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Post-harvest regulated gene expression and splicing efficiency in storage roots of sugar beet (Beta vulgaris L.)

Planta volume 227, pages 1321–1332 (2008)Cite this article

Abstract

Sixteen post-harvest upregulated genes from sugar beet comprising five novel sequences were isolated by subtractive cloning. Transcription profiles covering a period of up to 49 days after harvest under controlled storage conditions and in field clamps are reported. Post-harvest induced genes are involved in wound response, pathogen defense, dehydration stress, and detoxification of reactive oxygen species. An early induction of a cationic peroxidase indicates a response to post-harvest damage. Wound response reactions may also involve genes required for cell division such as a regulator of chromatin condensation and a precursor of the growth stimulating peptide phytohormone phytosulfokine-α. Surprisingly, also three putative non-protein coding genes were isolated. Two of these genes show intron specific and storage temperature dependent splicing of a precursor mRNA. The temperature dependent splicing of an intron containing sugar beet mRNA is also maintained in transgenic Arabidopsis thaliana. The storage induced genes are integrated into a model that proposes the response to several post-harvest stress conditions. Temperature regulated splicing may be a mechanism to sense seasonal temperature changes.

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Abbreviations

DEPC:

Diethyl pyrocarbonate

CaMV:

Cauliflower mosaic virus

EDTA:

Ethylenediamine tetraacetic acid

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

PCR:

Polymerase chain reaction

RACE:

Rapid amplification of cDNA ends

RT:

Reverse transcriptase

SDS:

Sodium dodecyl sulfate

References

  • Aalen RB, Opsahl-Ferstad HG, Linnestad C, Olsen OA (1994) Transcripts encoding an oleosin and a dormancy-related protein are present in both the aleurone layer and the embryo of developing barley (Hordeum vulgare L.) seeds. Plant J 5:385–396

    PubMed  Article  CAS  Google Scholar 

  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    PubMed  Article  CAS  Google Scholar 

  • Asard H, Kapila J, Verelst W, Berczi A (2001) Higher-plant plasma membrane cytochrome b561: a protein in search of a function. Protoplasma 217:77–93

    PubMed  Article  CAS  Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1988) Current protocols in molecular biology. Greene & Wiley Interscience, New York

    Google Scholar 

  • Barone LM, Shih C, Wasserman BP (1997) Mercury-induced conformational changes and identification of conserved surface loops in plasma membrane aquaporins from higher plants. Topology of PMIP31 from Beta vulgaris L. J Biol Chem 272:30672–30677

    PubMed  Article  CAS  Google Scholar 

  • Bellin D, Schulz B, Soerensen TR, Salamini F, Schneider K (2007) Transcript profiles at different growth stages and tap-root zones identify correlated developmental and metabolic pathways of sugar beet. J Exp Bot 58:699–715

    PubMed  Article  CAS  Google Scholar 

  • Bellin D, Werber M, Theis T, Schulz B, Weisshaar B, Schneider K (2002) EST sequencing, annotation and macroarray transcriptome analysis identify preferentially root-expressed genes in sugar beet. Plant Biol 4:700–710

    Article  CAS  Google Scholar 

  • Bernier F, Berna A (2001) Germins and germin-like proteins: plant do-all proteins. But what do they do exactly? Plant Physiol Biochem 39:545–554

    Article  CAS  Google Scholar 

  • Bosemark NO (1993) Genetics and breeding. In: Cooke DA, Scott RK (eds) The sugar beet crop: science into practice. Chapman & Hall, London, pp 67–119

    Google Scholar 

  • Carazo-Salas RE, Guarguaglini G, Gruss OJ, Segref A, Karsenti E, Mattaj IW (1999) Generation of GTP-bound Ran by RCC1 is required for chromatin-induced mitotic spindle formation. Nature 400:178–181

