Developmental and inducible accumulation of gene transcripts involved in alkaloid biosynthesis in opium poppy

Facchini, Peter J.; Park, Sang Un

doi:10.1016/s0031-9422(03)00292-9

Cited by 112 publications

(93 citation statements)

References 32 publications

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“…Marianne) plants were cultivated in growth chambers as described previously (8 (8). P. somniferum and Papaver bracteatum cell cultures were treated with an elicitor prepared from Botrytis cinerea mycelia as described previously (8).…”

Section: Methodsmentioning

confidence: 99%

“…A full-length cDNA encoding TNMT was identified by screening the EST data base in silico using the tBLASTn algorithm (42) in DNATools 6.0. CNMT cDNAs from P. somniferum (8), T. flavum (29), and C japonica (6) were used as query sequences. Total RNA was isolated using the method of Cheng and Seemann (43).…”

Section: Methodsmentioning

confidence: 99%

“…The recently reported molecular cloning of cDNAs encoding CNMT from P. somniferum (8), T. flavum (29), and C. japonica (6) provided query sequences to identify a full-length cDNA encoding opium poppy TNMT in an EST data base by in silico mining. We previously used this approach to identify cDNAs encoding opium poppy NCS isoforms with T. flavum NCS as the query sequence (4).…”

Section: Identification Of a Tnmt Cdna And Phylogenetic Analysis-mentioning

confidence: 99%

“…Norcoclaurine 6-O-methyltransferase (6OMT) and coclaurine N-methyltransferase (CNMT) convert (S)-norcoclaurine to (S)-N-methylcoclaurine (6 -8). The cytochrome P450-dependent monooxygenase (S)-N-methylcoclaurine-3Ј-hydroxylase (CYP80B3) catalyzes the 3Ј-hydroxylation of (S)-N-methylcoclaurine (9,10) prior to the formation of (S)-reticuline by 3Ј-hydroxy-N-methylcoclaurine 4Ј-O-methyltransferase (4ЈOMT) (8,11,12). (S)-Reticuline represents the last common intermediate in the biosynthesis of both sanguinarine and morphine (13).…”

mentioning

confidence: 99%

“…1). CNMT has been purified (28), and corresponding cDNAs have been isolated from Coptis japonica (6), Thalictrum flavum (29), and P. somniferum (8). TNMT protein has been isolated from Eschscholzia californica and Corydalis vaginans cell cultures (30) and purified from Sanguinaria canadensis cell cultures (31).…”

mentioning

confidence: 99%

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Molecular Cloning and Characterization of Tetrahydroprotoberberine cis-N-Methyltransferase, an Enzyme Involved in Alkaloid Biosynthesis in Opium Poppy

Liscombe¹,

Facchini

2007

Journal of Biological Chemistry

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S-Adenosyl-L-methionine:tetrahydroprotoberberine cis-Nmethyltransferase (EC 2.1.1.122) catalyzes the conversion of (S)-stylopine to the quaternary ammonium alkaloid, (S)-cis-Nmethylstylopine, as a key step in the biosynthesis of protopine and benzophenanthridine alkaloids in plants. A full-length cDNA encoding a protein exhibiting 45 and 48% amino acid identity with coclaurine N-methyltransferase from Papaver somniferum (opium poppy) and Coptis japonica, respectively, was identified in an elicitor-treated opium poppy cell culture expressed sequence tag data base. Phylogenetic analysis showed that the protein belongs to a unique clade of enzymes that includes coclaurine N-methyltransferase, the predicated translation products of the Arabidopsis thaliana genes, At4g33110 and At4g33120, and bacterial S-adenosyl-L-methionine-dependent cyclopropane fatty acid synthases. Expression of the cDNA in Escherichia coli produced a recombinant enzyme able to convert the protoberberine alkaloids stylopine, canadine, and tetrahydropalmatine to their corresponding N-methylated derivatives. However, the protoberberine alkaloids tetrahydroxyberbine and scoulerine, and simple isoquinoline, benzylisoquinoline, and pavine alkaloids were not accepted as substrates, demonstrating the strict specificity of the enzyme. The apparent K m values for (R,S)-stylopine and S-adenosyl-L-methionine were 0.6 and 11.5 M, respectively. TNMT gene transcripts and enzyme activity were detected in opium poppy seedlings and all mature plant organs and were induced in cultured opium poppy cells after treatment with a fungal elicitor. The enzyme was detected in cell cultures of other members of the Papaveraceae but not in species of related plant families that do not accumulate protopine and benzophenanthridine alkaloids.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%