Production Of Secondary Metabolites Research Articles

Effects of appropriate low-temperature treatment on the yield and quality of pigmented potato (Solanum tuberosum L.) tubers

Temperature is one of the important environmental factors affecting plant growth, yield and quality. Moreover, appropriately low temperature is also beneficial for tuber coloration. The red potato variety Jianchuanhong, whose tuber color is susceptible to temperature, and the purple potato variety Huaxinyangyu, whose tuber color is stable, were used as experimental materials and subjected to 20 °C (control check), 15 °C and 10 °C treatments during the whole growth period. The effects of temperature treatment on the phenotype, the expression levels of structural genes related to anthocyanins and the correlations of each indicator were analyzed. The results showed that treatment at 10 °C significantly inhibited the potato plant height, and the chlorophyll content and photosynthetic parameters in the leaves were reduced, and the enzyme activities of SOD and POD were significantly increased, all indicating that the leaves were damaged. Treatment at 10 °C also affected the tuberization of Huaxinyangyu and reduced the tuberization and coloring of Jianchuanhong, while treatment at 15 °C significantly increased the stem diameter, root-to-shoot ratio, yield and content of secondary metabolites, especially anthocyanins. Similarly, the expression of structural genes were enhanced in two pigmented potatoes under low-temperature treatment conditions. In short, proper low temperature can not only increase yield but also enhance secondary metabolites production. Previous studies have not focused on the effects of appropriate low-temperature treatment during the whole growth period of potato on the changes in metabolites during tuber growth and development, these results can provide a theoretical basis and technical guidance for the selection of pigmented potatoes with better nutritional quality planting environment and the formulation of cultivation measures.

Influence of Abiotic and Biotic Elicitors on Organogenesis, Biomass Accumulation, and Production of Key Secondary Metabolites in Asteraceae Plants.

The medicinal plants of the Asteraceae family are a valuable source of bioactive secondary metabolites, including polyphenols, phenolic acids, flavonoids, acetylenes, sesquiterpene lactones, triterpenes, etc. Under stressful conditions, the plants develop these secondary substances to carry out physiological tasks in plant cells. Secondary Asteraceae metabolites that are of the greatest interest to consumers are artemisinin (an anti-malarial drug from Artemisia annua L.-sweet wormwood), steviol glycosides (an intense sweetener from Stevia rebaudiana Bert.-stevia), caffeic acid derivatives (with a broad spectrum of biological activities synthesized from Echinacea purpurea (L.) Moench-echinacea and Cichorium intybus L.-chicory), helenalin and dihydrohelenalin (anti-inflammatory drug from Arnica montana L.-mountain arnica), parthenolide ("medieval aspirin" from Tanacetum parthenium (L.) Sch.Bip.-feverfew), and silymarin (liver-protective medicine from Silybum marianum (L.) Gaertn.-milk thistle). The necessity to enhance secondary metabolite synthesis has arisen due to the widespread use of these metabolites in numerous industrial sectors. Elicitation is an effective strategy to enhance the production of secondary metabolites in in vitro cultures. Suitable technological platforms for the production of phytochemicals are cell suspension, shoots, and hairy root cultures. Numerous reports describe an enhanced accumulation of desired metabolites after the application of various abiotic and biotic elicitors. Elicitors induce transcriptional changes in biosynthetic genes, leading to the metabolic reprogramming of secondary metabolism and clarifying the mechanism of the synthesis of bioactive compounds. This review summarizes biotechnological investigations concerning the biosynthesis of medicinally essential metabolites in plants of the Asteraceae family after various elicitor treatments.

Analysis of light recipe, seeding density, and fertilization effects on secondary metabolite accumulation and growth-defense responses in Brassicaceae microgreens