    PubMed  Article  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

    PubMed  Article  CAS  Google Scholar 

  • Colon-Carmona A, You R, Haimovitch-Gal T, Doerner P (1999) Technical advance: spatio-temporal analysis of mitotic activity with a labile cyclin-GUS fusion protein. Plant J 20:503–508

    PubMed  Article  CAS  Google Scholar 

  • Deblaere R, Bytebier B, De Greve H, Deboeck F, Schell J, Van Montagu M, Leemans J (1985) Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucleic Acids Res 13:4777–4788

    PubMed  Article  CAS  Google Scholar 

  • Douglas SE (1995) DNA Strider. An inexpensive sequence analysis package for the Macintosh. Mol Biotechnol 3:37–45

    PubMed  Article  CAS  Google Scholar 

  • Elliott MC, Weston GD (1993) Biology and physiology of the sugar-beet plant. In: Cooke DA, Scott RK (eds) The sugar beet crop: science into practice. Chapman & Hall, London, pp 37–66

    Google Scholar 

  • Felsheim RF, Das A (1992) Structure and expression of a heat-shock protein 83 gene of Pharbitis nil. Plant Physiol 100:1764–1771

    PubMed  CAS  Article  Google Scholar 

  • Fujimoto S, Matsunaga S, Yonemura M, Uchiyama S, Azuma T, Fukui K (2004) Identification of a novel plant MAR DNA binding protein localized on chromosomal surfaces. Plant Mol Biol 56:225–239

    PubMed  Article  CAS  Google Scholar 

  • Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the ExPASy server. In: Walker JM (ed) The proteomics protocols handbook. Humana Press Inc, New Jersey, pp 571–607

    Google Scholar 

  • Gonzalez MC, Roitsch T, Cejudo FJ (2005) Circadian and developmental regulation of vacuolar invertase expression in petioles of sugar beet plants. Planta 222:386–395

    PubMed  Article  CAS  Google Scholar 

  • Haagenson DM, Klotz KL, McGrath JM (2006) Sugarbeet sucrose synthase genes differ in organ-specific and developmental expression. J Plant Physiol 163:102–106

    PubMed  Article  CAS  Google Scholar 

  • Hatton D, Gray JC (1999) Two MAR DNA-binding proteins of the pea nuclear matrix identify a new class of DNA-binding proteins. Plant J 18:417–429

    PubMed  Article  CAS  Google Scholar 

  • Hennig L (1999) WinGene/WinPep: user-friendly software for the analysis of amino acid sequences. Biotechniques 26:1170–1172

    PubMed  CAS  Google Scholar 

  • Herwig R, Schulz B, Weisshaar B, Hennig S, Steinfath M, Drungowski M, Stahl D, Wruck W, Menze A, O’Brien J, Lehrach H, Radelof U (2002) Construction of a ‘unigene’ cDNA clone set by oligonucleotide fingerprinting allows access to 25 000 potential sugar beet genes. Plant J 32:845–857

    PubMed  Article  Google Scholar 

  • Hesse H, Willmitzer L (1996) Expression analysis of a sucrose synthase gene from sugar beet (Beta vulgaris L.). Plant Mol Biol 30:863–872

    PubMed  Article  CAS  Google Scholar 

  • Huang YJ, To KY, Yap MN, Chiang WJ, Suen DF, Chen SC (2001) Cloning and characterization of leaf senescence up-regulated genes in sweet potato. Physiol Plant 113:384–391

    PubMed  Article  CAS  Google Scholar 

  • Iglesias R, Perez Y, de Torre C, Ferreras JM, Antolin P, Jimenez P, Rojo MA, Mendez E, Girbes T (2005) Molecular characterization and systemic induction of single-chain ribosome-inactivating proteins (RIPs) in sugar beet (Beta vulgaris) leaves. J Exp Bot 56:1675–1684

    PubMed  Article  CAS  Google Scholar 

  • Jaggard KW, Clark CJA, May MJ, McCullagh S, Draycott AP (1997) Changes in the weight and quality of sugarbeet (Beta vulgaris) roots in storage clamps on farms. J Agr Sci 129:287–301

