Cloning and expression analysis of RcF3H in Rubus chingii
[Objective]Flavanone 3-hydroxylase(F3H)is a key enzyme in the synthesis of plant flavonoids.This study aims to investigate the relationship between RcF3H gene and flavonoid metabolism in Rubus chingii and analyze its biological function,so as to provide reference for further exploring the mechanism of RcF3H in the process of flavonoid accumulation in R.chingii.[Method]RcF3H gene was cloned from R.chingii,and bioinformatics analysis was performed,including physicochemical properties,homology comparison,phylogenetic tree and promoter cis-acting elements.At the same time,this study detected the expression level of RcF3H gene in different tissues,fruit growth period and exogenous methyl jasmonate(MeJA)induction.[Result]RcF3H gene,belonging to the 2-oxoglutarate-dependent dioxygenase superfamily,was located on the first chromosome,and its fragment length was 1 098 bp.RcF3H gene consisted of 1 exon and encoded 365 amino acids.RcF3H belonged to the hydrophilic stable protein.Subcellular localization predicted that the protein was located in the cytoplasm.The secondary and tertiary structure of RcF3H was mainly composed ofα-helix and irregular curl,which had the closest genetic relationship with dicotyledons such as Rubus coreanus and Arabidopsis thaliana.Analysis of promoter cis-acting elements showed that the promoter region of RcF3H gene contained multiple cis-acting elements,and its function mainly focused on responding to hormones and stress.RcF3H gene was highly expressed in fruits,and had obvious tissue specificity.In addition,the expression of RcF3H gene in different ripening stages of fruits was consistent with the accumulation of flavonoids,and the expression increased under the treatment of 100 μmol·L-1 MeJA.These results suggested that RcF3H gene is the key enzyme to promoting flavonoid synthesis.[Conclusion]RcF3H gene responds to MeJA stimulation and may act as a positive regulator of flavonoid biosynthesis in R.chingii,affecting flavonoid accumulation.[Ch,8 fig.2 tab.28 ref.]
万方数据
维普
