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A mitochondrial alkaline/neutral invertase isoform (A/N-InvC) functions in developmental energy-demanding processes in Arabidopsis

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Abstract

Recent findings demonstrate that alkaline/neutral invertases (A/N-Invs), enzymes that catalyze the breakdown of sucrose into glucose and fructose, are essential proteins in plant life. The fact that different isoforms are present in multiple locations makes them candidates for the coordination of metabolic processes. In the present study, we functionally characterized the encoding gene of a novel A/N-Inv (named A/N-InvC) from Arabidopsis, which localizes in mitochondria. A/N-InvC is expressed in roots, in aerial parts (shoots and leaves) and flowers. A detailed phenotypic analysis of knockout mutant plants (invc) reveals an impaired growth phenotype. Shoot growth was severely reduced, but root development was not affected as reported for A/N-InvA mutant (inva) plants. Remarkably, germination and flowering, two energy demanding processes, were the most affected stages. The effect of exogenous growth regulators led us to suggest that A/N-InvC may be modulating hormone balance in relation to the radicle emergence. We also show that oxygen consumption is reduced in inva and invc in comparison with wild-type plants, indicating that both organelle isoenzymes may play a fundamental role in mitochondrion functionality. Taken together, our results emphasize the involvement of mitochondrial A/N-Invs in developmental processes and uncover the possibility of playing different roles for the two isoforms located in the organelle.

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Abbreviations

ABA:

Abscisic acid

A/N-Inv:

Alkaline/neutral invertase

A/N-InvA or A/N-InvC:

At-A/N-InvA or At-A/N-InvC: Arabidopsis alkaline/neutral invertase A, or C

GA:

Gibberellic acid 3

Inv:

Invertase

inva or invc :

Arabidopsis mutant lacking alkaline/neutral invertase A or C

Suc:

Sucrose

wt:

Arabidopsis wild-type plants, ecotype Columbia 0

References

  • Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P, Stevenson DK, Zimmerman J, Barajas P, Cheuk R, Gadrinab C, Heller C, Jeske A, Koesema E, Meyers CC, Parker H, Prendis L, Ansari Y, Choy N, Deen H, Geralt M, Hazari N, Hom E, Karnes M, Mulholland C, Ndubaku R, Schmidt I, Guzman P, Aguilar-Henonin L, Schid M, Weigel D, Carter DE, Marchand T, Risseeuw E, Brogden D, Zeko A, Crosby WL, Berry CC, Ecker JR (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657

    Article  PubMed  Google Scholar 

  • Amor Y, Haigler CH, Johnson S, Wainscott M, Delmer DP (1995) A membrane-associated form of sucrose synthase and its potential role in synthesis of cellulose and callose in plants. Proc Natl Acad Sci USA 92:9353–9357

    Article  PubMed  CAS  Google Scholar 

  • Barratt DHP, Derbyshire P, Findlay K, Pike M, Wellner N, Lunn J, Feil R, Simpson C, Maule AJ, Smith AM (2009) Normal growth of Arabidopsis requires cytosolic invertase but not sucrose synthase. Proc Natl Acad Sci USA 106:13124–13129

    Article  PubMed  CAS  Google Scholar 

  • Bartoli CG, Yu J, Gómez F, Fernández L, McIntosh L, Foyer CH (2006) Inter-relationships between light and respiration in the control of ascorbic acid synthesis and accumulation in Arabidopsis thaliana leaves. J Exp Bot 57:1621–1631

    Article  PubMed  CAS  Google Scholar 

  • Boyes D, Zayed AM, Ascenzi R, McCaskill AJ, Hoffman NE, Davis KR, Görlach J (2001) Growth stage-based phenotypic analysis of Arabidopsis: a model for high throughput functional genomics in plants. Plant Cell 13:1499–1510

    PubMed  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  • Camacho-Pereira J, Meyer LE, Machado LB, Oliveira MF, Galina A (2009) Reactive oxygen species production by potato tuber mitochondria is modulated by mitochondrially bound hexokinase activity. Plant Physiol 149:1099–1110

    Article  PubMed  CAS  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  PubMed  CAS  Google Scholar 

  • Cormack BP, Valdivia RH, Falkow S (1996) FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173:33–38

    Article  PubMed  CAS  Google Scholar 

  • Dai H, Lo YS, Jane WN, Lee LW, Chiang KS (1998) Population heterogeneity of higher-plant mitochondria in structure and function. Eur J Cell Biol 75:198–209

    Article  PubMed  CAS  Google Scholar 

  • Estelle M, Somerville C (1987) Auxin-resistant mutants of Arabidopsis thaliana with an altered morphology. Mol Gen Genet 206:200–206

    Article  CAS  Google Scholar 

  • Giraud E, Van Aken O, Ho LH, Whelan J (2009) The transcription factor ABI4 is a regulator of mitochondrial retrograde expression of ALTERNATIVE OXIDASE1a. Plant Physiol 150:1286–1296

