New Deoxyenhygrolides from Plesiocystis pacifica Provide Insights into Butenolide Core Biosynthesis

Marine myxobacteria present a virtually unexploited reservoir for the discovery of natural products with diverse biological functions and novel chemical scaffolds. We report here the isolation and structure elucidation of eight new deoxyenhygrolides (1–8) from the marine myxobacterium Plesiocystis pacifica DSM 14875T. The herein described deoxyenhygrolides C–J (1–8) feature a butenolide core with an ethyl residue at C-3 of the γ-lactone in contrast to the previously described derivatives, deoxyenhygrolides A and B, which feature an isobutyl residue at this position. The butenolide core is 2,4-substituted with a benzyl (1, 2 and 7), benzoyl (3 and 4) or benzyl alcohol (5, 6 and 8) moiety in the 2-position and a benzylidene (1–6) or benzylic hemiketal (7 and 8) in the 4-position. The description of these new deoxyenhygrolide derivatives, alongside genomic in silico investigation regarding putative biosynthetic genes, provides some new puzzle pieces on how this natural product class might be formed by marine myxobacteria.

Methylation Profiling of Biosynthetic Genes Reveals The Role of D-Galacturonic Acid Reductase In Ascorbic Acid Accumulation In Tomato Fruit

Ascorbic acid (AsA) is an important nutrient component contributing to major flavor value of tomato fruit and human health. Although transcription regulation of AsA biosynthetic genes have been well demonstrated, epigenetic modification underlying AsA accumulation remains unclear. In this study, we exposed immature tomato fruits to a methyltransferase inhibitor (5-azacytidine) and detected the impacts on AsA accumulation. Inhibition of DNA methylation enhanced AsA accumulation in tomato leaves and fruits. We further isolated a AsA biosynthetic gene, SlGalUR5, which encodes a D-galacturonic acid reductase. SlGalUR5 showed reduced DNA methylation levels and higher transcription levels in Slmet1 mutant while have converse pattern in Sldml2 mutant. 5-azacytidine treatment significantly decreased DNA methylation levels of SlGalUR5 in fruits. Conversely, transcription profiles of SlGalUR5 and enzyme activity of GalUR were enhanced in 5-azacytidine–treated fruits. Our finding revealed a new insight into epigenome modification of SlGalUR5 involved in ascorbic acid accumulation and provide a potential means of increasing AsA levels for tomato breeding.

Expression of Ethylene Biosynthetic Genes in the Gynoecium and Receptacle Associated With Sepal Abscission During Senescence in Delphinium Grandiflorum

Delphinium flowers are highly sensitive to ethylene and its sepals abscise during senescence, which is associated with increases in 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) and ACC oxidase (ACO) activities and ethylene production in gynoecium and receptacle. Three ACS genes (DgACS1, DgACS2, and DgACS3) and three ACO genes (DgACO1, DgACO2, and DgACO3) were cloned from Delphinium grandiflorum cv. Super Grand Blue. To investigate the contribution of these genes to ethylene production, their expression was analyzed in these genes in the gynoecium and receptacle during natural senescence and following ethylene exposure and pollination. Ethylene production in the gynoecium and receptacle increased during natural flower senescence. The transcript levels of the ACS and ACO genes in these organs, excluding DgACS2 in the receptacle, increased during senescence. Exposure to ethylene accelerated sepal abscission and more strongly increased ethylene production in the receptacle than in the gynoecium. DgACS1 transcript levels in the gynoecium and DgACS2 and DgACO3 transcript levels in the receptacle were increased by ethylene exposure. Pollination accelerated sepal abscission and increased ethylene production in the gynoecium and receptacle. Pollination slightly affected ACS and ACO transcript levels in the gynoecium, whereas DgACO3 transcript level in the receptacle were markedly increased. These results reveal that ACS and ACO gene expression is differently regulated in the gynoecium and receptacle, and some of these genes are more strongly upregulated by ethylene exposure and pollination in the receptacle than in the gynoecium, suggesting the significance of the receptacle to sepal abscission.

Carotenoid Biosynthetic Genes in Cabbage: Genome-Wide Identification, Evolution, and Expression Analysis

Carotenoids are natural functional pigments produced by plants and microorganisms and play essential roles in human health. Cabbage (Brassica oleracea L. var. capitata L.) is an economically important vegetable in terms of production and consumption. It is highly nutritious and contains β-carotene, lutein, and other antioxidant carotenoids. Here, we systematically analyzed carotenoid biosynthetic genes (CBGs) on the whole genome to understand the carotenoid biosynthetic pathway in cabbage. In total, 62 CBGs were identified in the cabbage genome, which are orthologs of 47 CBGs in Arabidopsis thaliana. Out of the 62 CBGs, 46 genes in cabbage were mapped to nine chromosomes. Evolutionary analysis of carotenoid biosynthetic orthologous gene pairs among B. oleracea, B. rapa, and A. thaliana revealed that orthologous genes of B. oleracea underwent a negative selection similar to that of B. rapa. Expression analysis of the CBGs showed functional differentiation of orthologous gene copies in B. oleracea and B. rapa. Exogenous phytohormone treatment suggested that ETH, ABA, and MeJA can promote some important CBGs expression in cabbage. Phylogenetic analysis showed that BoPSYs exhibit high conservatism. Subcellular localization analysis indicated that BoPSYs are located in the chloroplast. This study is the first to study carotenoid biosynthesis genes in cabbage and provides a basis for further research on carotenoid metabolic mechanisms in cabbage.

