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Abstract
Physalis angulata is a renowned traditional Chinese medicine for the treatment of various conditions. Physalin is the major type of bioactive constituents conferring medicinal properties of P. angulata. Despite the medicinal importance, the pathways leading to physalin are largely unknown. In this study, we employed a transcriptomic approach to identify a Pa24ISO gene from P. angulata. Through heterologous expression in yeast, Pa24ISO was revealed to catalyze an isomerization reaction in converting 24-methylenecholesterol to 24-methyldesmosterol. Real-time PCR analysis showed that the abundance of Pa24ISO transcripts correlated with the accumulation pattern of physalin B in different tissues of P. angulata. A direct role of Pa24ISO in channeling of 24-methylenecholesterol for physalin B biosynthesis was illustrated by suppressing the gene in P. angulata via the VIGS approach. Down-regulation of Pa24ISO led to reduced levels of 24-methyldesmosterol and physalin B, accompanied with an increase of campesterol content in P. angulata. The results supported that 24ISO is involved in physalin biosynthesis in plants.
Highlights
Physalis angulata L, popularly known as Kuzhi in Chinese, has been prescribed for centuries for treatments of a variety of diseases, such as fever, malaria, liver disorders, and diabetes (Abe et al, 2006; Damu et al, 2007; Reyes-Reyes et al, 2013)
To investigate whether 24ISO is involved in physalin biosynthesis in vivo, here, we reported the cDNA cloning and functional analysis of a 24ISO gene, designated as
The identity of physalin B was determined by comparing its MS/MS fragmented pattern (Supplementary Figure 7) with that published in a previous literature (Huang, 2014)
Summary
Physalis angulata L, popularly known as Kuzhi in Chinese, has been prescribed for centuries for treatments of a variety of diseases, such as fever, malaria, liver disorders, and diabetes (Abe et al, 2006; Damu et al, 2007; Reyes-Reyes et al, 2013). Crude extracts of P. angulata are reported to show antinociceptive (Choi and Hwang, 2003), anti-breast cancer (Hsieh et al, 2006), and anti-inflammatory activities (Bastos et al, 2008). The medicinal effects of P. angulata are mainly due to the presence of polyoxygenated steroids with an ergosterol backbone possessing a C-22, 26-lactone (Damu et al, 2007). Concerning modifications of their side chains, especially the cleavage of C13-C14 bond and the formation of a 16,24-carboncyclic bond, these steroid lactones can be classified as physalin and withanolide (Figure 1A; Huang et al, 2020; Xia et al, 2021). Of the P. angulata physalin, physalin B has received the most interest due to its diversely pharmacological activities.
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