Optimisation of indole acetic acid production by Neopestalotiopsis aotearoa endophyte isolated from Thymus vulgaris and its impact on seed germination of Ocimum basilicum.

BACKGROUND: Microbial growth during plant tissue culture is a common problem that causes significant losses in the plant micro-propagation system. Most of these endophytic microbes have the ability to propagate through horizontal and vertical transmission. On the one hand, these microbes provide a rich source of several beneficial metabolites. RESULTS: The present study reports on the isolation of fungal species from different in vitro medicinal plants (i.e., Breynia disticha major, Breynia disticha, Duranta plumieri, Thymus vulgaris, Salvia officinalis, Rosmarinus officinalis, and Ocimum basilicum l) cultures. These species were tested for their indole acetic acid (IAA) production capability. The most effective species for IAA production was that isolated from Thymus vulgaris plant (11.16 µg/mL) followed by that isolated from sweet basil plant (8.78 µg/mL). On screening for maximum IAA productivity, medium, "MOS + tryptophan" was chosen that gave 18.02 µg/mL. The macroscopic, microscopic examination and the 18S rRNA sequence analysis indicated that the isolate that given code T4 was identified as Neopestalotiopsis aotearoa (T4). The production of IAA by N. aotearoa was statistically modeled using the Box-Behnken design and optimized for maximum level, reaching 63.13 µg/mL. Also, IAA extract was administered to sweet basil seeds in vitro to determine its effect on plant growth traits. All concentrations of IAA extract boosted germination parameters as compared to controls, and 100 ppm of IAA extract exhibited a significant growth promotion effect for all seed germination measurements. CONCLUSIONS: The IAA produced from N. aotearoa (T4) demonstrated an essential role in the enhancement of sweet basil (Ocimum basilicum) growth, suggesting that it can be employed to promote the plant development while lowering the deleterious effect of using synthetic compounds in the environment.

The Nurr1 ligand indole acetic acid hydrazide loaded onto ZnFe2O4 nanoparticles suppresses proinflammatory gene expressions in SimA9 microglial cells.

The nuclear receptor-related factor 1 (Nurr1), an orphan nuclear receptor in microglia, has been recognized as a major player in attenuating the transcription of the pro-inflammatory genes to maintain CNS homeostasis. In this study, we investigate Nurr1 trans-repression activity by targeting this receptor with one of the indole derivatives 3-Indole acetic acid hydrazide (IAAH) loaded onto zinc iron oxide (ZnFe2O4) NPs coated with PEG. XRD, SEM, FTIR, UV-Vis spectroscopy, and DLS were used to characterize the synthesized IAAH-NPs. The anti-inflammatory properties of IAAH-NPs on LPS-stimulated SimA9 microglia were assayed by measuring pro-inflammatory cytokine gene expressions and protein levels using RT-PCR and ELISA, respectively. As a result, IAAH-NPs showed an ability to suppress pro-inflammatory genes, including IL-6, IL-1ß, and TNF-α in LPS-stimulated SimA9 via targeting Nurr1. The current study suggests that ZnFe2O4 NPs as a delivery system can increase the efficiency of cellular uptake and enhance the IAAH ability to inhibit the pro-inflammatory cytokines. Collectively, we demonstrate that IAAH-NPs is a potential modulator of Nurr1 that combines nanotechnology as a delivery system to suppress neuroinflammation in CNS which opens a window for possible ambitious neuroprotective therapeutic approaches to neuro disorders.

Impaired Indole Acetic Acid and Reduced Indole-Producing Bacteria Contribute to Swainsonine-Induced Liver Inflammation.

