Skip to main content
Log in

Molecular characterization of a hydroxymethylglutaryl-CoA reductase gene from mulberry (Morus alba L.)

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

The genus Morus consists of trees and shrubs, which are distributed in temperate and subtropical regions. Commonly known as mulberry, a few of the Morus species are valued for their foliage, which constitutes the chief feed for mulberry silkworms. Steroids and isoprenoid compounds present in the foliage not only add nutritive factors to the feed but also contribute greatly to silkworm health and silk production. Mevalonate synthesis, which is the first step in isoprenoid biosynthesis, is catalyzed by the enzyme hydroxymethylglutaryl-CoA reductase (HMGR). A genomic clone, Mahmg1, was isolated from Morus alba and its expression characterized in mulberry and transgenic tobacco. In mulberry, Mahmg1 transcripts were highest in young leaves and flowers. The promoter region of the Mahmg1 gene was fused to the β-glucuronidase (GUS) reporter gene and the fusion introduced into tobacco. In imbibed embryos, GUS expression was limited to the cotyledons, epicotyl, and root elongation zone. Later, GUS staining was observed in floral tissues, guard cells, and the heads of trichomes on the stem and petioles. Mahmg1::GUS activity increased 3–4-fold by treatment with 100 μM abscisic acid and 15–80-fold in dark-grown versus light-grown seedlings. These results show that expression of the Mahmg1 gene is differentially regulated by developmental and environmental cues, suggesting that its HMGR isozyme a may provide a precursor for synthesis of specific isoprenoids during mulberry growth and development.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Anderson, J.A., Huprikar, S.S., Kochian, L.V., Lucas, W.J. and Gaber, R.F. 1992. Functional expression of a probable Arabidopsis thaliana potassium channel in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 89: 3763–3740.

    Google Scholar 

  • Aoyagi, K., Beyou, A., Moon, K., Fang, L. and Ulrich, T. 1993. Isolation and characterization of cDNAs encoding wheat 3-hydroxy-3-methylglutaryl coenzyme A reductase. Plant Physiol. 102: 623–628.

    Google Scholar 

  • Bevan, M. 1984. Binary Agrobacterium vectors for plant cell transformation. Nucl. Acids Res. 12: 8711–8721.

    Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248–254.

    Google Scholar 

  • Burnett, R.J., Maldonado-Mendoza, I.E., McKnight, T.D. and Nessler, C.L. 1993. Expression of a 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Camptotheca acuminata is differentially regulated by wounding and methyl jasmonate. Plant Physiol. 103: 41–48.

    Google Scholar 

  • Chappell, J. 1995. The biochemistry and molecular biology of isoprenoid metabolism. Plant Physiol. 107: 1–6.

    Google Scholar 

  • Chappell, J., VonLanken, C. and Vogeli, U. 1991. Elicitor inducible 3-hydroxy-3-methylglutaryl coenzyme A reductase activity is required for sesquiterpene accumulation in tobacco cell suspension cultures. Plant Physiol. 97: 693–698.

    Google Scholar 

  • Chappell, J., Wolf, F., Proulx, J., Cuellar, R. and Saunders C. 1995. Is the reaction catalyzed by 3-hydroxy-3-methylglutaryl coenzyme A reductase a rate-limiting step for isoprenoid biosynthesis in plants. Plant Physiol. 109: 1337–1343.

    Google Scholar 

  • Choi, D., Ward, B.L. and Bostock, R.M. 1992. Differential induction and suppression of potato 3-hydroxy-3-methylglutaryl coenzyme A reductase genes in response to Phytophthora infestans and to its elicitor arachidonic acid. Plant Cell 4: 1333–1344.

    Google Scholar 

  • Chye, M.L., Tan, C.T. and Chua, N.H. 1992. Three genes encode 3-hydroxy-3-methylglutaryl coenzyme A reductase in Hevea brasiliensis: hmg1 and hmg3 are differentially expressed. Plant Mol. Biol. 19: 473–484.

