Abstract
We have previously shown that mRNA and protein encoded by the Pvlea-18 gene from Phaseolus vulgaris L., a member of a new family of late embryogenesis-abundant (LEA) proteins, accumulate in dark-grown bean seedlings not only in response to water deficit but also during optimal irrigation. In this work, we studied Pvlea-18 gene transcriptional regulation by using transgenic Arabidopsis thaliana plants containing a chimeric gene consisting of the Pvlea-18 promoter region and the 3′-nos terminator fused to the GUS gene-coding region. We demonstrate that the chimeric gene is active during Arabidopsis normal development under well-irrigated conditions, and that it is further induced in response to ABA and dehydration treatments. Replacing the 3′-nos terminator with the Pvlea-18 3′ region led to an additional increase in expression during development and in response to dehydration, but not in response to exogenous ABA. These results reveal an enhancer effect of the Pvlea-18 3′ region, which showed to be higher specifically under dehydration. The small decrease in Pvlea-18 promoter expression observed when transgenic plants treated with fluridone (an ABA biosynthesis inhibitor) were subjected to dehydration suggests that the Pvlea-18 gene dehydration response is predominantly ABA-independent. Finally, we present evidence indicating that Pvlea-18 gene expression is negatively regulated during etiolated growth, particularly in roots, in contrast to the expression pattern observed during normal development.
Similar content being viewed by others
References
Abe, H., Yamaguchi-Shinozaki, K., Urao, T., Iwasaki, T., Hosokawa, D. and Shinozaki, K. 1997. Role of Arabidop-sis MYC and MYB homologs in drought-and abscisic acid-regulated gene expression. Plant Cell 9: 1859–1868..514
Baker, J., Steele, C. and Dure, L. III. 1988. Sequence and characterization of 6 Lea proteins and their genes from cotton. Plant Mol. Biol. 11: 277–291.
Bailey-Serres, J. and Dawe, R.K. 1996. Both 5′ and 3′ sequences of maize adh1 mRNA are required for enhanced translation under low-oxygen conditions. Plant Physiol. 112: 685–695.
Bechtold. N., Ellis, J. and Pelletier, G. 1993. In anta Agrobac-terium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. CR Acad. Sci. Paris Sci. Vie/Life Sci. 316: 1194–1199.
Benfey, P.N. and Chua, N.-H. 1990. The cauliflower mosaic virus 35S promoter: combinatorial regulation of transcription in plants. Science 250: 959–966.
Biedenkapp, H., Borgmeyer, U., Sipper, A.E. and Klempnauer, K.-H. 1988. Viral myb oncogene encodes a sequence-specific DNA binding activity. Nature 335: 835–837.
Bradford, M.M. 1976. A rapid and sensitive method for the quanti-tation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 278–293.
Bray, E.A. 1993. Molecular responses to water deficit. Plant Physiol. 103: 1035–1040.
Bray, E.A. 1991. Regulation of gene expression by endogenous ABA during drought stress. In: W.J. Davies and H.G. Jones (Eds.) Abscisic Acid: Physiology and Biochemistry, Bios Scientific Publishers, Lancaster LA1 4YQ, UK, pp. 81–96.
Busk, P. K. and Pagès, M. 1998. Regulation of abscisic acid-induced transcription. Plant Mol. Biol. 37: 425–435.
Chan, M.T. and Yu, S.M. 1998. The 3′ untranslated region of a rice alpha-amylase gene functions as a sugar-dependent mRNA stability determinant. Proc. Natl. Acad. Sci. USA 95: 6543–6547.
Church, G.M. and Gilbert, M. 1984. Genomic sequencing. Proc. Natl. Acad. Sci. USA 81: 1991–1995.
Close, T. 1997. Dehydrins: a commonality in the response of plants to dehydration and low temperature. Physiol. Plant. 100: 291–296.
