Advertisement

Theoretical and Applied Genetics

, Volume 113, Issue 6, pp 965–976 | Cite as

A branched-chain amino acid aminotransferase gene isolated from Hordeum vulgare is differentially regulated by drought stress

  • M. Malatrasi
  • M. Corradi
  • J. T. Svensson
  • T. J. Close
  • M. Gulli
  • N. Marmiroli
Original Paper

Abstract

Differential display was used to isolate cDNA clones showing differential expression in response to ABA, drought and cold in barley seedling shoots. One drought-regulated cDNA clone (DD12) was further analyzed and found to encode a branched-chain amino acid aminotransferase (HvBCAT-1). A genomic clone was isolated by probing the Morex BAC library with the cDNA clone DD12 and the structure of Hvbcat-1 was elucidated. The coding region is interrupted by six introns and contains a predicted mitochondrial transit peptide. Hvbcat1 was mapped to chromosome 4H. A comparison was made to rice and Arabidopsis genes to identify conserved structural patterns. Complementation of a yeast (Saccharomyces cerevisiae) double knockout strain revealed that HvBCAT-1 can function as the mitochondrial (catabolic) BCATs in vivo. Transcript levels of Hvbcat-1, increased in response to drought stress. As the first enzyme in the branched-chain amino acid (BCAA) catabolic pathway, HvBCAT-1 might have a role in the degradation of BCAA. Degradation of BCAA could serve as a detoxification mechanism that maintains the pool of free branched-chain amino acids at low and non toxic levels, under drought stress conditions.

Keywords

Drought Stress Relative Water Content Branch Chain Amino Acid Oregon Wolfe Barley Branch Chain Amino Acid Level 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We are grateful to prof. R. Lill and U. Muehlenhoff (University of Munchen, Germany) for the generous gift of the Δbat2/gal-bat1 yeast strain and to prof. Tiziana Lodi (University of Parma, Italy) for the gift of the plasmid pYeDP10 for yeast transformation. This work has been supported by NATO Grant (CLG 978261) to N. Marmiroli, by project “Biotecnologie Vegetali” (MIPA) to N. Marmiroli, by CNR-Agenzia 2000 project to M. Gulli and in part by NSF DBI-0321756, “Coupling Expressed Sequences and Bacterial Artificial Chromosome Resources to Access the Barley Genome” to T.J. Close.