    Google Scholar 

  • Cramer, C.L., Weissenborn, D., Cottingham, C.K., Denbow, C.J., Eisenback, J.D., Radin, D.N. and Yu, X. 1993. Regulation of defense-related gene expression during plant-pathogen interactions. J. Nematol. 25: 507–518.

    Google Scholar 

  • Daraselia, N.D., Tarchevskaya, S. and Narita, J.O. 1996. The promoter for tomato 3-hydroxy-3-methylglutaryl coenzyme A reductase gene 2 has unusual regulatory elements that direct high-level expression. Plant Physiol. 112: 727–733.

    Google Scholar 

  • Denbow, C.J., Lang, S. and Cramer, C.L. 1996. The N-terminal domain of tomato 3-hydroxy-3-methylglutaryl coenzyme A reductase. J. Biol. Chem. 271: 9710–9715.

    Google Scholar 

  • Donald, R.G.K., Schindler, U., Batschauer, A. and Cashmore, A.R. 1990. The plant G box promoter sequence activates transcription in Saccharomyces cerevisiae and is bound in vitro by a yeast activity similar to GBF, the plant G box binding factor. EMBO J. 9: 1727–1735.

    Google Scholar 

  • du Jardin, P., Harvengt, L., Kirsch, F., Van-Quy, L., Nguyen-Quoc, B. and Yelle, S. 1997. Sink-cell-specific activity of a potato ADP-glucose pyrophosphorylase β-subunit promoter in transgenic potato and tomato plants. Planta 203: 133–139.

    Google Scholar 

  • Enjuto, M., Balcells, L., Campos, N., Caelles, C., Arro, M. and Boronat, A. 1994. Arabidopsis thaliana contains two differentially expressed 3-hydroxy-3-methylglutaryl-CoA reductase genes, which encode microsomal forms of the enzyme. Proc. Natl. Acad. Sci. USA 91: 927–931.

    Google Scholar 

  • Enjuto, M., Lumbreras, V., Marin, C. and Boronat, A. 1995. Expression of the Arabidopsis HMG2 gene, encoding 3-hydroxy-3-methylglutaryl Coenzyme A reductase, is restricted to meristematic and floral tissues. Plant Cell 7: 517–527.

    Google Scholar 

  • Gertler, F.B., Chiu, C.Y., Richter-Mann, L. and Chin D.J. 1988. Developmental and metabolic regulation of Drosophila melanosgaster 3-hydroxy-3-methylglutaryl coenzyme A reductase. Mol. Cell Biol. 8: 2713–2721.

    Google Scholar 

  • Hirano, H. 1982. Varietal differences of leaf protein profiles in mulberry. Phytochemistry 21: 1513–1518.

    Google Scholar 

  • Horsch, R.B., Fry, J.E., Hoffmann, N.L., Eichholtz, D., Rogers, S.G. and Fraley, R.T. 1985. A simple and general method for transferring genes in to plants. Science 227: 1229–1231.

    Google Scholar 

  • Jain, A.K., Dandin, S.B. and Sengupta, K. 1990. In vitro propagation through axillary bud multiplication in different mulberry genotypes. Plant Cell Rep. 8: 737–740.

    Google Scholar 

  • Jain, A.K. and Datta, R.K. 1992. Shoot organogenesis and plant regeneration in mulberry (Morus bombycis Koidz): factors influencing morphological potential in callus cultures. Plant Cell Tiss. Org. Cult. 29: 43–50.

    Google Scholar 

  • Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.

    Google Scholar 

  • Korth, K.L., Stemer, B.A., Bhattacharyya, M.K. and Dixon, R.A. 1997. HMG-CoA reductase gene families that differentially accumulate transcripts in potato tubers are developmentally expressed in floral tissues. Plant Mol. Biol. 33: 545–551.