    Article  Google Scholar 

  • Jones-Rhoades MW, Bartel DP, Bartel B (2006) MicroRNAS and their regulatory roles in plants. Annu Rev Plant Biol 57:19–53

    PubMed  Article  CAS  Google Scholar 

  • Kloos DU, Oltmanns H, Dock C, Stahl D, Hehl R (2002) Isolation and molecular analysis of six taproot expressed genes from sugar beet. J Exp Bot 53:1533–1534

    PubMed  Article  CAS  Google Scholar 

  • Klotz KL, Haagenson DM (2008) Wounding, anoxia and cold induce sugarbeet sucrose synthase transcriptional changes that are unrelated to protein expression and activity. J Plant Physiol 165:423–434

    PubMed  Article  CAS  Google Scholar 

  • Köhler U, Donath M, Mendel RR, Cerff R, Hehl R (1996) Intron-specific stimulation of anaerobic gene expression and splicing efficiency in maize cells. Mol Gen Genet 251:252–258

    PubMed  Article  Google Scholar 

  • Lagrimini LM, Burkhart W, Moyer M, Rothstein S (1987) Molecular cloning of complementary DNA encoding the lignin-forming peroxidase from tobacco: molecular analysis and tissue-specific expression. Proc Natl Acad Sci USA 84:7542–7546

    PubMed  Article  CAS  Google Scholar 

  • Lin Q, Chen ZC, Antoniw JF, White RF (1991) Isolation and characterization of a cDNA clone encoding the anti-viral protein from Phytolacca americana. Plant Mol Biol 17:609–614

    PubMed  Article  CAS  Google Scholar 

  • Lorbiecke R, Sauter M (2002) Comparative analysis of PSK peptide growth factor precursor homologs. Plant Sci 163:321–332

    Article  CAS  Google Scholar 

  • Marchler-Bauer A, Anderson JB, Derbyshire MK, DeWeese-Scott C, Gonzales NR, Gwadz M, Hao L, He S, Hurwitz DI, Jackson JD, Ke Z, Krylov D, Lanczycki CJ, Liebert CA, Liu C, Lu F, Lu S, Marchler GH, Mullokandov M, Song JS, Thanki N, Yamashita RA, Yin JJ, Zhang D, Bryant SH (2007) CDD: a conserved domain database for interactive domain family analysis. Nucleic Acids Res 35:D237–D240

    PubMed  Article  CAS  Google Scholar 

  • Mastrangelo AM, Belloni S, Barilli S, Ruperti B, Di Fonzo N, Stanca AM, Cattivelli L (2005) Low temperature promotes intron retention in two e-cor genes of durum wheat. Planta 221:705–715

    PubMed  Article  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  • Nielsen KK, Bojsen K, Roepstorff P, Mikkelsen JD (1994) A hydroxyproline-containing class IV chitinase of sugar beet is glycosylated with xylose. Plant Mol Biol 25:241–257

    PubMed  Article  CAS  Google Scholar 

  • Oltmanns H, Kloos DU, Brieß W, Pflugmacher M, Stahl DJ, Hehl R (2006) Taproot promoters cause tissue specific gene expression within the storage root of sugar beet. Planta 224:485–495

    PubMed  Article  CAS  Google Scholar 

  • Park SW, Vepachedu R, Sharma N, Vivanco JM (2004) Ribosome-inactivating proteins in plant biology. Planta 219:1093–1096

    PubMed  Article  CAS  Google Scholar 

  • Passardi F, Cosio C, Penel C, Dunand C (2005) Peroxidases have more functions than a Swiss army knife. Plant Cell Rep 24:255–265

    PubMed  Article  CAS  Google Scholar 

  • Qi X, Harn C, Mu HH, Wasserman BP (1996) Nucleotide sequences of three cDNAs encoding major intrinsic proteins from Beta vulgaris L. (Accession Nos. U60147, U60148 and U60149) (PGR96–074). Plant Physiol 112:861

