bacterial growth during pathogen infection supports a

关键词： Acarbose Arabidopsis defense responses invertase inhibitor protein invertase plant-microbe interactions Pseudomonas syringae.

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Vacuolar invertase genes SbVIN1 and SbVIN2 are differently associated with stem and grain traits in sorghum (Sorghum bicolor)

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The Crop Journal 2020年第2期8卷 299-312页

作者： Yunhua Chi Kimani Wilson Zhiquan Liu Xiaoyuan Wu Li Shang Limin Zhang Haichun Jing Huaiqing Hao Key Laboratory of Plant Resources Institute of BotanyChinese Academy of SciencesBeijing 100093China University of Chinese Academy of Sciences Beijing 100049China Engineering Laboratory for Grass-based Livestock Husbandry Chinese Academy of SciencesBeijing 100093China

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.

关键词： Sorghum(Sorghum bicolor) Vacuolar invertase SbVIN1 SbVIN2 Enzymatic properties Association analysis

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Tomato Ovary-to-Fruit Transition is Characterized by a Spatial Shift of mRNAs for Cell Wall invertase and its Inhibitor with the Encoded Proteins Localized to Sieve Elements

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Molecular Plant 2015年第2期8卷 315-328页

作者： William M. Palmer School of Environmental and Life Sciences The University of Newcastle NSW 2308 Callaghan Australia Australia-China Research Centre for Crop Improvement The University of Newcastle NSW 2308 Callaghan Australia Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 300 Fenglin Road Shanghai 200032 China These authors contributed equally to the article. Present address: College of Life Sciences. Central China Normal University Wuhan 430079 China

Central to understanding fruit development is to elucidate the processes mediating a successful transition from pre-pollination ovaries to newly set fruit, a key step in establishing fruit yield potential. In tomato, cell wall invertase （CWIN） LIN5 and its inhibitor INH1 are essential for fruit growth. However, the molecular and cellular basis by which they exert their roles in ovary-to-fruit transition remains unknown. To address this issue, we conducted a study focusing on ovaries and fruitlets at 2 days before and 2 days after anthesis, respectively. In situ hybridization analyses revealed that LIN5 and INH1 exhibited a dispersed expression in ovaries compared with their phloem-specific expression in fruitlets. Remarkably, LIN5 and INH1 proteins were immunologically co-localized to cell walls of sieve elements （SEs） in ovaries immediately prior to anthesis and in young fruitlets, but were undetectable in provascular bundles of younger ovaries. A burst in CWlN activity occurred during ovary-to-fruit transition. Interestingly, the ovaries, but not the fruit- lets, exhibited high expression of a defective invertase, SldeCWIN1, an ortholog of which is known to enhance inhibition of INH on CWlN activity in tobacco. Imaging of a fluorescent symplasmic tracer indicated an apoplasmic phloem unloading pathway operated in ovaries, contrary to the previously observed symplasmic unloading pathway in fruit pericarp. These new data indicate that （1） a phloem-specific patterning of the CWIN and INH mRNAs is induced during ovary-to-fruit transition, and （2） LIN5 protein functions specifically in walls of SEs and increases its activity during ovary-to-fruit transition, probably to facilitate phloem unloading and to generate a glucose signal positively regulating cell division, hence fruit set.

关键词： fruit set invertase invertase inhibitor phloem unloading sugar signaling

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Sugar Input, Metabolism, and Signaling Mediated by invertase： Roles in Development, Yield Potential, and Response to Drought and Heat

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Molecular Plant 2010年第6期3卷 942-955页

作者： Yong-Ling Ruan Ye Jin Yue-Jian Yang Guo-Jing Li John S. Boyer School of Environmental and Life Sciences University of Newcastle Callaghan NSW 2308 Australia Australia-China Research Centre for Crop Improvement University of Newcastle Callaghan NSW 2308 Australia College of Life Sciences Central China Normal University Wuhan 430079 China China-Australia Research Centre for Crop Improvement Zhejiang Academy of Agricultural Sciences Hangzhou 310021 China College of Earth Ocean and Environment and College of Agriculture and Natural Resources University of Delaware Lewes DE 19958 USA

