Identification of a CYP84 Family of Cytochrome P450-Dependent Mono-Oxygenase Genes in <i>Brassica napus</i> and Perturbation of Their Expression for Engineering Sinapine Reduction in the Seeds

Nair, Ramesh; Joy, Richard W.; Kurylo, Eugen; Shi, Xiaohong; Schnaider, Joan; Datla, Raju; Keller, W. A.; Selvaraj, Gopalan

doi:10.1104/pp.123.4.1623

Cited by 47 publications

(48 citation statements)

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“…Sinapine synthesis, a distinct hallmark of crucifers, occurs only in seed tissue, and is a subject of our interest (Nair et al, 2000). Although the CYP98A3 promoter was active in the silique walls of transgenic Arabidopsis, it was not active in seeds.…”

Section: Developmental Regulation Of Cyp98a3 Commonality and Dissimimentioning

confidence: 99%

“…All tissues were frozen in liquid nitrogen and stored at Ϫ80°C. All DNA and RNA extractions and analyses were as in Nair et al (2000). A Molecular Dynamics PhosporImager:SI and ImageQuaNT program (Amersham, Sunnyvale, CA) were used for quantitating the RNA signal intensity according to the supplier's instructions.…”

Section: Dna and Rna Analysismentioning

confidence: 99%

“…First-strand cDNA was synthesized using 10 g of total RNA as described in Nair et al (2000). PCR was set in a 50-L reaction volume containing 2.5 L of the cDNA, 1ϫ Pfu polymerase buffer (Stratagene, La Jolla, CA), 200 m of each dNTP, and 50 pmol of each of the gene-specific primers (5Ј-gcggatccg-atgtcgtggtttctaatagcggtgg-3Ј and 5Ј-gcgaattcattacatatcgtaaggcacgcgt-3Ј) to amplify the CYP98A3 ORF.…”

Section: Reverse Transcriptase-pcrmentioning

confidence: 99%

“…In most cases the biosynthetic routes are unknown and even in some of the well-studied pathways many aspects remain uncertain. Phenylpropanoid metabolism generates phenolic intermediates and end products that include lignin monomers, flavonoids, isoflavonoids, lignans, tannins, quinones, and sinapate esters (Strack, 1997; Dixon and Steele, 1999;Nair et al, 2000). Lignin constitutes approximately 15% to 30% of the dry weight in woody plants, and contributes about 30% of the organic carbon in plant biomass in general (Lewis and Yamamoto, 1990; Douglas, 1996; Boudet, 2000).…”

mentioning

confidence: 99%

“…Sinapine is an antinutritional factor in canola (Brassica napus), an economically important oilseed crop, and consequently a target for elimination by breeding and biotechnology Velasco and Mö llers, 1998;Nair et al, 2000). Previously, we had characterized F5H genes from canola and showed that their transgenic suppression can result in up to 40% reduction in sinapine (Nair et al, 2000). We undertook the work presented here to investigate the elusive coumarate 3-hydroxylase (C3H) in Arabidopsis to gain an additional handle for metabolic engineering of sinapate ester synthesis in canola, a close relative of Arabidopsis.…”

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Arabidopsis CYP98A3 Mediating Aromatic 3-Hydroxylation. Developmental Regulation of the Gene, and Expression in Yeast

