Abstract
The Arabidopsis thaliana genes kin1 and cor6.6 belong to the same family and were expressed at higher levels following low temperature and ABA treatments. In an attempt to elucidate the mechanism of gene regulation by low temperature, the relationship between low-temperature- and abscisic acid (ABA)-induced gene expression and possible differential expression of the two genes, we have cloned a 5.3 kb genomic fragment harboring kin1 and cor6.6 and their respective 5′ sequences. The putative promoters of both genes were fused to the β-glucuronidase (GUS) coding sequence and GUS expression was analysed in transgenic tobacco and Arabidopsis plants. The cor6.6 promoter produced a higher basal level of expression than the kin1 promoter in transgenic tobacco. Enzyme assays of inducible GUS activity in transgenic Arabidopsis and tobacco plants showed that GUS activity directed by both kin1 and cor6.6 promoters was significantly induced by ABA, dehydration and osmoticum, but not by low temperature. Northern analysis revealed, in contrast, that GUS mRNA was significantly induced in these transgenic plants by low temperature. Further analysis showed that, at low temperature, GUS protein synthesis from the induced GUS mRNA was inhibited. Together these results reveal induction of kin1 and cor6.6 transcription by low temperature, exogenous ABA and dehydration. However, low-temperature expression is dramatically reduced at the translational level.
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Baker SS, Wilhelm KS, Thomashow MF: The 5′-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold-, drought- and ABA-regulated gene expression. Plant Mol Biol 24: 701–713 (1994).
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254 (1976).
Capell B, Dörffling K: Low temperature-induced changes of abscisic acid contents in barley and cucumber leaves in relation to their water status. J Plant Physiol 135: 571–575 (1989).
Chen HH, Li PH, Brenner ML: Involvement of abscisic acid in potato cold acclimation. Plant Physiol 71: 362–365 (1983).
Dallaire S, Houde M, Gagné Y, Saini HS, Boileau S, Chevrier N, Sarhan F: ABA and low temperature induce freezing tolerance via distinct regulatory pathways in wheat. Plant Cell Physiol 35: 1–9 (1994).
Davies PL, Hew CL: Biochemistry of fish antifreeze proteins. FASEB J 4: 2460–2468 (1990).
DureIII L, Crouch M, Harada J, Ho T-HD, Mundy J, Quatrano R, Thomas T, Sung ZR: Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol Biol 12: 475–486 (1989).
Gilmour SJ, Thomashow MF: Cold acclimation and cold-regulated gene expression in ABA mutants of Arabidopsis thaliana. Plant Mol Biol 17: 1233–1240 (1991).
Gilmour SJ, Artus NN, Thomashow MF: cDNA sequence analysis and expression of two cold-regulated genes of Arabidopsis thaliana. Plant Mol Biol 18: 13–21 (1992).
Guiltinan MJ, Marcotte WR, Quatrano RS: A plant leucine zipper protein that recognizes an abscisic acid response element. Science 250: 267–271 (1990).
Guy CL: Cold acclimation and freezing stress tolerance: role of protein metabolism. Annu Rev Plant Physiol Plant Mol Biol 41: 187–223 (1990).
Guy CL, Haskell D: Detection of polypeptides associated with the cold acclimation process in spinach. Electrophoresis 9: 787–796 (1988).
Hahn M, Walbot V: Effects of cold-treatment on protein synthesis and mRNA levels in rice leaves. Plant Physiol 91: 930–938 (1989).
Hajela RK, Horvath DP, Gilmour SJ, Thomashow MF: Molecular cloning and expression of cor (cold regulated) genes in Arabidopsis thaliana. Plant Physiol 93: 1246–1252 (1990).
Horvath DP, McLarney BK, Thomashow MF: Regulation of Arabidopsis thaliana L. (Heyn) cor78 in response to low temperature. Plant Physiol 103: 1047–1053 (1993).
Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).
Kurkela S, Borg-Franck M: Structure and expression of kin2, one of two cold- and ABA-induced genes of Arabidopsis thaliana. Plant Mol Biol 19: 689–692 (1992).
Kurkela S, Franck M: Cloning and characterization of a cold- and ABA-inducible Arabidopsis gene. Plant Mol Biol 15: 137–144 (1990).
Lalk I, Dörffling K: Hardening, abscisic acid, proline and freezing resistance in two winter wheat varieties. Physiol Plant 63: 287–292 (1985).
Lång V, Palva ET: The expression of a rab-related gene, rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 20: 951–962 (1992).
