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Bioformulations with Trichoderma strains offer cost-effective and sustainable options for integrated disease management and plant nutrition. Therefore, this study aimed to selected isolates of Trichoderma with antagonistic and plant growth-promoting potential, specifically in the early development stage of soybean seedlings. Trichoderma isolated from the soybean rhizosphere were screened to assess their antagonistic activity against three phytopathogens; as well as their capability for indole-3-acetic acid (IAA) synthesis and phosphate solubilization. Three promising were further examined for their tolerance to various in vitro conditions and agrochemicals. Experiments were performed to assess the effect of single inoculation and co-inoculation of strains on the growth and biochemical responses during early soybean development. Nine isolates showed effective antagonism against phytopathogens. Among them, T. reesei GT-31 and T. longibrachiatum GT-32 showed the highest IAA production, with 175.8 and 119.6 μg/mL, respectively, while the T. viride GT-8 showed the highest phosphate solubilization activity after 10 days of growth (285.6 μg P/mL). These strains displayed robust growth under various conditions and agrochemical treatments. The co-inoculation of three strains resulted in even higher dry shoot and root weights, increasing by 50.3% and 48.8%, respectively, compared to non-inoculated seedlings. Co-inoculated plants also exhibited elevated chlorophyll (31.9%), carotenoids (24.9%), flavonoids (13.2%), and phenolic compounds (42.3%). The results suggest that employing the three strains with beneficial mechanisms for plants could significantly enhance the growth and early development of soybean seedlings. Overall, these findings highlight the potential of these novel Trichoderma strains to enhance plant growth and offer benefits in soybean crops, providing a sustainable strategy for agriculture. [ABSTRACT FROM AUTHOR]

The purpose of this present study is to explore and decipher copper tolerant plant growth promoting bacteria from dolomite contaminated soil. Lysinibacillus sphaericus (NJCT1) isolated from tea garden soil near Kaljani river bank has been found to exhibit multifaceted characteristics and has shown its ability to tolerate copper up-to 40 mg/ml. Further, its ability to tolerate other heavy metals were also assessed and the result revealed that it can tolerate iron, zinc and aluminum upto 100 mg/ml, while for lead and arsenic it showed tolerance upto 10 mg/ml. GC-MS analysis showed that L. sphaericus (NJCT1) produced 16 compounds, among them, tromethamine, 2-methylbutanoic acid, 2,4,6-(1H,3H,5H)-pyrimidinetrione, 2-piperidinone, 3,5,5-trimethylhexyl acetate and cis-3-hexenyl propionate were found to exhibit various important biological properties. Also, this isolate was positive for phosphate solubilization and is capable of producing ammonia, HCN, siderophore, and 17.12 µg/ml of indole acetic acid. In seed germination, seeds treated with NJCT1 demonstrated 100% germination rate with 111-time increase in seedling vigor index. Further, in pot study, significant increase in all the growth parameters of NJCT1 treated Vigna radiata and Phaseolus vulgaris gave a strong indication that this strain can be a promising candidate for reclamation and revitalization of dolomite and copper ore affected tea garden soil. [ABSTRACT FROM AUTHOR]

Wild legumes are widely distributed and better adapted to extreme agro-climatic conditions because of their soil microbiota. In the present study, Root Nodulating Bacteria (RNB) were isolated from wild legumes and analysed for their abiotic stress tolerant and Plant Growth Promoting Traits (PGPT). Rhizobial strains were identified based on their 16S rRNA sequences. A total of 14 rhizobial species were characterized based, of which six were Rhizobium (AIS3, AIS9, AIR14, LUMCR3, LUMVRW4, LUMVRW8), four belongs to Burkholderia (AIS12, MOLKCS15, CHMP9, CHMP10) and one each of genera Cupriavidus (LUMMD12) , Herbaspirillum (LUMVRW4) , Paraburkholderia (CHMP1) and Ralstonia (LUMDes9). Stress tolerance analysis found most of the strains are to be tolerant to alkaline pH (9–11) where highest tolerance exhibited by Rhizobium sp. (strain AIS9) and Burkholderia territorii (strain MOKCS15). Seven Rhizobial strains were able to thrive at lower temperature stress (10 and 20 °C); while, strains AIS9, CHMP1 and LUMMD21 were able to grow at higher temperature (40 and 50 °C). Most of the isolate could tolerate salinity stress up to 1.5 %; but, only isolates AIS9, AIS12 and LUMMD12 could tolerate 2 and 2.5 % salt concentrations. The PGPT analyses found LUMCR3 having the highest IAA production potentiality (102.5 µg/ml); while, isolate MOKCS15 exhibited highest phosphate solubilizing activity (4.1 Phosphate Solubilization Index). The RNB isolates in the present study exhibited both stress tolerance and PGP traits indicating their potential use as biofertilizer under extreme agronomic conditions to improve productivity. [Display omitted] • A total 14 rhizobial RNB isolated from the root nodules of 8 wild legumes from Nagaland. • Rhizobial strains were found to have abiotic stress (pH, salt, temperature) tolerance abilities. • Rhizobial isolates also showed IAA production and PSB traits. • First report nodulating ability of Burkholderia sp. isolated from Aeschynomene americana nodules. • First report on RNB from Vigna nepalensis (Herbaspirillum sp. and Rhizobium sp). [ABSTRACT FROM AUTHOR]