et al. 2002

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The general phenylpropanoid pathways generate a wide array of aromatic secondary metabolites that range from monolignols, which are ubiquitous in all plants, to sinapine, which is confined to crucifer seeds. The biosynthesis of these compounds involves hydroxylated and methoxylated cinnamyl acid, aldehyde, or alcohol intermediates. Of the three enzymes originally proposed to hydroxylate the 4-, 3-, and 5-positions of the aromatic ring, cinnamate 4-hydroxylase (C4H), which converts trans-cinnamic acid to p-coumaric acid, is the best characterized and is also the archetypal plant P450 monooxygenase. Ferulic acid 5-hydroxylase (F5H), a P450 that catalyzes 5-hydroxylation, has also been studied, but the presumptive 3-hydroxylase converting p-coumarate to caffeate has been elusive. We have found that Arabidopsis CYP98A3, also a P450, could hydroxylate p-coumaric acid to caffeic acid in vivo when expressed in yeast (Saccharomyces cerevisiae) cells, albeit very slowly. CYP98A3 transcript was found in Arabidopsis stem and silique, resembling both C4H and F5H in this respect. CYP98A3 showed further resemblance to C4H in being highly active in root, but differed from F5H in this regard. In transgenic Arabidopsis, the promoters of CYP98A3 and C4H showed wound inducibility and a comparable developmental regulation throughout the life cycle, except in seeds, where the CYP98A3 promoter construct was inactive while remaining active in silique walls. Within stem and root tissue, the gene product and the promoter activity of CYP98A3 were most abundant in lignifying cells. Collectively, these studies show involvement of CYP98A3 in the general phenylpropanoid metabolism, and suggest a downstream function for CYP98A3 relative to the broader and upstream role of C4H.Plants synthesize thousands of secondary metabolites from offshoots of primary metabolism (Croteau et al., 2000). In most cases the biosynthetic routes are unknown and even in some of the well-studied pathways many aspects remain uncertain. Phenylpropanoid metabolism generates phenolic intermediates and end products that include lignin monomers, flavonoids, isoflavonoids, lignans, tannins, quinones, and sinapate esters (Strack, 1997; Dixon and Steele, 1999;Nair et al., 2000). Lignin constitutes approximately 15% to 30% of the dry weight in woody plants, and contributes about 30% of the organic carbon in plant biomass in general (Lewis and Yamamoto, 1990; Douglas, 1996; Boudet, 2000). Thus, lignin assembly places a huge demand on phenylpropanoid supply. Lignification is considered a biochemical adaptation to provide mechanical strength and "non-seeping" water transport channels as plants adopted terrestrial habitats. The biosynthetic pathways appear to have been further diversified and recruited to supply metabolites for a variety of other end uses such as attraction of pollinators for promoting sexual propagation, pest deterrence, pathogen resistance, UV radiation protection, and allelopathic exclusion of potentially competing plants (Dixon et al., 1996). The inherent ...

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Section: Developmental Regulation Of Cyp98a3 Commonality and Dissimimentioning

confidence: 99%

Section: Dna and Rna Analysismentioning

confidence: 99%

Section: Reverse Transcriptase-pcrmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Arabidopsis CYP98A3 Mediating Aromatic 3-Hydroxylation. Developmental Regulation of the Gene, and Expression in Yeast

et al. 2002

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Biosynthesis of Phenylpropanoids and Related Compounds (FromAPRVolume 40)

Petersen

Hans

Matern

2018

Annual Plant Reviews Online

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Phenolic compounds are ubiquitous in the plant kingdom. A main pathway for the formation of these compounds starts with the aromatic amino acidsl‐phenylalanine and – to a lesser extent –l‐tyrosine. In the general phenylpropanoid pathway, these are transformed to coenzyme A‐activated 4‐coumaric acid by phenylalanine ammonia‐lyase, cinnamic acid 4‐hydroxylase and 4‐coumarate CoA‐ligase. 4‐Coumaroyl‐CoA gives rise to a large number of different natural products, e.g. flavonoids, lignans, coumarins, tannins, hydroxycinnamic acid esters and amides as well as lignin monomers. This review gives an insight into recent findings concerning the general phenylpropanoid pathway as well as the biosyntheses of hydroxycinnamoyl conjugates, phenolic aroma and fragrance compounds, lignans, coumarins and gallo‐/ellagitannins.

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Biosynthesis of Phenylpropanoids and Related Compounds

Petersen

Hans

Matern

Biochemistry of Plant Secondary Metabolism

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Identification of a CYP84 Family of Cytochrome P450-Dependent Mono-Oxygenase Genes in Brassica napus and Perturbation of Their Expression for Engineering Sinapine Reduction in the Seeds

Cited by 47 publications

References 61 publications

Arabidopsis CYP98A3 Mediating Aromatic 3-Hydroxylation. Developmental Regulation of the Gene, and Expression in Yeast

Arabidopsis CYP98A3 Mediating Aromatic 3-Hydroxylation. Developmental Regulation of the Gene, and Expression in Yeast

Biosynthesis of Phenylpropanoids and Related Compounds (FromAPRVolume 40)

Biosynthesis of Phenylpropanoids and Related Compounds

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