Plant Symbionts Research Articles

Preferential nitrogen and carbon exchange dynamics in Mucoromycotina “fine root endophyte”-plant symbiosis

Mucoromycotina "fine root endophyte" (MFRE) fungi are an understudied group of plant symbionts that regularly co-occur with arbuscular mycorrhizal fungi. The functional significance of MFRE in plant nutrition remains underexplored, particularly their role in plant nitrogen (N)assimilation from the variety of sources typically found in soils. Using four 15N-labeled N sources to track N transfer between MFRE and Plantago lanceolata, applied singly and in tandem, we investigated N source discrimination, preference, and transfer to host plants by MFRE. We traced movement of 14C from plants to MFRE to determine the impact of N source type on plant carbon (C)allocation to MFRE. We found that MFRE preferentially transferred N derived from glycine and ammonium to plant hosts over that derived from nitrate and urea, regardless of other N sources present. MFRE mycelium supplied with glycine and ammonium contained more plant-derived carbon than those supplied with other N sources. We show that the MFRE directly assimilates and metabolizes organic compounds, retaining C to meet its own metabolic requirements and transferring N to plant hosts. Our findings highlight diversity in the function of endomycorrhizal associations, with potentially profound implications for our understanding of the physiology and ecology of plant-fungal symbioses.

Transcriptomic responses of Solanum tuberosum cv. Pirol to arbuscular mycorrhiza and potato virus Y (PVY) infection

Arbuscular mycorrhizal fungi (AMF) serve as both plant symbionts and allies in resisting pathogens and environmental stresses. Mycorrhizal colonization of plant roots can influence the outcomes of plant-pathogen interactions by enhancing specific host defense mechanisms. The transcriptional responses induced by AMF in virus-infected plants remain largely unexplored. In the presented study, we employed a comprehensive transcriptomic approach and qPCR to investigate the molecular determinants underlying the interaction between AMF and potato virus Y (PVY) in Solanum tuberosum L. Our primary goal was to identify the symbiosis- and defense-related determinants activated in mycorrhizal potatoes facing PVY. Through a comparative analysis of mRNA transcriptomes in experimental treatments comprising healthy and PVY-infected potatoes colonized by two AMF species, Rhizophagus regularis or Funneliformis mosseae, we unveiled the overexpression of genes associated with mycorrhiza, including nutrient exchange, lipid transfer, and cell wall remodeling. Furthermore, we identified several differentially expressed genes upregulated in all mycorrhizal treatments that encoded pathogenesis-related proteins involved in plant immune responses, thus verifying the bioprotective role of AMF. We investigated the relationship between mycorrhiza levels and PVY levels in potato leaves and roots. We found accumulation of the virus in the leaves of mycorrhizal plants, but our studies additionally showed a reduced PVY content in potato roots colonized by AMF, which has not been previously demonstrated. Furthermore, we observed that a virus-dependent reduction in nutrient exchange could occur in mycorrhizal roots in the presence of PVY. These findings provide an insights into the interplay between virus and AMF.

Transcriptomic Response of Rhizobium leguminosarum to Acidic Stress and Nutrient Limitation Is Versatile and Substantially Influenced by Extrachromosomal Gene Pool.

Multipartite genomes are thought to confer evolutionary advantages to bacteria by providing greater metabolic flexibility in fluctuating environments and enabling rapid adaptation to new ecological niches and stress conditions. This genome architecture is commonly found in plant symbionts, including nitrogen-fixing rhizobia, such as Rhizobium leguminosarum bv. trifolii TA1 (RtTA1), whose genome comprises a chromosome and four extrachromosomal replicons (ECRs). In this study, the transcriptomic responses of RtTA1 to partial nutrient limitation and low acidic pH were analyzed using high-throughput RNA sequencing. RtTA1 growth under these conditions resulted in the differential expression of 1035 to 1700 genes (DEGs), which were assigned to functional categories primarily related to amino acid and carbohydrate metabolism, ribosome and cell envelope biogenesis, signal transduction, and transcription. These results highlight the complexity of the bacterial response to stress. Notably, the distribution of DEGs among the replicons indicated that ECRs played a significant role in the stress response. The transcriptomic data align with the Rhizobium pangenome analysis, which revealed an over-representation of functional categories related to transport, metabolism, and regulatory functions on ECRs. These findings confirm that ECRs contribute substantially to the ability of rhizobia to adapt to challenging environmental conditions.

