Abstract
Dioscorea zingiberensis is a medicinal herb containing a large amount of steroidal saponins, which are the major bioactive compounds and the primary storage form of diosgenin. The CYP72A gene family, belonging to cytochromes P450, exerts indispensable effects on the biosynthesis of numerous bioactive compounds. In this work, a total of 25 CYP72A genes were identified in D. zingiberensis and categorized into two groups according to the homology of protein sequences. The characteristics of their phylogenetic relationship, intron–exon organization, conserved motifs and cis-regulatory elements were performed by bioinformatics methods. The transcriptome data demonstrated that expression patterns of DzCYP72As varied by tissues. Moreover, qRT-PCR results displayed diverse expression profiles of DzCYP72As under different concentrations of jasmonic acid (JA). Likewise, eight metabolites in the biosynthesis pathway of steroidal saponins (four phytosterols, diosgenin, parvifloside, protodeltonin and dioscin) exhibited different contents under different concentrations of JA, and the content of total steroidal saponin was largest at the dose of 100 μmol/L of JA. The redundant analysis showed that 12 DzCYP72As had a strong correlation with specialized metabolites. Those genes were negatively correlated with stigmasterol and cholesterol but positively correlated with six other specialized metabolites. Among all DzCYP72As evaluated, DzCYP72A6, DzCYP72A16 and DzCYP72A17 contributed the most to the variation of specialized metabolites in the biosynthesis pathway of steroidal saponins. This study provides valuable information for further research on the biological functions related to steroidal saponin biosynthesis.
Highlights
Cytochromes P450 (CYPs) are a superfamily of heme-containing enzymes that mainly function as monooxygenases in all kingdoms of life
The published CYP72A protein data of A. thaliana were used as the reference sequence against the genomic information of D. zingiberensis to identify DzCYP72A proteins, and 25 sequences were identified as CYP72A proteins (Table S1)
Based on the protein sequence of all DzCYP72As, we found that all DzCYP72A proteins contained motif 1, implying that motif 1 might be one of conserved motifs among all CYP72A proteins in D. zingiberensis
Summary
Cytochromes P450 (CYPs) are a superfamily of heme-containing enzymes that mainly function as monooxygenases in all kingdoms of life. The genes of CYPs in the plant kingdom are usually larger than those in the kingdoms of animals and microorganisms, accounting for approximately 1% of the protein-coding genes [1]. CYPs are involved in the biosynthesis of diverse specialized metabolites, such as phytohormones, fatty acids and flavonoids, playing a crucial role in plant growth and development [2]. The CYPs are mainly composed of single-family clans and multiple-family clans based on phylogenetic classification [3]. The CYP72A gene subfamily, an important component of the multiple-family clan in CYPs, catalyzes numerous crucial reactions in the biosynthesis pathway of many specialized metabolites. Gibberellins (GAs), especially GAs with low bioactivity, exert positive influences on seed dormancy. AtCYP72A9 has been confirmed to encode bioactive GA
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