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. 1995 Sep;109(1):73–85. doi: 10.1104/pp.109.1.73

Transgenic analysis of a hybrid poplar wound-inducible promoter reveals developmental patterns of expression similar to that of storage protein genes.

J B Hollick 1, M P Gordon 1
PMCID: PMC157565  PMID: 7480333

Abstract

The wound-inducible win3 multigene family from hybrid poplars (Populus trichocarpa x Populus deltoides) encodes proteins with structural similarities with Kunitz-type protease inhibitors (H.D. Bradshaw Jr., J.B. Hollick, T.J. Parsons, H.R.G. Clarke, M.P. Gordon [1990] Plant Mol Biol 14: 51-59), and at least one member, win3.12, is transcribed de novo in the injured and uninjured leaves of wounded trees (J.B. Hollick, M.P. Gordon [1993] Plant Mol Biol 22: 561-572). A previous study demonstrated that 1352 bp of 5' flanking DNA from the win3.12 gene confers local wound-regulated expression of the beta-glucuronidase gene in transgenic tobacco (Nicotiana tabacum cv Xanthi n.c.) (J.B. Hollick, M.P. Gordon [1993] Plant Mol Biol 22: 561-572). We extend this transgenic analysis here by examining the developmental regulation and systemic wound induction conferred by the same transgene construct in tobacco. Biochemical and histochemical surveys of beta-glucuronidase activity are described for four, independent transgenic lines. The observed spatial and temporal expression patterns coincide with dormant storage tissues and with previously described expression patterns for both seed and vegetative storage protein genes. Developmental northern blot analysis of win3 RNA levels in poplar seeds confirms that proper temporal expression of the reporter gene is maintained during tobacco seed maturation. These results demonstrate that a putative Kunitz-type protease inhibitor can be wound inducible in addition to being expressed in developing seeds.

