Abstract
We have isolated two cDNA clones (Mshsp18-1; Mshsp18-2) from alfalfa (Medicago sativa L.) which encode for small heat shock proteins (HSPs) belonging to the hsp17 subfamily. The predicted amino acid sequences of the two alfalfa proteins are 92% identical and a similar degree of homology (90%) can be detected between Mshsp 18-2 and the pea hsp 17. In comparison to various members of small HSPs from soybean amino acid sequence similarities of 80–86% were identified. The alfalfa HSPs share a homologous stretch of amino acids in the carboxy terminal region with hsp22, 23, 26 from Drosophila. This region contains the GVLTV motif which is characteristic of several members of small HSPs. At room temperature alfalfa hsp 18 mRNAs were not detectable in root and leaf tissues but northern analysis showed a low level of expression in microcallus suspension (MCS). The transcription of Mshsp 18 genes is induced by elevated temperature, CdCl2 treatment and osmotic shock in cultured cells. In alfalfa somatic embryos derived from MCS a considerable amount of hsp 18 mRNA can be detected during the early embryogenic stages under normal culture conditions. The differential expression of these genes during embryo development suggests a specific functional role for HSPs in plant cells at the time of the developmental switch in vitro.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bensaude O, Babinet C, Morange M, Jacob F: Heat shock proteins, first major products of zygotic gene activity in mouse embryos. Nature 305: 331–333 (1983).
Bienz M: Xenopus hsp70 genes are constitutively expressed in injected oocytes. EMBO J 3: 2477–2483 (1984).
Bond U, Schlesinger MJ: Heat shock proteins and development. Adv Genet 24: 1–29 (1987).
Cathala G, Savouret JF, Mendez B, West BL, Karin M, Martial JA, Baster JD: A method for isolation of intact, translationally active ribonucleic acid. DNA 2: 329–335 (1983).
Cheney CM, Shearn A: Developmental regulation of Drosophila imaginal disc proteins: synthesis of a HSP under non-heat-shock conditions. Devel Biol 95: 325–330 (1983).
Cheng MY, Hartl FU, Martin J, Pollock RA, Kalousek F, Neupert W, Hallberg EM, Hallberg RL, Horwich AL: Mitochondrial heat-shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria. Nature 337: 620–625 (1989).
Czarnecka E, Gurley WB, Nagao RT, Mosqueva LA, Key JL: DNA sequence and transcript mapping of a soybean gene encoding a small HSP. Proc Natl Acad Sci USA 82: 3726–3730 (1985).
De Loose M, Alliotte T, Gheysen G, Genetello L, Gielen J, Soetaert P, Van Montagu M, Inzé D: Primary structure of a hormonally regulated B-glucanase of Nicotiana plumbaginifolia. Gene 70: 13–23 (1988).
Ellis RJ: Molecular chaperones: the plant connection. Science 250: 954–959 (1990).
Ellis RJ, Hemmingsen SM: Molecular chaperons: proteins essential for the biogenesis of some macromolecular structures. TIBS 14: 339–342 (1989).
Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–267 (1984).
Howarth C: Heat shock proteins in Sorghum bicolor and Pennisetum americanum II. Stored RNA in sorghum seed and its relationship to HSP synthesis during germination. Plant Cell Envir 13: 57–64 (1990).
Ingolia TD, Craig EA: Four small Drosophila HSPs are related to each other and to mammalian crystallin. Proc Natl Acad Sci USA 79: 2360–2364 (1982).
Kanabus J, Pikaard CS, Cherry JH: Heat shock proteins in tobacco cell suspension during growth cycle. Plant Physiol 75: 639–644 (1984).
Kurtz S, Rossi J, Petko L, Lindquist S: An ancient developmental induction: HSPs induced in sporulation and oogenesis. Science 231: 1154–1157 (1986).
Lindquist S, Craig EA: The heat-shock proteins. Annu Rev Genet 22: 631–677 (1988).
Mansfield MA, Key JL: Synthesis of the low molecular weight HSPs in plants. Plant Physiol 84: 1007–1017 (1987).
Mason PJ, Hall LMC, Gausz J: The expression of heat shock genes during normal development in Drosophila melanogaster. Mol Gen Genet 194: 73–78 (1984).
Nagao RT, Czarnecka E, Gurley WB, Schöffl F, Key JL: Genes for low-molecular-weight HSPs of soybeans: Sequence analysis of a multigene family. Mol Cell Biol 5: 3417–3428 (1985).
Neumann D, Nover L, Parthier B, Rieger R, Scharf KD, Wollgiehn R, Nieden U: Heat shock and other stress response systems in plants. Biol Zentbl 108: 1–155 (1989).
Pitto J, Lo Schiavo F, Guiliano G, Terzi M: Analysis of the heat-shock protein pattern during somatic embryogenesis of carrot. Plant Mol Biol 2: 231–237 (1983).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Schenk RU, Hildebrandt AC: Medium and techniques for induction and growth of monocotyledonous and dicotyledonous plant cell cultures. Can J Bot 29: 199–204 (1972).
Schöffl F, Raschke E, Nagao RT: The DNA sequence analysis of soybean heat-shock genes and identification of possible regulatory promoter elements. The EMBO J 3: 2491–2497 (1984).
Schöffl F, Baumann G, Raschke E: The expression of heat shock genes. A model for the environmental stress response. In: Goldberg B, Verna DPS (eds) Temporal and Spatial Regulations of Plant Genes, pp. 253–273. Springer-Verlag, Berlin (1988).
Schuster G, Even D, Kloppstech K, Ohad J: Evidence for protection by heat-shock proteins against photoinhibition during heat shock. EMBO J 7: 1–6 (1988).
Stuart DA, Strickland SG: Somatic embryogenesis from cell cultures of Medicago sativa. I. The role of amino acid additions to the gene regeneration medium. Plant Sci Lett 34: 165–174 (1984).
Sung ZR: Development states of embryogenic cultures. In: Terzi M, Pitto L, Sung ZR (eds) Somatic Embryogenesis, pp. 117–121 IPRA Roma (1985).
Wu CH, Caspar T, Browse J, Lindquist S, Somerville C: Characterization of an hsp70 cognate gene family in Arabidopsis. Plant Physiol 88: 731–740 (1988).
Zimmermann JL, Petri W, Meselson M: Accumulation of a specific subset of D. melanogaster heat shock mRNAs in normal development without heat shock. Cell 32: 1161–1170 (1983).
Zimmermann JL, Apuya N, Darwish K, O'Caroll C: Novel regulation of heat shock genes during carrot somatic embryo development. Plant Cell 1: 1137–1146 (1989).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Györgyey, J., Gartner, A., Németh, K. et al. Alfalfa heat shock genes are differentially expressed during somatic embryogenesis. Plant Mol Biol 16, 999–1007 (1991). https://doi.org/10.1007/BF00016072
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00016072