Introduction: Serratia marcescens is most commonly known as an opportunistic pathogen causing nosocomial infections. It, however, was shown to infect a wide range of hosts apart from vertebrates such as insects or plants as well, being either pathogenic or growth-promoting for the latter. Despite being extensively studied in terms of virulence mechanisms during human infections, there has been little evidence of which factors determine S. marcescens host specificity. On that account, we analyzed S. marcescens pangenome to reveal possible specificity factors. Methods: We selected 73 high-quality genome assemblies of complete level and reconstructed the respective pangenome and reference phylogeny based on core genes alignment. To find an optimal pipeline, we tested current pangenomic tools and obtained several phylogenetic inferences. The pangenome was rich in its accessory component and was considered open according to the Heaps' law. We then applied the pangenome-wide associating method (pan-GWAS) and predicted positively associated gene clusters attributed to three host groups, namely, humans, insects, and plants. Results: According to the results, significant factors relating to human infections included transcriptional regulators, lipoproteins, ABC transporters, and membrane proteins. Host preference toward insects, in its turn, was associated with diverse enzymes, such as hydrolases, isochorismatase, and N- acetyltransferase with the latter possibly exerting a neurotoxic effect. Finally, plant infection may be conducted through type VI secretion systems and modulation of plant cell wall synthesis. Interestingly, factors associated with plants also included putative growth-promoting proteins like enzymes performing xenobiotic degradation and releasing ammonium irons. We also identified overrepresented functional annotations within the sets of specificity factors and found that their functional characteristics fell into separate clusters, thus, implying that host adaptation is represented by diverse functional pathways. Finally, we found that mobile genetic elements bore specificity determinants. In particular, prophages were mainly associated with factors related to humans, while genetic islands-with insects and plants, respectively. Discussion: In summary, functional enrichments coupled with pangenomic inferences allowed us to hypothesize that the respective host preference is carried out through distinct molecular mechanisms of virulence. To the best of our knowledge, the presented research is the first to identify specific genomic features of S. marcescens assemblies isolated from different hosts at the pangenomic level. [ABSTRACT FROM AUTHOR]

Environmental changes pose significant challenges to sustainable agriculture, adversely affecting crop production and soil fertility. Factors such as drought, salinity, pathogens, and soil type exert their influence on the behavior of fodder crops by altering their morphological, biochemical, and molecular mechanisms, ultimately leading to reduced yields and productivity. Consequently, there is a pressing need to develop mitigation strategies aimed at enhancing the tolerance of forage crops to both biotic and abiotic stresses, addressing a critical challenge in sustaining their growth. In recent times, the use of biofertilizers has emerged as an environmentally friendly alternative to chemical fertilizers, holding promise for sustainable horticultural, agricultural, and forestry production systems. Notably, endophytic microorganisms play a pivotal role in promoting plant growth through direct or indirect mechanisms. Additionally, endophytic bacteria actively regulate gene expression responsible for the production of antioxidant enzymes, various phytohormones, siderophores, and ROS scavenging enzymes, all of which contribute to supporting the growth of host plants even in extreme environments. Consequently, there is a growing focus on understanding and validating the mechanisms through which beneficial plant endophytes interact to combat both biotic and abiotic stresses. This review emphasizes the potential of endophytes as biofertilizers, biocontrol agents, and contributors to the mitigation of abiotic and biotic stresses, all of which play crucial roles in maintaining the development of forage crops and soil fertility.

Compared to rifampicin (600 mg/day), standard doses of rifabutin (300 mg/day) have a lower risk of drug–drug interactions due to induction of cytochrome P450 3A4 (CYP3A4) or P-glycoprotein (Pgp/ABCB1) mediated by the pregnane X receptor (PXR). However, clinical comparisons with equal rifamycin doses or in vitro experiments respecting actual intracellular concentrations are lacking. Thus, the genuine pharmacological differences and the potential molecular mechanisms of the discordant perpetrator effects are unknown. Consequently, the cellular uptake kinetics (mass spectrometry), PXR activation (luciferase reporter gene assays), and impact on CYP3A4 and Pgp/ABCB1 expression and activity (polymerase chain reaction, enzymatic assays, flow cytometry) were evaluated in LS180 cells after treatment with different rifampicin or rifabutin concentrations for variable exposure times and eventually normalized to actual intracellular concentrations. In addition, inhibitory effects on CYP3A4 and Pgp activities were investigated. While rifampicin is poorly taken up by LS180 cells, it strongly activates PXR and leads to enhanced expression and activity of CYP3A4 and Pgp. In contrast, rifabutin is a significantly less potent and less efficient PXR activator and gene inducer, despite sixfold to eightfold higher intracellular accumulation. Finally, rifabutin is a potent inhibitor of Pgp (IC50 = 0.3 µM) compared to rifampicin (IC50 = 12.9 µM). Together, rifampicin and rifabutin significantly differ by their effects on the regulation and function of CYP3A4 and Pgp, even when controlled for intracellular concentrations. Rifabutin's concurrent Pgp inhibitory action might partly compensate the inducing effects, explaining its weaker clinical perpetrator characteristics. [ABSTRACT FROM AUTHOR]

Manipuri black aromatic rice, 'Chakhao', is a nutrient rich unique local rice cultivar of Manipur, India. Rhizosphere bacteria may have potential as antifungal agent and seedling growth promoter. Hence, the current investigation was aimed at screening, for the above traits, using bacteria, Bacillus spp., isolated from the soil of Rhizosphere of six Chakhao rice cultivars. Altogether, 323 bacterial isolates were obtained from the rhizospheric soils of 6 different cultivars of Chakhao, namely Chakhao Amubi (CA), Chakhao Poireiton (CP), Chakhao Sempak (CS), Chakhao Angoubi (CAng), Chakhao Angangbi (CR) and Chakhao Wairi (CW). All the rhizobacterial isolates were screened for antifungal activity against 5 rice fungal pathogens viz. Rhizoctonia solani (RS), Fusarium oxysporum (FO), Curvularia oryzae (CO), Pyricularia oryzae (PO) and Aspergillus niger (AN). The isolates were also subjected to PGP (plant growth promotion) assays such as ammonia, indole acetic acid (IAA), siderophore and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase production, and phosphate solubilization. Of 323 isolates obtained, 64 were found to exhibit antifungal activity while 69 showed PGP traits. Twenty-five (25) isolates were found to be positive for both antifungal and PGP traits. Of 25 bioactive isolates, 4 (CR12, CW11, CA2 and CP2) potent isolates were shortlisted for further studies. The shortlisted potent isolates were subjected to quantitative estimation of PGP activities like phosphate solubilization, IAA and siderophore production. The strains could produce significant amount of IAA and siderophore and solubilize phosphate. Molecular characterization by 16S rDNA sequence analyses revealed the identity of the isolates: CR12: Bacillus subtilis (Accn. No.OM866257), CW11: Bacillus paralicheniformis (Accn. No.OM868047), CA2: Bacillus sp. CCMB1014 (Accn. No.OM868070) and CP2: Bacillus licheniformis (Accn. No.OM892495). Seedling vigor assays of the shortlisted potent strains were carried out on Chakhao Amubi rice. The isolates exhibited higher seedling vigor indices (CR12: 668.00, CA2: 618.66, CP2: 510.92 and CW11: 478.91) over the control (164.48). These rhizospheric strains have the potential to be developed as bioinoculants or biostimulants for enhancing Chakhao rice growth. [ABSTRACT FROM AUTHOR]

