Representational Difference Analysis of Transcripts Involved in Jervine Biosynthesis.

Magdalena Szeliga,Mirosław Tyrka,Joanna Ciura

doi:10.3390/life10060088

Abstract

Veratrum-type steroidal alkaloids (VSA) are the major bioactive ingredients that strongly determine the pharmacological activities of Veratrum nigrum. Biosynthesis of VSA at the molecular and genetic levels is not well understood. Next-generation sequencing of representational difference analysis (RDA) products after elicitation and precursor feeding was applied to identify candidate genes involved in VSA biosynthesis. A total of 12,048 contigs with a median length of 280 bases were received in three RDA libraries obtained after application of methyl jasmonate, squalene and cholesterol. The comparative analysis of annotated sequences was effective in identifying candidate genes. GABAT2 transaminase and hydroxylases active at C-22, C-26, C-11, and C-16 positions in late stages of jervine biosynthesis were selected. Moreover, genes coding pyrroline-5-carboxylate reductase and enzymes from the short-chain dehydrogenases/reductases family (SDR) associated with the reduction reactions of the VSA biosynthesis process were proposed. The data collected contribute to better understanding of jervine biosynthesis and may accelerate implementation of biotechnological methods of VSA biosynthesis.

Highlights

Veratrum-type steroidal alkaloids (VSA) are valuable bioactive compounds important for the pharmaceutical industry [1]
The results provide evidence that cholesterol and methyl jasmonate efficiently stimulate jervine accumulation in Veratrum plants
Methyl jasmonate has been reported to elicit the production of secondary metabolites in plants, including steroidal alkaloids in Solanum

Summary

Introduction

Veratrum-type steroidal alkaloids (VSA) are valuable bioactive compounds important for the pharmaceutical industry [1]. Natural sources of VSA are limited to the Melanthiaceae family and mainly the Veratrum genus [2]. Over 40 Veratrum species are distributed all over the Northern Hemisphere, only two species (Veratrum nigrum L and V. album) occur naturally in Europe as a natural source of VSA [3,4,5,6]. V. nigrum (2n = 16) belongs to a critically endangered species and a few isolated populations have been reported in Poland [4,7,8]; understanding of VSA biosynthesis is essential for effective biosynthesis of jervine in heterologous systems.

Methods

Results

Conclusion