Planta

, Volume 229, Issue 5, pp 1077–1086

Identification of a Polygonum cuspidatum three-intron gene encoding a type III polyketide synthase producing both naringenin and p-hydroxybenzalacetone

  • Lan-Qing Ma
  • Yan-Wu Guo
  • Dong-Yao Gao
  • Dong-Ming Ma
  • You-Nian Wang
  • Guo-Feng Li
  • Ben-Ye Liu
  • Hong Wang
  • He-Chun Ye
Original Article

Abstract

Benzalacetone synthase (BAS) is a member of the plant-specific type III PKS superfamily that catalyzes a one-step decarboxylative condensation of 4-coumaroyl-CoA with malonyl-CoA to produce p-hydroxybenzalacetone. In our recent work (Ma et al. in Planta 229(3):457–469, 2008), a three-intron type III PKS gene (PcPKS2) was isolated from Polygonum cuspidatum Sieb. et Zucc. Phylogenetic and functional analyses revealed this recombinant PcPKS2 to be a BAS. In this study, another three-intron type III PKS gene (PcPKS1) and its corresponding cDNA were isolated from P. cuspidatum. Sequence and phylogenetic analyses demonstrated that PcPKS1 is a chalcone sythase (CHS). However, functional and enzymatic analyses showed that recombinant PcPKS1 is a bifunctional enzyme with both, CHS and BAS activity. DNA gel blot analysis indicated that there are two to four CHS copies in the P. cuspidatum genome. RNA gel blot analysis revealed that PcPKS1 is highly expressed in the rhizomes and in young leaves, but not in the roots of the plant. PcPKS1 transcripts in leaves were inducible by pathogen infection and wounding. BAS is thought to play a crucial role in the construction of the C6–C4 moiety found in a variety of phenylbutanoids, yet so far phenylbutanoids have not been isolated from P. cuspidatum. However, since PcPKS1 and PcPKS2 (Ma et al. in Planta 229(3):457–469, 2008) have been identified in P. cuspidatum, it is possible that such compounds are also produced in that plant, albeit in low concentrations.

Keywords

Benzalacetone synthase Chalcone synthase Gene evolution Phenylbutanoid biosynthesis Polygonum Type III polyketide synthase 