Colmenero-Flores, J.M., Moreno, L.P., Smith, C.E. and Co-varrubias, A.A. 1999. Pvlea-18, a member of a new late-embryogenesis-abundant protein family that accumulates during water stress and in the growing regions of well-irrigated bean seedlings. Plant Physiol. 120: 93–103.
Colmenero-Flores, J.M., Campos, F., Garciarrubio, A. and Covarru-bias, A.A. 1997. Characterization of cDNA clones responsive to water deficit from Phaseolus vulgaris: a new late embryogenesis abundant protein. Plant Mol. Biol. 35: 393–405.
Dean, C., Favreau, M., Bond-Nutter, D, Bedbrook, J. and Dunsmuir, P. 1989. Sequences downstream of translation start regulate quantitative expression of two petunia rbcS genes. Plant Cell 1: 201–208.
Dellaporta, S.L., Wood, J. and Hicks, J. 1983. A plant DNA Minipreparation: Version II. Plant Mol. Biol. Rep. 1: 19–22.
Dietrich, R.A., Radke, S.E. and Harada, J.J. 1992. Downstream DNA sequences are required to activate a gene expressed in the root cortex of embryos and seedlings. Plant Cell 4: 1371–1382.
Dure, L. III. 1993. Structural motifs in Lea proteins. In: T.J. Close and E.A. Bray (Eds.) Plant Responses to Cellular Dehydra-tion during Environmental Stress, American Society of Plant Physiologists, Rockville, MD, pp. 91–103.
Esau, K. 1977. Anatomy of Seed Plants. John Wiley, New York, pp. 215–242.
Fu, H., Kim, S.Y. and Park, W.D. 1995. High-level tuber expression and sucrose inducibility of a potato Sus4 sucrose synthase gene require 5′ and 3′ flanking sequences and the leader intron. Plant Cell 7: 1387–1394.
Galau, G.A., Hughes, D.W. and Dure, L. 1986. Abscisic acid induction of cloned cotton late embryogenesis abundant (lea)mRNAs. Plant Mol. Biol. 7: 155–170.
Garciarrubio, A., Legaria, J.P. and Covarrubias, A.A. 1997. Abscisic acid inhibits germination of mature Arabidopsis seeds by limiting the availability of energy and nutrients. Planta 203: 182–187.
Graber, J.H., Cantor, C.R., Mohr and Smith, T.F. 1999. In silico detection of control signals: mRNA 3 0-end-processing sequences in diverse species. Proc. Natl. Acad. Sci. USA 96: 14055–14060.
Guan, L.M., Zhao, J. and Scandalios, J.G. 2000. Cis-elements and trans-factors that regulate expression of the maize Cat1 antioxidant gene in response to ABA and osmotic stress: H2 O2 is the likely intermediary signaling molecule for the response. Plant J. 22: 87–95.
Guiltinan, M. J., Marcotte, W.R. and Quatrano, R.S. 1990. A plant leucine zipper protein that recognizes an abscisic acid response element. Science 250: 267–271.
Gutiérrez, R.A., Macintoch, G.C. and Green, P.J. 1999. Current perspectives on mRNA stability in plants: multiple levels and mechanisms of control. Trends Plant Sci. 4: 429–438.
Hattori, T., Vasil, V., Rosenkrans, L., Hannah, L.C., McCarthy, D.R. and Vasil, I.K. 1992. The viviparous-1 gene and abscisic acid activate the C1 regulatory gene for anthocyanin biosynthesis during seed maturation in maize. Genes Dev. 6: 609–618.
Hong, S.W., Jon, J.H., Kwak, J.M. and Nam, J.H. 1997. Identification of a receptor-like protein kinase gene rapidly induced by abscisic acid, dehydration, high salt and cold treatments in Arabidopsis thaliana. Plant Physiol. 63: 531–535.
Imai, R., Chang, L., Ohta, A., Bray, E.A. and Takagi, M. 1995. A lea-class gene of tomato confers salt and freezing tolerance when expressed in Sacharomyces cerevisiae. Gene 170: 243–248.