References

  1. Abe H, Urao T, Ito T, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Arabidopsis AtMYC2 (HLH) and AtMYB2 (MYB) function as transcriptional activators in abscisic acid signaling. Plant Cell 15:63–78PubMedCrossRefGoogle Scholar
  2. Altschul SF, Madden TL, Schäffer AA, Zhang JH, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 2:3389–3402CrossRefGoogle Scholar
  3. Barr HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficit in leaves. Aust J Biol Sci 1515:413–428Google Scholar
  4. Bate N, Twell D (1998) Functional architecture of a late pollen promoter: pollen-specific transcription is developmentally regulated by multiple stage-specific and co-dependent activator elements. Plant Mol Biol 37:859–869PubMedCrossRefGoogle Scholar
  5. Berger BJ, English S, Chan G, Knodel MH (2003) Methionine regeneration and aminotransferase in Bacillus subtilis, Bacillus cereus, and Bacillus anthracis. J Bacteriol 185:2418–2431PubMedCrossRefGoogle Scholar
  6. Blum A (1996) Crop responses to drought and the interpretation of adaptation. Plant Growth Regul 20:135–148CrossRefGoogle Scholar
  7. Busk PK, Jensen AB, Pages M (1997) Regulatory elements in vivo in the promoter of the abscisic acid responsive gene rab17 from maize. Plant J 11:1285–1295PubMedCrossRefGoogle Scholar
  8. Campbell MA, Patel JK, Meyers JL, Myrick LC, Gustin JL (2001) Genes encoding for branched-chain amino acid aminotransferase are differentially expressed in plants. Plant Physiol Biochem 39:855–860CrossRefGoogle Scholar
  9. Close TJ, Wanamaker SI, Caldo RA, Turner SM, Ashlock DA, Dickerson JA, Wing RA, Muehlbauer GJ, Kleinhofs A, Wise RP (2004) A new resource for cereal genomics: 22 K barley GeneChip comes of age. Plant Physiol 134:960–968PubMedCrossRefGoogle Scholar
  10. Costa JM, Corey A, Hayes PM, Jobet C, Kleinhofs A, Kopisch-Obusch A, Kramer SF, Kudrna D, Li M, Riera-Lizarazu O, Sato K, Szucs P, Toojinda T, Vales MI, Wolfe RI (2001) Molecular mapping of the Oregon Wolfe Barleys: a phenotypically polymorphic doubled-haploid population. Theor Appl Genet 103:415–424CrossRefGoogle Scholar
  11. Diebold R, Schuster J, Daschner K, Binder S (2002) The branched-chain amino acid transaminase gene family in Arabidopsis encodes plastid and mitochondrial proteins. Plant Physiol 129:540–550PubMedCrossRefGoogle Scholar
  12. Di Martino C, Delfine S, Pizzuto R, Loreto F, Fuggi A (2003) Free amino acids and glycine in leaf osmoregulation of spinach responding to increasing salt stress. New Phytol 158:455–463CrossRefGoogle Scholar
  13. Dunn MA, White AJ, Vural S, Hughes MA (1998) Identification of promoter elements in a low-temperature-responsive gene (blt4.9) from barley (Hordeum vulgare L.). Plant Mol Biol 38:551–564PubMedCrossRefGoogle Scholar
  14. Eden A, Benvenisty N (1999) Involvement of branched-chain amino acid aminotransferase (Bcat1/Eca39) in apoptosis. FEBS Lett 457:255–261PubMedCrossRefGoogle Scholar
  15. Fujiki Y, Sato T, Ito M, Watanabe A (2000) Isolation and chracterization of cDNA clones for E1β and E2 subunits of the branched-chain α-ketoacid deydrogense complex in Arabidopsis. J Biol Chem 275:6007–6013PubMedCrossRefGoogle Scholar
  16. Fujiki Y, Ito M, Itoh T, Nishida I, Watanabe A (2002) Activation of the promoters of Arabidopsis genes for the branched-chain a-keto acid dehydrogenase complex in transgenic tobacco BY-2 cells under sugar starvation. Plant Cell Physiol 43:275–280PubMedCrossRefGoogle Scholar
  17. Gerbling H, Gerhardt B (1989) Peroxisomal degradation of brached-chain 2-oxo-acids. Plant Physiol 91:1387–1392PubMedCrossRefGoogle Scholar
  18. Higgins D, Thompson J, Gibson T, Thompson JD, Higgins DG, Gibson TJ (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedGoogle Scholar
  19. Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database:1999. Nucleic Acids Res 27:297–300PubMedCrossRefGoogle Scholar
  20. Huang N, Sutliff TD, Litts JC, Rodriguez RL (1990) Classification and characterization of the rice alpha-amylase multigene family. Plant Mol Biol 14:655–668PubMedCrossRefGoogle Scholar
  21. Hwang YS, Karrer EE, Thomas BR, Chen L, Rodriguez RL (1998) Three cis-elements required for rice alpha-amylase Amy3D expression during sugar starvation. Plant Mol Biol 36:331–341PubMedCrossRefGoogle Scholar
  22. Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168PubMedGoogle Scholar
  23. Kaplan F, Kopka J, Haskell DW, Zhao W, Schiller C, Gatzke N, Sung DY, Guy CL (2004) Exploring the temperature–stress metabolome of Arabidopsis. Plant Physiol 136:4159–4168PubMedCrossRefGoogle Scholar