Secondary metabolite biosynthetic diversity in Arctic Ocean metagenomes

Polyketide synthases (PKSs) and non-ribosomal peptide synthetases (NRPSs) are mega enzymes responsible for the biosynthesis of a large fraction of natural products (NPs). Molecular markers for biosynthetic genes, such as the ketosynthase (KS) domain of PKSs, have been used to assess the diversity and distribution of biosynthetic genes in complex microbial communities. More recently, metagenomic studies have complemented and enhanced this approach by allowing the recovery of complete biosynthetic gene clusters (BGCs) from environmental DNA. In this study, the distribution and diversity of biosynthetic genes and clusters from Arctic Ocean samples (NICE-2015 expedition), was assessed using PCR-based strategies coupled with high-throughput sequencing and metagenomic analysis. In total, 149 KS domain OTU sequences were recovered, 36 % of which could not be assigned to any known BGC. In addition, 74 bacterial metagenome-assembled genomes were recovered, from which 179 BGCs were extracted. A network analysis identified potential new NP families, including non-ribosomal peptides and polyketides. Complete or near-complete BGCs were recovered, which will enable future heterologous expression efforts to uncover the respective NPs. Our study represents the first report of biosynthetic diversity assessed for Arctic Ocean metagenomes and highlights the potential of Arctic Ocean planktonic microbiomes for the discovery of novel secondary metabolites. The strategy employed in this study will enable future bioprospection, by identifying promising samples for bacterial isolation efforts, while providing also full-length BGCs for heterologous expression.

Whole-Genome Identification and Comparative Expression Analysis of Anthocyanin Biosynthetic Genes in Brassica napus

Anthocyanins contribute to most colors of plants and play protective roles in response to abiotic stresses. Brassica napus is widely cultivated worldwide as both an oilseed and a vegetable. However, only several high anthocyanin-containing cultivars have been reported, and the mechanisms of anthocyanin accumulation have not been well-elucidated in B. napus. Here, the phenotype, comparative whole-genome identification, and gene expression analysis were performed to investigate the dynamic change of the anthocyanin content and the gene expression patterns of anthocyanin biosynthetic genes (ABGs) in B. napus. A total of 152 ABGs were identified in the B. napus reference genome. To screen out the critical genes involved in anthocyanin biosynthesis and accumulation, the RNA-seq of young leaves of two B. napus lines with purple leaves (PL) or green leaves (GL), and their F1 progeny at 41, 91, and 101 days were performed to identify the differentially expressed genes. The comparative expression analysis of these ABGs indicated that the upregulation of TT8 together with its target genes (such as DFR, ANS, UFGT, and TT19) might promote the anthocyanin accumulation in PL at the early developmental stage (41–91 days). While the downregulation of those ABGs and anthocyanin degradation at the late developmental stage (91–101 days) might result in the decrease in anthocyanin accumulation. Our results would enhance the understanding of the regulatory network of anthocyanin dynamic accumulation in B. napus.

β-Cyclocitral, a Master Regulator of Multiple Stress-Responsive Genes in Solanum lycopersicum L. Plants

β-cyclocitral (βCC), a major apocarotenoid of β-carotene, enhances plants’ defense against environmental stresses. However, the knowledge of βCC’s involvement in the complex stress-signaling network is limited. Here we demonstrate how βCC reprograms the transcriptional responses that enable Solanum lycopersicum L. (tomato) plants to endure a plethora of environmental stresses. Comparative transcriptome analysis of control and βCC-treated tomato plants was done by generating RNA sequences in the BGISEQ-500 platform. The trimmed sequences were mapped on the tomato reference genome that identifies 211 protein-coding differentially expressed genes. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis and their enrichment uncovered that only upregulated genes are attributed to the stress response. Moreover, 80% of the upregulated genes are functionally related to abiotic and biotic stresses. Co-functional analysis of stress-responsive genes revealed a network of 18 genes that code for heat shock proteins, transcription factors (TFs), and calcium-binding proteins. The upregulation of jasmonic acid (JA)-dependent TFs (MYC2, MYB44, ERFs) but not the JA biosynthetic genes is surprising. However, the upregulation of DREB3, an abscisic acid (ABA)-independent TF, validates the unaltered expression of ABA biosynthetic genes. We conclude that βCC treatment upregulates multiple stress-responsive genes without eliciting JA and ABA biosynthesis.