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Genes for alkaline/neutral invertase in rice: alkaline/neutral invertases are located in plant mitochondria and also in plastids

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Abstract

Two cDNA clones (OsNIN1 and OsNIN3) encoding an alkaline/neutral invertase localized in organelles were identified from rice. The deduced amino acid sequences of these cDNA clones showed high homology to other plant alkaline/neutral invertases. Semi-quantitative reverse transcription polymerase chain reaction revealed that the expression of OsNIN1 was constitutive and independent of organ difference, although its expression level was low. Analyses using five types of web software for the prediction of protein localization in the cell, Predotar, PSORT, Mitoprot, TargetP, and ChloroP, strongly supported the possibility that OsNIN1 is transported into the mitochondria and that OsNIN3 is transported into plastids. Transient expression of fusion proteins combining the amino terminal region of these two proteins with sGFP demonstrated that N-OsNIN1::GFP and N-OsNIN3::GFP fusion proteins were transported into the mitochondria and plastids, respectively. We expressed the OsNIN1 protein in vitro and revealed that the translated protein had an invertase activity. These results clearly indicate that some of alkaline/neutral invertases are located in plant organelles, mitochondria and plastids, and that they might have a novel physiological function in plant organelles.

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Abbreviations

CTAB:

Cetyl trimethyl ammonium bromide

GFP:

Green fluorescent protein

RT-PCR:

Reverse transcription polymerase chain reaction

SUS:

Sucrose synthase

UDP:

Uridine 5′-diphosphate

References

  • Chen JQ, Black CC (1992) Biochemical and immunological properties of alkaline invertase isolated from sprouting soybean hypocotyls. Arch Biochem Biophys 295:61–69

    Article  PubMed  CAS  Google Scholar 

  • Chiu W, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a reporter in plants. Curr Biol 6:325–330

    Article  PubMed  CAS  Google Scholar 

  • Claros MG, Vincens P (1996) Computational method to predict mitochondrially imported proteins and their targeting sequences. Eur J Biochem 241:779–786

    Article  PubMed  CAS  Google Scholar 

  • Elliott KJ, Butler WO, Dickinson CD, Konno Y, Vedvick TS, Fitzmaurice L, Mirkov TE (1993) Isolation and characterization of fruit vacuolar invertase genes from two tomato species and temporal differences in mRNA levels during fruit ripening. Plant Mol Biol 21:515–524

    Article  PubMed  CAS  Google Scholar 

  • Emanuelson O, Nielsen H, von Heijne G (1999) ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites. Protein Sci 8:978–984

    Article  Google Scholar 

  • Emanuelson O, Nielsen H, Brunak S, von Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016

    Article  CAS  Google Scholar 

  • Eschrich W (1980) Free space invertase, its possible role in phloem unloading. Ber Dtsch Bot Ges 93:363–378

    CAS  Google Scholar 

  • Gallagher JA, Pollock CJ (1998) Isolation and characterization of a cDNA clone from Lolium termulentum L. encoding for a sucrose hydrolytic enzyme which shows alkaline/neutral invertase activity. J Exp Bot 49:789–795

    Article  CAS  Google Scholar 

  • Gerrits N, Turk SCHJ, van Dun KPM, Hulleman SHD, Visser RGF, Weisbeek PJ, Smeekens SCM (2001) Sucrose metabolism in plastids. Plant Physiol 125:926–934

    Article  PubMed  CAS  Google Scholar 

  • Handa H, Kobayashi-Uehara A, Murayama S (2001) Characterization of a wheat cDNA encoding mitochondrial ribosomal protein L11: quantitative tissue-specific differences in its expression. Mol Genet Genomics 265:569–575

    Article  PubMed  CAS  Google Scholar 

  • Haouazine-Takvorian N, Tymowska-Lalanne Z, Takvorian A, Tregear J, Lejeune B, Lecharny A, Kreis M (1997) Characterization of two members of the Arabidopsis thaliana gene family, Atβfruct3 and Atβfruct4, coding for vacuolar invertases. Gene 197:239–251

    Article  PubMed  CAS  Google Scholar 

  • Ji X, Van den Ende W, Van Laere A, Cheng S, Benett 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 

  • Koch KE (1996) Carbohydrate modulated gene expression in plants. Annu Rev Plant Physiol Plant Mol Biol 47:509–540

    Article  PubMed  CAS  Google Scholar 

  • Lee HS, Sturm A (1996) Purification and characterization of neutral and alkaline invertase from carrot. Plant Physiol 112:1513–1522

