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Molecular cloning and characterization of the CHS gene family in turmeric (Curcuma longa Linn.)

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Abstract

Three chalcone synthase (CHS) genes were isolated from Curcuma longa Linn. using TAIL-PCR. ClCHS1 and ClCHS2 were 1,460 and 1,407 bp in length, respectively, containing 1,191 bp open reading frame (ORF) that encodes 396 amino acids, whereas ClCHS3 was 1,394 bp in length containing 1,170 bp ORF that encodes 389 amino acids. The structure of all three genes comprise two exons and one intron which are consistent with the other CHS gene family. Southern blot analysis using a ClCHS conserved fragment revealed the ClCHS genes belong to a gene family. Phylogenetic analysis showed the three putative ClCHS proteins to be closely related to DCS (diketide-CoA synthase) protein, a product of CHS-like gene in C. longa, which condenses malonyl-CoA with feruloyl-CoA or coumaroyl-CoA as the substrate in curcuminoid synthesis. These results suggest that the interaction of substrate and enzyme between the three putative ClCHS proteins and DCS might be highly similar. Homology modeling and docking analysis were consistent, indicating that the same substrate (coumaroyl-CoA) can be used in the putative ClCHS1 and ClCHS2 proteins. However, the putative ClCHS3 protein seems to have seven amino acids deletion in a loop involved in the binding site formation, suggesting that the binding site with coumaroyl-CoA might be altered.

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Abbreviations

AIC:

Akaike information criterion

BAS:

Benzalacetone synthase

CHS:

Chalcone synthase

ClCHS:

Curcuma longa chalcone synthase

CURS:

Curcumin synthase

DCS:

Diketide-CoA synthase

DDBJ:

DNA data bank of Japan

NCBI:

National Center for Biotechnology Information

PDB:

Protein data bank

STS:

Stilbene synthase

TAIL-PCR:

Thermal asymmetric interlaced polymerase chain reaction

VPS:

Valerophenone synthase

2-PS:

2-pyrone synthase CHS

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Acknowledgments

This work was financially supported by Kasetsart University Research and Development Institute (KURDI), Kasetsart University, Thailand. We also would like to thank Vichien Keeratinijakal, Kasetsart University, Thailand for providing plant materials.

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Authors and Affiliations

  1. Department of Genetics, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand

    Sirinrat Wannapinpong, Amara Thongpan & Surin Peyachoknagul

  2. Center for Advanced Studies in Tropical Natural Resources, National Research University-Kasetsart University, (CASTNAR, NRU-KU), Bangkok, 10900, Thailand

    Kornsorn Srikulnath, Kiattawee Choowongkomon & Surin Peyachoknagul

  3. Institute of Food Research and Product Development, Kasetsart University, Bangkok, 10900, Thailand

    Sirinrat Wannapinpong

  4. Laboratory of Animal Cytogenetics and Comparative Genomics, Department of Genetics, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand

    Kornsorn Srikulnath

  5. Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand

    Kiattawee Choowongkomon

Authors
  1. Sirinrat Wannapinpong
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  2. Kornsorn Srikulnath
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  3. Amara Thongpan
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  4. Kiattawee Choowongkomon
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  5. Surin Peyachoknagul
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Correspondence to Surin Peyachoknagul.

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Supplementary Table 1

Gene name and protein accession numbers of species used in the sequence alignment and phylogenetic tree reconstruction (DOC 83 kb)

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Wannapinpong, S., Srikulnath, K., Thongpan, A. et al. Molecular cloning and characterization of the CHS gene family in turmeric (Curcuma longa Linn.). J. Plant Biochem. Biotechnol. 24, 25–33 (2015). https://doi.org/10.1007/s13562-013-0232-8

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  • DOI: https://doi.org/10.1007/s13562-013-0232-8

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