Abstract
We have characterized a genomic clone containing the potato pathogenesis-related genes STH-2 and STH-21. The two genes are found 4 kb apart on the same chromosome and their sequences are highly similar. They present the same transcriptional orientation and are both interrupted by a single intron. A chimaeric gene consisting of 1015 bp of 5′-flanking sequence and part of the first exon of STH-2 fused to the bacterial β-glucuronidase gene was highly-expressed in tubers of transgenic potato plants after wounding and elicitor treatments. The levels of activity observed in these transgenic plants parallel those observed for the accumulation of STH-2 mRNAs under similar conditions. This indicates that cis-acting elements necessary for the proper activation of the gene are present within 1 kb of 5′-flanking sequences. Functional analysis of 5′ deletions of the STH-2/GUS constructs by transient expression in leaf protoplasts revealed the presence of an upstream regulatory sequence between -135 and -52 which contains a TGAC motif, and a possible negative regulatory region between -52 and -28. A factor present in nuclear extracts of wounded potato tubers was found to bind specifically to nucleotides located between -135 to -105, suggesting that this region contains important cis-regulatory elements.
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Barrat DHP, Clark JA: Proteins arising during the late stages of embryogenesis in Pisum sativum L. Planta 184: 14–23 (1991).
Benfey PN, Ren L, Chua NH: The CaMV 35S enhancer contains at least two domains which can confer developmental and tissue-specific expression patterns. EMBO J 8: 2195–2202 (1989).
Breiteneder H, Pettenburger K, Bito A, Valenta R, Kraft D, Rumpold H, Scheiner O, Breitenbach M: The gene coding for the major birch pollen allergen BetvI, is highly homologous to a pea resistance response gene. EMBO J 8: 1935–1938 (1989).
Camirand A, St-Pierre B, Marineau C, Brisson N: Occurrence of a copia-like transposable element in one of the introns of the potato starch phosphorylase gene. Mol Gen Genet 224: 33–39 (1990).
Chiang CC, Hadwiger LA: Cloning and characterization of a disease resistance response gene in pea inducible by Fusarium solani. Mol Plant-Microbe Interact 3: 78–85 (1990).
Constabel CP, Brisson N: The defense-related STH-2 gene product of potato shows race-specific accumulation after inoculation with low concentrations of Phytophthora infestans zoospores. Planta 188: 289–295 (1992).
Constabel CP, Matton DP, Brisson N: Concurrent synthesis and degradation of alcohol dehydrogenase in elicitor-treated and wounded potato tubers. Plant Physiol 94: 887–981 (1990).
Coolbear T, Threlfall DR: A comparison of the sites of phytoalexins accumulation and of biosynthetic activity in potato tuber tissue inoculated with biotic elicitors. Phytochemistry 24: 2219–2224 (1985).
Crowell DN, John MJ, Russell D, Amasino RM: Characterisation of a stress-induced developmentally regulated gene family from soybean. Plant Mol Biol 18: 459–466 (1992).
De Block M: Genotype-independent leaf disc transformation of potato (Solanum tuberosum) using Agrobacterium tumefaciens. Theor Appl Genet 76: 767–774 (1988).
Dehbi M, Mbiguino A, Beauchemin M, Chatelain G, Bédard P-A: Transcriptional activation of the CEF-4/9E3 cytokine gene by pp60v-src. Mol Cell Biol 12: 1490–1499 (1992).
Ditta G, Stanfield S, Corbin D, Helsinski DR: Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77: 7347–7351 (1980).
Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).
Fristensky B, Horovitz D, Hadwiger LA: cDNA sequences for pea disease resistance response genes. Plant Mol Biol 11: 713–715 (1988).
Fritzemeier KH, Cretin C, Kombrink E, Rohwer F, Taylor J, Scheel D, Hahlbrock K: Transient induction of phenylalanine ammonia-lyase and 4-coumarate: CoA ligase mRNAs in potato leaves infected with virulent or avirulent races of Phytophthora infestans. Plant Physiol 85: 34–41 (1987).
Hahlbrock K, Scheel D: Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Plant Mol Biol 40: 347–369 (1989).
Horsch RB, Fry JE, Hoffmann NL, Eicholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).
Jefferson RA, Burgess SM, Hirsh D: β-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc Natl Acad Sci USA 83: 8447–8451 (1986).
Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).
Jouanneau J-P, Lapous D, Guern J: In plant protoplasts, the spontaneous expression of defense reactions and the responsiveness to exogenous elicitors are under auxin control. Plant Physiol 96: 459–466 (1991).
Katagiri F, Lam E, Chua N-H: Two tobacco DNA-binding proteins with homology to the nuclear factor CREB. Nature 340: 727–730 (1989).
Kombrink E, Schröder M, Hahlbrock K: Several ‘pathogenesis-related’ proteins in potato are 1,3-β-glucanases and chitinases. Proc Natl Acad Sci USA 85: 782–786 (1988).