Although there have been large improvements to crop yield over time, this has not been accompanied by an increase in human nutritional wellbeing. In fact, there are worsening health crises associated with over- and under-consumption of particular food groups, resulting in negative human health outcomes. One solution to this is to utilize controlled environment agriculture to produce microgreens that have a high density of valuable Secondary Metabolites (SMs) such as antioxidants, phenolics, or pigment molecules that are associated with positive human health outcomes. However, optimal growth recipes to produce microgreens and their valuable nutritional compounds are not well known due to much species-specific variation, as well as biological tradeoffs between biomass and SM production. This is known as the growth defense hypothesis which describes how plants have a finite carbon budget from which to allocate to growth, or defensive, processes. To further research in this regard, this project used climate chambers to grow three species of Brassicaceae microgreens (kohlrabi, mustard, and radish) in highly controlled environmental conditions, where we used five Light Recipes, two fertilization levels, and two seeding density treatments. Our results showed that there were significant differences in SM production of these microgreens due to changes in incident light, as well as significant interactions between Fertilizer and Light Recipe for all SMs except Anthocyanins. For example, for all three species, the High Far-Red Light Recipe had the significantly highest Phenolic concentration, but with lower values of the other four SMs. The low-intensity 24-V high efficiency LEDs had the significantly highest Trolox Equivalent Antioxidant Capacity (TEAC) concentrations, while the High Red recipe had the highest Ferric Reducing Antioxidant Power (FRAP) and Flavonoid concentrations. For Anthocyanins, there were less clear patterns, with the No Green or High Red recipes having generally higher concentrations, but not always significantly. We did find some evidence supporting the growth-defense hypothesis, where our higher biomass values were negatively correlated with SM concentrations. We also found significant differences between the concentrations of SMs in leaves and stems for kohlrabi and mustard microgreens. Finally, we found some significant relationships between increasing Fertilizer dosage and SMs, for instance that Flavonoids and FRAP concentrations increased with Fertilizer application, while Anthocyanin decreased, and Phenolic and TEAC had little effect. In conclusion, we found that there were significant relationships for Light Recipe and Fertilizer, and oftentimes their interaction, for the accumulation of SMs in Brassicaceae microgreens, which can inform microgreen production environments.

Influence of Citrus sunki and Poncirus trifoliata Root Extracts on Metabolome of Phytophthora parasitica.

Phytophthora parasitica is an oomycete pathogen that infects a broad range of crops of worldwide economic interest; among them are citrus species. In general, some Citrus and the rootstocks of related genera offer considerable resistance against P. parasitica; therefore, understanding the mechanisms involved in the virulence of this pathogen is crucial. In this work, P. parasitica secondary metabolite production was studied using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) and ultrahigh-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC/ESI-Q-TOF-MS) combined with chemometric tools, and its metabolic profile was evaluated under the influence of Citrus sunki (a highly susceptible host) and Poncirus trifoliata (a resistant genotype) extracts. The root extracts of Citrus sunki had an influence on the growth and hyphae morphology, and the root extracts of P. trifoliata had an influence on the zoospore behavior. In parallel, the spatial distribution of several metabolites was revealed in P. parasitica colonies using MALDI-MSI, and the metabolite ion of m/z 246 was identified as the protonated molecule of Arg-Ala. The MALDI-MSI showed variations in the surface metabolite profile of P. parasitica under the influence of the P. trifoliata extract. The P. parasitica metabolome analysis using UHPLC-ESI-Q-TOF-MS resulted in the detection of Arg-Gln (m/z 303.1775), as well as L-arginine (m/z 175.1191) and other unidentified metabolites. Significant variations in this metabolome were detected under the influence of the plant extracts when evaluated using UHPLC-ESI-Q-TOF-MS. Both techniques proved to be complementary, offering valuable insights at the molecular level when used to assess the impact of the plant extracts on microbial physiology in vitro. The metabolites identified in this study may play significant roles in the interaction or virulence of P. parasitica, but their functional characterization remains to be analyzed. Overall, these data confirm our initial hypotheses, demonstrating that P. parasitica has the capabilities of (i) recognizing host signals and altering its reproductive programing and (ii) distinguishing between hosts with varying responses in terms of reproduction and the production of secondary metabolites.

Arohynapene A Produced by <i>Penicillium steckii</i> JB-NW-2-1 Isolated from <i>Avicennia marina</i> (Forssk.) Vierh and Its Cytotoxic Activities

Mangrove-associated endophytic fungi are producers of secondary metabolites in unique and diverse structures with interesting biological activities such as antiviral, antifungal, antibacterial, anti-inflammatory, and cytotoxic agents. Endophytes play an important role in the physiological activities of the host plants, influencing the improvement of resistance to stress, insects, nematodes, and diseases. In this study, arohynapene A, a polyketide compound, was successfully isolated from the mangrove-derived fungus Penicillium steckii JB-NW-2-1 obtained from mangrove plant Avicennia marina (Forssk) Vierh. The structure was determined by a spectroscopic method including IR, MS, 1D-, and 2D-NMR techniques. This compound was evaluated for cytotoxic activities using resazurin assay against four cancer cells, HeLa cervical, MCF-7 breast cancer, B16-F10 melanoma, and A549 lung adenocarcinoma. The results showed no significant activities against all cancer cells tested (IC50 > 500 µM).