Molecular characterization of plant growth-promoting Trichoderma from Saudi Arabia.

Fungi in the genus Trichoderma are widespread in the environment, mainly in soils. They are used in agriculture because of their mycoparasitic potential; Trichoderma have the ability to increase plant health and provide protection against phytopathogens, making them desirable plant symbionts. We isolated, identified, and characterized Trichoderma from different regions of Saudi Arabia and evaluated the ability of Trichoderma to promote plant growth. Morphological and molecular characterization, along with phylogenetic studies, were utilized to differentiate between Trichoderma species isolated from soil samples in the Abha and Riyadh regions, Saudi Arabia. Then, plant growth-promoting traits of the isolated Trichoderma species were assessed. Eight Trichoderma isolates were characterized via morphological and molecular analysis; six (Trichoderma koningiopsis, Trichoderma lixii, Trichoderma koningii, Trichoderma harzianum, Trichoderma brevicompactum, and Trichoderma velutinum) were from Abha and two (T. lixii and T. harzianum) were from Riyadh. The isolated Trichoderma strains belonged to three different clades (Clade 1: Harzianum, Clade 2: Brevicompactum, and Clade 3: Viride). The Trichoderma isolates varied in plant growth-promoting traits. Seeds treated with most isolates exhibited a high percentage of germination, except seeds treated with the T3-T. koningii isolate. 100% germination was reported for seeds treated with the T4-T. harzianum and T6-T. brevicompactum isolates, while seeds treated with the T1-T. koniniopsis and T5-T. lixii isolates showed 91.1% and 90.9% germination, respectively. Seeds treated with the T8-T. velutinum, T2-T. lixii, and T7-T. harzianum isolates had germination rates of 84.1%, 82.2%, and 72.7%, respectively. The Trichoderma isolate T5-T. lixii stimulated tomato plant growth the most, followed by T7-T. harzianum, T8-T. velutinum, T4-T. harzianum, T1-T. koniniopsis, T2-T. lixii, and T6-T. brevicompactum; the least effective was T3-T. koningii. A maximum fresh weight of 669.33mg was observed for the T5-T. lixii-treated plants. The Abha region had a higher diversity of Trichoderma species than the Riyadh region, and most isolated Trichoderma spp. promoted tomato growth.

Developing endophytic Penicillium oxalicum as a source of lignocellulolytic enzymes for enhanced hydrolysis of biorefinery relevant pretreated rice straw.

Endophytic fungi, as plant symbionts, produce an elaborate array of enzymes for efficient disintegration of lignocellulosic biomass into constituent monomeric sugars, making them novel source of lignocellulolytic CAZymes with immense potential in future biorefineries. The present study reports lignocellulolytic enzymes production potential of an endophytic halotolerant Penicillium oxalicum strain isolated from Citrus limon, under submerged and solid-state fermentation (SmF & SSF, respectively), in the presence and absence of salt (1M NaCl). The comparative QTOF-LC/MS-based exoproteome analysis of the culture extracts unveiled differential expression of CAZymes, with the higher abundance of GH6 and GH7 family cellobiohydrolase in the presence of 1M salt. The strain improvement program, employing cyclic mutagenesis and diploidization, was utilized to develop hyper-cellulase producing mutant strains of P. oxalicum. The enzyme production of the developed strain (POx-M35) was further enhanced through statistical optimization of the culture conditions utilizing glucose mix disaccharides (GMDs) as an inducer. This optimization process resulted in the lignocellulolytic cocktail that contained high titers (U/mL) of endoglucanase (EG) (146.16), cellobiohydrolase (CBHI) (6.99), β-glucosidase (β-G) (26.21), xylanase (336.05) and FPase (2.02 U/mL), which were 5.47-, 5.54-, 8.55-, 4.96-, and 4.39-fold higher when compared to the enzyme titers obtained in wild HP1, respectively. Furthermore, the lignocellulolytic cocktails designed by blending secretome produced by mutant POx-M35 with xylanases (GH10 and GH11) derived from Malbranchea cinnamomea resulted in efficient hydrolysis of unwashed acid pretreated (UWAP) rice straw slurry and mild alkali deacetylated (MAD) rice straw. This study underscores the potential of bioprospecting novel fungus and developing an improved strain for optimized production and constitution of lignocellulolytic cocktails that can be an important determinant in advancing biomass conversion technologies.