Liver inflammation could be elicited by swainsonine in livestock, affecting the development of agriculture and animal husbandry. Our previous study showed an important role of bile acids (BAs) in swainsonine-induced hepatic inflammation. However, its pathogenesis, particularly the roles of a comprehensive profile of liver and serum metabolites and microbial-derived indole metabolites, has not been clarified. This study aimed to demonstrate the mechanisms linking the indole-producing bacteria and indole metabolites to swainsonine-induced hepatic inflammation by combining Targeted 500 metabolomics and quantitative analysis of indole metabolites. Swainsonine significantly disturbed the liver and serum metabolomes in mice. Genus Akkermansia alleviating inflammation and genus Lactobacillus producing indole metabolites were significantly declined. Indole acetic acid (IAA) was the only reduced aryl hydrocarbon receptor (AHR) ligand in this study. Analogously, some bacteria causing liver damage markedly increased. These findings suggested that indole-producing bacteria and indole metabolites may be potential triggers of swainsonine-induced hepatic inflammation.

Drought-Tolerant Bacteria and Arbuscular Mycorrhizal Fungi Mitigate the Detrimental Effects of Drought Stress Induced by Withholding Irrigation at Critical Growth Stages of Soybean (Glycine max, L.).

Considering current global climate change, drought stress is regarded as a major problem negatively impacting the growth of soybeans, particularly at the critical stages R3 (early pod) and R5 (seed development). Microbial inoculation is regarded as an ecologically friendly and low-cost-effective strategy for helping soybean plants withstand drought stress. The present study aimed to isolate newly drought-tolerant bacteria from native soil and evaluated their potential for producing growth-promoting substances as well as understanding how these isolated bacteria along with arbuscular mycorrhizal fungi (AMF) could mitigate drought stress in soybean plants at critical growth stages in a field experiment. In this study, 30 Bradyrhizobium isolates and 30 rhizobacterial isolates were isolated from the soybean nodules and rhizosphere, respectively. Polyethylene glycol (PEG) 6000 was used for evaluating their tolerance to drought, and then the production of growth promotion substances was evaluated under both without/with PEG. The most effective isolates (DTB4 and DTR30) were identified genetically using 16S rRNA gene. A field experiment was conducted to study the impact of inoculation with DTB4 and DTR30 along with AMF (Glomus clarum, Funneliformis mosseae, and Gigaspora margarita) on the growth and yield of drought-stressed soybeans. Our results showed that the bioinoculant applications improved the growth traits (shoot length, root length, leaf area, and dry weight), chlorophyll content, nutrient content (N, P, and K), nodulation, and yield components (pods number, seeds weight, and grain yield) of soybean plants under drought stress (p ≤ 0.05). Moreover, proline contents were decreased due to the bioinoculant applications under drought when compared to uninoculated treatments. As well as the count of bacteria, mycorrhizal colonization indices, and the activity of soil enzymes (dehydrogenase and phosphatase) were enhanced in the soybean rhizosphere under drought stress. This study's findings imply that using a mixture of bioinoculants may help soybean plants withstand drought stress, particularly during critical growth stages, and that soybean growth, productivity, and soil microbial activity were improved under drought stress.

Production of Exopolysaccharides and Indole Acetic Acid (IAA) by Rhizobacteria and Their Potential against Drought Stress in Upland Rice.

Peatlands are marginal agricultural lands due to highly acidic soil conditions and poor drainage systems. Drought stress is a big problem in peatlands as it can affect plants through poor root development, so technological innovations are needed to increase the productivity and sustainability of upland rice on peatlands. Rhizobacteria can overcome the effects of drought stress by altering root morphology, regulating stress-responsive genes, and producing exopolysaccharides and indole acetic acid (IAA). This study aimed to determine the ability of rhizobacteria in upland rice to produce exopolysaccharides and IAA, identify potential isolates using molecular markers, and prove the effect of rhizobacteria on viability and vigor index in upland rice. Rhizobacterial isolates were grown on yeast extract mannitol broth (YEMB) medium for exopolysaccharides production testing and Nutrient Broth (NB)+L-tryptophan medium for IAA production testing. The selected isolates identify using sequence 16S rRNA. The variables observed in testing the effect of rhizobacteria were germination ability, vigour index, and growth uniformity. EPS-1 isolate is the best production of exopolysaccharides (41.6 mg/ml) and IAA (60.83 ppm). The isolate EPS-1 was identified as Klebsiella variicola using 16S rRNA sequencing and phylogenetic analysis. The isolate EPS-1 can increase the viability and vigor of upland rice seeds. K. variicola is more adaptive and has several functional properties that can be developed as a potential bioagent or biofertilizer to improve soil nutrition, moisture and enhance plant growth. The use of rhizobacteria can reduce dependence on the use of synthetic materials with sustainable agriculture.

