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Biologia Plantarum

, Volume 56, Issue 3, pp 545–550 | Cite as

Chalcone isomerase in grape vine: gene expression and localization in the developing fruit

  • W. Wang
  • H. -L. Wang
  • S. -B. Wan
  • J. -H. Zhang
  • P. Zhang
  • J. -C. Zhan
  • W. -D. Huang
Article

Abstract

Chalcone isomerase (CHI, EC 5.5.1.6) is an entrance enzyme in the flavonoid biosynthesis, which catalyzes the conversion of chalcones to flavanones. In this study, the full-length CHI cDNA from grape vine (Vitis vinifera L.) was cloned, the recombinant protein was purified and the polyclonal antibody was prepared. Using these tools, the expression and tissue localization of CHI in developing grape berry was analyzed by RT-PCR, gel blot hybridization and immunohistochemical techniques. The expression of CHI was dependent on developmental stage, and CHI protein was mainly distributed in vascular bundles throughout all the stages of berry development, which suggested that flavonoids in the berry might have been partially synthesized in situ.

Additional key words

antibody Escherichia coli; flavonoids immunohistochemical technique RT-PCR Vitis vinifera 

Abbreviations

BSA

bovine serum albumin

CHI

chalcone isomerase

CHS

chalcone synthase

DAF

day after full bloom

EDTA

ethylene diamine tetraacetic acid

IPTG

isopropyl-β-D-thiogalactoside

RT-PCR

reverse transcription-polymerase chain reaction

SDS

sodium dodecyl sulfate

Tris

tris(hydroxymethyl)amino methane

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • W. Wang
    • 1
    • 3
  • H. -L. Wang
    • 1
    • 2
  • S. -B. Wan
    • 1
    • 4
  • J. -H. Zhang
    • 1
  • P. Zhang
    • 1
  • J. -C. Zhan
    • 1
  • W. -D. Huang
    • 1
    • 2
  1. 1.Center for Research and Development of Viticulture and EnologyChina Agricultural UniversityBeijingP.R. China
  2. 2.Beijing Key Laboratory of Viticulture and EnologyBeijingChina
  3. 3.The Institute of ForestryThe Chinese Academy of ForestryBeijingP.R. China
  4. 4.School of Life ScienceShanghai UniversityShanghaiP.R. China

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