Evolution of the Ergot Alkaloid Biosynthetic Gene Cluster Results in Divergent Mycotoxin Profiles in Claviceps purpurea Sclerotia

Research into ergot alkaloid production in major cereal cash crops is crucial for furthering our understanding of the potential toxicological impacts of Claviceps purpurea upon Canadian agriculture and to ensure consumer safety. An untargeted metabolomics approach profiling extracts of C. purpurea sclerotia from four different grain crops separated the C. purpurea strains into two distinct metabolomic classes based on ergot alkaloid content. Variances in C. purpurea alkaloid profiles were correlated to genetic differences within the lpsA gene of the ergot alkaloid biosynthetic gene cluster from previously published genomes and from newly sequenced, long-read genome assemblies of Canadian strains. Based on gene cluster composition and unique polymorphisms, we hypothesize that the alkaloid content of C. purpurea sclerotia is currently undergoing adaptation. The patterns of lpsA gene diversity described in this small subset of Canadian strains provides a remarkable framework for understanding accelerated evolution of ergot alkaloid production in Claviceps purpurea.

ERYTHRINA L. (FABACEAE): UMA ABORDAGEM BIBLIOMÉTRICA

Erythrina L. stand out in folk medicine, environmental restoration, landscaping and other types of uses. These characteristics led to improvement the scientific research of the Erythrina genus. Therefore, this article aimed to perform a bibliometric analysis of Erythrina species distributed in Brazil. The search was based on the accepted name and synonyms of Erythrina species, with natural distribution in Brazil, being sampled the articles that contain this taxon in the title, subtitle, abstract or keywords. Data were categorized into study classes. Most publications were related to biochemical properties. This predominance comes from the discovery of alkaloid production by Erythrina species. E. velutina was the most common species, surpassing E. crista-galli scientific production. The rise of E. velutina in more recent research may be a result of the saturation of E. crista-galli research field. In relation to the other classes of study, little is known about the biology of the species, reinforcing the continuity of the scientific investigation and consequently, filling the gaps of studies with the Erythrina genus.

Machine learning discovery of missing links that mediate alternative branches to plant alkaloids

Engineering the microbial production of secondary metabolites is limited by the known reactions of correctly annotated enzymes in sequence databases. To expand the range of biosynthesis pathways, machine learning is herein demonstrated for the discovery of missing link enzymes, using benzylisoquinoline alkaloid production as a model application with potential to revolutionize the paradigm of sustainable biomanufacturing. Bacterial studies utilize a tetrahydropapaveroline pathway, whereas plants are reported to contain a more stable norcoclaurine pathway, which is exploited in yeast. However, committed aromatic precursors are currently produced by microbial enzymes that remain elusive in plants. Accordingly, the machine learning enzyme selection algorithm is first applied to clarify the early missing links in plant alkaloid pathways. Characterization of predicted sequences via metabolomics reveals distinct oxidases and carboxy-lyases, which complete a plant gene-only benzylisoquinoline alkaloid pathway from tyrosine. Synergistic application of aryl acetaldehyde producing enzymes results in enhanced production through hybrid norcoclaurine and tetrahydropapaveroline pathways. Transplantation of features into homologous enzyme templates leads to the highest levels of bacterial norcoclaurine and N-methylcoclaurine. Mechanism-directed isotope tracing patterns confirm alternative flux branches from aromatic precursors to alkaloids. This machine learning-driven workflow can be adapted to numerous pathways.

Different carbon sources and their concentrations change alkaloid production and gene expression in Catharanthus roseus shoots in vitro

To compare the effects of different carbon sources on physiological aspects, especially medicinal alkaloid biosynthesis and related gene expression in Catharantus roseus (L.) G.Don, we employed sucrose and sorbitol with two concentrations (87.64 mM, the equimolar concentration of sucrose in MS basal medium, and 150 mM) on the plant’s shoots in vitro in presence of 100 μM methyl jasmonate. The production of plant alkaloids including vincristine, vinblastine, ajmalicine, vindoline and catharantine and their biosynthetic and regulatory gene expression was measured. Both treatments had incremental effects on alkaloid production, upregulated the mitogen-activated protein kinase3 (MAPK3) and a downstream responsive transcription factor, ORCA3, which resulted in elevated transcript contents of the important genes in terpenoid indol alkaloids biosynthetic pathway including peroxidase1 (PRX1), geissoschizine synthase (GS), strictosidine synthase (STR) and deacetylvindoline acetyltransferase (DAT). Defensive responses such as antioxidant enzymes (catalase, peroxidase and superoxide dismutase) activities and non-enzymatic metabolites (total phenolics, flavonoids and carotenoids) contents increased under both treatments but the effects of sorbitol were stronger. Reduced fresh weight and chlorophylls contents, increased malondialdehyde (MDA) and carotenoid contents were shown after a week under all employed treatments. It seems that replacement of sucrose with sorbitol and also, increased concentrations of both carbon sources via increasing osmotic pressure make stressful conditions for the plant especially in longer times.