Recent advances in understanding the regulation of plant secondary metabolite biosynthesis by ethylene-mediated pathways.

Plants produce a large repertoire of secondary metabolites. The pathways that lead to the biosynthesis of these metabolites are majorly conserved in the plant kingdom. However, a significant portion of these metabolites are specific to certain groups or species due to variations in the downstream pathways and evolution of the enzymes. These metabolites show spatiotemporal variation in their accumulation and are of great importance to plants due to their role in development, stress response and survival. A large number of these metabolites are in huge industrial demand due to their potential use as therapeutics, aromatics and more. Ethylene, as a plant hormone is long known, and its biosynthetic process, signaling mechanism and effects on development and response pathways have been characterized in many plants. Through exogenous treatments, ethylene and its inhibitors have been used to manipulate the production of various secondary metabolites. However, the research done on a limited number of plants in the last few years has only started to uncover the mechanisms through which ethylene regulates the accumulation of these metabolites. Often in association with other hormones, ethylene participates in fine-tuning the biosynthesis of the secondary metabolites, and brings specificity in the regulation depending on the plant, organ, tissue type and the prevailing conditions. This review summarizes the related studies, interprets the outcomes, and identifies the gaps that will help to breed better varieties of the related crops and produce high-value secondary metabolites for human benefits.

CdgB Regulates Morphological Differentiation and Toyocamycin Production in Streptomyces diastatochromogenes 1628.

Bis (3',5')-cyclic diguanylic acid (c-di-GMP) is a ubiquitous second messenger that controls several metabolic pathways in bacteria. In Streptomyces, c-di-GMP is associated with morphological differentiation, which is related to secondary metabolite production. In this study, we identified and characterized a diguanylate cyclase (DGC), CdgB, from Streptomyces diastatochromogenes 1628, which may be involved in c-di-GMP synthesis, through genetic and biochemical analyses. To further investigate the role of CdgB, the cdgB-deleted mutant strain Δ-cdgB and the cdgB-overexpressing mutant strain O-cdgB were constructed by genetic engineering. A phenotypic analysis revealed that the O-cdgB colonies exhibited reduced mycelium formation, whereas the Δ-cdgB colonies displayed wrinkled surfaces and shriveled mycelia. Notably, O-cdgB demonstrated a significant increase in the toyocamycin (TM) yield by 47.3%, from 253 to 374 mg/L, within 10 days. This increase was accompanied by a 6.7% elevation in the intracellular concentration of c-di-GMP and a higher transcriptional level of the toy cluster within four days. Conversely, Δ-cdgB showed a lower c-di-GMP concentration (reduced by 6.2%) in vivo and a reduced toyocamycin production (decreased by 28.9%, from 253 to 180 mg/L) after 10 days. In addition, S. diastatochromogenes 1628 exhibited a slightly higher inhibitory effect against Fusarium oxysporum f. sp. cucumerinum and Rhizoctonia solani compared to Δ-cdgB, but a lower inhibition rate than that of O-cdgB. The results imply that CdgB provides a foundational function for metabolism and the activation of secondary metabolism in S. diastatochromogenes 1628.

Light as an elicitor for enhanced production of secondary metabolites in plant cell, tissue, and organ cultures

Light as an elicitor for enhanced production of secondary metabolites in plant cell, tissue, and organ cultures

Isolation, Characterization, and Expression Analysis of NAC Transcription Factor from Andrographis paniculata (Burm. f.) Nees and Their Role in Andrographolide Production.

