Combination of transcriptomic and proteomic approaches helps to unravel the protein composition of Chelidonium majus L. milky sap.

Robert Nawrot,Hans-Peter Mock,Rico Lippmann,Lothar Altschmied,Jakub Barylski

doi:10.1007/s00425-016-2566-7

Abstract

Main conclusion A novel annotated Chelidonium majus L. transcriptome database composed of 23,004 unique coding sequences allowed to significantly improve the sensitivity of proteomic C. majus assessments, which showed novel defense-related proteins characteristic to its latex. To date, the composition of Chelidonium majus L. milky sap and biosynthesis of its components are poorly characterized. We, therefore, performed de novo sequencing and assembly of C. majus transcriptome using Illumina technology. Approximately, 119 Mb of raw sequence data was obtained. Assembly resulted in 107,088 contigs, with N50 of 1913 bp and N90 of 450 bp. Among 34,965 unique coding sequences (CDS), 23,004 obtained CDS database served as a basis for further proteomic analyses. The database was then used for the identification of proteins from C. majus milky sap, and whole plant extracts analyzed using liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS) approach. Of about 334 different putative proteins were identified in C. majus milky sap and 1155 in C. majus whole plant extract. The quantitative comparative analysis confirmed that C. majus latex contains proteins connected with response to stress conditions and generation of precursor metabolites and energy. Notable proteins characteristic to latex include major latex protein (MLP, presumably belonging to Bet v1-like superfamily), polyphenol oxidase (PPO, which could be responsible for browning of the sap after exposure to air), and enzymes responsible for anthocyanidin, phenylpropanoid, and alkaloid biosynthesis.Electronic supplementary materialThe online version of this article (doi:10.1007/s00425-016-2566-7) contains supplementary material, which is available to authorized users.

Highlights

IntroductionElectronic supplementary material The online version of this article (doi:10.1007/s00425-016-2566-7) contains supplementary material, which is available to authorized users
Electronic supplementary material The online version of this article contains supplementary material, which is available to authorized users.Greater celandine (Chelidonium majus L.) (Fig. 1), an herbaceous perennial plant, belongs to Papaveraceae family, which is an important source of biologically active substances
With the purpose of understanding C. majus transcriptome, RNA was extracted from the young stem of 6-week-old C. majus plant with exuding milky sap (Fig. 1c) and sequenced with Illumina paired-end sequencing technology generating approximately 119 Mb of raw sequence data from 133,550,365 reads with an average length of almost 100 bp (Fig. 3; Table 1)

Summary

Introduction

Electronic supplementary material The online version of this article (doi:10.1007/s00425-016-2566-7) contains supplementary material, which is available to authorized users. Greater celandine (Chelidonium majus L.) (Fig. 1), an herbaceous perennial plant, belongs to Papaveraceae family, which is an important source of biologically active substances. Milky sap and extracts of greater celandine are used in traditional medicine to treat papillae, warts, and condylomas, which are symptoms of human papilloma virus (HPV) infections. C. majus extracts are used to treat liver disorders and fight fever (Hiller et al 1998). The medicinal and pharmaceutical interest in this plant is based on its ability to synthesize alkaloids, flavonoids, or phenolic acids (Colombo and Bosisio 1996). It is often suspected that many of these substances may be either synthesized or stored in laticifers of the plant.

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