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A novel type III polyketide synthase encoded by a three-intron gene from Polygonum cuspidatum

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Abstract

A type III polyketide synthase cDNA and the corresponding gene (PcPKS2) were cloned from Polygonum cuspidatum Sieb. et Zucc. Sequencing results showed that the ORF of PcPKS2 was interrupted by three introns, which was an unexpected finding because all type III PKS genes studied so far contained only one intron at a conserved site in flowering plants, except for an Antirrhinum majus chalcone synthase gene. Besides the unusual gene structure, PcPKS2 showed some interesting characteristics: (1) the CHS “gatekeepers” Phe215 and Phe265 are uniquely replaced by Leu and Cys, respectively; (2) recombinant PcPKS2 overexpressed in Escherichia coli efficiently afforded 4-coumaroyltriacetic acid lactone (CTAL) as a major product along with bis-noryangonin (BNY) and p-hydroxybenzalacetone at low pH; however, it effectively yielded p-hydroxybenzalacetone as a dominant product along with CTAL and BNY at high pH. Beside p-hydroxybenzalacetone, CTAL and BNY, a trace amount of naringenin chalcone could be detected in assays at different pH. Furthermore, 4-coumaroyl-CoA and feruloyl-CoA were the only cinnamoyl-CoA derivatives accepted as starter substrates. PcPKS2 did not accept isobutyryl-CoA, isovaleryl-CoA or acetyl-CoA as substrate. DNA gel blot analysis indicated that there are two to four PcPKS2 copies in the P. cuspidatum genome. RNA gel blot analysis revealed that PcPKS2 is highly expressed in the rhizomes and in young leaves, but not in the roots of the plant. PcPKS2 transcripts in leaves were induced by pathogen infection, but not by wounding.

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Abbreviations

ALS:

Aloesone synthase

BAS:

Benzalacetone synthase

CHS:

Chalcone synthase

CoA:

Coenzyme A

CTAS:

p-Coumaroyl triacetic acid synthase

LC-ESIMS:

Liquid chromatography electron spray ionization mass spectrometry

OKS:

Octaketide synthase

PKS:

Polyketide synthase

STS:

Stilbene synthase

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Acknowledgments

The authors wish to thank Ms. Hong Yan, State Key Laboratory of Plant Physiology and Biochemistry, China Agricultural University, for help with the HPLC experiments.

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Authors and Affiliations

  1. Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, The Chinese Academy of Sciences, Nanxincun 20#, Xiangshan, Haidian District, 100093, Beijing, People’s Republic of China

    Lan-Qing Ma, Hai-Yan Shen, Gao-Bin Pu, Hua-Hong Wang, Cai-Yan Lei, Hong Wang, Guo-Feng Li, Ben-Ye Liu & He-Chun Ye

  2. College of Plant Science, Jilin University, Xi An Road 5333#, Lüyuan District, 130062, Changchun, People’s Republic of China

    Lan-Qing Ma & Xiao-Bin Pang

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  1. Lan-Qing Ma
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Correspondence to Ben-Ye Liu or He-Chun Ye.

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Ma, LQ., Pang, XB., Shen, HY. et al. A novel type III polyketide synthase encoded by a three-intron gene from Polygonum cuspidatum . Planta 229, 457–469 (2009). https://doi.org/10.1007/s00425-008-0845-7

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