The legume Oxytropis sericea hosts a fungal endophyte, Alternaria oxytropis, which produces secondary metabolites (SM), including the toxin swainsonine. Polyketide synthase (PKS) and non-ribosomal peptide synthase (NRPS) enzymes are associated with biosynthesis of fungal SM. To better understand the origins of the SM, an unannotated genome of A. oxytropis was assessed for protein sequences similar to known PKS and NRPS enzymes of fungi. Contigs exhibiting identity with known genes were analyzed at nucleotide and protein levels using available databases. Software were used to identify PKS and NRPS domains and predict identity and function. Confirmation of sequence for selected gene sequences was accomplished using PCR. Thirteen PKS, 5 NRPS, and 4 PKS-NRPS hybrids were identified and characterized with functions including swainsonine and melanin biosynthesis. Phylogenetic relationships among closest amino acid matches with Alternaria spp. were identified for seven highly conserved PKS and NRPS, including melanin synthesis. Three PKS and NRPS were most closely related to other fungi within the Pleosporaceae family, while five PKS and PKS-NRPS were closely related to fungi in the Pleosporales order. However, seven PKS and PKS-NRPS showed no identity with fungi in the Pleosporales or the class Dothideomycetes, suggesting a different evolutionary origin for those genes.
Curtoviruses are transmitted by the beet leafhopper Circulifer tenellus, in a circulative (nonpropagative) manner. Curtoviruses are phloem-limited and are acquired by the vector during feeding. Sap-feeding insects harbor endosymbionts which can help provide essential nutrients required for the insects' survival. Candidatus Sulcia muelleri is an endosymbiont present in the beet leafhopper identified during this study. A housekeeping gene, groel, was identified from the endosymbiont. The groel gene sequence from this strain of Ca. S. muelleri differs from all other strains published in NCBI, suggesting the presence of a new strain, which was named S. muelleri beet leafhopper (SMBLH). A GroEL-homolog protein produced from groel was found in different vectors with circulative transmission. Analysis of nucleotide and translated sequences, using alignment, phylogenetic trees, and predicted secondary and tertiary structures showed that SMBLH GroHp has similarities to Escherichia coli GroEL and the GroEL-homolog proteins from Hamiltonella and Buchnera, endosymbionts of whiteflies and aphids, respectively. GroHp and GroEL were expressed as fusion proteins. Electron microscopy analyses indicate that purified expressed GroHp and GroEL proteins demonstrate correct folding.
Background: The marine environment hosts the vast majority of living species and marine microbes that produce natural products with great potential in providing lead compounds for drug development. With over 70% of Earth’s surface covered in water and the high interaction rate associated with liquid environments, this has resulted in many marine natural product discoveries. Our improved understanding of the biosynthesis of these molecules, encoded by gene clusters, along with increased genomic information will aid us in uncovering even more novel compounds. Results: We introduce MariClus (https://www.mariclus.com), an online user-friendly platform for mining and visualizing marine gene clusters. The first version contains information on clusters and the predicted molecules for over 500 marine-related prokaryotes. The user-friendly interface allows scientists to easily search by species, cluster type or molecule and visualize the information in table format or graphical representation. Conclusions: This new online portal simplifies the exploration and comparison of gene clusters in marine species for scientists and assists in characterizing the bioactive molecules they produce. MariClus integrates data from public sources, like GenBank, MIBiG and PubChem, with genome mining results from antiSMASH. This allows users to access and analyze various aspects of marine natural product biosynthesis and diversity.
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