  24. Kispal G, Steiner H, Court DA, Rolinski B, Lill R (1996) Mitochondrial and cytosolic branched-chain amino acid transaminases from yeast, homologs of myc oncogene-regulated Eca39 protein. J Biol Chem 271:24458–24464PubMedCrossRefGoogle Scholar
  25. Kosambi DD (1944) The estimation of map distances from recombination values. Ann Eugen 12:172–175Google Scholar
  26. Lafontaine D, Tollervey D (1996) One-step PCR mediated strategy for the construction of conditionally expressed and epitope tagged yeast proteins. Nucleic Acid Res 24:3469–3472PubMedCrossRefGoogle Scholar
  27. Lawlor DW, Cornic G (2002) Photosynthetic carbon assimilation and associated metabolism in relation to water deficit in higher plants. Plant Cell Environ 25:275–294PubMedCrossRefGoogle Scholar
  28. Liang P, Pardee AB (1992) Differential display of eukaryotic messenger RNA by means of polymerase chain reaction. Science 257:967–971PubMedGoogle Scholar
  29. Malatrasi M, Close TJ, Marmiroli N (2002) Identification and mapping of a putative stress response regulator gene in barley. Plant Mol Biol 50:141–150CrossRefGoogle Scholar
  30. Manly K, Cudmore J, Meer J (2001) Map manager QTX, cross-platform software for genetic mapping. Mamm Genome 12:930–932PubMedCrossRefGoogle Scholar
  31. Reddy AR, Chaitanya KV, Vivekanandan M (2004) Drought-induced responses of photosynthesis and antioxidant metabolism in higher plants. J Plant Physiol 161:1189–1202CrossRefGoogle Scholar
  32. Rizhsky L, Liang H, Shuman J, Shulaev V, Davletova S, Mittler R (2004) When defense pathways collide. The response of Arabidopsis to a combination of drought and heat stress. Plant Physiol 134:1683–1696PubMedCrossRefGoogle Scholar
  33. Rodriguez EM, Svensson JT, Malatrai M, Choi D-W, Close TJ (2005) Barley Dhn13 encodes a KS-type dehydrin with costitutive and stress responsive expression. Theor Appl Genet 110:852–858PubMedCrossRefGoogle Scholar
  34. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Plainview, NYGoogle Scholar
  35. Schuster J, Binder S (2005) The mitochondrial branched-chain aminotransferase (AtBCAT-1) is capable to initiate degradation of leucine, isoleucine and valine in almost all tissue of Arabidopsis thaliana. Plant Mol Biol 57:241–254PubMedCrossRefGoogle Scholar
  36. Simpson SD, Nakashima K, Narusaka Y, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Two different novel cis-acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark-induced senescence. Plant J 33:259–270PubMedCrossRefGoogle Scholar
  37. Singh BK (1999) Biosynthesis of valine, leucine and isoleucine. In: Singh BK (ed) Plant amino acids: biochemistry and biotechnology. Marcel Dekker, New York, pp 227–247Google Scholar
  38. Taylor NL, Heazlewood JL, Day DA, Millar AH (2004) Lipoic acid-dependent oxidative catabolism of alpha-keto acids in mitochondria provides evidence for branched-chain amino acid catabolism in Arabidopsis. Plant Physiol 134:838–848PubMedCrossRefGoogle Scholar
  39. Teulat B, Borries C, This D (2001) New QTLs identified for plant water status, water-soluble carbohydrate and osmotic adjustement in a barley population grown in a growth-chamber under two water regimes. Theor Appl Genet 103:161–170CrossRefGoogle Scholar
  40. Teulat B, Zoumarou-Wallis N, Rotter B, Ben Salem M, Bahri H, This D (2003) QTL for relative water content in field-grown barley and their stability across Mediterranean environments. Theor Appl Genet 108:181–188PubMedCrossRefGoogle Scholar
  41. Urao T, Yamaguchi-Shinozaki K, Urao S, Shinozaki K (1993) An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant Cell 5:1529–1539PubMedCrossRefGoogle Scholar
  42. Verwoerd TC, Dekker BMM, Hoekema A (1989) A small scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17:2362PubMedGoogle Scholar
  43. Yu Y, Tomkins JP, Waugh R, Frisch DA, Kudrna D, Kleinhofs A, Brueggeman RS, Muehlbauer GJ, Wise RP, Wing RA (2000) A bacterial artificial chromosome library for barley (Hordeum vulgare L.) and the identification of clones containing putative resistance genes. Theor Appl Genet 101:1093–1099CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • M. Malatrasi
    • 1
    • 2
  • M. Corradi
    • 1
  • J. T. Svensson
    • 2
  • T. J. Close
    • 2
  • M. Gulli
    • 1
  • N. Marmiroli
    • 1
  1. 1.Dipartimento di Scienze Ambientali, Sez. Genetica e Biotecnologie AmbientaliUniversità di ParmaParmaItaly
  2. 2.Department of Botany and Plant SciencesUniversity of CaliforniaRiversideUSA

Personalised recommendations