Indole-3-acetic acid induced cardiogenesis impairment in in-vivo zebrafish via oxidative stress and downregulation of cardiac morphogenic factors.

Plant growth regulators (PGRs) are increasingly used to promote sustainable agriculture, but their unregulated use raises concerns about potential environmental risks. Indole-3-acetic acid (IAA), a commonly used PGR, has been the subject of research on its developmental toxicity in the in-vivo zebrafish model. IAA exposure to zebrafish embryos caused oxidative stress, lipid peroxidation, and cellular apoptosis. The study also revealed that critical antioxidant genes including sod, cat, and bcl2 were downregulated, while pro-apoptotic genes such as bax and p53 were upregulated. IAA exposure also hampered normal cardiogenesis by downregulating myl7, amhc, and vmhc genes and potentially influencing zebrafish neurobehavior. The accumulation of IAA was confirmed by HPLC analysis of IAA-exposed zebrafish tissues. These findings underscore the need for further study on the potential ecological consequences of IAA use and the need for sustainable agricultural practices.

Isolation and characterization of PGPR obtained from different arsenic-contaminated soil samples and their effect on photosynthetic characters of maize grown under arsenic stress.

Contamination of the environment due to speedup of anthropogenic activities has become a serious threat to modern humanity. Among the contaminants, the new emerging concern is the heavy metal (HM) contamination in the environment. Because the persistence and harmfulness of heavy metals affect the ecosystem and the health of plants, animals, and humans, they are the most toxic substances in the environment. Among them, Arsenic (As) emerged as major environmental constraint leading to enormous negative effects on the plant, animal, and human health. Even in minute quantity, As is known to cause various critical diseases in humans and toxicity in plants. Research was performed to observe the capability of plant growth-promoting strains of bacteria in enhancing Zea mays (L.) growth in arsenic polluted soil. Total 30 bacterial strains were isolated from the polluted soils, screened for plant growth promotion potential and arsenic tolerance. Eighteen isolates showed resistance to different levels of sodium arsenate (ranging from 0 to 50 mM) in agar plate using LB media. Of 18 isolates, 83.3% produced IAA, methyl red, and hydrogen cyanide; 55.5% exhibited catalase activity; 61.1% showed siderophore production; 88.8% showed phosphate solubilization; and 44.4% showed oxidase, Voges proskauer activity, and KOH solubility. The most efficient isolates SR3, SD5, and MD3 with significant arsenic tolerance and plant growth-promoting (PGP) activity were examined via sequencing of amplified 16S rRNA gene. Isolates of bacteria, i.e., SR3, SD5, and MD3, showing multiple PGP-traits were identified as Bacillus pumilus (NCBI accession number: OR459628), Paenibacillus faecalis (NCBI accession number: OR461560), and Pseudochrobactrum asaccharolyticum (NCBI accession number: OR458922), respectively. Maize seeds treated with these PGPR strains were grown in pots contaminated with 50 ppm and 100 ppm sodium arsenate. Compared to untreated arsenic stressed plants, bacterial inoculation P. asaccharolyticum (MD3) resulted 20.54%, 18.55%, 33.45%, 45.08%, and 48.55% improvement of photosynthetic pigments (carotenoid content, chlorophyll content, stomatal conductance (gs), substomatal CO2, and photosynthetic rate), respectively. Principal component analysis explained that first two components were more than 96% of the variability for each tested parameter. The results indicate that in comparison to other isolates, P. asaccharolyticum isolate can be used as efficient agent for improving maize growth under arsenic polluted soil.