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Selected References

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  1. Allen R. L., Lonsdale D. M. Molecular characterization of one of the maize polygalacturonase gene family members which are expressed during late pollen development. Plant J. 1993 Feb;3(2):261–271. doi: 10.1111/j.1365-313x.1993.tb00177.x. [DOI] [PubMed] [Google Scholar]
  2. Bagga S., Sutton D., Kemp J. D., Sengupta-Gopalan C. Constitutive expression of the beta-phaseolin gene in different tissues of transgenic alfalfa does not ensure phaseolin accumulation in non-seed tissue. Plant Mol Biol. 1992 Sep;19(6):951–958. doi: 10.1007/BF00040527. [DOI] [PubMed] [Google Scholar]
  3. Beachy R. N., Chen Z. L., Horsch R. B., Rogers S. G., Hoffmann N. J., Fraley R. T. Accumulation and assembly of soybean beta-conglycinin in seeds of transformed petunia plants. EMBO J. 1985 Dec 1;4(12):3047–3053. doi: 10.1002/j.1460-2075.1985.tb04044.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boller T., Kende H. Hydrolytic enzymes in the central vacuole of plant cells. Plant Physiol. 1979 Jun;63(6):1123–1132. doi: 10.1104/pp.63.6.1123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Bradshaw H. D., Jr, Hollick J. B., Parsons T. J., Clarke H. R., Gordon M. P. Systemically wound-responsive genes in poplar trees encode proteins similar to sweet potato sporamins and legume Kunitz trypsin inhibitors. Plant Mol Biol. 1990 Jan;14(1):51–59. doi: 10.1007/BF00015654. [DOI] [PubMed] [Google Scholar]
  7. Breyne P., Gheysen G., Jacobs A., Van Montagu M., Depicker A. Effect of T-DNA configuration on transgene expression. Mol Gen Genet. 1992 Nov;235(2-3):389–396. doi: 10.1007/BF00279385. [DOI] [PubMed] [Google Scholar]
  8. Budar F., Thia-Toong L., Van Montagu M., Hernalsteens J. P. Agrobacterium-Mediated Gene Transfer Results Mainly in Transgenic Plants Transmitting T-DNA as a Single Mendelian Factor. Genetics. 1986 Sep;114(1):303–313. doi: 10.1093/genetics/114.1.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bustos M. M., Begum D., Kalkan F. A., Battraw M. J., Hall T. C. Positive and negative cis-acting DNA domains are required for spatial and temporal regulation of gene expression by a seed storage protein promoter. EMBO J. 1991 Jun;10(6):1469–1479. doi: 10.1002/j.1460-2075.1991.tb07667.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clarke H. R., Davis J. M., Wilbert S. M., Bradshaw H. D., Jr, Gordon M. P. Wound-induced and developmental activation of a poplar tree chitinase gene promoter in transgenic tobacco. Plant Mol Biol. 1994 Aug;25(5):799–815. doi: 10.1007/BF00028875. [DOI] [PubMed] [Google Scholar]
  11. Coleman G. D., Englert J. M., Chen THH., Fuchigami L. H. Physiological and Environmental Requirements for Poplar (Populus deltoides) Bark Storage Protein Degradation. Plant Physiol. 1993 May;102(1):53–59. doi: 10.1104/pp.102.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DeWald D. B., Sadka A., Mullet J. E. Sucrose Modulation of Soybean Vsp Gene Expression Is Inhibited by Auxin. Plant Physiol. 1994 Feb;104(2):439–444. doi: 10.1104/pp.104.2.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Downing W. L., Mauxion F., Fauvarque M. O., Reviron M. P., de Vienne D., Vartanian N., Giraudat J. A Brassica napus transcript encoding a protein related to the Künitz protease inhibitor family accumulates upon water stress in leaves, not in seeds. Plant J. 1992 Sep;2(5):685–693. [PubMed] [Google Scholar]
  14. Fujiwara T., Lessard P. A., Beachy R. N. Seed-specific repression of GUS activity in tobacco plants by antisense RNA. Plant Mol Biol. 1992 Dec;20(6):1059–1069. doi: 10.1007/BF00028893. [DOI] [PubMed] [Google Scholar]
  15. Gendloff E. H., Bowen B., Buchholz W. G. Quantitation of chloramphenicol acetyl transferase in transgenic tobacco plants by ELISA and correlation with gene copy number. Plant Mol Biol. 1990 Apr;14(4):575–583. doi: 10.1007/BF00027503. [DOI] [PubMed] [Google Scholar]
  16. Hattori T., Yoshida N., Nakamura K. Structural relationship among the members of a multigene family coding for the sweet potato tuberous root storage protein. Plant Mol Biol. 1989 Nov;13(5):563–572. doi: 10.1007/BF00027316. [DOI] [PubMed] [Google Scholar]
