Probing ergot alkaloid biosynthesis: identification of advanced intermediates along the biosynthetic pathway

1988 ◽  
Vol 110 (6) ◽  
pp. 1970-1971 ◽  
Author(s):  
Alan P. Kozikowski ◽  
Jiang Ping. Wu ◽  
Masaaki. Shibuya ◽  
Heinz G. Floss
Keyword(s):  
Ergot Alkaloid ◽  
Biosynthetic Pathway ◽  
Alkaloid Biosynthesis

scholarly journals Identifying Genes Involved in Alkaloid Biosynthesis in Vinca minor through Transcriptomics and Gene Co-Expression Analysis

Biomolecules ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1595
Author(s):  
Emily Amor Stander ◽  
Liuda Johana Sepúlveda ◽  
Thomas Dugé de Bernonville ◽  
Inês Carqueijeiro ◽  
Konstantinos Koudounas ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Biosynthetic Pathway ◽  
Fluorescent Proteins ◽  
Light Exposure ◽  
Indole Alkaloids ◽  
High Light ◽  
Alkaloid Biosynthesis ◽  
Vinca Minor ◽  
Young Leaves ◽  
Derivatives Of

The lesser periwinkle Vinca minor accumulates numerous monoterpene indole alkaloids (MIAs) including the vasodilator vincamine. While the biosynthetic pathway of MIAs has been largely elucidated in other Apocynaceae such as Catharanthus roseus, the counterpart in V. minor remains mostly unknown, especially for reactions leading to MIAs specific to this plant. As a consequence, we generated a comprehensive V. minor transcriptome elaborated from eight distinct samples including roots, old and young leaves exposed to low or high light exposure conditions. This optimized resource exhibits an improved completeness compared to already published ones. Through homology-based searches using C. roseus genes as bait, we predicted candidate genes for all common steps of the MIA pathway as illustrated by the cloning of a tabersonine/vincadifformine 16-O-methyltransferase (Vm16OMT) isoform. The functional validation of this enzyme revealed its capacity of methylating 16-hydroxylated derivatives of tabersonine, vincadifformine and lochnericine with a Km 0.94 ± 0.06 µM for 16-hydroxytabersonine. Furthermore, by combining expression of fusions with yellow fluorescent proteins and interaction assays, we established that Vm16OMT is located in the cytosol and forms homodimers. Finally, a gene co-expression network was performed to identify candidate genes of the missing V. minor biosynthetic steps to guide MIA pathway elucidation.

scholarly journals Transcriptome Analysis of Aconitum carmichaelii and Exploration of the Salsolinol Biosynthetic Pathway

2019 ◽  
Author(s):  
Yuxia Yang ◽  
Ping Hu ◽  
Xianjian Zhou ◽  
Ping Wu ◽  
Xinxin Si ◽  
...  
Keyword(s):  
Transcriptome Analysis ◽  
Theoretical Basis ◽  
Biosynthetic Pathway ◽  
Target Genes ◽  
Shikimate Pathway ◽  
Biosynthesis Pathway ◽  
Alkaloid Biosynthesis ◽  
Diterpenoid Alkaloids ◽  
Protein Databases ◽  
Aconitum Carmichaelii

Abstract Background Aconitum carmichaelii has been used in traditional Chinese medicine for treating various diseases for several thousand years. The biosynthetic pathway of some alkaloids such as C19-diterpenoid alkaloids has been reported, but pathways in different varieties of A. carmichaelii remain unknown. Herein, we performed transcriptome analysis of varieties A. carmichaelii and characterized the biosynthetic pathway of salsolinol. The results expand our knowledge of alkaloids biosynthesis, and provide a theoretical basis for analysing differences in alkaloids biosynthesis patterns in different varieties. Results A total of 56 million raw reads (8.28 G) and 55 million clean reads (8.24 G) were obtained from two varieties (Z175 and R184) leaf transcriptomes, respectively, and 176,793 unigenes were annotated using six protein databases. This yielded 6,873 differentially expressed genes (DEGs) in the two varieties, of which 281 are involved in the salsolinol biosynthetic pathway, including 158 and 75 related to glycolysis and the shikimate pathway, respectively. Furthermore, 843 DEGs were found to be involved in the formation of C19-diterpenoid alkaloids, with 34 differed between the two varieties. These target genes were analysed to explore differences in C19-diterpenoid alkaloid biosynthesis in Z175 and R184. In addition, 322 DEGs encoding transcription factors potentially related to alkaloid accumulation were identified. Conclusions The biosynthesis pathway for C19-diterpenoid alkaloids and salsolinol included 34 and 24 DEGs in Z175 and R184, respectively. Thus, genes involved in alkaloid biosynthesis and accumulation differ between varieties. The mechanisms underlying the differences and their relevance require further exploration.

A bifunctional old yellow enzyme from Penicillium roqueforti is involved in ergot alkaloid biosynthesis

2017 ◽  
Vol 15 (38) ◽  
pp. 8059-8071 ◽  
Author(s):  
Nina Gerhards ◽  
Shu-Ming Li
Keyword(s):  
Ergot Alkaloid ◽  
Penicillium Roqueforti ◽  
Alkaloid Biosynthesis ◽  
Old Yellow Enzyme

Bifunctional FgaOx3Pr3catalyses the formation of festuclavine in the presence of EasG or FgaFS and enhances the activity of several chanoclavine-I dehydrogenases tremendously.

scholarly journals Towards the elimination of ergot alkaloid biosynthesis genes in Neotyphodium Coenophialum

2007 ◽  
Vol 13 ◽  
pp. 477-479
Author(s):  
S. Florea ◽  
C. Machado ◽  
D. Zhang ◽  
D.G. Panaccione ◽  
C.L. Schardl
Keyword(s):  
Tall Fescue ◽  
Ergot Alkaloid ◽  
Festuca Arundinacea ◽  
Ergot Alkaloids ◽  
Loxp Site ◽  
Alkaloid Biosynthesis ◽  
Neotyphodium Coenophialum ◽  
Type Locus ◽  
Dimethylallyltryptophan Synthase ◽  
Recombination System

Neotyphodium coenophialum strain e19 from tall fescue cv. Kentucky 31 carries dmaW1 and dmaW2, two gene homologues that encode dimethylallyltryptophan synthase, the enzyme for the first step in ergot-alkaloid biosynthesis. In our effort to disrupt both homologues and ultimately obtain marker-free mutants, we are using a marker-exchange strategy employing the Cre/ loxP site-specific recombination system. Of 1522 transformants obtained and screened, three were likely dmaW2 disruptants because they gave no PCR product from the wild-type locus, but yielded the larger PCR fragment from the disruption construct. The putative dmaW2-knockouts were also transformed with pKAES186, a plasmid with a cassette containing the cre and ble genes in between loxP sequences. The transformants obtained were screened for the presence of hph, cre and ble genes. The preliminary results indicate a loop-out of the hph gene. The transformants inoculated into endophyte-free tall fescue preserved their compatibility with the plant. The fungus grown from these plants will be further analysed for the presence of hph, cre and ble genes. Keywords: Cre/LoxP, dimethylallyltryptophan synthase, dmaW, Epichloë, ergot alkaloids, Festuca arundinacea, gene knockouts, Lolium arundinaceum, Neotyphodium coenophialum, tall fescue

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