invertase （INV） hydrolyzes sucrose into glucose and fructose, thereby playing key roles in primary metabolism and plant development. Based on their pH optima and sub-cellular locations, INVs are categorized into cell wall, cytoplasmic, and vacuolar subgroups, abbreviated as CWlN, CIN, and VlN, respectively. The broad importance and implications of INVs in plant development and crop productivity have attracted enormous interest to examine INV function and regulation from multiple perspectives. Here, we review some exciting advances in this area over the last two decades, focusing on （1） new or emerging roles of INV in plant development and regulation at the post-translational level through interaction with inhibitors, （2） cross-talk between INV-mediated sugar signaling and hormonal control of development, and （3） sugar- and INV-mediated responses to drought and heat stresses and their impact on seed and fruit set. Finally, we discuss major questions arising from this new progress and outline future directions for unraveling mechanisms underlying INV-mediated plant development and their potential applications in plant biotechnology and agriculture.

关键词： Crop yield drought heat stress invertase reproductive development seed and fruit set sugar metabolism sugar signaling.

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Genome-Wide Identification and Expression Profiling Suggest that invertase Genes Function in Silique Development and the Response to Sclerotinia sclerotiorum in Brassica napus

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Phyton-International Journal of Experimental Botany 2020年第2期89卷 253-273页

作者： Jingsen Liu Jinqi Ma Ai Lin Chao Zhang Bo Yang Liyuan Zhang Lin Huang Jiana Li Chongqing Rapeseed Engineering Research Center College of Agronomy and BiotechnologyChongqing400715China Academy of Agricultural Sciences Southwest UniversityChongqing400715China

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 *** this study,we identified 27 members of the BnaINV family in Brassica *** 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 *** also analyzed the expression of the BnaINVs in different tissues and developmental stages in the *** cultivar Zhongshuang 11 using *** 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 *** used WGCNA(weighted coexpression network analysis)to uncover BnaINV regulatory ***,we explored the expression patterns of several BnaINV genes in cultivars with long(Zhongshuang 4)and short(Ningyou 12)*** results suggest that BnaINVs play important roles in the growth and development of rapeseed siliques and the defense response against *** findings could facilitate the breeding of high-yielding *** cultivars with strong disease resistance.

关键词： Brassica napus invertase Sclerotinia WGCNA RNA-seq

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Effects of Iron and Aluminum Oxides and Kaolinite on Adsorption and Activities of invertase *1

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Pedosphere 1998年第3期8卷 251-260页

作者： HUANGQIAOYUN JIANGMINGHUA 等 DepartmentofResources EnvironmentandAgrochemistryHuazhongAgricultualUniversiyWuhan430070（China） Dep

Experiments were conducted to study the influences of synthetic bayerite, non crystalline aluminum oxide (N AlOH), goethite, non crystalline iron oxide (N FeOH) and kaolinite on the adsorption, activity, kinetics and thermal stability of invertase. Adsorption of invertase on iron, aluminum oxides fitted Langmuir equation. The amount of invertase held on the minerals followed the sequence kaolinite > goethite > N AlOH > bayerite > N FeOH. No correlation was found between enzyme adsorption and the specific surface area of minerals examined. The differences in the surface structure of minerals and the arrangement of enzymatic molecules on mineral surfaces led to the different capacities of minerals for enzyme adsorption. The adsorption of invertase on bayerite, N AlOH, goethite, N FeOH and kaolinite was differently affected by pH. The order for the activity of invertase adsorbed on minerals was N FeOH > N AlOH > bayerite > reak goethite > kaolinite. The inhibition effect of minerals on enzyme activity was kaolinite > crystalline oxides > non crystalline oxides. The pH optimum of iron oxide and aluminum oxide invertase complexes was similar to that of free enzyme (pH 4.0), whereas the pH optimum of kaolinite inv ertase complex was one pH unit higher than that of free enzyme. The affinity to substrate and the maximum reaction velocity as well as the thermal stability of combined invertase were lower than those of the free enzyme.

关键词： enzyme activity enzyme adsorption invertase kaolinite oxides

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Using a Membrane Reactor for Sucrose Hydrolysis： The Effect of Reactor＇s Design on invertase Stability

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Journal of Environmental Science and Engineering(A) 2013年第9期2卷 582-592页