  17. Hildmann T., Ebneth M., Peña-Cortés H., Sánchez-Serrano J. J., Willmitzer L., Prat S. General roles of abscisic and jasmonic acids in gene activation as a result of mechanical wounding. Plant Cell. 1992 Sep;4(9):1157–1170. doi: 10.1105/tpc.4.9.1157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hollick J. B., Gordon M. P. A poplar tree proteinase inhibitor-like gene promoter is responsive to wounding in transgenic tobacco. Plant Mol Biol. 1993 Jul;22(4):561–572. doi: 10.1007/BF00047398. [DOI] [PubMed] [Google Scholar]
  19. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jofuku K. D., Goldberg R. B. Kunitz trypsin inhibitor genes are differentially expressed during the soybean life cycle and in transformed tobacco plants. Plant Cell. 1989 Nov;1(11):1079–1093. doi: 10.1105/tpc.1.11.1079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Khandjian E. W. UV crosslinking of RNA to nylon membrane enhances hybridization signals. Mol Biol Rep. 1986;11(2):107–115. doi: 10.1007/BF00364822. [DOI] [PubMed] [Google Scholar]
  22. Langheinrich U., Tischner R. Vegetative storage proteins in poplar : induction and characterization of a 32- and a 36-kilodalton polypeptide. Plant Physiol. 1991 Nov;97(3):1017–1025. doi: 10.1104/pp.97.3.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mason H. S., DeWald D. B., Mullet J. E. Identification of a methyl jasmonate-responsive domain in the soybean vspB promoter. Plant Cell. 1993 Mar;5(3):241–251. doi: 10.1105/tpc.5.3.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mason H. S., Dewald D. B., Creelman R. A., Mullet J. E. Coregulation of soybean vegetative storage protein gene expression by methyl jasmonate and soluble sugars. Plant Physiol. 1992 Mar;98(3):859–867. doi: 10.1104/pp.98.3.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ohto M. A., Nakamura-Kito K., Nakamura K. Induction of Expression of Genes Coding for Sporamin and beta-Amylase by Polygalacturonic Acid in Leaf-Petiole Cuttings of Sweet Potato. Plant Physiol. 1992 Jun;99(2):422–427. doi: 10.1104/pp.99.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Peach C., Velten J. Transgene expression variability (position effect) of CAT and GUS reporter genes driven by linked divergent T-DNA promoters. Plant Mol Biol. 1991 Jul;17(1):49–60. doi: 10.1007/BF00036805. [DOI] [PubMed] [Google Scholar]
  27. Pearce G., Johnson S., Ryan C. A. Purification and characterization from tobacco (Nicotiana tabacum) leaves of six small, wound-inducible, proteinase isoinhibitors of the potato inhibitor II family. Plant Physiol. 1993 Jun;102(2):639–644. doi: 10.1104/pp.102.2.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rackis J. J., Anderson R. L. Isolation of four soybean trypsin inhibitors by DEAE-cellulose chromatography. Biochem Biophys Res Commun. 1964 Mar 26;15(3):230–235. doi: 10.1016/0006-291x(64)90151-2. [DOI] [PubMed] [Google Scholar]
  29. Renckens S., De Greve H., Van Montagu M., Hernalsteens J. P. Petunia plants escape from negative selection against a transgene by silencing the foreign DNA via methylation. Mol Gen Genet. 1992 May;233(1-2):53–64. doi: 10.1007/BF00587561. [DOI] [PubMed] [Google Scholar]
  30. Ryan C. A., Huisman O. C. Chymotrypsin inhibitor I from potatoes: a transient protein component in leaves of young potato plants. Nature. 1967 Jun 3;214(5092):1047–1049. doi: 10.1038/2141047a0. [DOI] [PubMed] [Google Scholar]
  31. Staswick P. E. Developmental regulation and the influence of plant sinks on vegetative storage protein gene expression in soybean leaves. Plant Physiol. 1989 Jan;89(1):309–315. doi: 10.1104/pp.89.1.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Twell D., Yamaguchi J., Wing R. A., Ushiba J., McCormick S. Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Dev. 1991 Mar;5(3):496–507. doi: 10.1101/gad.5.3.496. [DOI] [PubMed] [Google Scholar]
  33. Walling L., Drews G. N., Goldberg R. B. Transcriptional and post-transcriptional regulation of soybean seed protein mRNA levels. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2123–2127. doi: 10.1073/pnas.83.7.2123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Warner S. A., Scott R., Draper J. Isolation of an asparagus intracellular PR gene (AoPR1) wound-responsive promoter by the inverse polymerase chain reaction and its characterization in transgenic tobacco. Plant J. 1993 Feb;3(2):191–201. doi: 10.1046/j.1365-313x.1993.t01-11-00999.x. [DOI] [PubMed] [Google Scholar]
  35. Wenzler H., Mignery G., Fisher L., Park W. Sucrose-regulated expression of a chimeric potato tuber gene in leaves of transgenic tobacco plants. Plant Mol Biol. 1989 Oct;13(4):347–354. doi: 10.1007/BF00015546. [DOI] [PubMed] [Google Scholar]

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