It is evident that legume root nodules can accommodate rhizobial and non-rhizobial bacterial endophytes. Our recent nodule microbiome study in peanuts described that small nodules can harbor diverse bacterial endophytes. To understand their functional role, we isolated 87 indigenous endophytes from small nodules of field-grown peanut roots and characterized them at molecular, biochemical, and physiological levels. The amplified 16S rRNA genes and phylogenetic analysis of these isolates revealed a wide variety of microorganisms related to the genera Bacillus, Burkholderia, Enterobacter, Herbaspirillum, Mistsuaria, Pantoea, Pseudomonas, and Rhizobia. It was observed that 37% (100% identity) and 56% (>99% identity) of the isolates matched with the amplified sequence variants (ASVs) from our previous microbiome study. All of these isolates were tested for stress tolerance (high temperature, salinity, acidic pH) and phosphate (P) solubilization along with ammonia (NH3), indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylate deaminase (ACCD), and siderophore production. The majority (78%) of the isolates were found to be halotolerant, thermotolerant, and acidophilic, and a few of them showed a significant positive response to the production of IAA, NH3, siderophore, ACCD, and P-solubilization. To evaluate the plant growth promotion (PGP) activity, plant and nodulation assays were performed in the growth chamber conditions for the selected isolates from both the non-rhizobial and rhizobial groups. However, these isolates appeared to be non-nodulating in the tested conditions. Nonetheless, the isolates 2 (Pantoea), 17 (Burkholderia), 21 (Herbaspirillum), 33o (Pseudomonas), and 77 (Rhizobium sp.) showed significant PGP activity in terms of biomass production. Our findings indicate that these isolates have potential for future biotechnological applications through the development of biologicals for sustainable crop improvement. [ABSTRACT FROM AUTHOR]

Naturally, all plants harbor a diversity of endophytes and can produce a variety of compounds which positively affect the growth of their host plant. Endophytes can contribute in providing plant supplements, conflicting with pathogens, or directly influencing the growth and development of plants through synthesis of phytohormones. The present study aimed to explore the diversity of endophytic fungi from different parts of the three healthy halophytic plants i.e., Solanum surattense Burm. f., Chenopodium album L. and Zizyphus nummularia (Burm. f). As sa result, twenty-two endophytic fungi mostly belonging to ascomycetes were isolated from 92 samples with an overall colonization frequency of 23.02%. The endophytic fungi showed a relatively high percentage of colonization frequency (25%) in roots as compared to other parts. The isolated fungal strains were tested for salt tolerance and for their ability to produce plant hormone indole-3-acetic acid (IAA). Out of 12 different strains, five strains have salt tolerance potential and eight strains have shown to produce IAA in culture media. Further the isolated strains were also screened for their growth promoting activities such as 1-aminocyclopropane1-carboxylate deaminase, flavonoids, phenol, solubilize phosphate and to produce siderophores. [ABSTRACT FROM AUTHOR]

Abstract Law enforcement agencies struggle with criminals using end-to-end encryption (E2EE). A recent policy paper states: “while encryption is vital and privacy and cyber security must be protected, that should not come at the expense of wholly precluding law enforcement”. The main argument is that E2EE hampers attribution and prosecution of criminals who rely on encrypted communication - ranging from drug syndicates to child sexual abuse material (CSAM) platforms. This statement - in policy circles dubbed ‘going dark’ - is not yet supported by empirical evidence. That is why, in our work, we analyse public court data from the Netherlands to show to what extent law enforcement agencies and the public prosecution service are impacted by the use of E2EE in bringing cases to court and their outcome. Our results show that in cases brought to court, the Dutch courts appear to be as successful in convicting offenders who rely on E2EE as those who do not. Our data do not permit us to draw conclusions on the effect of E2EE on criminal investigations.

Multidrug resistance (MDR) is the cell resistance to a number of drugs. Proteins from the ABC-transporter family are the most studied factors of the MDR phenomenon. According to the literature Pgp, MRP1, BCRP, and LRP are proteins involved in the development of the MDR. The authors detected the expression of these proteins in untreated patients with acute myeloid leukemia (other than M3). Expression of these proteins was observed in 64.3%, 46.4%, 64.3%, and 42.9% of patients, respectively. In all non-responders blast cells expressed more than one marker of MDR. Expression of 3 and 4 markers was found in 80% of non-responders while in responders 1—2 markers were expressed in 83%.

The indiscriminate and rampant use of pesticides has raised serious concerns regarding their toxic impact on non-target organisms which underlines need for the development of an effective antidote. Metabolic activation of organophosphate pesticides by the phase I enzyme, cytochrome P450 plays a key role in influencing pesticide-toxicity. In this study, we have investigated the effect of environmentally relevant malathion concentration (100 μg/L) alone and in combination with vitamin B12 on the expression of genes related to xenobiotic metabolism such as CYP enzymes, PgP and the key oxidative stress responsive transcription factor, Nrf2 in zebra fish liver and brain. Expressions of Nrf2-trasncribed antioxidant genes and their activities were also measured. Administration of vitamin B12 successfully revived motor functions by modulation of AchE activity. Mechanistically, vitamin B12 was demonstrated to alleviate oxidative stress which was accompanied by decreased phase-I enzyme cyp3c1 and increased pgp expressions. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungal biocides are preferred for environment friendly sustainable management of insect pests due to their host specificity and harmlessness to non-target insects. Plant growth promotion (PGP) functions of the entomofungi are also important attributes but hitherto insignificantly explored. Therefore, virulence of 17 natural fungal entomocides (Cordyceps, Beauveria, Metarhizium, Nomuraea, Fusarium, Verticillium, Trichoderma and Paecilomyces spp.) were evaluated for pathogenicity against five rice pests (brown plant hopper (Nilaparvata lugens) and green leaf hopper (Nephotettix virescens) nymphs, leaf folder (Cnaphalocrosis medinalis) and yellow stem borer (Scirpophaga incertulas) larvae and swarming caterpillar (Spodoptera mauritia), respectively), and PGP traits of the potent leaf folder pathogens. Among the fungi, only the leaf folder pathogens (3 isolates each of Beauveria and Metarhizium spp.) infected > 50% (80–90%) larvae but other fungi were ineffective as infected < 50% (0–47%) insects. Besides, the leaf folder pathogens exhibited diverse PGP traits such as organic/inorganic phosphate solubilization (104.7–236.4 µg/ml), and siderophore, ammonia, hydrogen cyanide (HCN), indole production etc. Restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD), simple sequences repeat (SSR) and internal transcribed spacers (ITS) analysis ascertained strain identity and genetic (inter and intra-specific) diversity among the potent biocides Beauveria and Metarhizium spp. The virulent natural fungal pathogens of rice pests with polyvalent PGP traits may be prospected for rice growth promotion and biocontrol of leaf folder. [ABSTRACT FROM AUTHOR]