    Article  PubMed  CAS  Google Scholar 

  • Millar AH, Day DA, Whelan J (2004) Mitochondrial biogenesis and function in Arabidopsis. In: Somerville CR, Meyerowitz EM (eds) The Arabidopsis book. Amer Soc Plant Biologists, Rockville, DOI 10.1199/tab.0009, http://www.aspb.org/publications/arabidopsis/

  • Murayama S, Handa H (2000) Isolation and characterization of cDNAs encoding mitochondrial uncoupling proteins in wheat: wheat UCP genes are not regulated by low temperature. Mol Gen Genet 264:112–118

    Article  PubMed  CAS  Google Scholar 

  • Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8:4321–4325

    Article  PubMed  CAS  Google Scholar 

  • Nakai K, Kanehisa M (1991) Export system for predicting protein localization site in gram negative bacteria. Proteins 11:95–110

    Article  PubMed  CAS  Google Scholar 

  • Ohto M Onai K, Furukawa Y, Aoki E, Araki T, Nakamura K (2001) Effects of sugar on vegetative development and floral transition in Arabidopsis. Plant Physiol 127:252–261

    Article  Google Scholar 

  • Roitsch T, Bittner M, Godt DE (1995) Induction of apoplastic invertases of Chenopodium rubrum by d-glucose and a glucose analog and tissue-specific expression suggest a role in sink-source regulation. Plant Physiol 108:285–294

    Article  PubMed  CAS  Google Scholar 

  • Rolland F, Moore B, Sheen J (2002) Sugar sensing and signaling in plants. Plant Cell S185–S205, supplement 2002

  • Ross HA, McRae D, Davies HV (1996) Sucrolytic enzyme activities in cotyledons of the faba bean. Plant Physiol 111:329–338

    PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Sheen J, Zhou L, Jang J-C (1999) Sugars as signaling molecules. Curr Opin Plant Biol 2:410–418

    Article  PubMed  CAS  Google Scholar 

  • Small I, Peeters N, Legeai F, Lurin C (2004) Predotar: a tool for rapidly screening proteomes for N-terminal targeting sequences. Proteomics 4:1581–1590

    Article  PubMed  CAS  Google Scholar 

  • Somogyi M (1952) Notes on sugar determination. J Biol Chem 195:19–23

    CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Sturm A, Chrispeels MJ (1990) cDNA cloning of carrot extracellular β-furactosidase and its expression in response to wounding and bacterial infection. Plant Cell 2:1107–1119

    Article  PubMed  CAS  Google Scholar 

  • Sturm A, Hess D, Lee HS, Lienhard S (1999) Neutral invertase is a novel type of sucrose-cleaving enzyme. Physiol Plant 107:159–165

    Article  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 

  • Van den Ende W, Van Laere A (1995) Purification and properties of a neutral invertase from the roots of Cichorium intybus. Physiol Plant 93:241–248

    Article  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Vorster DJ, Botha FC (1998) Partial purification and characterization of sugarcane neutral invertase. Phytochemistry 49:651–655

    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 

Download references

Acknowledgments

We thank Drs. Yasuo Niwa and Hirokazu Kobayashi (University of Shizuoka, Shizuoka, Japan) for the generous gift of the enhanced GFP expression vector. We express our appreciation to Dr. Hiroyuki Kawahigashi (National Institute of Agrobiological Sciences, Tsukuba, Japan) for providing us with the RNA samples for quantitative RT-PCR. We also thank Drs. Midori Yoshida, Akira Kawakami (National Agricultural Research Center for Hokkaido Region, Sapporo, Japan), and Dr. Hanae Kaku (Meiji University, Kawasaki, Japan) for their valuable suggestions. This work was supported in part by a Grant-in-Aid for Scientific Research (C) from the Ministry of Education, Science and Culture of Japan (No. 17580012).

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Authors and Affiliations

  1. Research Team for Crop Cold Tolerance, National Agricultural Research Center for Hokkaido Region, 1 Hitsuji-ga-oka, Toyohira-ku, Sapporo, 062-8555, Japan

    Seiji Murayama

  2. Plant Genome Research Unit, National Institute of Agrobiological Sciences, 2-1-2 Kan-non-dai, Tsukuba, 305-8602, Japan

    Hirokazu Handa

  3. Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Ten-noh-dai, Tsukuba, 305-8572, Japan

    Hirokazu Handa

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  1. Seiji Murayama
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  2. Hirokazu Handa
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Correspondence to Hirokazu Handa.

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Murayama, S., Handa, H. Genes for alkaline/neutral invertase in rice: alkaline/neutral invertases are located in plant mitochondria and also in plastids. Planta 225, 1193–1203 (2007). https://doi.org/10.1007/s00425-006-0430-x

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  • DOI: https://doi.org/10.1007/s00425-006-0430-x

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