cDNA cloning and expression pattern of Cinnamate-4-Hydroxylase in the Korean black raspberry

Abstract

Cinnamate-4-hydroxylase (C4H) is a key enzyme in the phenylpropanoid pathway, which is responsible for synthesizing a variety of secondary metabolites that participate in development and adaptation. In this study, we isolated a full-length cDNA of the C4H gene from the Korean black raspberry (Rubus sp.) and found that this gene existed as a single gene. By comparing the deduced amino acid sequence of Rubus sp. C4H with other sequences reported previously we determined that this sequence was highly conserved among widely divergent plant species. In addition, quantitative real time PCR studies indicated that the C4H gene had a differential expression pattern during fruit development, where gene expression was first detected in green fruit and was then remarkably reduced in yellow fruit, followed by an increase in red and black fruit. To investigate the two peaks in expression observed during fruit development and ripening, we measured the flavonoid content. The content of the major flavanol of Korean black raspberry fruits was determined to be highest at the beginning of fruit development, followed by a gradually decrease according to the developmental stages. In contrast, the content of anthocyanins during the progress of ripening was dramatically increased. Our results suggest that the C4H gene in Korean black raspberry plays a role during color development at the late stages of fruit ripening, whereas the expression of C4H gene during the early stages may be related to the accumulation of flavanols. [BMB reports 2008; 41(7): 529-536]

Identification of a novel anthocyanin synthesis pathway in the fungus Aspergillus sydowii H-1

“…C4H and CHS are vital functional genes in existing studies regarding the process of anthocyanin genes [6,8,75]. However, CHS and C4H could be assembled only by the de novo method, which suggests that the reference genome, CBS 593.65 [35], may not contain these two genes.…”

“…Finally, Pg and Mv are synthesized from Cy and Dp, respectively, with the help of O-methyltransferase (OMT). Among the genes involved in anthocyanin synthesis, C4H is one of the core genes of the phenylpropanoid pathway and mediates the synthesis of secondary metabolites such as anthocyanins [6] and artemisinin [7]. CHS links the phenylpropanoid pathway and the flavonoid pathway as well as plays an important role in the biosynthesis of anthocyanins [8].…”

Advances in functional research of antioxidants and organoleptic traits in berry crops

“…On the other hand, RiPAL2 was associated with late flower and fruit development [54]. The expression analysis of C4H during fruit ripening in the Korean black raspberry showed a two-phase pattern: comparison with metabolite levels indicates that the early expression of Rubus C4H gene seems to be related with the synthesis of flavan-3-ols in green fruits, while the peak at late ripening should be associated with the biosynthesis of anthocyanins [12]. The characterization of the raspberry 4CL family identified three genes, each with a distinct developmental pattern of expression in various organs.…”

“…The pathway starts from the amino acid phenylalanine and leads to the synthesis of phenylpropanoid precursors of lignin and flavonoid compounds by the action of phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H) and 4-coumarate CoA ligase (4CL) enzymes. The expression analysis of C4H during fruit ripening in the Korean black raspberry showed a two-phase pattern: comparison with metabolite levels indicates that the early expression of Rubus C4H gene seems to be related with the synthesis of flavan-3-ols in green fruits, while the peak at late ripening should be associated with the biosynthesis of anthocyanins [12]. 1) [51], providing one prominent example of secondary metabolic route producing both antioxidant and volatile compounds.…”

Identification of upregulated genes in laminarin-treated poplar (Populus alba × P. tremula var. glandulosa) suspension cells by suppression subtractive hybridization and cDNA microarray

“…A gene for a PDR-type ABC transporter (PO06006E09) was upregulated 5 h after treatment. Trans-cinnamate 4-hydroxylase, C4H (PO05002B09), is involved in synthesizing various secondary metabolites that participate in plant development and adaptation (BAEK et al, 2008). Induction of C4H as a plant defence response to fungal infection is well documented (SCHMIDT et al, 2004).…”