    Article  PubMed  CAS  Google Scholar 

  • Graham JW, Williams TC, Morgan M, Fernie AR, Ratcliffe RG, Sweetlove LJ (2007) Glycolytic enzymes associate dynamically with mitochondria in response to respiratory demand and support substrate channelling. Plant Cell 19:3723–3738

    Article  PubMed  CAS  Google Scholar 

  • Gupta KJ, Stoimenova M, Werner MK (2005) In higher plants, only root mitochondria, but not leaf mitochondria reduce nitrite to NO, in vitro and in situ. J Exp Bot 56:2601–2609

    Article  PubMed  CAS  Google Scholar 

  • Holdsworth MJ, Bentsink L, Soppe WJ (2008) Molecular networks regulating Arabidopsis seed maturation, after-ripening, dormancy and germination. New Phytol 179:33–54

    Article  PubMed  CAS  Google Scholar 

  • Ji X, Van den Ende W, Van Laere A, Cheng S, Bennett J (2005) Structure, evolution, and expression of the two invertase gene families of rice. J Mol Evol 60:615–634

    Article  PubMed  CAS  Google Scholar 

  • Jia L, Zhang B, Mao C, Li J, Wu Y, Wu P, Wu Z (2008) OsCYT-INV1 for alkaline/neutral invertase is involved in root cell development and reproductivity in rice (Oryza sativa L.). Planta 288:51–59

    Article  Google Scholar 

  • Koch KE (2004) Sucrose metabolism regulatory mechanisms and pivotal roles in sugar sensing and plant development. Curr Opin Plant Biol 7:235–246

    Article  PubMed  CAS  Google Scholar 

  • Lammens W, Roy KL, Schroeven L, Laere AV, Rabijins A, Van den Ende W (2009) Structural insights into glycoside hydrolase family 32 and 68 enzymes: functional implications. J Exp Bot 60:727–740

    Article  PubMed  CAS  Google Scholar 

  • Lo YS, Hsiao LJ, Jane WN, Charng YC, Dai H, Chiangy KS (2004) GFP-targeted mitochondria show heterogeneity of size, morphology, and dynamics in transgenic Nicotiana tabacum L. plants in vivo. Int J Plant Sci 165:949–955

    Article  Google Scholar 

  • Lou Y, Gou JY, Xue HW (2007) PIP5K9, an Arabidopsis phosphatidylinositol monophosphate kinase, interacts with a cytosolic invertase to negatively regulate sugar mediated root growth. Plant Cell 19:163–181

    Article  PubMed  CAS  Google Scholar 

  • Murayama S, Handa H (2007) Genes for alkaline/neutral invertase in rice: alkaline/neutral invertases are located in plant mitochondria and also in plastids. Planta 225:1193–1203

    Article  PubMed  CAS  Google Scholar 

  • Qi X, Wu Z, Li J, Mo X, Wu S, Chu J, Wu P (2007) AtCYT-INV1, a neutral invertase, is involved in osmotic stress-induced inhibition on lateral root growth in Arabidopsis. Plant Mol Biol 64:575–587

    Article  PubMed  CAS  Google Scholar 

  • Rhoads DM (2011) Plant mitochondrial retrograde regulation. In: Kempken F (ed) Plant mitochondria. Springer, Berlin, pp 411–437

    Chapter  Google Scholar 

  • Rhoads DM, Subbaiah CC (2007) Mitochondrial retrograde regulation in plants. Mitochondrion 7:177–194

    Article  PubMed  CAS  Google Scholar 

  • Roitsch T, González MC (2004) Function and regulation of plant invertases: sweet sensations. Trends Plant Sci 9:606–613

    Article  PubMed  CAS  Google Scholar 

  • Smeekens S, Hanson J, Rolland F (2010) Sugar signals and molecular networks controlling plant growth. Curr Opin Plant Biol 13:274–279

    Article  PubMed  CAS  Google Scholar 

  • Stewart CN Jr, Via LE (1993) A rapid CTAB DNA isolation technique useful for RAPD’s fingerprinting and other PCR applications. BioTech 14:748–758

    CAS  Google Scholar 

  • Sturm A (1999) Invertases. Primary structures, functions and roles in plant development and sucrose partitioning. Plant Physiol 121:1–7

    Article  PubMed  CAS  Google Scholar 

  • Subbaiah CC, Palaniappan A, Duncan K, Rhoads DM, Huber SC, Sachs MM (2006) Mitochondrial localization and putative signaling function of sucrose synthase in maize. J Biol Chem 281:15625–15635

    Article  PubMed  CAS  Google Scholar 

  • Szarka A, Horemans N, Passarella S, Tarcsay A, Örsi F, Salgó A, Bánhegyi G (2008) Demonstration of an intramitochondrial invertase activity and the corresponding sugar transporters of the inner mitochondrial membrane in Jerusalem artichoke (Helianthus tuberosus L.) tubers. Planta 228:765–775