作者： Francesco di Addezio Ester Junko Yoriyaz Laura Cantarella Maria Cantarella Michele Vitolo Department of lndustrial and lnformation Engineering and Economics University of L 'Aquila L 'Aquila 67100 Italy Department of Biochemical and Pharmaceutical Technology University of Sao Paulo S6o Paulo 05508900 Brazil Department of Civil and Mechanical Engineering University of Cassino and of Lazio Meridionale Cassino 03043 Italy

invertase hydrolyses sucrose, produces inverted sugar syrup, which is used, mainly, as a food composition in industries. To carry out the hydrolysis properly, the invertase should be stable in the soluble form through a considerable reaction period and recovered afterwards. The chosen reactor was a CSTR-type （continuous stirred tank reactor-type） coupled with an UFM （ultrafiltration-membrane）, the so-called MR （membrane reactor）. The varied parameters were： sucrose concentration （10-300 mM）, temperature （5-65 ℃）, reaction volume （14 mL and 65 mL）, stirring （100-500 rpm）, volumetric feeding rate （2.2-12 mL/h） and UFM MWCO （molecular weight cut off）（10, 20, 30, 50 and 100 kDa）. The invertase kinetic constants （KM = 23.5 mM; Vmax = 2,758 μmolgluJmin-mgprot; Ea = 9.1 kcal/mol） and the temperature deactivation energy （Ead= 20 kcal/mol） were calculated. Moreover, the invertase was unstable as the MR capacity diminished and the agitation increased up to 500 rpm most likely due to the damaging effect of shearing forces （present inside the MR） on the invertase molecule. Finally, both the MWCO and the chemical nature of the UFM affected the invertase stability along the hydrolysis. The enzyme stability increased as the UFM cut-off decreased, the highest value being observed with the 10 kDa-UFM.

关键词： invertase sucrose membrane reactor.

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Research Progress in Plant invertase

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Agricultural Science & Technology 2017年第1期18卷 21-22,34页

作者： Yulin ZHANG Yuqiao CAO Nannan HU Sanmenxia Branch of Henan Tobacco Company Sanmenxia 472000 China College of Tobacco Science Henan Agricultural University Zhengzhou 450002 China Shanzhou Branch of Sanmenxia Tobacco Company Shanzhou 472100 China

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 ***, this article summarized theresearch progress of invertase in biological characteristics, molecular characteristics and expression regulation.

关键词： invertase Sucrose catabolism Expression regulation

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Functional characterization of an invertase gene,PstINV1 from Puccinia striiformis ***

Functional characterization of an invertase gene,PstINV1 fro...

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中国植物病理学会2015年学术年会

作者： CHANG Qing LIU Jie HUAI Bao-yu ZHENG Pei-jing LI Dan HUANG Li-li KANG Zhen-sheng College of Plant Protection State Key Laboratory of Crop Stress Biology for Arid AreasNorthwest A&F University College of Life Sciences State Key Laboratory of Crop Stress Biology for Arid AreasNorthwest A&F University

Puccinia striiformis ***（Pst） is the causal agent of wheat stripe rust which is one of the most serious diseases on wheat *** the mutation rate of Pst is really high and the virulence changes frequently,the pattern of Pst nutrition absorption is highly conserved as an obligate biotrophic *** can only take up hexoses from host cells,thus invertases,hydrolyzing sucrose into monosaccharide,are very important for Pst *** this paper,the ORF of an invertase gene PsINV from Pst was cloned for the first time and the amino acid sequence of PsINV showed a specific structure of rust *** expression studies showed that this gene was highly expressed during the haustoria *** expression of PsINV in Saccharomyces cerevisiae could rescue the defect of invertases,indicating that PsINV is an invertase *** enzymatic protein of PsINV was obtained by eukaryotic expression,and then the biochemical characteristics were *** of different PsINV domains clarified the relationship between the properties of this protein and its special *** down of PsINV by host induced gene silencing leaded to abnormal sporulation structures and eventually reduced urediospores on the infected wheat *** results demonstrate that PsINV plays a key role during Pst infection of wheat.

关键词： Puccinia striiformis West.f.sp.tritici invertase eukaryotic expression biochemical characteristics host induced gene silencing

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Production of invertases by Anamorphic(Aspergillus nidulans)and Teleomorphic(Emericela nidulans)Fungi under Submerged Fermentation Using Rye Flour as Carbon Source

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Advances in Microbiology 2013年第5期3卷 421-429页

作者： Juliana Nunes e Oliveira Alves Joao Atílio Jorge Luis Henrique Souza Guimaraes Department of Biology Faculty of PhylosophySciences and Letters of Ribeirao PretoUniversity of Sao PauloSao PauloBrazil

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.

关键词： invertase Fructofuranosidase Aspergillus Emericela

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