Transcriptome analysis of Fritillaria cirrhosa D. Don

<em>Fritillaria cirrhosa</em> 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 <em>F.</em> <em>cirrhosa</em>. Here, we performed full-length transcriptome analysis and discovered 48 <em>CYP450</em> genes belonging to 10 clans, 25 families, and 46 subfamilies. By combining phylogenetic trees, gene expression, and key <em>F. cirrhosa</em> ISA content analysis, we presumably identify seven <em>FcCYP</em> 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 <em>FcCYP</em> 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 <em>Nicotiana benthamiana</em> showed that the FcCYPs were mainly localized in the chloroplast. This study presents a systematic analysis of the CYP450 gene family in <em>F. cirrhosa</em> and provides a foundation for further functional characterization of the <em>CYPs</em> involved in ISA biosynthesis.