Nodulation Experiment by Cross-Inoculation of Nitrogen-Fixing Bacteria Isolated from Root Nodules of Several Leguminous Plants.

Root-nodule nitrogen-fixing bacteria are known for being specific to particular legumes. This study isolated the endophytic root-nodule bacteria from the nodules of legumes and examined them to determine whether they could be used to promote the formation of nodules in other legumes. Forty-six isolates were collected from five leguminous plants and screened for housekeeping (16S rRNA), nitrogen fixation (nifH), and nodulation (nodC) genes. Based on the 16S rRNA gene sequencing and phylogenetic analysis, the bacterial isolates WC15, WC16, WC24, and GM5 were identified as Rhizobium, Sphingomonas, Methylobacterium, and Bradyrhizobium, respectively. The four isolates were found to have the nifH gene, and the study confirmed that one isolate (GM5) had both the nifH and nodC genes. The Salkowski method was used to measure the isolated bacteria for their capacity to produce phytohormone indole acetic acid (IAA). Additional experiments were performed to examine the effect of the isolated bacteria on root morphology and nodulation. Among the four tested isolates, both WC24 and GM5 induced nodulation in Glycine max. The gene expression studies revealed that GM5 had a higher expression of the nifH gene. The existence and expression of the nitrogen-fixing genes implied that the tested strain had the ability to fix the atmospheric nitrogen. These findings demonstrated that a nitrogen-fixing bacterium, Methylobacterium (WC24), isolated from a Trifolium repens, induced the formation of root nodules in non-host leguminous plants (Glycine max). This suggested the potential application of these rhizobia as biofertilizer. Further studies are required to verify the N2-fixing efficiency of the isolates.

Identification and Evaluation of the Growth Promotion of Endophytic Bacteria on in vitro Potato Plants.

<b>Background and Objective:</b> Isolation and investigation of plant growth promoting bacteria on potato plants can provide significant information for the application of beneficial bacteria in potato production. This study aims to isolate and characterize endophytic bacteria isolated from potato roots. In addition, the potential application of endophytes in promoting potato growth under <i>in vitro</i> conditions was also investigated. <b>Materials and Methods:</b> The roots from 15 healthy potato plants were excised and surface sterilized by NaOCl and finally rinsed by sterilized water. The confirmed surface-sterilized roots were then aseptically cut into small fragments and spread onto the isolation media, followed by incubation at 27°C for up to 3 days. Six isolates that showed differences in colony morphology were selected for further investigation. All isolates were screened for IAA production, nitrogen fixation, and phosphate solubilization. <b>Results:</b> Five of the isolates were identified as <i>Bacillus</i> and isolate 30 was identified as <i>Paenibacillus alvei</i>. All isolates exhibited good IAA production. While Iso-27 had no nitrogen fixation activity, Iso-28 showed the highest level of nitrogen fixation activity (3.59 mg L¹), four isolates (Iso-9, Iso-10, Iso-11, Iso-28) could solubilize phosphate, ranging from 49.64 g L¹ to 67.98 mg L¹. After being inoculated with <i>in vitro</i> potato plants, isolates 9, 10, 28, 30, improved the stalk length, root number, fresh mass and dried mass of the potato plants. <b>Conclusion:</b> The four isolates can potentially be applied in <i>in vitro</i> potato culture.

Colon-Targeted Delivery of Indole Acetic Acid Helps Regulate Gut Motility by Activating the AHR Signaling Pathway.