Engineering a Sinorhizobium meliloti Chassis with Monopartite, Single Replicon Genome Configuration.

Multipartite bacterial genomes pose challenges for genome engineering and the establishment of additional replicons. We simplified the tripartite genome structure (3.65 Mbp chromosome, 1.35 Mbp megaplasmid pSymA, 1.68 Mbp chromid pSymB) of the nitrogen-fixing plant symbiont Sinorhizobium meliloti. Strains with bi- and monopartite genome configurations were generated by targeted replicon fusions. Our design preserved key genomic features such as replichore ratios, GC skew, KOPS, and coding sequence distribution. Under standard culture conditions, the growth rates of these strains and the wild type were nearly comparable, and the ability for symbiotic nitrogen fixation was maintained. Spatiotemporal replicon organization and segregation were maintained in the triple replicon fusion strain. Deletion of the replication initiator-encoding genes, including the oriVs of pSymA and pSymB from this strain, resulted in a monopartite genome with oriC as the sole origin of replication, a strongly unbalanced replichore ratio, slow growth, aberrant cellular localization of oriC, and deficiency in symbiosis. Suppressor mutation R436H in the cell cycle histidine kinase CckA and a 3.2 Mbp inversion, both individually, largely restored growth, but only the genomic rearrangement recovered the symbiotic capacity. These strains will facilitate the integration of secondary replicons in S. meliloti and thus be useful for genome engineering applications, such as generating hybrid genomes.

Arbuscular mycorrhizal diversity increases across a plant productivity gradient driven by soil nitrogen availability.

Arbuscular mycorrhizal fungi (AMF) are widespread obligate symbionts of plants. This dynamic symbiosis plays a large role in successful plant performance, given that AMF help to ameliorate plant responses to abiotic and biotic stressors. Although the importance of this symbiosis is clear, less is known about what may be driving this symbiosis, the plant's need for nutrients or the excess of plant photosynthate being transferred to the AMF, information critical to assess the functionality of this relationship. Characterizing the AMF community along a natural plant productivity gradient is a first step in understanding how this symbiosis may vary across the landscape. We surveyed the AMF community diversity at 12 sites along a plant productivity gradient driven by soil nitrogen availability. We found that AMF diversity in soil environmental DNA significantly increased along with the growth of the host plants Acer rubrum and A. saccharum., a widespread tree genus. These increases also coincided with a natural soil inorganic N availability gradient. We hypothesize photosynthate from the increased tree growth is being allocated to the belowground AMF community, leading to an increase in diversity. These findings contribute to understanding this complex symbiosis through the lens of AMF turnover and suggest that a more diverse AMF community is associated with increased host-plant performance.

Evaluation of the Colonization of Plants from Five Quercus Taxa Native to Greece by Tuber aestivum (Ascomycota, Pezizales).

Fungi of the genus Tuber are famous for their hypogeous ascomata (truffles), many of which possess noteworthy organoleptic properties. T. aestivum shows a wide geographic distribution, has many plant symbionts and is well adapted to various climatic conditions. In this study, five Quercus taxa native to Greece (i.e., Q. coccifera, Q. ilex, Q. ithaburensis subsp. macrolepis, Q. pubescens and Q. trojana subsp. trojana) were inoculated with spore suspensions obtained from a single ascoma of T. aestivum. The fungal colonization of oak roots was evaluated at three, seven and 12 months after inoculation; the respective colonization rates for each time period were as follows: low to medium (17-41%) for Q. pubescens, Q. ithaburensis subsp. macrolepis and Q. trojana subsp. trojana, medium to relatively high (58-80%) for Q. ithaburensis subsp. macrolepis, Q. ilex, Q. pubescens and Q. trojana subsp. trojana, and medium to high (45-87%) for all oak species examined. Positive correlations were assessed between the number of colonized root tips and the total root tips number, but no significant differences were detected between the inoculated plants and the respective control as regards plant growth. The ectomycorrhizae formed by T. aestivum with Q. ithaburensis subsp. macrolepis and Q. trojana subsp. trojana are described for the first time. The outcome of the study evidences the feasibility of generating the seedlings of various indigenous oak species (covering a large range of diverse habitats) successfully inoculated with autochthonous truffles to be readily used for cultivation purposes.