Lamb CJ, Lawton MA, Dron M, Dixon RA: Signals and transduction mechanisms for activation of plant defenses against microbial attack. Cell 56: 215–224 (1989).
Lipman DJ, Pearson WR: Improved tools for biological sequence analysis. Proc Natl Acad Sci USA 85: 2444–2448 (1988).
Logemann J, Mayer JE, Schell J, Willmitzer L: Differential expression of genes in potato tubers after wounding. Proc Natl Acad Sci USA 85: 1136–1140 (1988).
Maeda N, Smithies O: The evolution of multigene families: human haptoglobin genes. Annu Rev Genet 20: 81–108 (1986).
Magnien E, Dalschaert X, Devreux M: Different radiosensitivity of N. plumbaginifolia leaves and regeneration of protoplasts. Plant Sci Lett 9: 231–241 (1980).
Marineau C, Matton DP, Brisson N: Differential accumulation of potato tuber mRNAs during the hypersensitive response induced by arachidonic acid elicitor. Plant Mol Biol 9: 335–342 (1987).
Matton DP, Brisson N: Cloning, expression, and sequence conservation of pathogenesis-related gene transcripts of potato. Mol Plant-Microbe Interac 2: 325–331 (1989).
Matton DP, Bell B, Brisson N: Nucleotide sequence of a pathogenesis-related gene of potato. Plant Mol Biol 14: 863–865 (1990).
Matton DP, Constabel CP, Brisson N: Alcohol dehydrogenase gene expression in potato following elicitor and stress treatment. Plant Mol Biol 14: 775–783 (1990).
Meier I, Hahlbrock K, Somssich IE: Elicitor-inducible and constitutive in vivo DNA footprints indicate novel cis-acting elements in the promoter of a parsley gene encoding pathogenesis-related protein 1. Plant Cell 3: 309–315 (1991).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–477 (1962).
Pryor T: The origin and structure of fungal disease resistance genes in plants. Trends Genet 3: 157–161 (1987).
Revzin A: Gel electrophoresis assays for DNA-protein interactions. Bio Techniques 7: 346–355 (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Siebertz B, Logemann J, Willmitzer L, Schell J: Cis-analysis of the wound-inducible promoter wun1 in transgenic tobacco plants and histochemical localization of its expression. Plant Cell 1: 961–968 (1989).
Singh K, Dennis ES, Ellis JG, Llewellyn DJ, Tokuhisa JG, Wahleithner JA, Peacock WJ: OCSBF-1, a maize Ocs enhancer binding factor: isolation and expression during development. Plant Cell 2: 891–903 (1990).
Smith HO, Birnstiel ML: A simple method for DNA restriction mapping. Nucl Acids Res 3: 2380–2398 (1976).
Somssich IE, Schmelzer E, Kawalleck P, Hahlbrock K: Gene structure and in situ transcript localization of pathogenesis-related protein 1 in parsley. Mol Gen Genet 213: 93–98 (1988).
Stanford AC, Bevan MH, Northcote DH: Differential expression within a family of novel wound-induced genes in potato. Mol Gen Genet 215: 200–208 (1989).
Tabata T, Nakayama T, Mikami K, Iwabuchi M: HBP-1a and HBP-1b: leucine zipper-type transcription factors of wheat. EMBO J 10: 1459–1467 (1991).
Taylor JL, Fritzemeier K-H, Häuser I, Kombrink E, Rohwer F, Schröder M, Strittmatter G, Hahlbrock K: Structural analysis and activation by fungal infection of a gene encoding a pathogenesis-related protein in potato. Mol Plant-Microbe Interact 3: 72–77 (1990).
van de Löcht U, Meier I, Hahlbrock K, Somssich IE: A 125 bp promoter fragment is sufficient for strong elicitormediated gene activation in parsley. EMBO J 9: 2945–2950 (1990).
Walter MH, Liu J-W, Grand C, Lamb CJ, Hess D: Bean pathogenesis-related (PR) proteins deduced from elicitor-induced transcripts are members of a ubiquitous new class of conserved PR proteins including pollen allergens. Mol Gen Genet 222: 353–360 (1990).
Warner SAJ, Scott R, Draper J: Characterisation of a wound-induced transcript that shares similarity with a class of intracellular pathogenesis-related (PR) proteins. Plant Mol Biol 19: 555–561 (1992).
Willmitzer L, Wagner KG: The isolation of nuclei from tissue-cultured plant cells. Exp Cell Res 135: 69–77 (1981).
Wu CH, Caspar T, Browse CJ, Lundquist S, Somerville C: Characterization of an HSP70 cognate gene family in Arabidopsis. Plant Physiol 88: 731–740 (1988).
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Matton, D.P., Prescott, G., Bertrand, C. et al. Identification of cis-acting elements involved in the regulation of the pathogenesis-related gene STH-2 in potato. Plant Mol Biol 22, 279–291 (1993). https://doi.org/10.1007/BF00014935
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DOI: https://doi.org/10.1007/BF00014935