Abstract
Reducing the lignin content of trees could provide both economic and environmental benefits. To this end, the coumarate:coenzyme A ligase 1 gene (4CL1) was isolated from Pinus massoniana Lamb (Pm4CL1). The sequence of the full-length Pm4CL1 cDNA (accession no. FJ810495) contained an entire open reading frame (ORF) of 1,614 bp, which encoded a polypeptide of 537 amino acid residues. Tobacco (Nicotiana tabacum L.) as a model plant was used for functional characterization of the Pm4CL1 gene in transgenic plants. Results revealed that 4CL1 enzyme activity and lignin content in most antisense Pm4CL1 transgenic tobacco lines were decreased as compared to wild-type; the average 4CL1 enzyme activity was decreased by 48.75% and lignin content was decreased by 24.5%. In contrast, in the sense Pm4CL1 transgenic tobacco lines, average 4CL1 enzyme activity was increased by 72.3% and lignin content was increased by 27.6%. These results suggest that the Pm4CL1 gene from P. massoniana could be applied to regulate lignin biosynthesis in transgenic trees.
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References
Allina SM, Hadash AP, Theilmann DA et al (1998) 4-Coumarate:coenzyme A ligase in hybrid poplar. Plant Physiol 116:743–754
Atanassova R, Favet N, Martz F et al (1995) Altered lignin composition in transgenic tobacco expressing O-methyltransferase sequences in sense and antisense orientation. Plant J 6:465–477
Blee K, Choi JW, Connell AP et al (2001) Antisense and sense expression of cDNA coding for CYP73A15, a class II cinnamate 4-hydroxylase, leads to a delayed and reduced production of lignin in tobacco. Phytochemistry 57:1159–1166
Chen ZZ, Chen YC, Chou YH, Lin Y (2006) cDNA cloning and molecular characterization of 4-coumarate:coenzyme A ligase in Eucalyptus camaldulensis. Taiwan J Forest Sci 21:87–100
Dwivedi UN, Campbell WH, Yu J et al (1994) Modification of lignin biosynthesis in transgenic Nicotiana through expression of an antisense Omethyltransferase gene from Populus. Plant Mol Biol 26:61–71
Elkind Y, Edwards E, Mavandad M et al (1990) Abnormal plant development and down-regulation of phenylpropanoid biosynthesis in transgenic tobacco containing a heterologous phenylalanine ammonia-lyase gene. Proc Natl Acad Sci USA 87:9057–9061
Halpin C, Knight ME, Foxon GA et al (1994) Manipulation of lignin quality by the down-regulation of cinnamyl alcohol dehydrogenase. Plant J 6:339–350
Hamberger B, Hahlbrock K (2003) The 4-coumarate: CoA ligase gene family in Arabidopsis thaliana comprises one rare, sinapate-activating and three commonly occurring isoenzymes. PNAS 101:2209–2214
Hu WJ, Kawaoka A, Tsai CJ et al (1998) Compartmentalized expression of two structurally and functionally distinct 4-coumarate:CoA ligase genes in aspen (Populus tremuloides). Proc Natl Acad Sci USA 95:5407–5412
Hu WJ, Harding SA, Lung J et al (1999) Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees. Nat Biotechnol 17:808–812
Iiyama K, Wallis AFA (1990) Determination of lignin in herbaceous plants by an improved acetyl bromide procedure. J Food Sci Agric 51:145–161
Jiang H, Wood KV, Morgan JA (2005) Metabolic engineering of the phenylpropanoid pathway in Saccharomyces cerevisiae. Appl Environ Microbiol 71(6):2962–2969
Jin GQ, Qin GF, Liu WH et al (2009) Genetic analysis of crossing effects for growth traits of P. massoniana and selection of cross combinations. Front Forest China 4(1):101–106
Kajita S, Katayama Y, Omori S (1996) Alterations in the biosynthesis of lignin in transgenic plants with chimeric genes for 4-coumarate:coenzyme A ligase. Plant Cell Physiol 37:957–965
Keb-Llanes M, Gonzalez G, Chi-Manzanero B, Infante D (2002) A rapid and simple method for small scale DNA extraction in Agavaceae and other tropical plants. Plant Mol Biol Rep 20(3):299a–e
Knobloch KH, Hanlbrock K (1977) 4-Coumarate:CoA ligase from cell suspension cultures of Petroselinum hortense Hoffm. Partial purification, substrate specificity, and further properties. Arch Biochem Biophys 184:237–248
Lee D, Douglas CJ (1996) Two divergent members of a tobacco 4-coumarate:coenzyme A ligase (4CL) gene family. Plant Physiol 112:193–205
Lee D, Ellard M, Wanner LA et al (1995) The Arabidopsis thaliana 4-coumarate:CoA ligase (4CL) gene: stress and developmentally regulated expression and nucleotide sequence of its cDNA. Plant Mol Biol 28(5):871–884
Lu H, Zhao YL, Jiang XN (2004) Stable and specific expression of 4-coumarate: coenzyme A ligase gene (4CL1) driven by the xylemspecific Pto4CL1 promoter in the transgenic tobacco. Biotechnol Lett 26:1147–1152
Lu H, Zeng QY, Zhao YL et al (2003) Xylem-specific expression of a GRP1.8 promoter:4CL gene construct in transgenic tobacco. Plant Growth Regul 41:279–286
Ming D (1987) Population structure and dynamics of P. massoniana Lamb. on Mount Jinyun, Sichuan, China. Vegetatio 72(1):35–44
Pincon G, Maury S, Hoffmann L et al (2001) Repression of O-methyltransferase genes in transgenic tobacco affects lignin synthesis and plant growth. Phytochemistry 57:1167–1176
Piquemal J, Lapierre C, Myton K et al (1998) Down-regulation of cinnamoyl-CoA reductase induces significant changes of lignin profiles in transgenic tobacco plants. Plant J 13:71–83
Sewalt VJH, Ni W, Blount JW et al (1997) Reduced lignin content and altered lignin composition in transgenic tobacco down-regulated in expression of L-phenylalanine ammonia-lyase or cinnamate 4-hydroxylase. Plant Physiol 115:41–50
Voelker SL, Lachenbruch B, Meinzer FC et al (2010) Antisense down-regulation of 4CL expression alters lignification, tree growth, and saccharification potential of field-grown poplar. Plant Physiol 154:874–886
Voo KS, Whetten RW, O’Malley DM, Sederoff RR (1995) 4-Coumarate:coenzyme A ligase from loblolly pine xylem. Plant Physiol 108:85–97
Wagner A, Donaldson L, Kim H et al (2009) Suppression of 4-coumarate-CoA ligase in the coniferous gymnosperm Pinus radiate. Plant Physiol 149:370–383
Yang XP, Lu H, Chen XM, Jiang XN (2003) Anti-sense 4CL1 changes the content and ratio of lignin and cellulose in transformed tobacco plantlets. J Beijing Forest Univ 25(3):381–387
Zhang XH, Chiang VL (1997) Molecular cloning of 4-coumarate:coenzyme A ligase in loblolly pine and the roles of this enzyme in the biosynthesis of lignin in compression wood. Plant Physiol 113:65–74
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This work is supported by the agricultural biotechnology program of the Ministry of Agricultural and Rural Development, Vietnam.
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Van Huan, H., Van Giang, H., Van Thanh, N. et al. Identification and functional analysis of the Pm4CL1 gene in transgenic tobacco plant as the basis for regulating lignin biosynthesis in forest trees. Mol Breeding 29, 173–180 (2012). https://doi.org/10.1007/s11032-010-9535-9
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DOI: https://doi.org/10.1007/s11032-010-9535-9