Ingram, J. and Bartels, D. 1996. The molecular basis of dehydration tolerance in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47: 377–403.
Ishitani, M., Xiong, L., Stevenson, B. and Zhu, J.-K. 1997. Genetic analysis of osmotic and cold stress signal transduction in Arabidopsis: interactions and convergence of abscisic acid-dependent and abscisic acid-independent pathways. Plant Cell 9: 1935–1949.
Izawa, T., Foster, R. and Chua, N.-H. 1993. Plant bZIP protein DNA binding specificity. J. Mol. Biol. 230: 1131–1144.
Iwasaki, T., Yamaguchi-Shinozaki, K. and Shinozaki, K. 1995. Identification of a cis regulatory region of a gene in Arabidopsis thaliana whose induction by dehydration is mediated by abscisic acid and requires protein synthesis. Mol. Gen. Genet. 247: 391–398.
Jacobsen, J.V. and Close, T. 1991. Control of transient expression of chimaeric genes by gibberellic acid and abscisic acid in protoplast prepared from mature barley aleurone layers. Plant Mol. Biol. 16: 713–724.
Jefferson R.A., Kavanagh, T.A. and Bevan, M.W. 1987. Beta-glucoronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.
Jefferson, R.A., Burgess, S.M. and Hirsh, D. 1986. Beta-glucoronidase from Escherichia coli as a gene-fusion marker. Proc. Natl. Acad. Sci. USA 83: 8447–8451.
Khurana, J.P., Kochhar, A. and Tyagi, A.K. 1998. Photosensory perception and signal transduction in higher plants. Molecular genetic analysis. Crit. Rev. Plant Sci. 17: 465–539.
Lam, E. and Chua, N.-H. 1991. Tetramer of 21-base pair synthetic element confers seed expression and transcriptional enhance-ment in response to water stress and abscisic acid. J. Biol. Chem. 266: 17131–17135.
Larkin, J.C., Oppenheimer, D.G., Pollock, S. and Marks, M.D. 1993. Arabidopsis GLABROUS1 gene requires downstream sequences for function. Plant Cell 5: 1739–1748.
Leung, J. and Giraudat, J. 1998. Abscisic acid signal transduction. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49: 199–222.
McCully, M.E. 1994. Accumulation of high levels of potassium in the developing xylem elements in roots of soybean and some other dicotyledons. Protoplasma 183: 116–125.
Moller, S.G. and Chua N.-H. 1999. Interactions and intersections of plant signaling pathways. J. Mol. Biol. 293: 219–234.
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473–497.
Murre, C., McCaw, P.S. and Baltimore, D. 1989. A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, myoD, and myc proteins. Cell 56: 777–783.
Nakagoshi, H.N., Nagase, T., Kanei-Ishii, C., Ueno, Y. and Ishii, S. 1990. Binding of the c-myb proto-oncogene product to the simian virus 40 enhancer stimulates transcription. J. Biol. Chem. 265: 3479–3483.
Rouse, D.T., Marotta, R. and Parish, R.W. 1996. Promoter and ex-pression of an Arabidopsis thaliana dehydrin gene. FEBS Lett. 381: 252–256.
Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.
Shen, Q., Zhang, P. and Ho T.-H.D. 1996. Modular nature of abscisic acid (ABA) response complexes: composite promoter units that are necessary and sufficient for ABA-induction of gene expression in barley. Plant Cell 8: 1107–1119.
Shinozaki, K. and Yamaguchi-Shinozaki, K. 1996. Molecular responses to drought and cold stress. Curr. Opin. Biotechnol. 7: 161–167.
Skriver, K. and Mundy, J. 1990. Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 505–512.
Stockinger, E.J., Gilmour, S.J. and Thomashow, M.F. 1997. Ara-bidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE, a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. Proc. Natl. Acad. Sci. USA 94: 1035–1040.
