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Overexpression of Ps-CHI1, a homologue of the chalcone isomerase gene from tree peony (Paeonia suffruticosa), reduces the intensity of flower pigmentation in transgenic tobacco

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Abstract

Chalcone isomerase (CHI) is one of the key enzymes in the flavonoid and anthocyanin biosynthesis pathway catalyzing the stereospecific isomerization of chalcones into their corresponding (2S)-flavanones. In this study, to investigate the role of CHI in tree peony flower coloration mechanism, a CHI gene was isolated from Paeonia suffruticosa cv. Guan Shi Mo Yu, and designated as Ps-CHI1. The cDNA sequence of Ps-CHI1 was 924 bp in full length containing an opening reading frame of 654 bp that encoded a peptide of 217 amino acids with a predicted molecular mass of 23.3 kDa and a pI of 5.0. Sequence alignment and phylogenetic analysis revealed that Ps-CHI1 shared high homology with other type I higher plant CHI proteins. Quantitative real-time PCR analysis indicated that Ps-CHI1 showed the highest transcript abundance in petals, moderate levels in sepals, low levels in leaves and carpels and the lowest levels in stamens. Ps-CHI1 was expressed at the highest level in early stage of uncolored flowers and underwent moderate decreases in expression until there was little transcription could be detected when flowers fully opened and deeply pigmented. By inserting Ps-CHI1 into tobacco (Nicotiana tabacum L.) via Agrobacterium-mediated transformation, T1 transgenic plants were obtained. Phenotypes and pigment analysis revealed that they produced up to threefold total flavonols and flavones over wild-type control, and showed a remarkable reduction in anthocyanin content and flower color intensity. These results provide theoretical deduction supporting Ps-CHI1 as an important gene involved in tree peony flower pigmentation.

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Abbreviations

Bp:

Base pair

CaMV:

Cauliflower mosaic virus

cDNA:

Complementary DNA

CHI:

Chalcone isomerase

CTAB:

Cetyltrimethylammonium bromide

DNase:

Deoxyribonuclease

dNTP:

Deoxyribonucleoside 5′-triphosphate

MS:

Murashige and Skoog

EDTA:

Ethylenediaminetetraacetic acid

ORF:

Opening reading frame

PCR:

Polymerase chain reaction

PVPP:

Polyvinyl polypyrrolidone

RACE:

Rapid amplification of cDNA ends

RNase:

Ribonuclease

RT-PCR:

Reverse transcription polymerase chain reaction

5′-UTR:

5′-Untranslation region

3′-UTR:

3′-Untranslation region

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Acknowledgments

This research was supported by the National High Technology Research and Development Program of China (863 Program) (2011AA10020701) and National Natural Science Foundation of China (31201654).

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Authors and Affiliations

  1. State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of State Forestry Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing, 100091, People’s Republic of China

    Lin Zhou, Yan Wang, Lei Ren, Qianqian Shi, Baoqiang Zheng, Kun Miao & Xin Guo

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  1. Lin Zhou
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  2. Yan Wang
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  3. Lei Ren
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Correspondence to Yan Wang.

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Zhou, L., Wang, Y., Ren, L. et al. Overexpression of Ps-CHI1, a homologue of the chalcone isomerase gene from tree peony (Paeonia suffruticosa), reduces the intensity of flower pigmentation in transgenic tobacco. Plant Cell Tiss Organ Cult 116, 285–295 (2014). https://doi.org/10.1007/s11240-013-0403-2

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