Abstract
Dirigent and dirigent-like family proteins contain a number of proteins involved in lignification or in the response to pathogen infection and abiotic stress in plants. In the present study, a full-length cDNA sequence of a dirigent-like gene designated ScDir (GenBank Accession Number JQ622282) was obtained from sugarcane based on the stem full-length cDNA library. The ScDir gene was 819-bp long, including a 564-bp ORF encoding 187 amino acid residues. The protein N-terminus contained signal peptides at amino acid residues of 1–25 and transmembrane regions at 7–26 aa. A his-tagged ScDir protein with an estimated molecular mass of 27.4 kDa was expressed in Escherichia coli system. The expressed ScDir protein had increased the host cell’s tolerance to PEG and NaCl. When an endogenous GAPDH gene was used as internal control, results from real-time qPCR demonstrated that the ScDir mRNA amount in sugarcane stems was significantly higher than that in the roots, leaves and buds by 18.64 ± 0.48, 25,635.16 ± 2,966.03 and 721.50 ± 8.17-fold, respectively. The ScDir transcript levels in sugarcane seedling increased under H2O2, PEG or NaCl stress. The expression level of ScDir was significantly upregulated under PEG stress, and the highest level was observed at 12 h after stress. Thus, both the ScDir-hosted cell performance and the enhanced expressions in sugarcane imply that the ScDir gene is involved in the response to abiotic stresses of drought, salts and oxidation. The transcription of the ScDir gene is highly stem-specific, as revealed by real-time qPCR.
Key message A novel sugarcane Sc-Dir gene, DIRd subfamily, which is highly stalk-specific expression and involved in the response to artificial stresses of drought, salts, and oxidatives.
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References
Burlat V, Kwon M, Davin LB, Lewis NG (2001) Dirigent proteins and dirigent sites in lignifying tissues. Phytochemistry 57:883–897
Casu RE, Dimmock CM, Chapman SC, Grof CPL, McIntyre CL, Bonnett GD, Manners JM (2004) Identification of differentially expressed transcripts from maturing stem of sugarcane by in silico analysis of stem expressed sequence tags and gene expression profiling. Plant Mol Biol 54:503–517
Damaj MB, Kumpatla SP, Emani C, Beremand PD, Avutu S, Reddy AS, Rathore KS, Buenrostro-Nava MT, Curtis IS, Thomas TL, Mirkov TE (2010) Sugarcane DIRIGENT and O-methyltransferase promoters confer stem-regulated gene expression in diverse monocots. Planta 231:1439–1458
Davin LB, Lewis NG (2000) Dirigent proteins and dirigent sites explain the mystery of specificity of radical precursor coupling in lignan and lignin biosynthesis. Plant Physiol 123:453–462
Davin LB, Wang HB, Crowell AL, Bedgar DL, Martin DM, Sarkanen S, Lewis NG (1997) Stereoselective bimolecular phenoxy radical coupling by an auxiliary (dirigent) protein without an active center. Science 275:362–366
Gang DR, Costa MA, Fujita M, Dinkova-Kostova AT, Wang HB, Burlat V, Martin W, Sarkanen S (1999) Regiochemical control of monolignol radical coupling: a new paradigm for lignin and lignan biosynthesis. Chem Biol 6:143–151
Gao CQ, Liu GF, Wang YC, Jiang J, Yang CP (2010) Cloning and analysis of dirigent-like protein in gene from Tamarix androssowii. Bull Bot Res 30:81–86
Guo JL, Que YX, Liu JX, Zheng YF, Chen RK, Xu LP (2009) Construction of full-length cDNA library for sugarcane stem by optimized oligo-capping. Chin J Trop Crop 30:672–676
Gupta K, Agarwal PK, Reddy MK, Jha B (2010) SbDREB2A, an A-2 type DREB transcription factor from extreme halophyte Salicornia brachiata confers abiotic stress tolerance in Escherichia coli. Plant Cell Rep 29:1131–1137
Iskandar HM, Simpson RS, Casu RE, Bonnett GD, Maclean DJ, Manners JM (2004) Comparison of reference genes for quantitative real-time polymerase chain reaction analysis of gene expression in sugarcane. Plant Mol Biol Report 22:325–337
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT method. Methods 25:402–408
Que YX, Xu LP, Xu JS, Zhang JS, Zhang MQ, Chen RK (2009a) Selection of control genes in real-time qPCR analysis of gene expression in sugarcane. Chin J Trop Crop 30:274–278
Que YX, Yang ZX, Xu LP, Chen RK (2009b) Isolation and identification of differentially expressed genes in sugarcane infected by Ustilago scitaminea. Acta Agron Sin 3:452–458
Ralph S, Park JY, Bohlmann J, Mansfield SD (2006) Dirigent proteins in conifer defense: gene discovery, phylogeny, and differential wound- and insect-induced expression of a family of DIR and DIR-like genes in spruce (Picea spp.). Plant Mol Biol 60:21–40
Ralph SG, Jancsik S, Bohlmann J (2007) Dirigent proteins in conifer defense II: extended gene discovery, phylogeny, and constitutive and stress-induced gene expression in spruce (Picea spp.). Phytochemistry 68:1975–1991
Van der Weele CM, Spollen WG, Sharp RE, Baskin TI (2000) Growth of Arabidopsis thaliana seedlings under water deficit studied by control of water potential in nutrient—agar media. J Exp Bot 51:1555–1562
Wu RH, Wang LL, Wang Z, Shang HH, Liu X, Zhu Y, Qi DD, Deng X (2009) Cloning and expression analysis of a dirigent protein gene from the resurrection plant Boea hygrometrica. Prog Nat Sci 19:347–352
Zhang LJ, Fan JJ, Ruan YH, Guan YX (2004) Application of polyethylene glycol in the study of plant osmotic stress physiology. Plant Physiol Commun 40:361–364
Acknowledgments
This work was supported by the National Natural Science Foundation of China (Grant No. 30871581) and the earmarked fund for the Modern Agro-industry Technology Research System (CARS-20). The authors are thankful to Dr. Luguang Wu, Scientist on Sugarcane Molecular Biology in the University of Queensland, for his valuable suggestions while revising this manuscript.
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Communicated by P. Lakshmanan.
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Jin-long, G., Li-ping, X., Jing-ping, F. et al. A novel dirigent protein gene with highly stem-specific expression from sugarcane, response to drought, salt and oxidative stresses. Plant Cell Rep 31, 1801–1812 (2012). https://doi.org/10.1007/s00299-012-1293-1
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DOI: https://doi.org/10.1007/s00299-012-1293-1