Transcriptome Analysis of CYP450 Family Members in Fritillaria cirrhosa D. Don and Profiling of Key CYP450s Related to Isosteroidal Alkaloid Biosynthesis

Abstract

Fritillaria cirrhosa D. Don (known as Chuan-Bei-Mu in Chinese) can synthesize isosteroidal alkaloids (ISA) with excellent medicinal value, and its bulb has become an indispensable ingredient in many patented drugs. Members of the cytochrome P450 (CYP450) gene superfamily have been shown to play essential roles in regulating steroidal alkaloids biosynthesis. However, little information is available on the P450s in F. cirrhosa. Here, we performed full-length transcriptome analysis and discovered 48 CYP450 genes belonging to 10 clans, 25 families, and 46 subfamilies. By combining phylogenetic trees, gene expression, and key F. cirrhosa ISA content analysis, we presumably identify seven FcCYP candidate genes, which may be hydroxylases active at the C-22, C-23, or C-26 positions in the late stages of ISA biosynthesis. The transcript expression levels of seven FcCYP candidate genes were positively correlated with the accumulation of three major alkaloids in bulbs of different ages. These data suggest that the candidate genes are most likely to be associated with ISA biosynthesis. Finally, the subcellular localization prediction of FcCYPs and transient expression analysis within Nicotiana benthamiana showed that the FcCYPs were mainly localized in the chloroplast. This study presents a systematic analysis of the CYP450 gene family in F. cirrhosa and provides a foundation for further functional characterization of the CYPs involved in ISA biosynthesis.

Integrative analysis of the metabolome and transcriptome provides insights into the mechanisms of lignan biosynthesis in Herpetospermum pedunculosum (Cucurbitaceae)

“…The marker protein RFP-NLS SV40 was used to locate the proteins in the nucleus [ 52 ]. Instantaneous transformations and fluorescence signals were observed, as previously described [ 53 ].…”

Multi-Omics Techniques in Genetic Studies and Breeding of Forest Plants

“…However, given the relatively high error rate of TGS reads, highly accurate NGS RNA-seq reads are required to improve TGS-based transcriptome assembly accuracy [90]. At present, the high-quality and full-length transcriptome of forest plant species, including Larix kaempferi [91], Chosenia arbutifolia [92], Fritillaria cirrhosa [93], and Alsophila spinulosa [94] has been obtained.…”

Multi-Omics Techniques for Forest Plant Breeding

“…However, given the relatively high error rate of TGS reads, highly accurate NGS RNA-seq reads are required to improve TGS-based transcriptome assembly accuracy [85]. At present, the high-quality and full-length transcriptome of forest plant species, including Larix kaempferi [86], Chosenia arbutifolia [87], Fritillaria cirrhosa [88], and Alsophila spinulosa has been obtained [89].…”