Intestinal peristalsis is vital for gastrointestinal physiology and host homeostasis and is frequently dysregulated in intestinal disorders. Gut microbiota can regulate gut motility, especially through the tryptophan metabolism pathway. However, the role of indoles as microbial tryptophan metabolites in colonic function requires further exploration. Here, we show that the delivery of indole acetic acid (IAA) targeting the colon can improve gut motility by activating the aryl hydrocarbon receptor (AHR). To achieve colon-targeted delivery, Eudragit S-100 (ES) and chitosan (CS) were used as drug carriers. After optimisation, IAA-loaded ES-coated CS nanoparticles exhibited an encapsulation efficiency of 83% and a drug-loading capacity of 16%. These nanoparticles exhibited pH-dependent characteristics and remained stable in acidic conditions and the upper intestine. In simulated intestinal fluid (pH 7.4) and colonic lumen, considerable amounts of IAA were released after approximately 4 h. Compared with free IAA, the nanoparticles exerted enhanced therapeutic effects on gut movement disorders induced by loperamide. The efficacy of IAA treatment was attributable to the activation of the AHR signalling pathway and increased levels of AHR agonists. Furthermore, the oral administration of IAA-loaded nanoparticles promoted serotonin secretion and maintained the intestinal barrier function. The experimental outcomes demonstrate the efficiency of the proposed colon-specific delivery system and highlight the role of IAA, produced by gut microbiota metabolism, in regulating gut peristalsis through AHR activation.

Study of growth-improving and sporophore-inducing endobacteria isolated from Pleurotus pulmonarius.

Several Pleurotus species (oyster mushrooms) are commercially cultivated in India owing to the favorable tropical agro-climatic conditions. However, there are only a few studies on the microbiome of mushrooms, especially oyster mushrooms. The aim of this study was to assess the effect of endobacteria on mycelial growth, spawning, sporophore development, and proximate composition of P. pulmonarius. We isolated several bacterial strains from the sporophores of P. pulmonarius and assessed the in vitro production of indole acetic acid, ammonia, and siderophores. The selected bacteria were individually supplemented with spawn, substrate, or both for sporophore production. Three of 130 isolates were selected as mycelial growth-promoting bacteria in both solid and submerged fermentation. These bacterial isolates were identified through Gram staining, biochemical characterization, and 16S rRNA sequencing. Isolate PP showed 99.24% similarity with Priestia paraflexa, whereas isolates PJ1 and PJ2 showed 99.78% and 99.65% similarities, respectively, with Rossellomorea marisflavi. The bacterial supplementation with spawn, substrate, or both, increased the biological efficiency (BE) and nutrient content of the mushrooms. The bacterial supplementation with substrate augmented BE by 64.84%, 13.73%, and 27.13% using PJ2, PP, and PJ1, respectively; under similar conditions of spawn supplementation, BE was increased by 15.24%, 47.30%, 48.10%, respectively. Overall, the supplementation of endobacteria to improve oyster mushroom cultivation may open a new avenue for sustainable agricultural practices in the mushroom industry.

Planobacterium oryzisoli sp. nov., a novel bacterium isolated from roots of rice plant.

A Gram-negative, aerobic, short rod-shaped, non-motile, non-spore forming bacterium, designated strain GCR5T, was isolated from soil of paddy field. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GCR5T belongs to the genus Planobacterium and is related to Planobacterium taklimakanense NCTC 13490 T (96.1%, 16S rRNA gene sequence similarity). Colonies on R2A were white but they turn into bright yellow after exponential growth. They produce carotenoid pigment after 5-6 days of incubation, before that carotenoid pigment was not found. The major isoprenoid quinone was MK-6, and major cellular fatty acids were iso-C15:0, anteiso-C15:0 and iso-C17:0 3OH. Polar lipids include phosphatidylethanolamine, three unidentified phosphoglycolipids, three unidentified glycolipids, one unidentified aminophosphoglycolipid and five unidentified polar lipids. The strain GCR5T was found to have a 2,106,200 bp linear genome with G + C content of 43.7%. The ANI, dDDH and AAI values between the strain GCR5T and the type strains of phylogenetically related species were 60.2-71.1%, 19-24.3%, and 60.2-69.6%, respectively. The strain designated GCR5T produced indole acetic acid (IAA) in the presence of tryptophan only, and auxin responsive genes and tryptophan biosynthesis genes were found in its genome. Based on its polyphasic characteristics, strain GCR5T represents a novel species within the genus Planobacterium, for which the name Planobacterium oryzisoli sp. nov. was proposed. The type strain is GCR5T (= KCTC 82713 T = TISTR 2996 T = TBRC 15746 T).Repositories: The draft genome and 16S rRNA gene sequences of strain GCR5T have been deposited at GenBank/EMBL/DDBJ under accession numbers JADKYY000000000 and MN955408, respectively.

