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Planta

, Volume 230, Issue 5, pp 899–915 | Cite as

Expression balances of structural genes in shikimate and flavonoid biosynthesis cause a difference in proanthocyanidin accumulation in persimmon (Diospyros kaki Thunb.) fruit

  • Takashi Akagi
  • Ayako Ikegami
  • Yasuhiko Suzuki
  • Junya Yoshida
  • Masahiko Yamada
  • Akihiko Sato
  • Keizo YonemoriEmail author
Original Article

Abstract

Persimmon fruits accumulate a large amount of proanthocyanidin (PA) during development. Fruits of pollination-constant and non-astringent (PCNA) type mutants lose their ability to produce PA at an early stage of fruit development, while fruits of the normal (non-PCNA) type remain rich in PA until fully ripened. To understand the molecular mechanism for this difference, we isolated the genes involved in PA accumulation that are differentially expressed between PCNA and non-PCNA, and confirmed their correlation with PA content and composition. The expression of structural genes of the shikimate and flavonoid biosynthetic pathways and genes encoding transferases homologous to those involved in the accumulation of phenolic compounds were downregulated coincidentally only in the PCNA type. Analysis of PA composition using the phloroglucinol method suggested that the amounts of epigallocatechin and its 3-O-gallate form were remarkably low in the PCNA type. In the PCNA type, the genes encoding flavonoid 3′5′ hydroxylase (F3′5′H) and anthocyanidin reductase (ANR) for epigallocatechin biosynthesis showed remarkable downregulation, despite the continuous expression level of their competitive genes, flavonoid 3′ hydroxylation (F3′H) and leucoanthocyanidin reductase (LAR). We also confirmed that the relative expression levels of F3′5H to F3H, and ANR to LAR, were considerably higher, and the PA composition corresponded to the seasonal expression balances in both types. These results suggest that expressions of F35H and ANR are important for PA accumulation in persimmon fruit. Lastly, we tested enzymatic activity of recombinant DkANR in vitro, which is thought to be an important enzyme for PA accumulation in persimmon fruits.

Keywords

Diospyros Flavonoid Polymerisation Proanthocyanidin Shikimate pathway 

Abbreviations

4CL

4-Coumarate: coenzyme A ligase

ANR

Anthocyanidin reductase

ANS

Anthocyanidin synthase

CA

Catechin

C4H

Cinnamate-4-hydroxymate

CHI

Chalcone isomerase

CHS

Chalcone synthase

CS

Chorismate synthase

DAHPS

3-Deoxy-d-arabino-heptulosonate 7-phosphate synthase

DFR

Dihydroflavonol 4-reductase

DHD/SDH

3-Dehydroquinate dehydratase/shikimate 5-dehydrogenase

DHQS

3-Dehydroquinate synthase

EC

Epicatechin

EGC

Epigallocatechin

EGCG

Epigallocatechin gallate

EPSPS

5-Enolpyruvylshikimate-3-phosphte synthase

F3GalT

Flavonoid 3-O-galactosyltransferase

F3H

Flavanone 3-hydroxylase

F3′H

Flavanone 3′-hydroxylase

F3′5′H

Flavanone 3′5′-hydroxylase

GC

Gallocatechin

GST

Glutathione S-transferase

LAR

Leucoanthocyanidin reductase

PAL

Phenylalanine ammonia lyase

PA

Proanthocyanidin

PCNA

Pollination-constant and non-astringent mutants

SCPL

Serine carboxypeptidase like

SK

Shikimate kinase

SSH

Suppression subtractive hybridisation

Notes

Acknowledgments

We are grateful to Dr. Shozo Kobayashi (Department of Grape and Persimmon Research, National Institute of Fruit Tree Science, Japan) for helpful discussion, and Dr. Kentaro Inoue (Department of Plant Sciences, University of California, Davis, USA) for critical advices and providing the HPLC equipments, and Dr. Richard A. Dixon (Plant Biology Division, Samuel Roberts Noble Foundation, USA) for providing pMMtANR, which encodes N-terminal in-frame fusion of MtANR with a maltose-binding protein.

Supplementary material

425_2009_991_MOESM1_ESM.doc (148 kb)
Supplementary material (DOC 148 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Takashi Akagi
    • 1
  • Ayako Ikegami
    • 2
  • Yasuhiko Suzuki
    • 1
  • Junya Yoshida
    • 3
  • Masahiko Yamada
    • 4
  • Akihiko Sato
    • 5
  • Keizo Yonemori
    • 1
    Email author
  1. 1.Laboratory of Pomology, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Laboratory of Pomology, Department of Bioproduction SciencesIshikawa Prefectural UniversityNonoichiJapan
  3. 3.Agricultural Products Distribution DivisionTakamatsuJapan
  4. 4.Department of Citrus ResearchNational Institute of Fruit Tree ScienceKuchinotsuJapan
  5. 5.Grape and Persimmon Research StationNational Institute of Fruit Tree ScienceAkitsu, Higashi-HiroshimaJapan

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