Lång V, Mäntylä E, Welin B, Sundberg B, Palva ET: Alterations in water status, endogenous abscisic acid content, and expression of rab18 gene during the development of freezing tolerance in Arabidopsis thaliana. Plant Physiol 104: 1341–1349 (1994).
Lee SP, Chen TH-H: Molecular biology of plant cold hardiness development. In: Li PH, Christersson L (eds) Plant Cold Hardiness, pp. 1–29. PRC Press, Boca Raton, FL (1993).
Lin C, Thomashow MF: DNA sequence analysis of a complementary DNA for cold-regulated Arabidopsis gene cor15 and characterization of the COR15 polypeptide. Plant Physiol 99: 519–525 (1992).
Lundberg KS, Shoemaker DD, Adams MWW, Short JM, Sorge JA, Mathur EJ: High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus. Gene 108: 1–6 (1991).
Marcotte WR, Russell SH, Quatrano RS: Abscisic acid-responsive sequences from the Em gene of wheat. Plant Cell 1: 969–976 (1989).
McKendree WL, Ferl RJ: Functional elements of the Arabidopsis Adh promoter include the G-box. Plant Mol Biol 19: 859–862 (1992).
Mundy J, Yamaguchi-Shinozaki K, Chua N-H: Nuclear proteins bind concerved elements in the abscisic acid-responsive promoter of a rice rab gene. Proc Natl Acad Sci USA 87: 1406–1410 (1990).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
Neven LG, Haskell DW, Hofig A, Li Q-B, Guy CL: Characterization of a spinach gene responsive to low temperature and water stress. Plant Mol Biol 21: 291–305 (1993).
Nordin K, Heino P, Palva ET: Separate signal pathways regulate the expression of a low-temperature-induced gene in Arabidopsis thaliana (L.) Heynh. Plant Mol Biol 16: 1061–1071 (1991).
Nordin K, Vahala T, Palva ET: Differential expression of two related, low-temperature-induced genes in arabidopsis thaliana (L.) Heynh. Plant Mol Biol 21: 641–653 (1993).
Pellé R, Murphy NB: Northern hybridization: rapid and simple electrophoretic conditions. Nucl Acids Res 21: 2783–2784 (1993).
Schuler MA, Zielinski RE: Methods in plant molecular biology. Academic Press, San Diego, CA (1989).
Skriver K, Mundy J: Gene expression in response to abscisic acid and osmotic stress. Plant Cell 2: 503–512 (1990).
Snustad DP, Haas NA, Kopczak S, Silflow CD: The small genome of Arabidopsis contains at least nine expressed β-tubulin genes. Plant Cell 4: 549–556 (1992).
Sutton F, Ding X, Kenefick DG: Group 3 LEA gene HVA1 regulation by cold acclimation and deacclimation in two barley cultivars with varying freeze resistance. Plant Physiol 99: 338–340 (1992).
Thomashow MF: Molecular genetics of cold acclimation in higher plants. Adv Genet 28: 99–131 (1990).
Valvekens D, VanMontagu M, VanLijsebettens M: Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci USA 85: 5336–5540 (1988).
Yamaguchi-Shinozaki K, Shinozaki K: Characterization of the expression of a desiccation-responsive rd29 gene of Arabidopsis thaliana and analysis of its promoter in transgenic plants. Mol Gen Genet 236: 331–340 (1993).
Yamaguchi-Shinozaki K, Shinozaki K: A novel cis-acting element in an arabidopsis gene is involved in responsiveness to drought, low temperature, or high-salt stress. Plant Cell 6: 251–264 (1994).
Wang H, Qi M, Cutler AJ: A simple method of preparing plant samples for PCR. Nucl Acid Res 21: 4153–4154 (1993).
Wang H, Georges F, Pelcher LE, Saleem M, Cutler AJ: A 5.3 kb genomic sequence from Arabidopsis thaliana containing kin1 and cor6.6. Plant Physiol 104: 291–292 (1994).
Wilhelm KS, Thomashow MF: Arabidopsis thaliana cor15b, an apparent homologue of cor15a, is strongly responsive to cold and ABA, but not drought. Plant Mol Biol 23: 1073–1077 (1993).
Williams ME, Foster R, Chua NH: Sequences flanking the hexameric G-box core CACGTG affect the specificity of protein binding. Plant Cell 4: 485–496 (1992).
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Wang, H., Datla, R., Georges, F. et al. Promoters from kin1 and cor6.6, two homologous Arabidopsis thaliana genes: transcriptional regulation and gene expression induced by low temperature, ABA, osmoticum and dehydration. Plant Mol Biol 28, 605–617 (1995). https://doi.org/10.1007/BF00021187
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DOI: https://doi.org/10.1007/BF00021187