Optimization and reactor-scale production of plant growth regulators by Pleurotus eryngii.

The aims of the this study are to select the best cultivation type for plant growth regulator (PGR) production, to optimize PGR production with statistical experimental design, and to calculate bioprocess parameters and yield factors during PGR production by P. eryngii in flask and reactor scales. Submerged fermentation was the best cultivation type with 4438.67 ± 37.14, 436.95 ± 27.31, and 54.32 ± 3.21 mg/L of GA3, ABA, and IAA production values, respectively. The Plackett-Burman and Box-Behnken designs were used to determine effective culture parameters and interactive effects of the selected culture parameters on PGR production by Pleurotus eryngii under submerged fermentation. The statistical model is valid for predicting PGR production by P. eryngii. After these studies, maximum PGR production (7926.17 ± 334.09, 634.92 ± 12.15, and 55.41 ± 4.38 mg/L for GA3, ABA, and IAA, respectively) was reached on the 18th day of fermentation under optimized conditions. The optimum formula was 50 g/L fructose, 3 g/L NaNO3, and 1.5 g/L KH2PO4, 1 mg/L thiamine, incubation temperature 25 °C, initial medium pH 7.0, and an agitation speed of 150 rpm. The kinetics of PGR production was investigated in batch cultivation under 3-L stirred tank reactor conditions. Concentrations of GA3, ABA, and IAA of 10,545.00 ± 527.25, 872.32 ± 21.81, and 60.48 ± 3.48 mg/L were obtained at the reactor scale which were 4.1, 3.4, and 2.3 times higher than the initial screening values. The specific growth rate (µ), the volumetric (rp) and specific (Qp) PGR production rates, 486.11 mg/L/day and 107.43 mg/g biomass/day for GA3, confirmed the successful transfer of optimized conditions to the reactor scale. In the presented study, PGR production of P. eryngii is reported for the first time.

Flask and reactor scale production of plant growth regulators by Inonotus hispidus: optimization, immobilization and kinetic parameters.

The aims of the presented study are to compare submerged, static, and solid-state fermentation in the production of gibberellic acid (GA3), indole acetic acid (IAA), and abscisic acid (ABA) by Inonotus hispidus, to optimize with a statistical approach, and to determine the kinetic parameters under flask and reactor conditions. The maximum concentrations of GA3, (2478.85 ± 68.53 mg/L), ABA, (273.26 ± 6.17 mg/L) and IAA (30.67 ± 0.19 mg/L) were obtained in submerged conditions. After optimization, these values reached 2998.85 ± 28.85, 339.47 ± 5.50, and 34.56 ± 0.25 mg/L, respectively. Immobilization of fungal cells on synthetic fiber, polyurethane foam, and alginate beads resulted in an increase in plant growth regulators (PGR) production by 5.53%- 5.79% under optimized conditions. At the reactor scale, a significant increase was observed for GA3 concentration, 5441.54 mg/L, which was 2.14 and 1.45 times higher than non-optimized and optimized conditions in the flask scale, respectively. The maximum values for ABA and IAA were 390.39 and 44.79 mg/L, respectively. Although the specific growth rate (µ) decreases relatively from non-optimized flask conditions to optimized reactor conditions, it was observed that the PGR amounts produced per liter medium (rp) and per gram biomass (Qp) increased significantly. This is the first report on the synthesis of PGR by Inonotus hispidus which could be crucial for sustainable agriculture.