Abbreviations

ALS

Aloesone synthase

BAS

Benzalacetone synthase

CHS

Chalcone synthase

CoA

Coenzyme A

LC-ESIMS

Liquid chromatography-electron spray ionization mass spectrometry

OKS

Octaketide synthase

PKS

Polyketide synthase

STS

Stilbene synthase

References

  1. Abe I, Takahashi Y, Morita H, Noguchi H (2001) Benzalacetone synthase. A novel polyketide synthase that plays a crucial role in the biosynthesis of phenylbutanones in Rheum palmatum. Eur J Biochem 268:3354–3359PubMedCrossRefGoogle Scholar
  2. Abe I, Takahashi Y, Noguchi H (2002) Enzymatic formation of an unnatural C(6)-C(5) aromatic polyketide by plant type III polyketide synthases. Org Lett 4:3623–3626PubMedCrossRefGoogle Scholar
  3. Abe I, Sano Y, Takahashi Y, Noguchi H (2003) Site-directed mutagenesis of benzalacetone synthase. The role of the Phe215 in plant type III polyketide synthases. J Biol Chem 278:25218–25226PubMedCrossRefGoogle Scholar
  4. Abe I, Oguro S, Utsumi Y, Sano Y, Noguchi H (2005) Engineered biosynthesis of plant polyketides: chain length control in an octaketide-producing plant type III polyketide synthase. J Am Chem Soc 127:12709–12716PubMedCrossRefGoogle Scholar
  5. Akiyama T, Shibuya M, Liu HM, Ebizuka Y (1999) p-Coumaroyltriacetic acid synthase, a new homologue of chalcone synthase, from Hydrangea macrophylla var. thunbergii. Eur J Biochem 263:834–839PubMedCrossRefGoogle Scholar
  6. Arias JA, Dixon RA, Lamb CJ (1993) Dissection of the functional architecture of a plant defense gene promoter using a homologous in vitro transcription initiation system. Plant Cell 5:485–496PubMedCrossRefGoogle Scholar
  7. Austin MB, Noel JP (2003) The chalcone synthase superfamily of type III polyketide synthases. Nat Prod Rep 20:79–110PubMedCrossRefGoogle Scholar
  8. Beekwilder J, van der Meer IM, Sibbesen O, Broekgaarden M, Qvist I, Mikkelsen JD, Hall RD (2007) Microbial production of natural raspberry ketone. Biotechnol J 2:1270–1279PubMedCrossRefGoogle Scholar
  9. Blacklock BJ, Jaworski JG (2006) Substrate specificity of Arabidopsis 3-ketoacyl-CoA synthases. Biochem Biophys Res Commun 346:583–590PubMedCrossRefGoogle Scholar
  10. Borejsza-Wysocki W, Hrazdina G (1994) Biosynthesis of p-hydroxyphenylbutan-2-one in raspberry fruits and tissue culture. Phytochemistry 35:623–628CrossRefGoogle Scholar
  11. Borejsza-Wysocki W, Hrazdina G (1996) Aromatic polyketide synthases (purification, characterization, and antibody development to benzalacetone synthase from raspberry fruits). Plant Physiol 110:791–799PubMedGoogle Scholar
  12. Brown JW, Smith P, Simpson CG (1996) Arabidopsis consensus intron sequences. Plant Mol Biol 32:531–535PubMedCrossRefGoogle Scholar
  13. Buchanan BB, Gruissem W, Jones RL (2002) Biochemistry and molecular biology of plants. American Society of Plant Physiologists, WaldorfGoogle Scholar
  14. Durbin ML, McCaig B, Clegg MT (2000) Molecular evolution of the chalcone synthase multigene family in the morning glory genome. Plant Mol Biol 42:79–92PubMedCrossRefGoogle Scholar
  15. Ferrer JL, Jez JM, Bowman ME, Dixon RA, Noel JP (1999) Structure of chalcone synthase and the molecular basis of plant polyketide biosynthesis. Nat Struct Biol 6:775–784PubMedCrossRefGoogle Scholar
  16. Hartmann U, Sagasser M, Mehrtens F, Stracke R, Weisshaar B (2005) Differential combinatorial interactions of cis-acting elements recognized by R2R3-MYB, BZIP, and BHLH factors control light-responsive and tissue-specific activation of phenylpropanoid biosynthesis genes. Plant Mol Biol 57:155–171PubMedCrossRefGoogle Scholar
  17. Hegde VR, Pu H, Patel M, Black T, Soriano A, Zhao W, Gullo VP, Chan TM (2004) Two new bacterial DNA primase inhibitors from the plant Polygonum cuspidatum. Bioorg Med Chem Lett 14:2275–2277PubMedCrossRefGoogle Scholar
  18. James DW Jr, Lim E, Keller J, Plooy I, Ralston E, Dooner HK (1995) Directed tagging of the Arabidopsis FATTY ACID ELONGATION1 (FAE1) gene with the maize transposon Activator. Plant Cell 7:309–319PubMedCrossRefGoogle Scholar
  19. Jiang C, Schommer CK, Kim SY, Suh DY (2006) Cloning and characterization of chalcone synthase from the moss, Physcomitrella patens. Phytochemistry 67:2531–2540PubMedCrossRefGoogle Scholar
  20. Kajikawa M, Yamaoka S, Yamato KT, Kanamaru H, Sakuradani E, Shimizu S, Fukuzawa H, Ohyama K (2003a) Functional analysis of a beta-ketoacyl-CoA synthase gene, MpFAE2, by gene silencing in the liverwort Marchantia polymorpha L. Biosci Biotechnol Biochem 67:605–612PubMedCrossRefGoogle Scholar
  21. Kajikawa M, Yamato KT, Kanamaru H, Sakuradani E, Shimizu S, Fukuzawa H, Sakai Y, Ohyama K (2003b) MpFAE3 a beta-ketoacyl-CoA synthase gene in the liverwort Marchantia polymorpha L., is preferentially involved in elongation of palmitic acid to stearic acid. Biosci Biotechnol Biochem 67:1667–1674PubMedCrossRefGoogle Scholar
  22. Lewin B (2000) Gene VII. Oxford University Press Inc., New YorkGoogle Scholar
  23. Liu YG, Mitsukawa N, Oosumi T, Whittier RF (1995) Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J 8:457–463PubMedCrossRefGoogle Scholar
  24. Liu B, Falkenstein-Paul H, Schmidt W, Beerhues L (2003) Benzophenone synthase and chalcone synthase from Hypericum androsaemum cell cultures: cDNA cloning, functional expression, and site-directed mutagenesis of two polyketide synthases. Plant J 34:847–855PubMedCrossRefGoogle Scholar
  25. Ma LQ, Pang XB, Shen HY, Pu GB, Wang HH, Lei CY, Wang H, Li GF, Liu BY, Ye HC (2008) A novel type III polyketide synthase encoded by a three-intron gene from Polygonum cuspidatum. Planta 229(3):457–469PubMedCrossRefGoogle Scholar
  26. Morita H, Takahashi Y, Noguchi H, Abe I (2000) Enzymatic formation of unnatural aromatic polyketides by chalcone synthase. Biochem Biophys Res Commun 279:190–195PubMedCrossRefGoogle Scholar
  27. Schröder J (1997) A family of plant-specific polyketide synthases: facts and predictions. Trends Plant Sci 2:373–378CrossRefGoogle Scholar
  28. Sommer H, Saedler H (1986) Structure of the chalcone synthase gene of Antirrhinum majus. Mol Gen Genet 202:429–434CrossRefGoogle Scholar
  29. Todd J, Post-Beittenmiller D, Jaworski JG (1999) KCS1 encodes a fatty acid elongase 3-ketoacyl-CoA synthase affecting wax biosynthesis in Arabidopsis thaliana. Plant J 17:119–130PubMedCrossRefGoogle Scholar
  30. Trapp SC, Croteau RB (2001) Genomic organization of plant terpene synthases and molecular evolutionary implications. Genetics 158:811–832PubMedGoogle Scholar
  31. Xiao K, Xuan L, Xu Y, Bai D, Zhong D (2002) Constituents from Polygonum cuspidatum. Chem Pharm Bull (Tokyo) 50:605–608CrossRefGoogle Scholar
  32. Yi T, Zhang H, Cai Z (2007) Analysis of rhizoma Polygoni cuspidati by HPLC and HPLC-ESI/MS. Phytochem Anal 18:387–392PubMedCrossRefGoogle Scholar
  33. Zheng D, Hrazdina G (2008) Molecular and biochemical characterization of benzalacetone synthase and chalcone synthase genes and their proteins from raspberry (Rubus idaeus L.). Arch Biochem Biophys 470:139–145PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Lan-Qing Ma
    • 1
  • Yan-Wu Guo
    • 1
  • Dong-Yao Gao
    • 2
  • Dong-Ming Ma
    • 1
  • You-Nian Wang
    • 2
  • Guo-Feng Li
    • 1
  • Ben-Ye Liu
    • 1
  • Hong Wang
    • 1
  • He-Chun Ye
    • 1
  1. 1.Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of BotanyChinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.Key Laboratory of New Technology in Agricultural Application of BeijingBeijing University of AgricultureBeijingPeople’s Republic of China

Personalised recommendations