Het vertrouwensbeginsel: een heet hangijzer. Over het interstatelijke vertrouwensbeginsel bij het JIT in de SkyECC-operatie In december 2022 stelde de rechtbank Noord-Nederland de vraag of het interstatelijke vertrouwensbeginsel zonder meer van toepassing is op bewijs verkregen door de inzet van een opsporingsmiddel in het buitenland, in het kader van een JIT waarbij Nederland partner is. In deze bijdrage wordt bepleit dat die vraag ontkennend dient te worden beantwoord met het oog op het Stojkovic-arrest van het EHRM, nu sprake is van een dermate intensieve samenwerking in het kader van het JIT. Tevens wordt uiteengezet wat de gevolgen zijn van het loslaten van het vertrouwensbeginsel. [ABSTRACT FROM AUTHOR]

The Earth is undergoing alterations at a high speed, which causes problems such as environmental pollution and difficulty in food production. This is where halophytes are interesting, due to their high potential in different fields, such as remediation of the environment and agriculture. For this reason, it is necessary to deepen the knowledge of the development of halophytes and how plant growth-promoting bacteria (PGP) can play a fundamental role in this process. Therefore, in this work were tested the effects of five PGP bacteria on its rhizosphere and other endophytic bacteria at different concentrations of NaCl on seed germination, plant growth (0 and 171 mM) and cutting growth (0 mM) of Arthrocaulon macrostachyum. The growth promotion in this strict halophyte is highlighted due to the presence of PGP bacteria and the fact that no salt is needed. Thus, without salt, the bacterial strains Kocuria polaris Hv16, Pseudarthrobacter psychrotolerans C58, and Rahnella aceris RTE9 enhanced the biomass production by more than 60% in both stems and roots. Furthermore, germination was encouraged by more than 30% in the presence of both R. aceris RTE9 and K. polaris Hv16 at 171 mM NaCl; the latter also had a biocontrol effect on the fungi that grew on the seeds. Additionally, for the first time in cuttings of this perennial species, the root biomass was improved thanks to the consortium of K. polaris Hv16 and P. psychrotolerans C58. Finally, this study demonstrates the potential of PGPs for optimising the development of halophytes, either for environmental or agronomic purposes. [ABSTRACT FROM AUTHOR]

Law enforcement agencies struggle with criminals using end-to-end encryption (E2EE). A recent policy paper states: "while encryption is vital and privacy and cyber security must be protected, that should not come at the expense of wholly precluding law enforcement". The main argument is that E2EE hampers attribution and prosecution of criminals who rely on encrypted communication - ranging from drug syndicates to child sexual abuse material (CSAM) platforms. This statement - in policy circles dubbed 'going dark' - is not yet supported by empirical evidence. That is why, in our work, we analyse public court data from the Netherlands to show to what extent law enforcement agencies and the public prosecution service are impacted by the use of E2EE in bringing cases to court and their outcome. Our results show that in cases brought to court, the Dutch courts appear to be as successful in convicting offenders who rely on E2EE as those who do not. Our data do not permit us to draw conclusions on the effect of E2EE on criminal investigations. [ABSTRACT FROM AUTHOR]

Seeds harbor naturally occurring microbial endophytes that proliferate during seedling development; playing crucial roles in seedling growth, establishment, and protection against fungal pathogens. Resilient actinobacteria of wheat seeds have been explored in this study for their beneficial traits. Ten actinobacteria isolated from the surface‐sterilized seeds of wheat variety HD3117 were identified as nine species of Streptomyces and one of Nocardiopsis. Most isolates could grow at 42°C, 5% NaCl, and 10% poly ethylene glycol (PEG); exhibited variable hydrolytic enzyme production for amylase, cellulase, and protease. Few isolates produced indole acetic acid (9.0–18.9 µg ml−1) and could solubilize P (11.3–85.2 µg ml−1). The isolates were antagonistic against one or more fungal pathogens under test (Fusarium graminearum, Bipolaris sorokiniana, Alternaria sp., and Tilletia indica), of which Streptomyces sampsonii WSA20 inhibited all in dual culture assay. Priming of wheat seeds with the efficient isolate WSA20 led to effective colonization in the root zone and significantly improved germination, shoot and root length in seed germination assay. Significant protection was recorded in microcosm experiment where no symptoms of disease were observed. This study shows the significance of actinobacterial endophytes of wheat seeds in influencing seed germination and seedling growth while protecting from soil‐borne pathogens. It is original and suggests that the seed inhabiting efficient actinobacteria may be developed as efficient bioinoculant for sustainable farming system. [ABSTRACT FROM AUTHOR]

The inoculation of plants with plant-growth-promoting microorganisms (PGPM) (i.e., bacterial and fungal strains) is an emerging approach that helps plants cope with abiotic and biotic stresses. However, knowledge regarding their synergic effects on plants growing in metal-rich soils is limited. Consequently, the aim of this study was to investigate the biomass, ecophysiology, and metal accumulation of the facultative Ni-hyperaccumulator Alyssoides utriculata (L.) Medik. inoculated with single or mixed plant-growth-promoting (PGP) bacterial strain Pseudomonas fluorescens Migula 1895 (SERP1) and PGP fungal strain Penicillium ochrochloron Biourge (SERP03 S) on native serpentine soil (n = 20 for each treatment). Photosynthetic efficiency (Fv/Fm) and performance indicators (PI) had the same trends with no significant differences among groups, with Fv/Fms > 1 and PI up to 12. However, the aboveground biomass increased 4–5-fold for single and mixed inoculated plants. The aboveground/belowground dry biomass ratio was higher for plants inoculated with fungi (30), mixed (21), and bacteria (17). The ICP-MS highlighted that single and mixed inocula were able to double the aboveground biomass' P content. Mn metal accumulation significantly increased with both single and mixed PGP inocula, and Zn accumulation increased only with single PGP inocula, whereas Cu accumulation increased twofold only with mixed PGP inocula, but with a low content. Only Ni metal accumulation approached the hyperaccumulation level (Ni > 1000 mg/kg DW) with all treatments. This study demonstrated the ability of selected single and combined PGP strains to significantly increase plant biomass and plant tolerance of metals present in the substrate, resulting in a higher capacity for Ni accumulation in shoots. [ABSTRACT FROM AUTHOR]

