Tree Genetics & Genomes

, Volume 9, Issue 5, pp 1351–1360 | Cite as

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

  • Ya-nan Yang
  • Guang Zhao
  • Wen-quan Yue
  • Shao-ling Zhang
  • Chao Gu
  • Jun Wu
Original Paper

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.

Keywords

Pear Color mutant Anthocyanin Molecular cloning Gene expression 

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

Supplementary material

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Ya-nan Yang
    • 1
  • Guang Zhao
    • 1
  • Wen-quan Yue
    • 2
  • Shao-ling Zhang
    • 1
  • Chao Gu
    • 1
  • Jun Wu
    • 1
  1. 1.Centre of Pear Engineering Technology ResearchNanjing Agricultural UniversityNanjingPeople’s Republic of China
  2. 2.Changli Institute of PomologyHebei Academy of Agriculture and Forestry SciencesChangliPeople’s Republic of China

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