Compatibility study of Trichoderma sp. with Chemical Fungicides Commonly Used by Nepalese Farmers, Under In-Vitro Condition

The excessive use of synthetic fungicides has led to the emergence offungicide-resistant strains of pathogens, raising concerns about human health andenvironmental impact. Trichoderma spp., an endophytic and versatileopportunistic plant symbiont, has recently gained popularity as a biocontrolagent. Integrated use of Trichoderma with compatible fungicides gives betterdisease management and causes less harm to the environment in the long run.This study aimed to evaluate the compatibility of six fungicides commonly used byNepalese farmers at concentrations of 250, 500, and 1000 ppm with Trichodermasp. using the poisoned food technique in-vitro. The experiment was conducted ina completely randomized design with four replications for each treatment, anddata were taken at 24, 36, 48, 60, and 72 hours after incubation. Among thefungicides tested, Mancozeb was compatible at all concentrations from the startand exhibited the highest compatibility at 72 hours, with a growth inhibitionpercent of 0.39%. Metalaxyl+Mancozeb at 250 and 500 ppm, and Copperoxychloride at 250 ppm showed good compatibility. However, Carbendazim,Hexaconazole, and Carbendazim+Mancozeb resulted in 100% growth inhibition ofTrichoderma sp. throughout the experiment. While all treatments significantlyreduced mycelial diameter, Mancozeb and Metalaxyl+Mancozeb showed agradual decrease in growth inhibition percent over time, indicating increasedcompatibility. In contrast, Copper oxychloride demonstrated variable growthinhibition percent over time. So, the fungicides Mancozeb, Metalaxyl+Mancozeband Copper oxychloride at compatible concentrations can be used withTrichoderma sp. in integrated disease management to control soil and seed-bornepathogens.

In vitro antifungal effect of endophytic fungi of creole cucurbits of Yucatan

Endophytic fungi are plant symbiont microorganisms, diverse and abundant in tropical regions, with great potential as biocontrol agents of agricultural fungal diseases. The objective of the present work was to evaluate in vitro antifunfal activity of endophytic fungi associated with creole cultivars of Cucumis sativus and Lagenaria siceraria from Yucatán. The endophytic strains were isolated and identified from different organs, to be subjected to dual confrontations and antibiotic tests. Twenty species with antifungal effects against Corynespora cassiicola, Colletotrichum truncatum and Fusarium equiseti were identified, obtaining inhibition ranges of 17.72–100%, 16.51–91.55% and 4.64–91.89%, respectively. Aspergillus flavus, Lasiodiplodia pseudotheobromae and Trichoderma longibrachiatum exhibited the highest percentages of inhibition against the three phytopathogens. Likewise, these three species of endophytes demonstrated an antibiotic effect in their extracts, mainly inhibiting the growth of F. equiseti by up to 46.46%. The L. pseudotheobromae strain has characteristics that give it potential as an antifungal agent, so its applications and growth conditions should continue to be explored. This work represents a first effort to understand the diversity of endophytic fungi in native crops in the region.

Evolutionary history of arbuscular mycorrhizal fungi and genomic signatures of obligate symbiosis

BackgroundThe colonization of land and the diversification of terrestrial plants is intimately linked to the evolutionary history of their symbiotic fungal partners. Extant representatives of these fungal lineages include mutualistic plant symbionts, the arbuscular mycorrhizal (AM) fungi in Glomeromycota and fine root endophytes in Endogonales (Mucoromycota), as well as fungi with saprotrophic, pathogenic and endophytic lifestyles. These fungal groups separate into three monophyletic lineages but their evolutionary relationships remain enigmatic confounding ancestral reconstructions. Their taxonomic ranks are currently fluid.ResultsIn this study, we recognize these three monophyletic linages as phyla, and use a balanced taxon sampling and broad taxonomic representation for phylogenomic analysis that rejects a hard polytomy and resolves Glomeromycota as sister to a clade composed of Mucoromycota and Mortierellomycota. Low copy numbers of genes associated with plant cell wall degradation could not be assigned to the transition to a plant symbiotic lifestyle but appears to be an ancestral phylogenetic signal. Both plant symbiotic lineages, Glomeromycota and Endogonales, lack numerous thiamine metabolism genes but the lack of fatty acid synthesis genes is specific to AM fungi. Many genes previously thought to be missing specifically in Glomeromycota are either missing in all analyzed phyla, or in some cases, are actually present in some of the analyzed AM fungal lineages, e.g. the high affinity phosphorus transporter Pho89.ConclusionBased on a broad taxon sampling of fungal genomes we present a well-supported phylogeny for AM fungi and their sister lineages. We show that among these lineages, two independent evolutionary transitions to mutualistic plant symbiosis happened in a genomic background profoundly different from that known from the emergence of ectomycorrhizal fungi in Dikarya. These results call for further reevaluation of genomic signatures associated with plant symbiosis.