Ononis repens is a legume of the Fabaceae family with great ecological and medicinal importance that grows wild in the Maamora cork oak forest (Rabat, Morocco). The aim of this work was to analyze the diversity, phylogeny, and plant growth-promoting features of the plant root nodules microsymbionts. Out of twenty nodC-containing rhizobial bacteria, four strains, ORM6, ORM8.1, ORM 13.1, and ORM16, were selected for further molecular and phenotypic analyses. All four strains were able to metabolize a wide range of carbohydrates and amino acids as sole carbon and nitrogen sources, respectively, and grew in the presence of different pH, NaCl, and temperature conditions. They also solubilized inorganic phosphate and produced siderophores, and ORM16 was the only strain to produce indol acetic acid. The rrs gene sequence analysis showed that the four strains are members of the Mesorhizobium genus. The individual and concatenated sequences of the recA, glnII, and gyrB housekeeping genes revealed that the strains ORM6 and ORM 13.1 clustered with M. jarvisii ATCC 33,669T and M. intechii BD68 T, with which they share 93.3 and 93.7% similarity, respectively; that the strain ORM8.1 affiliated with a group of which the strain M. shonense AC39aT was the closest relative species with 84.3% similarity; and that the strain ORM16 clustered with M. australicum LMG24608T, with 80.5% similarity. These results suggest that the four O. repens-isolated strains could belong to three putative new genospecies. All four strains nodulated their original host as well as Astragalus gombiformis and A. armatus, but only the strain ORM16 was able to form nodules on Vachellia gummifera. Analysis of the symbiotic nodC gene phylogenies showed that the strains ORM6, ORM8.1, ORM 13.1, and ORM16 formed a group apart from the known symbiovars in the genus Mesorhizobium, for which the name maamori is proposed. [ABSTRACT FROM AUTHOR]

Serine/threonine kinase 11 (STK11) deletion and downregulation caused cancer progression, and were widely associated with drug resistance. Accurate screening of natural small molecules about anti-cancer and anti-drug resistance is the key to the development and utilization of natural product application, which could promote traditional Chinese medicine in the treatment of cancer. Dauricine, which is derived from the rhizome of Menispermum dauricum DC., has certain potential but unexplored mechanism for the treatment of cancer. The aim of this study was to screen and validate the role and mechanism of natural STK11 agonists with anti-drug resistance from plants in the treatment of NSCLC. A lentiviral STK11 overexpression cell model was employed for the screening of natural STK11 agonists. The efficacy of dauricine in the treatment of NSCLC was validated on PC-9 and HCC827 cells. In vivo validation of dauricine activity was performed using nude mouse models equipped with PC9 xenografts. To investigate the anti-resistant effects of dauricine, gefitinib-resistant PC9 cell models were constructed. As a natural agonist of STK11, it causes the activation of the STK11/AMPK pathway and inhibits the growth of PC-9 cells. Dauricine synergises the inhibitory effect with gefitinib on PC9. The up-regulation of STK11 protein expression by dauricine was demonstrated in vitro and in vivo , while restoring the sensitivity of PC9/GR to gefitinib by down-regulating the protein expression of Nrf2 and Pgp. Dauricine, a natural agonist of STK11, effectively inhibited NSCLC, and its combination treatment with gefitinib reversed drug-resistant NSCLC. [Display omitted] • Dauricine was found to be a natural STK11 agonist. • Dauricine activated STK11 and inhibited the growth of PC9 xenografts. • Dauricine regulated energy metabolism in PC9 cells. • Dauricine restored PC9/GR resistance by downregulating Nrf2 and Pgp expression. • Combination therapy with Dauricine improved gefitinib exposure in resistant tumors. [ABSTRACT FROM AUTHOR]

Endophytic actinobacteria aid in plant development and disease resistance by boosting nutrient uptake or producing secondary metabolites. For the first time, we investigated the culturable endophytic actinobacteria associated with ten epiphytic orchid species of Assam, India. 51 morphologically distinct actinobacteria were recovered from surface sterilized roots and leaves of orchids and characterized based on different PGP and antifungal traits. According to the 16S rRNA gene sequence, these isolates were divided into six families and eight genera, where Streptomyces was most abundant (n=29, 56.86%), followed by Actinomadura, Nocardia, Nocardiopsis, Nocardioides, Pseudonocardia, Microbacterium, and Mycolicibacterium. Regarding PGP characteristics, 25 (49.01%) isolates demonstrated phosphate solubilization in the range of 61.1±4.4 - 289.7±11.9 µg/ml, whereas 27 (52.94%) isolates biosynthesized IAA in the range of 4.0 ± 0.08 - 43.8 ± 0.2 µg/ml, and 35 (68.62%) isolates generated ammonia in the range of 0.9 ± 0.1 - 5.9 ± 0.2 µmol/ ml. These isolates also produced extracellular enzymes, viz. protease (43.13%), cellulase (23.52%), pectinase (21.56%), ACC deaminase (27.45%), and chitinase (37.25%). Out of 51 isolates, 27 (52.94%) showed antagonism against at least one test phytopathogen. In molecular screening, most isolates with antifungal and chitinase producing traits revealed the presence of 18 family chitinase genes. Two actinobacterial endophytes, Streptomyces sp. VCLA3 and Streptomyces sp. RVRA7 were ranked as the best strains based on PGP and antifungal activity on bonitur scale. GC-MS examination of ethyl acetate extract of these potent strains displayed antimicrobial compound phenol, 2,4-bis-(1,1-dimethylethyl) as the major metabolite along with other antifungal and plant growth beneficial bioactive chemicals. SEM analysis of fungal pathogen F. oxysporum (MTCC 4633) affected by Streptomyces sp. VCLA3 revealed significant destruction in the spore structure. An in vivo plant growth promotion experiment with VCLA3 and RVRA7 on chili plants exhibited statistically significant (p<0.05) improvements in all of the evaluated vegetative parameters compared to the control. Our research thus gives insight into the diversity, composition, and functional significance of endophytic actinobacteria associated with orchids. This research demonstrates that isolates with multiple plant development and broad-spectrum antifungal properties are beneficial for plant growth. They may provide a viable alternative to chemical fertilizers and pesticides and a sustainable solution for chemical inputs in agriculture. [ABSTRACT FROM AUTHOR]

