Abstract
Ribosome-inactivating proteins (RIPs) represent those proteins that universally depurinate conserved α-sarcin loops of large rRNAs. In this study, a 0.6-kb fragment of a 5′ flanking region preceding a curcin gene, encoding a type I RIP curcin, of Jatropha curcas L. endosperm was cloned, and its regulation of expression of the β-glucuronidase (GUS) reporter gene was investigated in transgenic tobacco. Analysis of GUS activities showed that the 0.6-kb flanking fragment of the curcin gene was sufficient to drive the GUS reporter gene expression in tobacco seed. The activity of this flanking fragment was analyzed at different stages of seed development. Histochemical localization of GUS activity indicated that the promoter was specifically active in the endosperm tissue of the dicotyledonous tobacco embryo. Moreover, this activity was first initiated at the heart-shaped embryonic stage during seed development.
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Abbreviations
- DAF:
-
Days after flowering
- GUS:
-
β-glucuronidase
- IC50 :
-
Half maximal inhibitory concentration
- DOFCORE:
-
Regulatory element of AAAG sequence
- DPBFCORE:
-
Regulatory element of ACACNNG sequence
- E-BOX:
-
Regulatory element of CANNTG sequence
- Prolamin box:
-
Regulatory element of TGCAAAG sequence
- ABRE:
-
Regulatory element of ACGTG sequence
References
Asai T, Tena G, Plotnikova J, Willmann MR, Chiu WL, Gomez-Gomez L et al (2002) MAP kinase signalling cascade in Arabidopsis innate immunity. Nature 415:977–983 doi:10.1038/415977a
Barbieri L, Battelli MG, Stirpe F (1993) Ribosome-inactivating proteins from plants. Biochim Biophys Acta 1154:237–282
Bradford MM (1976) 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 doi:10.1016/0003-2697(76)90527-3
Chen PY, Wang CK, Soong SC, To KY (2003) Complete sequence of the binary vector pBI121 and its application in cloning T-DNA insertion from transgenic plants. Mol Breed 11:287–293 doi:10.1023/A:1023475710642
Eulgem T, Rushton PJ, Schmelzer E, Hahlbrock K, Somssich IE (1999) Early nuclear events in plant defence signalling: rapid gene activation by RKY transcription factors. EMBO J 18:4689–4699 doi:10.1093/emboj/18.17.4689
Finkelstein RR, Lynch TJ (2000) The Arabidopsis abscisic acid response gene ABI5 encodes a basic leucine zipper transcription factor. Plant Cell 12:599–609
Hartmann U, Sagasser M, Mehrtens F, Stracke R, Weisshaar B (2005) Differential combinatorial interactions of cis-acting elements recognized by R2R3-MYB, BZIP, and BHLH factors control light-responsive and tissue-specific activation of phenylpropanoid biosynthesis genes. Plant Mol Biol 57:155–171 doi:10.1007/s11103-004-6910-0
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database. Nucleic Acids Res 27:297–300 doi:10.1093/nar/27.1.297
Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303:179–180 doi:10.1038/303179a0
Horsch RB, Fry JE, Hoffman NL, Eichholz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231 doi:10.1126/science.227.4691.1229
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Johnson CS, Kolevski B, Smyth DR (2002) TRANSPARENT TESTA GLABRA2, a trichome and seed coat development gene of Arabidopsis, encodes a WRKY transcription factor. Plant Cell 14:1359–1375 doi:10.1105/tpc.001404
Kim SY, Chung HJ, Thomas TL (1997) Isolation of a novel class of bZIP transcription factors that interact with ABA-responsive and embryo-specification elements in the Dc3 promoter using a modified yeast one-hybrid system. Plant J 11:1237–1251 doi:10.1046/j.1365-313X.1997.11061237.x
