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| Construction and functional analysis of a synthetic wound- and hormone-inducible promoter ocs/mas |
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Abstract Abstract The use of optimal regulatory sequences for simultaneous expression of the transgenes might play a significant role in engineering plants with increased disease and insect resistance. The plant expression vector pOMS-GUS, which contained the GUS gene under the control of a chimeric promoter based upon the mannopine synthase (mas) promoter and the octopine synthase (ocs) enhancer, was constructed. Used as control, another vector pMAS-GUS, carried the GUS gene driven by only the mas promoter. The two vectors were introduced into tobacco plants by Agrobacterium-mediated transformation. Fluorometric assays for GUS activity and reverse transcription–polymerase chain reaction (RT-PCR) analysis revealed that GUS gene expressed weakly with untreated transgenic tobacco while the level of GUS activity increased steadily after 1 h subjected to wounding. The expression of the mas and ocs/mas promoters was induced a further 1.8-fold and 5.7-fold, respectively. SA(1 mmol/L) or MJ(250 μmol/L) treatment also caused a large induction of the ocs/mas chimeric promoter;And the application of SA in combination with MJ(1 mmol/LSA & 250 μmol/L MJ) produced an additive effect that exceeded the wounding response. The results showed that the ocs/mas chimeric promoter is a strong inducible promoter that can be activated by various stresses. The chimeric promoter should have utility in development of disease and insect resistant transgenic crops.
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Received: 23 October 2008
Published: 28 July 2009
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| ［1］ Kiba A, Tomiyama H, Takahashi H, et al. Induction of resistance and expression of defense-related genes in tobacco leaves infiltrated with Ralstonia solanacearum.Plant Cell Physio, 2003, 44(3): 287~295
［2］ Lee S C, Kim S H, An S H, et al. Identification and functional expression of the pepper pathogen-induced gene,CAPIP2, involved in disease resistance and drought and salt stress tolerance. Plant Mol Biol, 2006, 62(1-2): 151~164
［3］ Silvar C, Merino F, Díaz J. Differential activation of defense-related genes in susceptible and resistant pepper cultivars infected with Phytophthora capsici. J Plant Physiol, 2008, 165(10): 1120~1124
［4］ Kasuga M, Liu Q, Miura S, et al. Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol, 1999, 17(3): 287~291
［5］ Sunilkumar G, Mohr L, Lopata-Finch E, et al. Developmental and tissue-specific expression of CaMV 35S promoter in cotton as revealed by GFP. Plant Mol Biol, 2002, 50(3):463~474
［6］ Liu J J, Ekramoddoullah A K, Piggott N, et al. Molecular cloning of a pathogen/wound-inducible PR10 promoter from Pinus monticola and characterization in transgenic Arabidopsis plants. Planta, 2005, 221(2): 159~169
［7］ Chatterjee M, Banerjee A K, Hannapel D J. A BELL1-like gene of potato is light activated and wound inducible. Plant Physiol, 2007, 145(4): 1435~1443
［8］ 柴玉荣，田芳，刘红涛，等．杜氏盐藻rbcS启动子的克隆和功能分析．中国生物工程杂志，2008，28(4): 47~52
Chai Y R, Tian F, Liu H T, et al. China Biotechnology, 2008, 28(4): 47~52
［9］ Venter M. Synthetic promoters: genetic control through cis engineering. Trends Plant Sci, 2007, 12(3): 118~124
［10］ Rushton PJ, Reinstadler A, Lipka V, et al. Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen- and wound-induced signaling. Plant Cell, 2002, 14(4): 749~762
［11］ Ni M, Cui D C, Einstein J, et al. Strength and tissue specificity of chimeric promoters derived from the octopine and mannopine synthase genes. Plant J, 1995, 7(4): 661~676
［12］ Feltkam PD, Masterson R, Starke J, et al. Analysis of the involvement of ocs-like bZIP-binding elements in the differential strength of the bidirectional mas1'2' promoter. Plant Physiol, 1994, 105(1): 259~268
［13］ Comai L, Moran P, Maslyar D. Novel and useful properties of a chimeric plant promoter combining CaMV 35S and MAS elements. Plant Mol Biol, 1990, 15(3): 373~381
［14］ Ni M, Cui D, Gelvin S B. Sequence-specific interactions of wound-inducible nuclear factors with mannopine synthase 2' promoter wound-responsive elements. Plant Mol Biol, 1996, 30(1): 77~96
［15］ Guevara-García A, Mosqueda-Cano G, Argüello-Astorga G, et al. Tissue-specific and wound- inducible pattern of expression of the mannopine synthase promoter is determined by the interaction between positive and negative cis-regulatory elements. Plant J, 1993, 4(3): 495~505
［16］ Zhang B, Singh K B. ocs element promoter sequences are activated by auxin and salicylic acid in Arabidopsis. Proc Natl Acad Sci USA, 1994, 91(7): 2507~2511
［17］ Chen W Q, Singh K B. The auxin, h?rogen peroxide and salicylic acid induced expression of the Arabidopsis GST6 promoter is mediated in part by an ocs element. Plant J, 1999, 19(6): 667~677
［18］ Fox PC, Vasil V, Vasil I K, et al. Multiple ocs-like elements required for efficient transcription of the mannopine synthase gene of T-DNA in maize protoplasts. Plant Mol Biol, 1992, 20(2): 219~233
［19］ Leisner S M, Gelvin S B. Structure of the octopine synthase upstream activator sequence. Proc Natl Acad Sci USA, 1988, 85(8): 2553~2557
［20］ Horsch R B, Fry J E, Hoffmann N L, et al. A simple and general methods for transferring genes into plants. Science, 1985, 227(4691): 1229~1231
［21］ Jefferson R A, Kavanagh T A, Bevan M W. GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J, 1987, 6(13): 3901~3907
［22］ 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, 72(1-2): 248~254
［23］ Guevara-García A, López-Ochoa L López-Bucio J, et al. A 42 b Pfragment of the pmas1' promoter containing an ocs-like element confers a developmental, wound and chemically inducible expression pattern. Plant Mol Biol, 1998, 38(5): 743~753
［24］ Kim S R, Kim Y, An G. Identification of methyl jasmonate and salicylic acid response elements from the nopaline synthase (nos) promoter. Plant Physiol, 1996, 103(1): 97~103
［25］ Kim Y J, Kim D G, Lee S H, et al. Wound-induced expression of the ferulate 5-hydroxylase gene in Camptotheca acuminata. Biochim Biophys Acta, 2006, 1760(2): 182~190
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