Glyprolines are Gly-Pro (GP)- or Pro-Gly (PG)-containing biogenic peptides. These peptides can act as neutrophil chemoattractants, or atheroprotective, anticoagulant, and neuroprotective agents. The Pro-Gly-Pro (PGP) tripeptide is an active factor of resistance to the biodegradation of peptide drugs. The synthetic Semax peptide, which includes Met-Glu-His-Phe (MEHF) fragments of adrenocorticotropic hormone and the C-terminal tripeptide PGP, serves as a neuroprotective drug for the treatment of ischemic stroke. Previously, we revealed that Semax mostly prevented the disruption of the gene expression pattern 24 h after a transient middle cerebral artery occlusion (tMCAO) in a rat brain model. The genes of this pattern were grouped into an inflammatory cluster (IC) and a neurotransmitter cluster (NC). Here, using real-time RT-PCR, the effect of other PGP-containing peptides, PGP and Pro-Gly-Pro-Leu (PGPL), on the expression of a number of genes in the IC and NC was studied 24 h after tMCAO. Both the PGP and PGPL peptides showed Semax-unlike effects, predominantly without changing gene expression 24 h after tMCAO. Moreover, there were IC genes (iL1b, iL6, and Socs3) for PGP, as well as IC (iL6, Ccl3, Socs3, and Fos) and NC genes (Cplx2, Neurod6, and Ptk2b) for PGPL, that significantly changed in expression levels after peptide administration compared to Semax treatment under tMCAO conditions. Furthermore, gene enrichment analysis was carried out, and a regulatory gene network was constructed. Thus, the spectra of the common and unique effects of the PGP, PGPL, and Semax peptides under ischemia–reperfusion were distinguished. [ABSTRACT FROM AUTHOR]

Salinity is one of the most serious problems causing desertification of agricultural lands and decreasing agricultural yield. Recently, one of the most promising approaches is the application of plant growth promoting (PGP) bacteria to ameliorate agricultural lands and increase agricultural yield. In the current study, it was aimed to evaluate the PGP activities of fifteen halophilic/halotolerant bacteria. The production of siderophore, HCN, IAA and ACC deaminase, nitrogen fixation and phosphate solubilisation of bacteria were determined either qualitatively or quantitatively. Besides, four bacterial isolates with the best PGP traits [Halomonas sp. (H2), Bacillus licheniformis (H3), Halobacillus sp. (H5), and Oceanobacillus sp. (H10)] among these fifteen bacterial isolates were inoculated to the wheat under saline condition (NaCl) (200 mM) stress in a hydroponic system. Under salt stress, all of the bacterial strains tested enhanced plant growth compared to the un-inoculated group. While the dry mass (%) of the plant under salt stress was 42.31, the plant dry mass inoculated by bacterial isolates ranged between 87.98 and 107.50. In the current study, ACC deaminase activity of Chromohalobacter sp. and plant (wheat) growth enhancement potential of Oceanobacillus sp. were reported for the first time. Consequently, the bacteria isolated from Sanlıurfa, Turkey have significant potential to ameliorate salt stress and increase plant yield. [ABSTRACT FROM AUTHOR]

Endophytic actinobacteria aid in plant development and disease resistance by boosting nutrient uptake or producing secondary metabolites. For the first time, we investigated the culturable endophytic actinobacteria associated with ten epiphytic orchid species of Assam, India. 51 morphologically distinct actinobacteria were recovered from surface sterilized roots and leaves of orchids and characterized based on different PGP and antifungal traits. According to the 16S rRNA gene sequence, these isolates were divided into six families and eight genera, where Streptomyces was most abundant (n=29, 56.86%), followed by Actinomadura, Nocardia, Nocardiopsis, Nocardioides, Pseudonocardia, Microbacterium, and Mycolicibacterium. Regarding PGP characteristics, 25 (49.01%) isolates demonstrated phosphate solubilization in the range of 61.1±4.4 - 289.7±11.9 µg/ml, whereas 27 (52.94%) isolates biosynthesized IAA in the range of 4.0 ± 0.08 - 43.8 ± 0.2 µg/ml, and 35 (68.62%) isolates generated ammonia in the range of 0.9 ± 0.1 - 5.9 ± 0.2 µmol/ml. These isolates also produced extracellular enzymes, viz. protease (43.13%), cellulase (23.52%), pectinase (21.56%), ACC deaminase (27.45%), and chitinase (37.25%). Out of 51 isolates, 27 (52.94%) showed antagonism against at least one test phytopathogen. In molecular screening, most isolates with antifungal and chitinase producing traits revealed the presence of 18 family chitinase genes. Two actinobacterial endophytes, Streptomyces sp. VCLA3 and Streptomyces sp. RVRA7 were ranked as the best strains based on PGP and antifungal activity on bonitur scale. GC-MS examination of ethyl acetate extract of these potent strains displayed antimicrobial compound phenol, 2,4-bis-(1,1-dimethylethyl) as the major metabolite along with other antifungal and plant growth beneficial bioactive chemicals. SEM analysis of fungal pathogen F. oxysporum (MTCC 4633) affected by Streptomyces sp. VCLA3 revealed significant destruction in the spore structure. An in vivo plant growth promotion experiment with VCLA3 and RVRA7 on chili plants exhibited statistically significant (p

