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Open AccessArticle

Representational Difference Analysis of Transcripts Involved in Jervine Biosynthesis

1
Department of Biotechnology and Bioinformatics, Faculty of Chemistry, Rzeszow University of Technology, Powstańców Warszawy 6 Ave, 35-959 Rzeszow, Poland
2
Department of Plant Physiology and Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland
*
Author to whom correspondence should be addressed.
Life 2020, 10(6), 88; https://doi.org/10.3390/life10060088
Received: 4 May 2020 / Revised: 10 June 2020 / Accepted: 12 June 2020 / Published: 19 June 2020
(This article belongs to the Section Plant Science and Engineering)
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. View Full-Text
Keywords: black false hellebore; cDNA RDA; gene annotation; Next Generation Sequencing (NGS); Veratrum nigrum L. black false hellebore; cDNA RDA; gene annotation; Next Generation Sequencing (NGS); Veratrum nigrum L.
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Szeliga, M.; Ciura, J.; Tyrka, M. Representational Difference Analysis of Transcripts Involved in Jervine Biosynthesis. Life 2020, 10, 88.

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