Swire-Clark, G.A. and Marcotte, R.W. 1999. The wheat LEA protein Em functions as an osmoprotective molecule in Sacharomyces cerevisiae. Plant Mol. Biol. 39: 117–128.
Takahashi, Y., Sakai, T., Ishida, S. and Nagata, T. 1995. Identification of auxin-responsive elements of parB and their expression in apices of shoot and root.Proc. Natl. Acad. Sci. USA 92: 6359–6363.
Tobin, E.M. and Silverthorne, J. 1985. Light regulation of gene expression in higher plants. Annu. Rev. Plant Physiol. 36: 569–593.
Valvekens, D., Van Montagu, M. and Van Lijsebettens, M. 1988. Agrobacterium tumefaciens-mediated transformation of Ara-bidopsis thaliana root explants by using kanamycin selection. Proc. Natl. Acad. Sci. USA 85: 5536–5540.
Vasil, V., Marcotte, W.R. Jr, Rosenkrans, L., Cocciolone, S.M. and Vasil, I.K. 1995. Overlap of viviparous1 (VP1) and abscisic acid response elements in the Em promoter: G-box elements are suf-ficient but not necessary for VP1 transactivation. Plant Cell 7: 1511–1518.
Venglat, S.P. and Sawhney, V.K. 1996. Benzylaminopurine induces phenocopies of floral meristem and organ identity mutans in wild-type Arabidopsis thaliana plants. Planta 3: 480–487.
von Arnim, G.A. and Deng, X.W. 1996. A role for transcriptional repression during light control of plant development. BioEssays 18: 905–910.
Walton, C.D. and Li, Y. 1995. Abscisic acid biosynthesis and metabolism. In: J.P. Davies (Ed.) Plant Hormones: Physiol-ogy, Biochemistry and Molecular Biology, Kluwer Academic Publishers, Boston, pp. 140–157.
Wang, M.L., Belmonte, S., Kim, U., Dolan, M., Morris, J.W. and Goodman, H.M. 1999. A cluster of ABA-regulated genes of Arabidopsis thaliana BAC T07M07. Genome Res. 9: 325–333.
Weatherwax, S.C., Ong, M.S., Degernhardt, J., Bray, E.A. and To-bin, E.M. 1996. The interaction of light and abscisic acid in the regulation of plant gene expression. Plant Physiol. 111: 363–370.
Xu, D., Duan, X., Wang, B.M., Hong, B., Ho, D.T.-H. and Wu, R. 1996. Expression of a late embryogenesis abundant protein gene, HVA1, from barley confers tolerance to water deficit and salt stress in transgenic rice. Plant Physiol. 110: 249–257.
Yamaguchi-Shinozaki, K. and Shinozaki, K. 1993. Characterization of the expression of a desiccation-responsive rd29 gene of Ara-bidopsis and analysis of its promoter in transgenic plants. Mol. Gen. Genet. 236: 331–340.
Yamaguchi-Shinozaki, K. and Shinozaki, K. 1994. A novel cis-acting element in an Arabidopsis gene is involved in responsive-ness to drought, low-temperature, or high-salt stress. Plant Cell 6: 251–264.
Ye, G.-N., Stone, D., Pang, S.-Z., Creely, W., González, K. and Hinchee, M. 1999.Arabidopsis ovule is target for Agrobacterium in planta vacuum infiltration transformation. Plant J. 19: 249–257.
Zeevaart, J.A.D. and Creelman, R.A. 1988. Metabolism and physi-ology of abscisic acid. Annu. Rev. Plant Physiol. Plant Mol. Biol. 39: 439–473.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Moreno-Fonseca, L.P., Covarrubias, A.A. Downstream DNA sequences are required to modulate Pvlea-18 gene expression in response to dehydration. Plant Mol Biol 45, 501–515 (2001). https://doi.org/10.1023/A:1010607223533
Issue Date:
DOI: https://doi.org/10.1023/A:1010607223533