Planta

, Volume 234, Issue 5, pp 945–958 | Cite as

Simultaneous post-transcriptional gene silencing of two different chalcone synthase genes resulting in pure white flowers in the octoploid dahlia

  • Sho Ohno
  • Munetaka Hosokawa
  • Misa Kojima
  • Yoshikuni Kitamura
  • Atsushi Hoshino
  • Fumi Tatsuzawa
  • Motoaki Doi
  • Susumu Yazawa
Original Article

Abstract

Garden dahlias (Dahlia variabilis) are autoallooctoploids with redundant genes producing wide color variations in flowers. There are no pure white dahlia cultivars, despite its long breeding history. However, the white areas of bicolor flower petals appear to be pure white. The objective of this experiment was to elucidate the mechanism by which the pure white color is expressed in the petals of some bicolor cultivars. A pigment analysis showed that no flavonoid derivatives were detected in the white areas of petals in a star-type cultivar ‘Yuino’ and the two seedling cultivars ‘OriW1’ and ‘OriW2’ borne from a red-white bicolor cultivar, ‘Orihime’, indicating that their white areas are pure white. Semi-quantitative RT-PCR showed that in the pure white areas, transcripts of two chalcone synthases (CHS), DvCHS1 and DvCHS2 which share 69% nucleotide similarity with each other, were barely detected. Premature mRNA of DvCHS1 and DvCHS2 were detected, indicating that these two CHS genes are silenced post-transcriptionally. RNA gel blot analysis revealed that small interfering RNAs (siRNAs) derived from CHSs were produced in these pure white areas. By high-throughput sequence analysis of small RNAs in the pure white areas with no mismatch acceptance, small RNAs were mapped to two alleles of DvCHS1 and two alleles of DvCHS2 expressed in ‘Yuino’ petals. Therefore, we concluded that simultaneous siRNA-mediated post-transcriptional gene silencing of redundant CHS genes results in the appearance of pure white color in dahlias.

Keywords

CHS Dahlia variabilis PTGS Pure white flower siRNA 

Abbreviations

3GT

Anthocyanidin 3-glucosyltransferase

ANS

Anthocyanidin synthase

bHLH

Basic helix-loop-helix

CHI

Chalcone isomerase

CHS

Chalcone synthase

DFR

Dihydroflavonol 4-reductase

F3H

Flavanone 3-hydroxylase

FLS

Flavonol synthase

FNS

Flavone synthase

HPLC

High-performance liquid chromatography

ORF

Open-reading frame

PTGS

Post-transcriptional gene silencing

siRNA

Small interfering RNA

SNPs

Single nucleotide polymorphisms

TLC

Thin layer chromatography

TGS

Transcriptional gene silencing

WDR

WD40 repeats

Supplementary material

425_2011_1456_MOESM1_ESM.docx (1.1 mb)
Supplementary material (DOCX 1,079 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Sho Ohno
    • 1
  • Munetaka Hosokawa
    • 1
  • Misa Kojima
    • 1
  • Yoshikuni Kitamura
    • 2
  • Atsushi Hoshino
    • 3
  • Fumi Tatsuzawa
    • 4
  • Motoaki Doi
    • 1
  • Susumu Yazawa
    • 1
  1. 1.Laboratory of Vegetable and Ornamental Horticulture, Graduate School of AgricultureKyoto UniversityKyotoJapan
  2. 2.Faculty of AgricultureShinshu UniversityNaganoJapan
  3. 3.Division of Molecular GeneticsNational Institute for Basic BiologyOkazakiJapan
  4. 4.Department of Agriculture and Life ScienceIwate UniversityIwateJapan

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