Enzymatic formation of β-citraurin from β-cryptoxanthin and Zeaxanthin by carotenoid cleavage dioxygenase4 in the flavedo of citrus fruit

Plant Physiol. 2013 Oct;163(2):682-95. doi: 10.1104/pp.113.223297. Epub 2013 Aug 21.

Abstract

In this study, the pathway of β-citraurin biosynthesis, carotenoid contents and the expression of genes related to carotenoid metabolism were investigated in two varieties of Satsuma mandarin (Citrus unshiu), Yamashitabeni-wase, which accumulates β-citraurin predominantly, and Miyagawa-wase, which does not accumulate β-citraurin. The results suggested that CitCCD4 (for Carotenoid Cleavage Dioxygenase4) was a key gene contributing to the biosynthesis of β-citraurin. In the flavedo of Yamashitabeni-wase, the expression of CitCCD4 increased rapidly from September, which was consistent with the accumulation of β-citraurin. In the flavedo of Miyagawa-wase, the expression of CitCCD4 remained at an extremely low level during the ripening process, which was consistent with the absence of β-citraurin. Functional analysis showed that the CitCCD4 enzyme exhibited substrate specificity. It cleaved β-cryptoxanthin and zeaxanthin at the 7,8 or 7',8' position. But other carotenoids tested in this study (lycopene, α-carotene, β-carotene, all-trans-violaxanthin, and 9-cis-violaxanthin) were not cleaved by the CitCCD4 enzyme. The cleavage of β-cryptoxanthin and zeaxanthin by CitCCD4 led to the formation of β-citraurin. Additionally, with ethylene and red light-emitting diode light treatments, the gene expression of CitCCD4 was up-regulated in the flavedo of Yamashitabeni-wase. These increases in the expression of CitCCD4 were consistent with the accumulation of β-citraurin in the two treatments. These results might provide new strategies to improve the carotenoid contents and compositions of citrus fruits.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carotenoids / metabolism*
  • Chromatography, High Pressure Liquid
  • Citrus / drug effects
  • Citrus / enzymology*
  • Citrus / genetics
  • Citrus / radiation effects
  • Cryptoxanthins
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • Dioxygenases / metabolism*
  • Ethylenes / pharmacology
  • Gene Expression Regulation, Plant / drug effects
  • Gene Expression Regulation, Plant / radiation effects
  • Genes, Plant / genetics
  • Green Fluorescent Proteins / metabolism
  • Light
  • Metabolic Networks and Pathways / drug effects
  • Metabolic Networks and Pathways / genetics
  • Metabolic Networks and Pathways / radiation effects
  • Molecular Sequence Data
  • Nicotiana / drug effects
  • Nicotiana / genetics
  • Nicotiana / metabolism
  • Nicotiana / radiation effects
  • Plant Leaves / drug effects
  • Plant Leaves / metabolism
  • Plant Leaves / radiation effects
  • Plants, Genetically Modified
  • Recombinant Fusion Proteins / metabolism
  • Subcellular Fractions / drug effects
  • Subcellular Fractions / metabolism
  • Subcellular Fractions / radiation effects
  • Xanthophylls / chemistry
  • Xanthophylls / metabolism*
  • Zeaxanthins
  • beta Carotene / analogs & derivatives*
  • beta Carotene / chemistry
  • beta Carotene / isolation & purification
  • beta Carotene / metabolism

Substances

  • Cryptoxanthins
  • DNA, Complementary
  • Ethylenes
  • Recombinant Fusion Proteins
  • Xanthophylls
  • Zeaxanthins
  • citraurin
  • beta Carotene
  • Green Fluorescent Proteins
  • Carotenoids
  • ethylene
  • Dioxygenases

Associated data

  • GENBANK/AB114648
  • GENBANK/AB114649
  • GENBANK/AB114650
  • GENBANK/AB114651
  • GENBANK/AB114653
  • GENBANK/AB114654
  • GENBANK/AB114655
  • GENBANK/AB219164
  • GENBANK/AB219169
  • GENBANK/AB219174
  • GENBANK/AB781688
  • GENBANK/AB781691