Abstract

Alkaloids in bulbs of Corydalis (C.) yanhusuo are the major pharmacologically active compounds in treatment of blood vessel diseases, tumors and various pains. However, due to the absence of gene sequences in C. yanhusuo, the genes involved in alkaloid biosynthesis and their expression during bulb development remain unknown. We therefore established the first transcriptome database of C. yanhusuo via Illumina mRNA-Sequencing of a RNA composite sample collected at Bulb initiation (Day 0), early enlargement (Day 10) and maturation (Day 30). 25,013,630 clean 90 bp paired-end reads were de novo assembled into 47,081 unigenes with an average length of 489 bp, among which 30,868 unigenes (65.56%) were annotated in four protein databases. Of 526 putative unigenes involved in biosynthesis o f various alkaloids, 187 were identified as the candidate genes involved in the biosynthesis of benzylisoquinoline alkaloids (BIAs), the only alkaloid type reported in C. yanhusuo untill now. BIAs biosynthetic genes were highly upregulated in the overall pathway during bulb development. Identification of alkaloid biosynthetic genes in C. yanhusuo provide insights on pathways and molecular regulation of alkaloid biosynthesis, to initiate metabolic engineering in order to improve the yield of interesting alkaloids and to identify potentially new alkaloids predicted from the transcriptomic information.

Highlights

  • Alkaloids in bulbs of Corydalis (C.) yanhusuo are the major pharmacologically active compounds in treatment of blood vessel diseases, tumors and various pains

  • For the first time, we presented the transcriptomic information for C. yanhusuo bulbs via the next-generation transcriptomic sequencing and bioinformatic analysis

  • Through blast searches against our transcriptomic database using known alkaloid biosynthetic genes as queries, the majority of benzylisoquinoline alkaloids (BIAs) enzymes involved in the biosynthetic pathway had homologs present in C. yanhusuo bulbs

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Summary

Introduction

Alkaloids in bulbs of Corydalis (C.) yanhusuo are the major pharmacologically active compounds in treatment of blood vessel diseases, tumors and various pains. About 60 alkaloids have been identified from dried C. yanhusuo bulbs[6,13,14,15], and all, except leonticine and coryphenanthrine, belong to benzylisoquinoline alkaloids (BIAs) These known BIAs, based on their chemical structures, were classified into six types including protoberberines, aporphines, protopines, isoquinobenzazepines, benzophenanthridines and bisbenzylisoquinolines[13]. Candidate genes involved in the biosynthesis of BIAs and other types of alkaloids that haven’t been isolated previously in C. yanhusuo, were identified by Tblastn searches of our assembled unigenes using known alkaloid biosynthetic genes as queries. In order to examine molecular regulation of alkaloid biosynthesis during bulb development, transcript abundances of these candidate genes obtained by the Illumina sequencing method were compared in C. yanhusuo bulbs at the initiation, enlargement and maturation stages, which showed the significant metabolomic difference[5]

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