Characterization of proteins in latex of the opium poppy (Papaver somniferum) using two-dimensional gel electrophoresis and microsequencing

Decker, Gabriele; Wanner, Gerhard; Zenk, Meinhard H.; Lottspeich, Friedrich

doi:10.1002/1522-2683(20001001)21:16<3500::aid-elps3500>3.3.co;2-f

Cited by 13 publications

(31 citation statements)

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“…It is thus necessary for identification of proteins from 2D-gels to obtain sequence data and not rely solely on peptide masses. Such analyses can be done by fragmentation of peptides measured by mass spectrometry or by Edman degradation, as is used for the identification of protein spots (Decker et al 2000). They are needed for C. roseus as a model plant for studies of the secondary metabolism.…”

Section: Discussionmentioning

confidence: 99%

Proteome analysis of the medicinal plant Catharanthus roseus

Jacobs¹,

Gaspari²,

Greef³

et al. 2005

Planta

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A proteomic approach is undertaken aiming at the identification of novel proteins involved in the alkaloid biosynthesis of Catharanthus roseus. The C. roseus cell suspension culture A11 accumulates the terpenoid indole alkaloids strictosidine, ajmalicine and vindolinine. Cells were grown for 21 days, and alkaloid accumulation was monitored during this period. After a rapid increase between day 3 and day 6, the alkaloid content reached a maximum on day 16. Systematic analysis of the proteome was performed by two-dimensional polyacrylamide gel electrophoresis. After day 3, the proteome started to change with an increasing number of protein spots. On day 13, the proteome changed back to roughly the same as at the start of the growth cycle. 88 protein spots were selected for identification by mass spectrometry (MALDI-MS/MS). Of these, 58 were identified, including two isoforms of strictosidine synthase (EC 4.3.3.2), which catalyzes the formation of strictosidine in the alkaloid biosynthesis; tryptophan synthase (EC 4.1.1.28), which is needed for the supply of the alkaloid precursor tryptamine; 12-oxophytodienoate reductase, which is indirectly involved in the alkaloid biosynthesis as it catalyzes the last step in the biosynthesis of the regulator jasmonic acid. Unique sequences were found, which may also relate to unidentified biosynthetic proteins.

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Section: Discussionmentioning

confidence: 99%

Proteome analysis of the medicinal plant Catharanthus roseus

Jacobs¹,

Gaspari²,

Greef³

et al. 2005

Planta

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show abstract

“…However, genomic resources have not been extensively developed for the model plants used to investigate alkaloid biosynthesis. Expressed sequence tags (ESTs) , DNA microarrays Ziegler et al 2005;Siminszky et al 2005), proteomics (Decker et al 2000;Ounaroon et al 2003;Jacobs et al 2005), and metabolomics (Choi et al 2004) have recently been used to isolate new genes and glean novel insights into alkaloid metabolism. It is notable that NCS (Liscombe et al 2005) and TNMT (Liscombe and Facchini 2006) were identiWed using the genomics tools described herein, and 4ЈOMT (Ziegler et al 2005) was isolated using similar means.…”

Section: Introductionmentioning

confidence: 99%

Gene transcript and metabolite profiling of elicitor-induced opium poppy cell cultures reveals the coordinate regulation of primary and secondary metabolism

Zulak

Cornish

Daskalchuk

et al. 2006

Planta

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Elicitor-induced sanguinarine accumulation in opium poppy (Papaver somniferum) cell cultures provides a responsive model system to profile modulations in gene transcripts and metabolites related to alkaloid biosynthesis. An annotated expressed sequence tag (EST) database was assembled from 10,224 random clones isolated from an elicitor-treated opium poppy cell culture cDNA library. The most abundant ESTs encoded defense proteins, and enzymes involved in alkaloid metabolism and S-adenosylmethionine-dependent methyl transfer. ESTs corresponding to 40 enzymes involved in the conversion of sucrose to sanguinarine were identified. A corresponding DNA microarray was probed with RNA from cell cultures collected at various time-points after elicitor treatment, and compared with RNA from control cells. Several diverse transcript populations were coordinately induced, with alkaloid biosynthetic enzyme and defense protein transcripts displaying the most rapid and substantial increases. In addition to all known sanguinarine biosynthetic gene transcripts, mRNAs encoding several upstream primary metabolic enzymes were coordinately induced. Fourier transform-ion cyclotron resonance-mass spectrometry was used to characterize the metabolite profiles of control and elicitor-treated cell cultures. Principle component analysis revealed a significant and dynamic separation in the metabolome, represented by 992 independent detected analytes, in response to elicitor treatment. Identified metabolites included sanguinarine, dihydrosanguinarine, and the methoxylated derivatives dihydrochelirubine and chelirubine, and the alkaloid pathway intermediates N-methylcoclaurine, N-methylstylopine, and protopine. Some of the detected analytes showed temporal changes in abundance consistent with modulations in the profiles of alkaloid biosynthetic gene transcripts.

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“…Kutchan 1995;Sato et al 2001;Park et al 2002), and latex chemistry (e.g. Decker et al 2000;Memelink 2004;Frick et al 2005).…”

Section: Introductionmentioning

confidence: 99%

EST database for early flower development in California poppy (Eschscholzia californica Cham., Papaveraceae) tags over 6000 genes from a basal eudicot

et al. 2006

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The Floral Genome Project (FGP) selected California poppy (Eschscholzia californica Cham. ssp. Californica) to help identify new florally-expressed genes related to floral diversity in basal eudicots. A large, non-normalized cDNA library was constructed from premeiotic and meiotic floral buds and sequenced to generate a database of 9,079 high quality Expressed Sequence Tags (ESTs). These sequences clustered into 5,713 unigenes, including 1,414 contigs and 4,299 singletons. Homologs of genes regulating many aspects of flower development were identified, including those for organ identity and development, cell and tissue differentiation, cell cycle control, and secondary metabolism. Over 5% of the transcriptome consisted of homologs to known floral gene families. Most are the first representatives of their respective gene families in basal eudicots and their conservation suggests they are important for floral development and/or function. App. 10% of the transcripts encoded transcription factors and other regulatory genes, including nine genes from the seven major lineages of the important MADS-box family of developmental regulators. Homologs of alkaloid pathway genes were also recovered, providing opportunities to explore adaptive evolution in secondary products. Furthermore, comparison of the poppy ESTs with the Arabidopsis genome provided support for putative Arabidopsis genes that previously lacked annotation. Finally, over 1,800 unique sequences had no observable homology in the public databases. The California poppy EST database and library will help bridge our understanding of flower initiation and development among higher eudicot and monocot model plants and provide new opportunities for comparative analysis of gene families across angiosperm species.

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Characterization of proteins in latex of the opium poppy (Papaver somniferum) using two-dimensional gel electrophoresis and microsequencing

Cited by 13 publications

References 0 publications

Proteome analysis of the medicinal plant Catharanthus roseus

Proteome analysis of the medicinal plant Catharanthus roseus

Gene transcript and metabolite profiling of elicitor-induced opium poppy cell cultures reveals the coordinate regulation of primary and secondary metabolism

EST database for early flower development in California poppy (Eschscholzia californica Cham., Papaveraceae) tags over 6000 genes from a basal eudicot

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