    Article  PubMed  CAS  Google Scholar 

  • Vargas WA, Salerno GL (2010) The Cinderella story of sucrose hydrolysis: alkaline/neutral invertases, from cyanobacteria to unforeseen roles in plant cytosol and organelles. Plant Sci 178:1–8

    Article  CAS  Google Scholar 

  • Vargas WA, Cumino A, Salerno GL (2003) Cyanobacterial alkaline/neutral invertases. Origin of sucrose hydrolysis in the plant cytosol? Planta 216:951–960

    PubMed  CAS  Google Scholar 

  • Vargas WA, Pontis HG, Salerno GL (2007) Differential expression of alkaline and neutral invertases in response to environmental stresses: characterization of an alkaline isoform as a stress-response enzyme in wheat leaves. Planta 226:1535–1545

    Article  PubMed  CAS  Google Scholar 

  • Vargas WA, Pontis HG, Salerno GL (2008) New insights on sucrose metabolism: evidence for an active A/N Inv in chloroplasts uncovers a novel component of the intracellular carbon trafficking. Planta 227:795–807

    Article  PubMed  CAS  Google Scholar 

  • Vargas WA, Martin ML, Salerno GL (2012) Myths and facts on cytosolic sucrose hydrolysis. In: “Sucrose: properties, biosynthesis and health implications” Nova Science Publishers (in press)

  • Welham T, Pike J, Horst I, Flemetakis E, Katinakis P, Kaneko T, Sato S, Tabata S, Perry J, Parniske M, Wang TL (2009) A cytosolic invertase is required for normal growth and cell development in the model legume, Lotus japonicas. J Exp Bot 60:3353–3365

    Article  PubMed  CAS  Google Scholar 

  • Winter H, Huber SC (2000) Regulation of sucrose metabolism in higher plants: localization and regulation of activity of key enzymes. Crit Rev Plant Sci 19:31–67

    Article  CAS  Google Scholar 

  • Xiang L, Le Roy K, Bolouri-Moghaddam MR, Vanhaecke M, Lammens W, Rolland F, Van den Ende W (2011) Exploring the neutral invertase-oxidative stress defence connection in Arabidopsis thaliana. J Exp Bot 62:3849–3862

    Article  PubMed  CAS  Google Scholar 

  • Yao SG, Kodama R, Wang H, Ichii M, Taketa S, Yoshida H (2009) Analysis of the rice SHORT-ROOT5 gene revealed functional diversification of plant neutral/alkaline invertase family. Plant Sci 176:627–634

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We are very thankful to C. Fernández and M. Vidal for technical assistance. This research was funded by grants from CONICET (PIP 134 and PIP 102), Universidad Nacional de Mar del Plata (EXA 553/11), and FIBA.

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Authors and Affiliations

  1. Instituto de Investigaciones en Biodiversidad y Biotecnología (INBIOTEC-CONICET) and Fundación para Investigaciones Biológicas Aplicadas (FIBA), C.C. 1348, 7600, Mar del Plata, Argentina

    Mariana L. Martín, Leandra Lechner & Graciela L. Salerno

  2. Facultad de Ciencias Exactas y Naturales, Instituto de Investigaciones Biológicas, Universidad Nacional de Mar del Plata-CONICET, Mar del Plata, Argentina

    Eduardo J. Zabaleta

Authors
  1. Mariana L. Martín
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  2. Leandra Lechner
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  3. Eduardo J. Zabaleta
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  4. Graciela L. Salerno
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Corresponding author

Correspondence to Graciela L. Salerno.

Electronic supplementary material

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425_2012_1794_MOESM1_ESM.jpg

Supplemental Fig. 1 Comparison of Arabidopsis A/N-InvA and A/N-InvC gene expression. Analysis was performed from the Arabidopsis GENEVESTIGATOR browser (www.genevestigator.com). (JPEG 210 kb)

425_2012_1794_MOESM2_ESM.jpg

Supplemental Fig. 2 A/N-InvC-knockout Arabidopsis plants (invc). a Structure of A/N-InvC gene (locus At3g06500) composed of six exons and five introns. The coding and untranslated regions are gray and black depicted, respectively. The 5´and 3´untranslated regions are represented as white boxes. The T-DNA insertion site is indicated as a triangle (SALK_080181 mutant line). b Genotypic characterization of invc by PCR. The primers used (CLP, LBP and CRP), according to http://signal.salk.edu/tdnaprimers.2.html, are indicated by arrows in (a). Wild-type (wt) corresponds to Col-0 plants. Amplification products were analyzed on 1 % agarose gels and visualized after ethidium bromide staining. (JPEG 52 kb)

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Martín, M.L., Lechner, L., Zabaleta, E.J. et al. A mitochondrial alkaline/neutral invertase isoform (A/N-InvC) functions in developmental energy-demanding processes in Arabidopsis. Planta 237, 813–822 (2013). https://doi.org/10.1007/s00425-012-1794-8

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