Four New Species and One New Record of Thelephora from China.

Species of the genus Thelephora (Thelephorales, Thelephoraceae) are ectomycorrhizal symbionts of coniferous and broad-leaved plants, and some of them are well-known edible mushrooms, making it an exceptionally important group ecologically and economically. However, the diversity of the species from China has not been fully elucidated. In this study, we conducted a phylogenetic analysis based on the internal transcribed spacer (ITS) regions, using Maximum Likelihood and Bayesian analyses, along with morphological observations of this genus. Four new species from China are proposed, viz., T. dactyliophora, T. lacunosa, T. petaloides, and T. pinnatifida. In addition, T. sikkimensis originally described from India is reported for the first time from China. Thelephora dactyliophora, T. pinnatifida, and T. sikkimensis are distributed in subtropical forests and mainly associated with plants of the families Fagaceae and Pinaceae. Thelephora lacunosa and T. petaloides are distributed in tropical to subtropical forests. Thelephora lacunosa is mainly associated with plants of the families Fagaceae and Pinaceae, while T. petaloides is mainly associated with plants of the family Fagaceae. Line drawings of microstructures, color pictures of fresh basidiomes, and detailed descriptions of these five species are provided.

Cytochrome P450 CYP736A12 is crucial for Trichoderma asperellum-induced alleviation of phoxim phytotoxicity and reduction of pesticide residue in tomato roots

Trichoderma can enhance the metabolism of organophosphate pesticides in plants, but the mechanism is unclear. Here, we performed high-throughput transcriptome sequencing of roots upon Trichoderma asperellum (TM) inoculation and phoxim (P) application in tomato (Solanum lycopersicum L.). A total of 4059 differentially expressed genes (DEGs) were obtained, including 2110 up-regulated and 1949 down-regulated DEGs in P vs TM+P. COG and KOG analysis indicated that DEGs were mainly enriched in signal transduction mechanisms. We then focused on the pesticide detoxification pathway and screened out cytochrome P450 CYP736A12 as a putative gene for functional analysis. We suppressed the expression of CYP736A12 in tomato plants by virus-induced gene silencing and analyzed tissue-specific phoxim residues, oxidative stress markers, glutathione pool, GST activity and related gene expression. Silencing CYP736A12 significantly increased phoxim residue and induced oxidative stress in tomato plants, by attenuating the TM-induced increased activity of antioxidant and detoxification enzymes, redox homeostasis and transcripts of detoxification genes including CYP724B2, GSH1, GSH2, GR, GPX, GST1, GST2, GST3, and ABC. The study revealed a critical mechanism by which TM promotes the metabolism of phoxim in tomato roots, which can be useful for further understanding the Trichoderma-induced xenobiotic detoxification and improving food safety.

The impacts of host association and perturbation on symbiont fitness

Symbiosis can benefit hosts in numerous ways, but less is known about whether interactions with hosts benefit symbionts—the smaller species in the relationship. To determine the fitness impact of host association on symbionts in likely mutualisms, we conducted a meta-analysis across 91 unique host-symbiont pairings under a range of spatial and temporal contexts. Specifically, we assess the consequences to symbiont fitness when in and out of symbiosis, as well as when the symbiosis is under suboptimal or varying environments and biological conditions (e.g., host age). We find that some intracellular symbionts associated with protists tend to have greater fitness when the symbiosis is under stressful conditions. Symbionts of plants and animals did not exhibit this trend, suggesting that symbionts of multicellular hosts are more robust to perturbations. Symbiont fitness also generally increased with host age. Lastly, we show that symbionts able to proliferate in- and outside host cells exhibit greater fitness than those found exclusively inside or outside cells. The ability to grow in multiple locations may thus help symbionts thrive. We discuss these fitness patterns in light of host-driven factors, whereby hosts exert influence over symbionts to suit their own needs.