It is evident that legume root nodules can accommodate rhizobial and non-rhizobial bacterial endophytes. Our recent nodule microbiome study in peanuts described that small nodules can harbor diverse bacterial endophytes. To understand their functional role, we isolated 87 indigenous endophytes from small nodules of field-grown peanut roots and characterized them at molecular, biochemical, and physiological levels. The amplified 16S rRNA genes and phylogenetic analysis of these isolates revealed a wide variety of microorganisms related to the genera Bacillus, Burkholderia, Enterobacter, Herbaspirillum, Mistsuaria, Pantoea, Pseudomonas, and Rhizobia. It was observed that 37% (100% identity) and 56% (>99% identity) of the isolates matched with the amplified sequence variants (ASVs) from our previous microbiome study. All of these isolates were tested for stress tolerance (high temperature, salinity, acidic pH) and phosphate (P) solubilization along with ammonia (NH3), indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylate deaminase (ACCD), and siderophore production. The majority (78%) of the isolates were found to be halotolerant, thermotolerant, and acidophilic, and a few of them showed a significant positive response to the production of IAA, NH3, siderophore, ACCD, and P-solubilization. To evaluate the plant growth promotion (PGP) activity, plant and nodulation assays were performed in the growth chamber conditions for the selected isolates from both the non-rhizobial and rhizobial groups. However, these isolates appeared to be non-nodulating in the tested conditions. Nonetheless, the isolates 2 (Pantoea), 17 (Burkholderia), 21 (Herbaspirillum), 33o (Pseudomonas), and 77 (Rhizobium sp.) showed significant PGP activity in terms of biomass production. Our findings indicate that these isolates have potential for future biotechnological applications through the development of biologicals for sustainable crop improvement.

The primary objective of this study was to examine the effect of gestational diabetes mellitus (GDM) on pelvic girdle pain (PGP) occurrence and symptom severity. Pregnant women who were with/without GDM, 20–40 years of age, and also in the second and third trimesters of pregnancy were included in the study. PGP provocation tests were administered to 187 pregnant women to determine the presence and severity of PGP. Based on the test results, the study subjects were divided into two groups; Group 1 (GDM+, PGP+; n:32) and Group 2 (GDM−, PGP+; n:35). Both groups were asked to fill in the Pelvic Girdle Questionnaire (PGQ). The relationship between the presence of GDM and the presence of PGP was found to be significant (p =.043). It was found the groups were similar in view of pain, and also in PGQ total/subscale scores (p >.05). Although GDM has no effect on symptom severity, it has been determined that it may relate to the development of PGP. Therefore, early interventions (nutrition, exercise, belt using, etc.) are recommended to prevent the development of PGP in pregnant women with a family history of diabetes, with a previous diagnosis of diabetes and/or with GDM detected in their previous pregnancies. Clinical Trial Number: 04769375 What is already known on this subject? Gestational diabetes mellitus and pelvic girdle pain are pathologies that develops secondary to pregnancy-related systemic and biomechanical changes. What do results on this study add? GDM may related the development of PGP. What are the implications of these findings for clinical practice and/or further research? Early interventions (nutrition, exercise, belt using, etc.) and strict control of pregnant women in view of PGP is recommended to prevent the development of PGP in pregnant women with a family history of diabetes, with previous diagnosis of diabetes and/or with GDM detected in their previous pregnancies. The evaluation of pregnant women for PGP before administering interventions, such as exercise and diet (both decrease the pro-inflammatory markers), following the diagnosis of GDM and the measurement of plasma anti- and pro-inflammatory marker values in the same time period will further reveal the relationship between GDM and PGP. [ABSTRACT FROM AUTHOR]

Assessment of compound permeability through a Caco-2 cell monolayer is a well-accepted model to evaluate its in-vivo permeability potential and transporter interaction. While this assay has commonly been conducted using a 24-well assay plate format, a miniaturised 96-well assay format is highly desirable to achieve greater capacity and higher efficiency. Previous attempts to convert this assay from 24-well to 96-well format at our lab, however, had met with varied efflux capacities and unacceptable efflux ratios for digoxin, a substrate of P-glycoprotein (Pgp), which indicated inadequate Pgp transporter expression in the 96-well format. These challenges in converting the assays were attributed to the heterogeneous and unstable nature of the Caco-2 cells. To overcome the challenges, single-cell sorting of Caco-2 cells was conducted by flow cytometry to obtain a more homogeneous and stable cell population. The sorted cells were then seeded to 96-well transwell plates and the Pgp expression under various cell culture conditions was monitored by a LC-MS/MS-based targeted proteomics method. Through cell sorting and direct Pgp expression measurement, Caco-2 cells with adequate and sustained Pgp expression in a 96-well format were obtained, which led to the successful development and implementation of a 96-well Caco-2 assay with significant efficiency gain and faster turnaround time than the historical 24-well assay. [ABSTRACT FROM AUTHOR]

Leukotriene A 4 hydrolase (LTA 4 H) is a bifunctional enzyme, with dual activities critical in defining the scale of tissue inflammation and pathology. LTA 4 H classically operates intracellularly, primarily within myeloid cells, to generate pro-inflammatory leukotriene B 4. However, LTA 4 H also operates extracellularly to degrade the bioactive collagen fragment proline-glycine-proline to limit neutrophilic inflammation and pathological tissue remodeling. While the dichotomous functions of LTA 4 H are dictated by location, the cellular source of extracellular enzyme remains unknown. We demonstrate that airway extracellular LTA 4 H concentrations are governed by the level of pulmonary vascular permeability and influx of an abundant repository of blood-borne enzyme. In turn, blood LTA 4 H originates from liver hepatocytes, being released constitutively but further upregulated during an acute phase response. These findings have implications for our understanding of how inflammation and repair are regulated and how perturbations to the LTA 4 H axis may manifest in pathologies of chronic diseases. [Display omitted] • LTA 4 H operates intracellularly to generate LTB 4 and extracellularly to degrade PGP • The previously unknown origin of extracellular LTA 4 H was shown to be liver hepatocytes • Constitutive LTA 4 H release into blood is enhanced during an acute phase response • Extracellular airway LTA 4 H levels are dictated by changes in vascular permeability Mincham et al. define pathways governing the bioavailability of extracellular leukotriene A 4 hydrolase (LTA 4 H). LTA 4 H's location defines its function, whereby it generates leukotriene B 4 intracellularly but degrades proline-glycine-proline extracellularly. Airway extracellular LTA 4 H concentrations are governed by changes in vascular permeability and influx of blood-borne enzyme, which is in turn secreted by liver hepatocytes. [ABSTRACT FROM AUTHOR]

