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.
Similar content being viewed by others
References
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 15:63–78
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 2:3389–3402
Barr HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficit in leaves. Aust J Biol Sci 1515:413–428
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 37:859–869
Berger BJ, English S, Chan G, Knodel MH (2003) Methionine regeneration and aminotransferase in Bacillus subtilis, Bacillus cereus, and Bacillus anthracis. J Bacteriol 185:2418–2431
Blum A (1996) Crop responses to drought and the interpretation of adaptation. Plant Growth Regul 20:135–148
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 11:1285–1295
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 39:855–860
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 134:960–968
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 103:415–424
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 129:540–550
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 158:455–463
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 38:551–564
Eden A, Benvenisty N (1999) Involvement of branched-chain amino acid aminotransferase (Bcat1/Eca39) in apoptosis. FEBS Lett 457:255–261
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 275:6007–6013
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 43:275–280
Gerbling H, Gerhardt B (1989) Peroxisomal degradation of brached-chain 2-oxo-acids. Plant Physiol 91:1387–1392
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 22:4673–4680
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database:1999. Nucleic Acids Res 27:297–300
Huang N, Sutliff TD, Litts JC, Rodriguez RL (1990) Classification and characterization of the rice alpha-amylase multigene family. Plant Mol Biol 14:655–668
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 36:331–341
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168
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 136:4159–4168
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 271:24458–24464
Kosambi DD (1944) The estimation of map distances from recombination values. Ann Eugen 12:172–175
Lafontaine D, Tollervey D (1996) One-step PCR mediated strategy for the construction of conditionally expressed and epitope tagged yeast proteins. Nucleic Acid Res 24:3469–3472
Lawlor DW, Cornic G (2002) Photosynthetic carbon assimilation and associated metabolism in relation to water deficit in higher plants. Plant Cell Environ 25:275–294
Liang P, Pardee AB (1992) Differential display of eukaryotic messenger RNA by means of polymerase chain reaction. Science 257:967–971
Malatrasi M, Close TJ, Marmiroli N (2002) Identification and mapping of a putative stress response regulator gene in barley. Plant Mol Biol 50:141–150
Manly K, Cudmore J, Meer J (2001) Map manager QTX, cross-platform software for genetic mapping. Mamm Genome 12:930–932
Reddy AR, Chaitanya KV, Vivekanandan M (2004) Drought-induced responses of photosynthesis and antioxidant metabolism in higher plants. J Plant Physiol 161:1189–1202
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 134:1683–1696
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 110:852–858
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Plainview, NY
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 57:241–254
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 33:259–270
Singh BK (1999) Biosynthesis of valine, leucine and isoleucine. In: Singh BK (ed) Plant amino acids: biochemistry and biotechnology. Marcel Dekker, New York, pp 227–247
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 134:838–848
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 103:161–170
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 108:181–188
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 5:1529–1539
Verwoerd TC, Dekker BMM, Hoekema A (1989) A small scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17:2362
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 101:1093–1099
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.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by P. Langridge
M. Malatrasi and M. Corradi equally contributed to this research paper.
Rights and permissions
About this article
Cite this article
Malatrasi, M., Corradi, M., Svensson, J.T. et al. A branched-chain amino acid aminotransferase gene isolated from Hordeum vulgare is differentially regulated by drought stress. Theor Appl Genet 113, 965–976 (2006). https://doi.org/10.1007/s00122-006-0339-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00122-006-0339-6