Strain-specific evolution and host-specific regulation of transposable elements in the model plant symbiont Rhizophagus irregularis.

Transposable elements (TEs) are repetitive DNA that can create genome structure and regulation variability. The genome of Rhizophagus irregularis, a widely studied arbuscular mycorrhizal fungus (AMF), comprises ∼50% repetitive sequences that include TEs. Despite their abundance, two-thirds of TEs remain unclassified, and their regulation among AMF life stages remains unknown. Here, we aimed to improve our understanding of TE diversity and regulation in this model species by curating repeat datasets obtained from chromosome-level assemblies and by investigating their expression across multiple conditions. Our analyses uncovered new TE superfamilies and families in this model symbiont and revealed significant differences in how these sequences evolve both within and between R. irregularis strains. With this curated TE annotation, we also found that the number of upregulated TE families in colonized roots is 4 times higher than in the extraradical mycelium, and their overall expression differs depending on the plant host. This work provides a fine-scale view of TE diversity and evolution in model plant symbionts and highlights their transcriptional dynamism and specificity during host-microbe interactions. We also provide Hidden Markov Model profiles of TE domains for future manual curation of uncharacterized sequences (https://github.com/jordana-olive/TE-manual-curation/tree/main).

Plant Growth Promoting Activity of Colletotrichum sp., an Endophyte of Helicteres isora L. and its Growth Optimization using Response Surface Methodology

Endophytes are the beneficial group of plant symbionts and are potent sources for producing novel metabolites with biotechnological applications. They are efficient sources for the synthesis of plant growth-promoting substances- Indole acetic acid, ammonia, solubilizing phosphates and thus open up sustainable agricultural opportunities. In the present investigation, twenty-two fungal endophytes were isolated from an ethnomedicinal plant-Helicteres isora L., collected from forests of Singbhum, Jhargram, India, and the isolate-Colletotrichum sp. HelS1, exhibited the most potent in vitro plant growth-promoting activities among all. HelS1 synthesized auxin (111.13 µg ml-1), ammonia, and solubilized phosphate (47.22 µg ml-1) in specific culture conditions. A treatment with fungal extract effectively increases the growth of the experimental plant (tomato seedlings) in terms of improvement in relative water contents, dry weight, nodal length, and pigment profiles compared to the untreated ones. There was an increase in root length by 27% compared to the control group. The isolate produced the maximum IAA after 7 days of incubation on Czepak Dox Broth supplemented with tryptophan (5 mg mL-1), sucrose (30 g L-1), and NaNO3 (2.2 g L-1) at pH 6.2. Fourier Transform Infrared Spectroscopic analysis of the crude fungal extract further confirmed the production of indole-like compounds. This investigation suggested that secondary metabolites of endophytes act as a potent plant growth inducer and can be utilized in bio-based crop management techniques.

In Vitro Module Editing Of NRPS Enables Production Of Highly Potent Gq -Signaling Inhibitor FR900359 Derived From Unculturable Plant Symbiont.

Heterotrimeric G proteins are key mediators in the signaling of G protein-coupled receptors (GPCR) that are involved in a plethora of important physiological processes and thus major targets of pharmaceutical drugs. The cyclic depsipeptides YM-254890 and FR900359 are strong and selective inhibitors of the Gq subfamily of G proteins. FR900359 was first reported to be produced by unculturable plant symbiont, however, a culturable FR900359 producer was discovered recently by the standard strategy, screening of the producing strain from the environment. As another strategy, we introduce herein the different way to supply natural compounds of unculturable microorganism origin. We therefore embarked on constructing an artificial biosynthetic gene cluster (BGC) for FR900359 with YM-254890 BGC as a template using "in vitro module editing" technology, first developed for the modification of type-I PKS BGCs, to edit YM-254890 BGC. The resulting artificial BGCs coding FR900359 were heterologously expressed in the Pseudomonas putida KT2440 host strain.

