Plant growth regulator pteridic acids Streptomyces producers
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Agilent Quadrupole Time-of-Flight LC/MS
ORGANISM(S): Streptomyces (ncbitaxon:1883)
SUBMITTER: 
Ling Ding
PROVIDER: MSV000090742 | MassIVE | Fri Nov 18 11:41:00 GMT 2022
REPOSITORIES: MassIVE
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Project description:Soil microbiota can confer fitness advantages to plants and increase crop resilience to drought and other abiotic stressors. However, there is little evidence on the mechanisms correlating a microbial trait with plant abiotic stress tolerance. Here, we report that a class of Streptomyces effectively alleviates the drought and salinity stress by producing new spiroketal polyketide pteridic acid H (1) and its isomer F (2). The bifunctional pteridic acid biosynthetic gene cluster (pta), which is also responsible for the biosynthesis of the known antimicrobial elaiophylin, was confirmed by bioinformatic analysis and in vivo CRISPR base editing. Pteridic acids H and F exhibited profound effects in promoting root growth in Arabidopsis at a concentration of 0.5 ng mL-1 (1.3 nM) under abiotic stress, indicating they are a new class of plant stress regulators. Phylogenetic and geographical distribution analysis revealed that the pta was mainly disseminated by vertical transmission and occasional horizontal gene transfer and is widely distributed in numerous Streptomyces in different environments. This discovery provides a new perspective for understanding plant-Streptomyces interactions and provides a novel, promising approach for utilising beneficial Streptomyces and their secondary metabolites in agriculture to mitigate the detrimental effects of climate change.
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Project description:Streptomyces rimosus is an industrial streptomycete, best known as a producer of oxytetracycline, one of the most widely used antibiotics. Despite the significant contribution of Streptomyces species to the pharmaceutical industry, most omics analyses have only been conducted on the model organism, Streptomyces coelicolor. In recent years, protein phosphorylation on serine, threonine and tyrosine (Ser/Thr/Tyr) has been shown to play a crucial role in the regulation of numerous cellular processes, including metabolic changes leading to antibiotic production and morphological changes. In this study, we performed a comprehensive quantitative (phospho)proteomic analysis during the growth of S. rimosus under conditions of oxytetracycline production and pellet fragmentation. LC-MS/MS analysis combined with phosphopeptide enrichment detected a total of 3,725 proteins, corresponding to 45.6% of the proteome and 417 phosphorylation sites from 230 phosphoproteins. Significant changes in abundance during three distinct growth phases were determined for 494 proteins and 98 phosphorylation sites. Functional analysis revealed changes in phosphorylation events of proteins involved in important cellular processes, including regulatory mechanisms, primary and secondary metabolism, cell division and stress response. About 80% of the phosphoproteins detected during submerged growth of S. rimosus have not yet been reported in streptomycetes, and 55 phosphoproteins were not reported in any prokaryote studied so far. This enabled the creation of a unique resource that provides novel insights into the dynamics of (phospho)proteins and reveals many potential regulatory events during antibiotic production in liquid culture of an industrially-important bacterium.
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Project description:Temperature passively affects many biological processes. It is therefore challenging to study dedicated temperature signalling pathways orchestrating plant thermomorphogenesis; a suite of elongation growth-based adaptations that enhance leaf cooling capacity. Following a chemical genetics approach, we screened a chemical library for compounds that restored abolished hypocotyl elongation in the pif4-2 deficient mutant background in the model plant Arabidopsis thaliana. The small aromatic compound ‘Heatin' (N'-[(2-hydroxy-1-naphthyl)methylene]-2-(1-naphthylamino)propanohydrazide), with 1-aminomethyl-2-naphthol, as minimal active moiety, was isolated as potent enhancer of elongation growth. Here we assessed the transcriptomic changes induced by the compound, by supplementing the MS-agar growth medium with Heatin compared to DMSO solvent, in whole seedlings of the Col-0 wild type background in control (22oC) and high ambient temperature conditions (27oC).
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Project description:In actinomycetes, antibiotic production is often associated with a morpho-physiological differentiation program that is regulated by complex molecular and metabolic networks. Many aspects of these regulatory circuits have been already elucidated and many others still deserve further investigations. In this regard, the possible role of many small open-reading frames(smORFs) in actinomycete morpho-physiological differentiation is still elusive. In Streptomyces coelicolor, inactivation of the smORF trpM (SCO2038) - whose product modulates L-tryptophan metabolism - impairs production of antibiotics and morphological differentiation. Indeed, it was demonstrated that TrpM is able to interact with PepA (SCO2179), a putative cytosol aminopeptidase playing a key role in antibiotic production and sporulation. In this work, a S. coelicolor trpM knock-in (Sco-trpMKI) mutant strain was generated by cloning trpM into overexpressing vector to further investigate the role of trpM in actinomycete growth and morpho-physiological differentiation. Results highlighted that trpM: i) stimulates growth and actinorhodin production; ii) decreases calcium-dependent antibiotic production; iii) has no effect on undecylprodigiosin production. Metabolic pathways influenced by trpM knock-in were investigated by combining two-difference in gel electrophoresis/nanoliquid chromatography coupled to electrospray linear ion trap tandem mass spectrometry (2D-DIGE/nanoLC-ESI-LIT- MS/MS) and by LC-ESI-MS/MS procedures, respectively. These analyses demonstrated that over-expression of trpM causes an over-representation of factors involved in protein synthesis and nucleotide metabolism as well as a down-representation of proteins involved in central carbon and amino acid metabolism. At the metabolic level, this corresponded to a differential accumulation pattern of different amino acids - including aromatic ones but tryptophan – and central carbon intermediates. PepA was also down-represented in Sco-trpMKI. The latter was produced as recombinant His- tagged protein and was originally proven having the predicted aminopeptidase activity. Altogether, these results highlight the stimulatory effect of trpM in S. coelicolor growth and actinorhodin biosynthesis, which are elicited through the modulation of various metabolic pathways and PepA representation, further confirming the complexity of regulatory networks that control antibiotic production in actinomycetes.
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