Andrographis paniculata (Burm. f.) Nees is an important medicinal plant known for its bioactive compound andrographolide. NAC transcription factors (NAM, ATAF1/2, and CUC2) play a crucial role in secondary metabolite production, stress responses, and plant development through hormonal signaling. In this study, a putative partial transcript of three NAC family genes (ApNAC83, ApNAC21 22 and ApNAC02) was used to isolate full length genes using RACE. Bioinformatics analyses such as protein structure prediction, cis-acting regulatory elements, and gene ontology analysis were performed. Based on in silico predictions, the diterpenoid profiling of the plant's leaves (five-week-old) and the real-time PCR-based expression analysis of isolated NAC genes under abscisic acid (ABA) treatment were performed. Additionally, the expression analysis of isolated NAC genes under MeJA treatment and transient expression in Nicotiana tabacum was performed. Full-length sequences of three members of the NAC transcription factor family, ApNAC83 (1102 bp), ApNAC21 22 (996 bp), and ApNAC02 (1011 bp), were isolated and subjected to the promoter and gene ontology analysis, which indicated their role in transcriptional regulation, DNA binding, ABA-activated signaling, and stress management. It was observed that ABA treatment leads to a higher accumulation of andrographolide and 14-deoxyandrographolide content, along with the upregulation of ApNAC02 (9.6-fold) and the downregulation of ApNAC83 and ApNAC21 22 in the leaves. With methyl jasmonate treatment, ApNAC21 22 expression decreased, while ApNAC02 increased (1.9-fold), with no significant change being observed in ApNAC83. The transient expression of the isolated NAC genes in a heterologous system (Nicotiana benthamiana) demonstrated their functional transcriptional activity, leading to the upregulation of the NtHMGR gene, which is related to the terpene pathway in tobacco. The expression analysis and heterologous expression of ApNAC21 22 and ApNAC02 indicated their role in andrographolide biosynthesis.

Exploring Indonesian Sponge-Associated Marine Aspergillus hortai: Characterization of Bioactive Compounds with Potential Anti-Escherichia coli Properties

Sponge-associated marine fungi are potential source for secondary metabolite compounds. The aim of this research was to investigate sponge-associated marine fungus as secondary metabolite producers against Escherichia coli. The fungus was isolated from Indonesian marine sponge Stylissa sp. and identified as Aspergillus hortai through a combination of morphological and molecular characteristics of ITS DNA and β-tubulin genes. The fungus was tested against E. coli using fungal broth and mycelial extracts. The optimized condition was achieved by fungal broth grown in corn meal broth at 6-days of shaking incubation. Fungal extract was produced using three liters of filtered fungal broth and extracted in ethyl acetate. The antibiotic activity of the extract is vulnerable to 45°C heat and basic or acidic conditions. Therefore, the extraction was done at pH 7 with evaporation at 40°C. The extract shows 7 major bands on TLC with 1 band shows activity against E. coli (Rf 0.81) on bioautogram. The band was observed as a yellow color and turned black in short-wave UV and did not show any fluorescence in long-wave UV. This research shows that sponge-associated marine fungi obtained from Indonesia has the potential as anti E. coli worth to be explored for searching new antibiotics.

Utilization of CRISPR-Cas genome editing technology in filamentous fungi: function and advancement potentiality.

Filamentous fungi play a crucial role in environmental pollution control, protein secretion, and the production of active secondary metabolites. The evolution of gene editing technology has significantly improved the study of filamentous fungi, which in the past was laborious and time-consuming. But recently, CRISPR-Cas systems, which utilize small guide RNA (sgRNA) to mediate clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas), have demonstrated considerable promise in research and application for filamentous fungi. The principle, function, and classification of CRISPR-Cas, along with its application strategies and research progress in filamentous fungi, will all be covered in the review. Additionally, we will go over general matters to take into account when editing a genome with the CRISPR-Cas system, including the creation of vectors, different transformation methodologies, multiple editing approaches, CRISPR-mediated transcriptional activation (CRISPRa) or interference (CRISPRi), base editors (BEs), and Prime editors (PEs).

Biological Control of Pseudomonas syringae in Tomato Using Filtrates and Extracts Produced by Alternaria leptinellae

Endophytic fungi offer promising alternatives for sustainable plant disease management strategies, often through the production of bioactive secondary metabolites. This study investigated the biocontrol potential of filtrates and extracts, produced under controlled conditions, from Alternaria leptinellae E138 against Pseudomonas syringae in tomato plants under greenhouse conditions. To understand the main mechanisms involved in biocontrol, the direct inhibition of bacterial growth and disruption of quorum sensing activity caused by metabolites were studied in vitro, as well as indirect mechanisms, such as their capacity to produce phytohormone-like substances, nutrient mobilization, and antioxidant activity, which can enhance plant growth and fitness. Moreover, a mass spectrometry analysis was used to tentatively identify the secondary metabolites present in the extract with antimicrobial properties, which could explain the biocontrol effects observed. Mycopriming assays, involving the direct treatment of tomato seeds with the fungal A. leptinellae E138 extracts, produced increased germination rates and seedling vigor in tomato seeds. As another treatment, postemergence application of the extracts in greenhouse conditions significantly improved plant health and resulted in a 41% decrease in disease severity. Overall, this study underscores the potential of A. leptinellae E138 extract as a plant growth promoter with biocontrol capabilities, offering promising avenues for sustainable plant disease management.