In Vitro Module Editing Of NRPS Enables Production Of Highly Potent Gq‐Signaling Inhibitor FR900359 Derived From Unculturable Plant Symbiont

AbstractHeterotrimeric G proteins are key mediators in the signaling of G protein‐coupled receptors (GPCR) that are involved in a plethora of important physiological processes and thus major targets of pharmaceutical drugs. The cyclic depsipeptides YM‐254890 and FR900359 are strong and selective inhibitors of the Gq subfamily of G proteins. FR900359 was first reported to be produced by unculturable plant symbiont, however, a culturable FR900359 producer was discovered recently by the standard strategy, screening of the producing strain from the environment. As another strategy, we introduce herein the different way to supply natural compounds of unculturable microorganism origin. We therefore embarked on constructing an artificial biosynthetic gene cluster (BGC) for FR900359 with YM‐254890 BGC as a template using “in vitro module editing” technology, first developed for the modification of type‐I PKS BGCs, to edit YM‐254890 BGC. The resulting artificial BGCs coding FR900359 were heterologously expressed in the Pseudomonas putida KT2440 host strain.

Bacillus sp. G2112 Detoxifies Phenazine-1-carboxylic Acid by N5 Glucosylation.

Microbial symbionts of plants constitute promising sources of biocontrol organisms to fight plant pathogens. Bacillus sp. G2112 and Pseudomonas sp. G124 isolated from cucumber (Cucumis sativus) leaves inhibited the plant pathogens Erwinia and Fusarium. When Bacillus sp. G2112 and Pseudomonas sp. G124 were co-cultivated, a red halo appeared around Bacillus sp. G2112 colonies. Metabolite profiling using liquid chromatography coupled to UV and mass spectrometry revealed that the antibiotic phenazine-1-carboxylic acid (PCA) released by Pseudomonas sp. G124 was transformed by Bacillus sp. G2112 to red pigments. In the presence of PCA (>40 µg/mL), Bacillus sp. G2112 could not grow. However, already-grown Bacillus sp. G2112 (OD600 > 1.0) survived PCA treatment, converting it to red pigments. These pigments were purified by reverse-phase chromatography, and identified by high-resolution mass spectrometry, NMR, and chemical degradation as unprecedented 5N-glucosylated phenazine derivatives: 7-imino-5N-(1'β-D-glucopyranosyl)-5,7-dihydrophenazine-1-carboxylic acid and 3-imino-5N-(1'β-D-glucopyranosyl)-3,5-dihydrophenazine-1-carboxylic acid. 3-imino-5N-(1'β-D-glucopyranosyl)-3,5-dihydrophenazine-1-carboxylic acid did not inhibit Bacillus sp. G2112, proving that the observed modification constitutes a resistance mechanism. The coexistence of microorganisms-especially under natural/field conditions-calls for such adaptations, such as PCA inactivation, but these can weaken the potential of the producing organism against pathogens and should be considered during the development of biocontrol strategies.

Fungal Endophytes - An Alternative Source of Bioactive Compounds from Plant Species for Prospective Applications – Review

Plant derived compounds have a strong antagonistic activity on pathogenic microorganisms using tremendous natural bioactive compounds accumulated in different parts of plants. Limonoids are important group of metabolically altered triterpene in Meliaceae. Out of 300 limonoids known today, Melia azedarach (melianone, meliarachin, meliacine), Melia dubia (nimbin, meliacarpin) and Azadirachta indica (azadirachtin, nimbolide) contributes about one-third of limonoids. Endophytic fungi are plant symbionts that act as an elicitor for the production of secondary metabolites which serves as an excellent source of drugs for various ailments. It has a great potential to synthesize several novel bioactive compounds which paved a way for commercial exploitation of secondary metabolites for pharmaceutical and agricultural sectors. Many dominant endophytic fungi isolated from these plants includes Colletotrichum sp, Trichoderma sp, Penicillium citrinum, Fusarium solani, Aspergillus sp, Nigrospora sp and Alternaria sp which acts on human pathogenic microorganisms such as Escherichia coli Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Candida albicans common causative agents for Urinary Tract Infections (UTI) predominantly occurring in women. This review aims to delineate the overview on sources of limonoids present in the leaves and the endophytic fungi isolated from these leaves of Meliaceae. The isolated endophytic fungi from the leaves exhibited antagonistic activity on UTI causing pathogens which often leads to infertility among young women. Thus, the sources of limonoids and isolation of endophytic fungi can be enhanced by reinforcing the production of commercial bioactive secondary metabolites that can be substantially utilized by pharmaceutical and agricultural industries to safeguard human beings.