    Google Scholar 

  • Learned, R.M. and Connolly, E.L. 1997. Light modulates the spatial patterns of 3-hydroxy-3-methylglutarylcoenzyme A reductase gene expression in Arabidopsis thaliana. Plant J. 11: 499–511.

    Google Scholar 

  • Liscum, L., Finer-Moore, J., Stroud, R.M., Luskey, K.L., Brown, M.S. and Goldstein, J.L. 1985. Domain structure of 3-hydroxy-3-methylglutaryl coenzyme A reductase, a glycoprotein of the endoplasmic reticulum. J. Biol. Chem. 260: 522–530.

    Google Scholar 

  • Ló pez-Meyer, M. and Nessler, C.L. 1997. Tryptophan decarboxylase is encoded by two autonomously regulated genes in Camptotheca acuminata which are differentially expressed during development and stress. Plant J. 11: 1167–1175.

    Google Scholar 

  • MacRobbie, E.A. 1998. Signal transduction and ion channels in guard cells. Phil. Trans. R. Soc. Lond. B Biol. Sci. 353: 1475–1488.

    Google Scholar 

  • Maldonado-Mendoza, I.E., Rebecca, M.V. and Nessler, C.L. 1997. Molecular characterization of three differentially expressed members of the Camptotheca acuminata 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene family. Plant Mol. Biol. 34: 781–790.

    Google Scholar 

  • Mascarenhas, J.P. 1993. Molecular mechanisms of pollen tube growth and differentiation. Plant Cell 5: 1303–1314.

    Google Scholar 

  • Muller-Rober, B., Ellenberg, J., Provart, N., Willmitzer, L., Busch, H., Becker, D., Dietrich, P., Hoth, S. and Hedrich, R. 1995. Cloning and electrophysiological analysis of KST1, an inward rectifying KC channel expressed in potato guard cells. EMBO J. 14: 2409–2416.

    Google Scholar 

  • Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473–497.

    Google Scholar 

  • Park, H., Denbow, C.J. and Cramer, C.L. 1992. Structure and nucleotide sequence of tomato HMG2 encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase. Plant Mol. Biol. 20: 327–331.

    Google Scholar 

  • Randall, S.K., Marshall, M.S. and Crowell, D.N. 1993. Protein isoprenylation in suspension-cultured tobacco cells. Plant Cell 5: 433–442.

    Google Scholar 

  • Rodriguez-Concepcion, M., Yalovsky, S. and Gruissem, W. 1999. Protein prenylation in plants: old friends and new targets. Plant Mol. Biol. 39: 865–870.

    Google Scholar 

  • Sanger, F., Nicklen, S. and Coulson, A.R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467.

    Google Scholar 

  • Stermer, B.A., Bianchini, G.M., Korth, K.L. 1994. Regulation of HMG-CoA reductase activity in plants. J. Lipid Res. 35: 1133–1140.

    Google Scholar 

  • Taylor, B. and Powell, A. 1982. Isolation of plant DNA and RNA. Focus 4: 4–6.

    Google Scholar 

  • Terryn, N., Aris, M.B., Engler, G., Tire, C., Villarroel, R., Van Montagu, M. and Inzé, D. 1993. rha1, a gene encoding a small GTP binding protein from Arabidopsis, is expressed primarily in developing guard cells. Plant Cell 5: 1761–1769.

    Google Scholar 

  • Yang, Z., Park, H., Lacy, G.H. and Cramer, C.L. 1991. Differential activation of potato 3-hydroxy-3-methylglutaryl coenzyme A reductase genes by wounding and pathogen challenge. Plant Cell 3: 397–405.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jain, A.K., Vincent, R.M. & Nessler, C.L. Molecular characterization of a hydroxymethylglutaryl-CoA reductase gene from mulberry (Morus alba L.). Plant Mol Biol 42, 559–569 (2000). https://doi.org/10.1023/A:1006336825226

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006336825226

Navigation