Advertisement

Cloning and Expression Analysis of 4-Coumarate CoA Ligase (4CL) Gene in Sugarcane Clones Varying in Lignin Content

  • Lakshmi KasirajanEmail author
  • Sara Charles
  • Kalaivaani Aruchamy
Research Article

Abstract

Sugarcane (Saccharum sp.) is one of the highest biomass producing plant, which is the best lignocellulosic feedstock for ethanol production. To achieve a more efficient conversion of biomass to ethanol, a better understanding of the main factors affecting biomass recalcitrance is needed. In the present study, the authors have cloned the partial coding sequence of 4-coumarate-CoA ligase (4CL) from an energy cane (EC 11010). The deduced amino acid sequence of this gene showed 98% similarity with the Zea mays protein and the phylogeny revealed clustering with the same gene from monocots. The quantitative expression of the 4CL gene among the four different sugarcane genotypes (EC 11010, IK 76-81, Co 86032 and CoC 671) showed higher level of transcripts in the energy cane EC 11010.

Keywords

Lignocellulosic biomass Secondary cell wall Energy canes Real time polymerase chain reaction 

Notes

Acknowledgements

This work has been supported by the Department of Science and Technology, Science Engineering and Research Board project No. SB/EMEQ-001/2013. The authors wish to thank Dr. M. N. Premachandran, Head, Division of Crop Improvement, and Dr. A. Selvi, Principle Scientist, Biotechnology Section and Head, Sugarcane Breeding Institute, Coimbatore for kindly providing sugarcane clones and extending laboratory facility for carrying out the work respectively.

Compliance with Ethical Standards

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

References

  1. 1.
    Bottcher A, Cesarino I, Brombini AS, Vicentini R, Mayer JLS, Vanholme R (2013) Lignification in sugarcane: biochemical characterization, gene discovery, and expression analysis in two genotypes contrasting for lignin content. Plant Physiol 163(4):1539–1557CrossRefGoogle Scholar
  2. 2.
    Wu X, Wu J, Luo Y, Bragg J, Anderson O, Vogel J (2013) Phylogenetic, molecular, and biochemical characterization of caffeic acid o-methyltransferase gene family in Brachypodium distachyon. Int J Plant Genomics. doi: 10.1155/2013/423189 Google Scholar
  3. 3.
    Ehlting J, Shin JJ, Douglas CJ (2001) Identification of 4-coumarate: coenzyme A ligase (4CL) substrate recognition domains. Plant J 27(5):455–465CrossRefGoogle Scholar
  4. 4.
    Costa MA, Bedgar DL, Moinuddin SG, Kim KW, Cardenas CL et al (2005) Characterization in vitro and in vivo of the putative multigene 4-coumarate:CoA ligase network in Arabidopsis: syringyl ligninand sinapate/sinapyl alcohol derivative formation. Phytochemistry 66(17):2072–2091CrossRefGoogle Scholar
  5. 5.
    Lindermayr C, Mollers B, Fliegmann J, Uhlmann A, Lottspeich F, Meimberg H (2002) Divergent members of a soybean (Glycine max L.) 4-coumarate: coenzyme A ligase gene family. Eur J Biochem 269(4):1304–1315CrossRefGoogle Scholar
  6. 6.
    CdeA Souza, Barbazuk B, Ralph SG, Bohlmann J, Hamberger B, Douglas CJ (2008) Genome-wide analysis of a land plant-specific acyl:coenzymeAsynthetase (ACS) gene family in Arabidopsis, poplar, rice and Physcomitrella. New Phytol 179(4):987–1003Google Scholar
  7. 7.
    Gui J, Shen J, Li L (2011) Functional characterization of evolutionarily divergent 4-coumarate: coenzyme A ligases in rice. Plant Physiol 157(2):574–586CrossRefGoogle Scholar
  8. 8.
    Allina SM, Pri-Hadash A, Theilmann DA, Ellis BE, Douglas CJ (1998) 4-coumarate: coenzyme A ligase in hybrid poplar: properties of native enzymes, cDNA cloning, and analysis of recombinant enzymes. Plant Physiol 116(2):743–754CrossRefGoogle Scholar
  9. 9.
    Hu WJ, Kawaoka A, Tsai CJ, Lung J, Osakabe K, Ebinuma H, Chiang VL (1998) Compartmentalized expression of two structurally and functionally distinct 4-coumarate: CoA ligase genes in aspen (Populus tremuloides). Proc Natl Acad Sci USA 95(9):5407–5412CrossRefGoogle Scholar
  10. 10.
    Harding SA, Leshkevich J, Chiang VL, Tsai CJ (2002) Differential substrate inhibition couples kinetically distinct 4-coumarate: coenzyme A ligases with spatially distinct metabolic roles in quaking aspen. Plant Physiol 128(2):428–438CrossRefGoogle Scholar
  11. 11.
    Steve R, Skaletsky JH (1998) Primer3. http://www.genome.wi.mit.edu/genome_software/other/primer3
  12. 12.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta DeltaC(T)) method. Methods 25(4):402–408CrossRefGoogle Scholar
  13. 13.
    Gao S, Yu HN, Xu RX, Cheng AX, Lou HX (2015) Cloning and functional characterization of a 4-coumarate CoA ligase from liverwort Plagiochasma appendiculatum. Phytochemistry 111:48–58CrossRefGoogle Scholar
  14. 14.
    Kao YY, Harding SA, Tsai CJ (2002) Differential expression of two distinct phenylalanine ammonia lyase genes in condensed tannin-accumulating and lignifying cells of quaking aspen. Plant Physiol 130(2):796–807CrossRefGoogle Scholar
  15. 15.
    Eudes A, Pollet B, Sibout R, Do CT, Seguin A, Lapierre C, Jouanin L (2006) Evidence for a role of AtCAD1 in lignification of elongating stems of Arabidopsis thaliana. Planta 225(1):23–39CrossRefGoogle Scholar
  16. 16.
    Cao Y, Hu SL, Huang SX, Ren P, Lu XQ (2012) Molecular cloning, expression pattern, and putative cis-acting elements of a 4-coumarate:CoA ligase gene in bamboo (Neosino calamusaffinis). Electron J Biotechnol 15(5). doi: 10.2225/vol15-issue5-fulltext-10
  17. 17.
    Hu WJ, Harding SA, Lung J, Popko JL, Ralph J, Stokke DD, Tsai CJ, Chiang VL (1999) Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees. Nat Biotechnol 17(8):808–812CrossRefGoogle Scholar
  18. 18.
    Hamberger B, Hahlbrock K (2004) The 4-coumarate:CoA ligase gene family in Arabidopsis thaliana comprises one rare, sinapate-activating and three commonly occurring isoenzymes. Proc Natl Acad Sci USA 101:2209–2214CrossRefGoogle Scholar
  19. 19.
    Souza GM et al (2011) The sugarcane genome challenge: strategies for sequencing a highly complex genome. Trop Plant Biol 4(3–4):145–156CrossRefGoogle Scholar

Copyright information

© The National Academy of Sciences, India 2017

Authors and Affiliations

  • Lakshmi Kasirajan
    • 1
    Email author
  • Sara Charles
    • 1
    • 2
  • Kalaivaani Aruchamy
    • 1
  1. 1.Genomics Laboratory, Division of Crop ImprovementICAR-Sugarcane Breeding InstituteCoimbatoreIndia
  2. 2.Karunya UniversityCoimbatoreIndia

Personalised recommendations