Rice is a symbol of life and a representation of prosperity in South Korea. However, studies on the diversity of the bacterial communities in the rhizosphere of rice plants are limited. In this study, four bundles of root samples were collected from the same rice field located in Goyang, South Korea. These were systematically analyzed to discover the diversity of culturable bacterial communities through culture-dependent methods. A total of 504 culturable bacteria were isolated and evaluated for their plant growth-promoting abilities in vitro. Among them, Arthrobacter sp. GN70 was selected for inoculation into the rice plants under laboratory and greenhouse conditions. The results showed a significantly positive effect on shoot length, root length, fresh plant weight, and dry plant weight. Moreover, scanning electron microscopic (SEM) images demonstrated the accumulation of bacterial biofilm networks at the junction of the primary roots, confirming the root-colonizing ability of the bacterium. The strain also exhibited a broad spectrum of in vitro antimicrobial activities against bacteria and fungi. Here, we first report the rice plant growth-promoting ability of the Arthrobacter species with the biofilm-producing and antimicrobial activities against plant and human pathogens. Genome analyses revealed features attributable to enhance rice plant growth, including the genes involved in the synthesis of plant hormones, biofilm production, and secondary metabolites. This study revealed that the rhizobacteria isolated from the roots of rice plants have dual potential to be utilized as a plant growth promoter and antimicrobial agent. [ABSTRACT FROM AUTHOR]

The present study aimed to demonstrate the use of two strains of Streptomyces albus (CAI-24 and KAI-27) and one strain of S. griseus (MMA-32) for plant growth-promotion (PGP) and improving pearl millet yield and nutrient content under greenhouse and field conditions. Two hybrids, a low-Fe (PA-9444; non-biofortified hybrid) and high-Fe (ICMH-1201; biofortified hybrid), treated with selected Streptomyces strains, significantly enhanced a range of traits including grain yield in the glasshouse (13–23%) and field (9–12%) over the control. In the greenhouse experiments an enhanced stover and grain nutrient concentrations were observed in ICMH-1201 (Fe 53% and 40%; Zn 15% and 10%; Ca 11% and 29%) over the control, while such nutrition augmentations were not found in PA-9444. The field harvested stover and grain nutrient concentrations were also increased over the control in both hybrids. A higher stover nutrient concentration was found in ICMH-1201 while PA-9444 had an increase in grain nutrient concentration indicating the significance of these Streptomyces strains' PGP role in the non-biofortified hybrid. Based on this study, strains KAI-27 and MMA-32 significantly improved shoot weight, root weight and grain yield while CAI-24 and MMA-32 improved nutrient concentrations including Fe contents (up to 49%) in grain as well as in stover. Further, the stover samples of pearl millet contained a higher Fe concentration (150–200%) compared to grain samples. This study confirms that the selected Streptomyces strains have the potential for enhancing PGP and stover and grain nutrient concentrations in pearl millet and can complement the existing conventional biofortification strategies. Biofortification of staple food crops with essential micronutrients has emerged as a sustainable solution to diminish malnutrition, especially in developing countries. This study showed PGP strains (Streptomyces) enhanced yield and nutritional traits (Fe, Zn) of pearl millet hybrids under greenhouse and field conditions. This study suggests the use of selected plant-growth promoters for enhancing stover and grain yield and its nutrient concentrations in pearl millet and can complement the existing conventional biofortification strategies. [ABSTRACT FROM AUTHOR]

Abstract Background Adenosine triphosphate binding cassette transporters such as P-glycoprotein (PGP) play an important role in drug pharmacokinetics by actively effluxing their substrates at barrier interfaces, including the blood-brain, blood-cerebrospinal fluid (CSF) and placental barriers. For a molecule to access the brain during fetal stages it must bypass efflux transporters at both the placental barrier and brain barriers themselves. Following birth, placental protection is no longer present and brain barriers remain the major line of defense. Understanding developmental differences that exist in the transfer of PGP substrates into the brain is important for ensuring that medication regimes are safe and appropriate for all patients. Methods In the present study PGP substrate rhodamine-123 (R123) was injected intraperitoneally into E19 dams, postnatal (P4, P14) and adult rats. Naturally fluorescent properties of R123 were utilized to measure its concentration in blood-plasma, CSF and brain by spectrofluorimetry (Clariostar). Statistical differences in R123 transfer (concentration ratios between tissue and plasma ratios) were determined using Kruskal-Wallis tests with Dunn’s corrections. Results Following maternal injection the transfer of R123 across the E19 placenta from maternal blood to fetal blood was around 20 %. Of the R123 that reached fetal circulation 43 % transferred into brain and 38 % into CSF. The transfer of R123 from blood to brain and CSF was lower in postnatal pups and decreased with age (brain: 43 % at P4, 22 % at P14 and 9 % in adults; CSF: 8 % at P4, 8 % at P14 and 1 % in adults). Transfer from maternal blood across placental and brain barriers into fetal brain was approximately 9 %, similar to the transfer across adult blood-brain barriers (also 9 %). Following birth when placental protection was no longer present, transfer of R123 from blood into the newborn brain was significantly higher than into adult brain (3 fold, p

Understanding plant-microbe interactions with the possibility to modulate the plant's microbiome is essential to design new strategies for a more productive and sustainable agriculture and to maintain natural ecosystems. Therefore, a key question is how to design bacterial consortia that will yield the desired host phenotype. This work was designed to identify the potential genomic features involved in the interaction between Micromonospora and known host plants. Seventy-four Micromonospora genomes representing diverse environments were used to generate a database of all potentially plant-related genes using a novel bioinformatic pipeline that combined screening for microbial-plant related features and comparison with available plant host proteomes. The strains were recovered in three clusters, highly correlated with several environments: plant-associated, soil/rhizosphere, and marine/mangrove. Irrespective of their isolation source, most strains shared genes coding for commonly screened plant growth promotion features, while differences in plant colonization related traits were observed. When Arabidopsis thaliana plants were inoculated with representative Micromonospora strains selected from the three environments, significant differences were in found in the corresponding plant phenotypes. Our results indicate that the identified genomic signatures help select those strains with the highest probability to successfully colonize the plant and contribute to its wellbeing. These results also suggest that plant growth promotion markers alone are not good indicators for the selection of beneficial bacteria to improve crop production and the recovery of ecosystems. [ABSTRACT FROM AUTHOR]