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Molecular cloning and gene expression differences of the anthocyanin biosynthesis-related genes in the red/green skin color mutant of pear (Pyrus communis L.)

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Abstract

To reveal the molecular mechanisms that led to the red/green color mutation of pear between the cultivar ‘Early red Doyenne du Comice’ and its green variant strain, the full-length cDNA of the seven anthocyanin biosynthesis genes (PAL, CHS, CHI, DFR, F3H, ANS, UFGT) was cloned in both cultivars. The accession number has been submitted to National Center for Biotechnology Information (NCBI) as KC460392, KC460393, KC460394, KC460395, KC460396, KC460397, and KC460398, respectively. However, there was no sequence difference between the color mutants, which means that the skin color change was not caused by mutation of any of these genes. Meanwhile, the expression levels of these seven genes were examined by quantitative real-time PCR (qRT-PCR). Results showed that most of the structural genes were up-regulated in the red-skinned cultivar during fruit development, but the CHI and UFGT genes were highly expressed only at an early stage. The expression levels of the transcription factors MYB10, bHLH, and WD40 were also investigated by qRT-PCR, and the MYB10 gene was found to be expressed at significantly higher levels in the red variety than in the green mutant at the early stage, while the expression levels of bHLH and WD40 were higher at a later stage. These data indicate that the expression difference of structural genes in the anthocyanin biosynthesis pathway led to the skin color change of the mutant. However, MYB10, bHLH, and WD40 do not appear to be the key transcription factors that regulate the biosynthesis of anthocyanin and determine the red/green color mutant.

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Abbreviations

ANS:

Anthocyanin synthase

bHLH:

Basic helix loop helix

CHI:

Chalcone isomerase

CHS:

Chalcone synthase

DAFB:

Days after full bloom

DFR:

Dihydroflavonol-4-reductase

F3H:

Flavanone 3-hydroxylase

PAL:

Phenylalanine ammonialyase

RACE:

Rapid amplification of cDNA ends

qRT-PCR:

Quantitative reverse transcription polymerase chain reaction

SE:

Standard error

TAIL-PCR:

Thermal asymmetric interlaced polymerase chain reaction

UFGT:

UDP-glucose: flavonoid-3-O-glucosyltransferase

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Acknowledgments

The work was financially supported by the National Public Benefit (Agricultural) Research Foundation of China (200903044), the National Science Foundation of China (30900974), and the Earmarked Fund for China Agriculture Research System (no. CARS-29).

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Authors and Affiliations

  1. Centre of Pear Engineering Technology Research, Nanjing Agricultural University, No. 1 Weigang Rd., Nanjing, Jiangsu, 210095, People’s Republic of China

    Ya-nan Yang, Guang Zhao, Shao-ling Zhang, Chao Gu & Jun Wu

  2. Changli Institute of Pomology, Hebei Academy of Agriculture and Forestry Sciences, Changli, Hebei, 066600, People’s Republic of China

    Wen-quan Yue

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  1. Ya-nan Yang
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  2. Guang Zhao
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  3. Wen-quan Yue
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  4. Shao-ling Zhang
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  5. Chao Gu
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  6. Jun Wu
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Corresponding author

Correspondence to Jun Wu.

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Communicated by A. Dandekar

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Yang, Yn., Zhao, G., Yue, Wq. et al. Molecular cloning and gene expression differences of the anthocyanin biosynthesis-related genes in the red/green skin color mutant of pear (Pyrus communis L.). Tree Genetics & Genomes 9, 1351–1360 (2013). https://doi.org/10.1007/s11295-013-0644-6

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  • DOI: https://doi.org/10.1007/s11295-013-0644-6

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