    Article  Google Scholar 

  • Reintanz B, Szyroki A, Ivashikina N, Ache P, Godde M, Becker D, Palme K, Hedrich R (2002) AtKC1, a silent Arabidopsis potassium channel alpha -subunit modulates root hair K+ influx. Proc Natl Acad Sci USA 99:4079–4084

    PubMed  Article  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  • Rosenkranz H, Vogel R, Greiner S, Rausch T (2001) In wounded sugar beet (Beta vulgaris L.) tap-root, hexose accumulation correlates with the induction of a vacuolar invertase isoform. J Exp Bot 52:2381–2385

    PubMed  Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Schulze-Lefert P (2004) Plant immunity: the origami of receptor activation. Curr Biol 14:R22–R24

    PubMed  Article  CAS  Google Scholar 

  • Simon P (1993) Diveristy and conservation of plant peroxidases. Plant Peroxidase Newsletter 1:4–7

    Google Scholar 

  • Stahl DJ, Kloos DU, Hehl R (2004) A sugar beet chlorophyll a/b binding protein promoter void of G-box like elements confers strong and leaf specific reporter gene expression in transgenic sugar beet. BMC Biotechnol 4:31

    PubMed  Article  CAS  Google Scholar 

  • van Loon LC, Rep M, Pieterse CM (2006) Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol 44:135–162

    PubMed  Article  CAS  Google Scholar 

  • Venema J, Tollervey D (1999) Ribosome synthesis in Saccharomyces cerevisiae. Annu Rev Genet 33:261–311

    PubMed  Article  CAS  Google Scholar 

  • Wiltshire JJJ, Cobb AH (2000) Bruising of sugar beet roots and consequential sugar loss: current understanding and research needs. Ann Appl Biol 136:159–166

    Article  Google Scholar 

  • Wu P, Brockenbrough JS, Metcalfe AC, Chen S, Aris JP (1998) Nop5p is a small nucleolar ribonucleoprotein component required for pre-18 S rRNA processing in yeast. J Biol Chem 273:16453–16463

    PubMed  Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by the KWS SAAT AG and a grant (0312859) from the Federal Ministry of Education and Research (BMBF) to the KWS SAAT AG. We would like to thank Elke Faurie, Maike Pflugmacher (PLANTA GmbH, Einbeck, Germany) and Jeanette Kurrasch (PLANTA GmbH) for excellent technical assistance and Norbert Käufer for stimulating discussions concerning splicing. We thank Sebastian Weißman and Thomas Zobel for isolating and sequencing of a genomic clone from Bvnpcg3, and Ralph Lisson for advice concerning primer design.

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  1. Dorothee U. Kloos

    Present address: Definiens AG, Trappentreustr. 1, 80339, München, Germany

Affiliations

  1. Institut für Genetik, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany

    Alexander Rotthues, Jeannette Kappler, Anna Lichtfuß, Dorothee U. Kloos & Reinhard Hehl

  2. PLANTA Angewandte Pflanzengenetik und Biotechnologie GmbH, Grimsehlstr. 31, 37555, Einbeck, Germany

    Dietmar J. Stahl

Authors
  1. Alexander Rotthues
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  2. Jeannette Kappler
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  3. Anna Lichtfuß
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  4. Dorothee U. Kloos
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  5. Dietmar J. Stahl
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  6. Reinhard Hehl
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Correspondence to Reinhard Hehl.

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Rotthues, A., Kappler, J., Lichtfuß, A. et al. Post-harvest regulated gene expression and splicing efficiency in storage roots of sugar beet (Beta vulgaris L.). Planta 227, 1321–1332 (2008). https://doi.org/10.1007/s00425-008-0704-6

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  • DOI: https://doi.org/10.1007/s00425-008-0704-6

Keywords

  • Differential gene expression
  • Post-harvest
  • Splicing
  • Storage
  • Stress response
  • Transcription