Composition and antimicrobial activity of Rosmarinus officinalis L. and Artemisia monosperma L. leaf essential oils and methanolic extracts from plants grown in normal and saline habitats in Egypt

The present work aimed to investigate the effect of salinity in natural habitats in Egypt on the main secondary metabolites of Rosmarinus officinalis L. and Artemisia monosperma L. plants compared to plants grown at normal conditions. Plants grown under salinity were collected from Egyptian Western Coastal region habitats irrigated with underground water. Results showed that salinity increased the essential oil percentage of R. officinalis L. by 52.7% and A. monosperma L by 0.29% in addition to the total phenolics and flavonoids content in dry leaves compared to control plants. GC/MS analysis of rosemary essential oils revealed that salinity decreased the amount of some major oil monoterpenes component as verbenone, with a slight effect on 1,8 cineole and increased Camphor, endo- Boreneol, and linalool in addition to the appearance of new specific components such as Chrysanthenone monoterpene ketone and Caryophyllene sesquiterpene, while, in the case of Artemisia, the GC/MS showed that Artemisia ketone, Camphor, β -phellandrene monoterpenes andα-Bisabolol sesquiterpenewere the major oil components; salinity decreased Camphor and β -phellandrene content and increased artemisia ketone and α-Bisabolol oil content. About 11 new oil constituents were detected such as ( +)-2-Bornanone and Sesquisabinene hydrate. Mineral ions (N, K+, Ca+2, P, and Mg+2) uptake by R. officinalis and A. monosperma decreased in plants grown under salinity, while Na content increased compared to corresponding controls. Results demonstrated that both plants could tolerate the high salinity level in natural Western Coastal region soil which promoted more production of valuable secondary metabolites. The antimicrobial effect of R. officinalis L. and A. monosperma L. leaf methanolic extracts, results showed that R. officinalis extracts had an inhibitory response against all tested gram-positive and negative bacteria, in addition to the yeast (Candida albicans), whereas there was no any inhibitory effect concerning A. monosperma L extract on the tested species.

Climate change and the future of medicinal plants research

Due to their pharmaceutical properties and secondary metabolites, medicinal plants have played a crucial role in alleviating human suffering from various illnesses, disorders, and diseases. Along with other natural and artificial disasters, rapid climate change is one of the critical causes of the decline in wild medicinal plant species. Therefore, considering their significance in traditional medicine practices and economic value, investigating the potential consequences of climate change on medicinal plants is specifically relevant. This review aimed to investigate the diversity of species of medicinal plants within current climate scenarios. Additionally, it assesses the potential implications of climate change on the projected distribution of these species in subsequent scenarios and evaluates the possible impacts of these changes on the trajectory of future research in medicinal plants. The distribution and life cycles of all vegetation, including medicinal plants, are significantly impacted by climate change. Additionally, future climate scenarios have been shown to affect the physiological performance of all vegetation worldwide. Various aspects related to climate change and its repercussions on medicinal plants are explored including the impact of increased carbon dioxide (CO2) and ozone (O3) levels, the effect of low temperature (Cold), climate warming, drought on the production of secondary metabolites, impact of threats on medicinal plants, and phenological changes. Each of these environmental factors influences the productivity and quality of different products and components of medicinal plants, either positively or negatively. Consequently, there is excellent complexity surrounding how climate change affects medicinal plants. Thus, for human survival on Earth, future researchers should carefully examine the interactions of various direct and indirect causes and their corresponding effects. Keywords: Secondary metabolites, Climate change, medicinal plants, Phenology.

Simultaneous Impact of Rhizobacteria Inoculation and Leaf-Chewing Insect Herbivory on Essential Oil Production and VOC Emissions in Ocimum basilicum.

