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

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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.

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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

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Authors and Affiliations

  1. Laboratory of Vegetable and Ornamental Horticulture, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan

    Sho Ohno, Munetaka Hosokawa, Misa Kojima, Motoaki Doi & Susumu Yazawa

  2. Faculty of Agriculture, Shinshu University, Nagano, 399-4598, Japan

    Yoshikuni Kitamura

  3. Division of Molecular Genetics, National Institute for Basic Biology, Myodaiji, Okazaki, Aichi, 444-8585, Japan

    Atsushi Hoshino

  4. Department of Agriculture and Life Science, Iwate University, Iwate, 020-8550, Japan

    Fumi Tatsuzawa

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  1. Sho Ohno
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  2. Munetaka Hosokawa
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  3. Misa Kojima
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  4. Yoshikuni Kitamura
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  5. Atsushi Hoshino
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  6. Fumi Tatsuzawa
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  7. Motoaki Doi
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  8. Susumu Yazawa
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Correspondence to Munetaka Hosokawa.

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Ohno, S., Hosokawa, M., Kojima, M. et al. Simultaneous post-transcriptional gene silencing of two different chalcone synthase genes resulting in pure white flowers in the octoploid dahlia. Planta 234, 945–958 (2011). https://doi.org/10.1007/s00425-011-1456-2

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