Komari T, Hiei Y, Saito Y, Murai N, Kumashiro T (1996) Vectors carrying two separate T-DNAs for co-transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers. Plant J 10:165–174 doi:10.1046/j.1365-313X.1996.10010165.x
Li J, Brader G, Palva ET (2004) The WRKY70 transcription factor: a node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense. Plant Cell 16:319–331 doi:10.1105/tpc.016980
Lin J, Chen Y, Xu Y, Yan F, Tang L, Chen F (2003) Cloning and expression of curcin, a ribosome-inactivating protein from the seeds of Jatropha curcas. Acta Bot Sin 45:858–863
McElroy D, Brettell RIS (1994) Foreign gene expression in transgenic cereals. Trends Biotechnol 12:62–67 doi:10.1016/0167-7799(94)90102-3
Mourgue M, Delphaut J, Baret R, Kassab R (1961) Study of the toxicity and localization of toxalbumin (curcin) in the seeds of Jatropha curcas Linn. Bull Soc Chim Biol (Paris) 43:517–531
Nishiuchi T, Shinshi H, Suzuki K (2004) Rapid and transient activation of transcription of the ERF3 gene by wounding in tobacco leaves: possible involvement of NtWRKYs and autorepression. J Biol Chem 279:55355–55361 doi:10.1074/jbc.M409674200
Robatzek S, Somssich IE (2002) Targets of AtWRKY6 regulation during plant senescence and pathogen defense. Genes Dev 16:1139–1149 doi:10.1101/gad.222702
Sambrook J, Russell D (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA
Simpson SD, Nakashima K, Narusaka Y, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Two different novel cis-acting elements of erd1, a clpA homologous Arabidopsis gene function in induction by dehydration stress and dark-induced senescence. Plant J 33:259–270 doi:10.1046/j.1365-313X.2003.01624.x
Stirpe F (2004) Ribosome-inactivating proteins. Toxicon 44:371–383 doi:10.1016/j.toxicon.2004.05.004
Stirpe F, Pession-Brizzi A, Lorenzoni E (1976) Studies on the proteins from the seeds of Croton toglium and of Jatropha curcas. Biochem J 156:1–6
Sun C, Palmqvist S, Olsson H, Boren M, Ahlandsberg S, Jansson C (2003) A novel WRKY transcription factor, SUSIBA2, participates in sugar signaling in barley by binding to the sugar-responsive elements of the isoI promoter. Plant Cell 15:2076–2092 doi:10.1105/tpc.014597
Wang XD, Wang ZP, Zou YP (1996) An improved procedure for the isolation of nuclear DNA from leaves of wild grapevine dried with silica gel. Plant Mol Biol Rep 14:369–373 doi:10.1007/BF02673369
Wu C, Washida H, Onodera Y, Harada K, Takaiwa F (2000) Quantitative nature of the Prolamin-box, ACGT and AACA motifs in a rice glutelin gene promoter: minimal cis-element requirements for endosperm-specific gene expression. Plant J 23:415–421 doi:10.1046/j.1365-313x.2000.00797.x
Yanagisawa S (2000) Dof1 and Dof2 transcription factors are associated with expression of multiple genes involved in carbon metabolism in maize. Plant J 21:281–288 doi:10.1046/j.1365-313x.2000.00685.x
Yanagisawa S, Schmidt RJ (1999) Diversity and similarity among redognition sequences of Dof transcription factors. Plant J 17:209–214 doi:10.1046/j.1365-313X.1999.00363.x
Acknowledgments
This work was supported by the General Program of National Natural Science Foundation of China (No. 30670204), China International Science and Technology Cooperation Project (No. 2004DFB00300, No. 2006DFB63400), and the Key Project of Chinese Ministry of Education (No. 104151).
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Qin, X., Zhang, J., Shao, C. et al. Isolation and Characterization of a Curcin Promoter from Jatropha curcas L. and Its Regulation of Gene Expression in Transgenic Tobacco Plants. Plant Mol Biol Rep 27, 275–281 (2009). https://doi.org/10.1007/s11105-008-0078-8
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DOI: https://doi.org/10.1007/s11105-008-0078-8