Inoculation with rhizobacteria and feeding by herbivores, two types of abiotic stress, have been shown to increase the production of secondary metabolites in plants as part of the defense response. This study explored the simultaneous effects of inoculation with Bacillus amyloliquefaciens GB03 (a PGPR species) and herbivory by third-instar Spodoptera frugiperda larvae on essential oil (EO) yield and volatile organic compound (VOC) emissions in Ocimum basilicum plants. The density of glandular trichomes was also examined, given that they are linked to EO production and VOC emission. Herbivory increased EO content, but inoculation on its own did not. When combined, however, the two treatments led to a 10-fold rise in EO content with respect to non-inoculated plants. VOC emissions did not significantly differ between inoculated and non-inoculated plants, but they doubled in plants chewed by the larvae with respect to their undamaged counterparts. Interestingly, no changes were observed in VOC emissions when the treatments were tested together. In short, the two biotic stressors elicited differing plant defense responses, mainly when EO was concerned. PGPR did not stimulate EO production, while herbivory significantly enhanced it and increased VOC emissions. The combined treatment acted synergistically, and in this case, PGPR inoculation may have had a priming effect that amplified plant response to herbivory. Peltate trichome density was higher in inoculated plants, those damaged by larvae, and those subjected to the combination of both treatments. The findings highlight the intricate nature of plant defense mechanisms against various stressors and hint at a potential strategy to produce essential oil through the combined application of the two stressors tested here.

Isolation of Secondary Metabolites From Marine Actinobacterium of Microbispora sp.T3S11 and Their Antibacterial Activities.

Introduction Marine actinobacteria are promising sources of novel bioactive compounds due to their distinct ecological niches and diverse secondary metabolite production capabilities. Among these,Microbisporasp. T3S11 is notable for its unique spore chain structure, which allows for both morphological and genetic identification. Despite its potential, little is understood about the secondary metabolites produced by this strain. In this study, we hope to fill this gap by extracting and analyzing the antibacterial activities of secondary metabolites fromMicrobisporasp. T3S11, which will be the first time its bioactive compound profile is investigated. Aim To evaluate the antibacterial activity of secondary metabolites isolated from the marine actinobacteriumMicrobisporasp. T3S11. Materials and methods The antibacterial assays were carried out on agar plates containing the appropriate media for each pathogen. Sterile filter paper disks were impregnated with secondary metabolites extracted fromMicrobisporasp. T3S11 and placed on the surface of agar plates inoculated with the appropriate pathogens. Similarly, disks containing tetracycline were used as a positive control. The plates were then incubated at the appropriate temperature for each pathogen, and the zones of inhibition around the disks were measured to determine the extracted metabolites' antibacterial activity. Result Secondary metabolites had antimicrobial activity againstStreptococcus mutans,Klebsiella pneumonia, and methicillin-resistantStaphylococcus aureus(MRSA). The inhibition ofS. mutanswas 7.5 mm and 8.5 mm at 75 μg/mL and 100 μg/mL, respectively.Klebsiella pneumoniazones measured 7 mm and 7.5 mm, while MRSA zones measured 7.6 mm and 8.5 mm at the same concentrations. Tetracycline, the standard antibiotic, had larger inhibition zones: 22 mm forS. mutansandKlebsiella pneumoniaand 16 mm for MRSA, indicating variable susceptibility. Conclusion We conclude that the secondary metabolites extracted fromMicrobisporasp. T3S11exhibits high antibacterial activity. This could be attributed to the presence of various active compounds.

Bacterial growth-mediated systems remodelling of Nicotiana benthamiana defines unique signatures of target protein production in molecular pharming.

The need for therapeutics to treat a plethora of medical conditions and diseases is on the rise and the demand for alternative approaches to mammalian-based production systems is increasing. Plant-based strategies provide a safe and effective alternative to produce biological drugs but have yet to enter mainstream manufacturing at a competitive level. Limitations associated with batch consistency and target protein production levels are present; however, strategies to overcome these challenges are underway. In this study, we apply state-of-the-art mass spectrometry-based proteomics to define proteome remodelling of the plant following agroinfiltration with bacteria grown under shake flask or bioreactor conditions. We observed distinct signatures of bacterial protein production corresponding to the different growth conditions that directly influence the plant defence responses and target protein production on a temporal axis. Our integration of proteomic profiling with small molecule detection and quantification reveals the fluctuation of secondary metabolite production over time to provide new insight into the complexities of dual system modulation in molecular pharming. Our findings suggest that bioreactor bacterial growth may promote evasion of early plant defence responses towards Agrobacterium tumefaciens (updated nomenclature to Rhizobium radiobacter). Furthermore, we uncover and explore specific targets for genetic manipulation to suppress host defences and increase recombinant protein production in molecular pharming.

Streptomyces sungeiensis SD3 as a Microbial Chassis for the Heterologous Production of Secondary Metabolites.

We present the newly isolated Streptomyces sungeiensis SD3 strain as a promising microbial chassis for heterologous production of secondary metabolites. S. sungeiensis SD3 exhibits several advantageous traits as a microbial chassis, including genetic tractability, rapid growth, susceptibility to antibiotics, and metabolic capability supporting secondary metabolism. Genomic and transcriptomic sequencing unveiled the primary metabolic capabilities and secondary biosynthetic pathways of S. sungeiensis SD3, including a previously unknown pathway responsible for the biosynthesis of streptazone B1. The unique placement of S. sungeiensis SD3 in the phylogenetic tree designates it as a type strain, setting it apart from other frequently employed Streptomyces chassis. This distinction makes it the preferred chassis for expressing biosynthetic gene clusters (BGCs) derived from strains within the same phylogenetic or neighboring phylogenetic clade. The successful expression of secondary biosynthetic pathways from a closely related yet slow-growing strain underscores the utility of S. sungeiensis SD3 as a heterologous expression chassis. Validation of CRISPR/Cas9-assisted genetic tools for chromosomal deletion and insertion paved the way for further strain improvement and BGC refactoring through rational genome editing. The addition of S. sungeiensis SD3 to the heterologous chassis toolkit will facilitate the discovery and production of secondary metabolites.

Climate-induced changes in essential oil production and terpene composition in alpine aromatic plants

Alpine regions are biodiversity hotspots for endemic species with unique adaptations to harsh climatic conditions. These adaptations often involve specialized behaviours and metabolism including the production of secondary metabolites that protects plants from environmental stresses. This study examines the impacts of elevated CO2 (eCO2) and combined effects of elevated CO2 and temperature (eCO2+T) on two alpine aromatic herbs, namely Angelica glauca and Nardostachys jatamansi. These species were raised in open-top chambers (OTCs) and treated with elevated CO2 (600 ppm) and temperature (+2.0 °C from control) during 2018 to 2020. The primary (total soluble sugar, starch, total soluble protein, amino acid and proline contents), secondary metabolites (Phenolics, essential oil yield, and components using GC-MS) and antioxidant activities (superoxide dismutase and peroxidase) were measured. Total soluble sugar increased under eCO2, while starch content, total amino acid and total soluble protein initially increased but later declined in response to the treatments. Proline content and antioxidant activities were higher under eCO2 and eCO2+T treatments. Total phenolic content showed both increasing (in A. glauca) and decreasing (in N. jatamansi) responses to eCO2 and eCO2+T treatments. GC–MS analysis showed decreased monoterpenes in CO2 and temperature treatments, but some sesquiterpenes showed mixed responses with both increases and decreases. These findings suggest that changes in climate can alter the composition of secondary metabolites in alpine herbs, potentially affecting their therapeutic properties and suitability for use in herbal medicine. This study provides valuable insights into the species-specific physiological responses of plants to changing climate.

Antibacterial activity of endophytic fungi associated with Aspidosperma subincanum Mart. against Gram-positive bacteria

Endophytic fungi are microorganisms that inhabit plant tissues, without causing damage to the host. When in association with medicinal plants, they are able to prompt the production of secondary metabolites with important biological activities. The aim of this study was to evaluate the antibacterial activity of endophytic fungi associated with Aspidosperma subincanum against Gram positive bacteria of clinical relevance and perform the chemical characterization of the extract with the highest antibacterial activity. Endophytic fungi isolated from leaves of A. subincanum were submitted to ethanolic extraction. The antibacterial activity of the extracts was evaluated against S. aureus ATCC 25923, using the microdilution method. From a total of 116 extracts, only 4 (3.44%) were considered positive and then evaluated against B. cereus. The ethanolic endophytic extract AM59.1-98 was able to completely inhibit the growth of both microorganisms and was selected for chemical characterization, which indicated the presence of saponins and tannins. Further studies have to be conducted to confirm if the chemical substances detected are those responsible for its antibacterial activity. The current research provides, for the first time, substantial results concerning the antibacterial activity of endophytes associated with A. subincanum as an alternative path in the search for new drugs.