Concentration-dependent mechanisms of fluoranthene uptake by ryegrass.

Fluoranthene (Flu) uptake by plants is affected by plant growth and environmental concentration. Although plant growth processes, including substance synthesis and antioxidant enzyme activities, have been reported to regulate Flu uptake, their contributions have been poorly evaluated. Moreover, the effect of Flu concentration is little known. Here, low concentrations (0, 1, 5, and 10 mg/L) and high concentrations (20, 30, and 40 mg/L) of Flu were set to compare the changes in Flu uptake by ryegrass (Lolium multiflorum Lam.). Indices of plant growth (biomass, root length, root area, root tip number, and photosynthesis and transpiration rates), substance synthesis (indole acetic acid [IAA] content), and antioxidant enzyme activities (superoxide dismutase [SOD], peroxidase [POD], and catalase [CAT]) were recorded to unravel the mechanism of Flu uptake. Findings suggested that the Langmuir model fitted Flu uptake by ryegrass well. Flu absorption capacity in the root was stronger than that that in the leaf. Flu bioconcentration and translocation factors increased then reduced with the increase in Flu concentration and reached the maximum value under 5 mg/L Flu treatment. Plant growth and IAA content had the same pattern as before bioconcentration factor (BCF). SOD and POD activities increased then decreased with Flu concentration and reached their highest levels under 30 and 20 mg/L Flu treatments, respectively, whereas CAT activity decreased continuously and reached its lowest level under 40 mg/L Flu treatment. Variance partitioning analysis indicated that IAA content had the greatest significant effect on Flu uptake under low-concentration Flu treatments, whereas antioxidant enzyme activities had the greatest significant effect on Flu uptake under high-concentration Flu treatments. Revealing the concentration-dependent mechanisms of Flu uptake could provide a basis for regulating pollutant accumulation in plants.

Developmental delay and non-phenylketonuria (PKU) hyperphenylalaninemia in DNAJC12 deficiency: Case and approach.

BACKGROUND: Hyperphenylalaninemia is a biomarker for several monogenic neurotransmitter disorders where the body cannot metabolise phenylalanine to tyrosine. Biallelic pathogenic variants in DNAJC12, co-chaperone of phenylalanine, tyrosine, and tryptophan hydroxylases, leads to hyperphenylalaninemia and biogenic amines deficiency. METHODS AND RESULTS: A male firstborn to non-consanguineous Sudanese parents had hyperphenylalaninemia 247 µmol/L [reference interval (RI) < 200 µmol/L] at newborn screening. Dried blood spot dihydropteridine reductase (DHPR) assay and urine pterins were normal. He had severe developmental delay and autism spectrum disorder without a notable movement disorder. A low phenylalanine diet was introduced at two years without any clinical improvements. Cerebrospinal fluid (CSF) neurotransmitters at five years demonstrated low homovanillic acid (HVA) 0.259 µmol/L (reference interval (RI) 0.345-0.716) and 5-hydroxyindoleaetic acid (5HIAA) levels 0.024 µmol/L (reference interval (RI) 0.100-0.245). Targeted neurotransmitter gene panel analysis identified a homozygous c.78 + 1del variant in DNAJC12. At six years, he was commenced on 5-hydroxytryptophan 20 mg daily, and his protein-restricted diet was liberalised, with continued good control of phenylalanine levels. Sapropterin dihydrochloride 7.2 mg/kg/day was added the following year with no observable clinical benefits. He remains globally delayed with severe autistic traits. CONCLUSIONS: Urine, CSF neurotransmitter studies, and genetic testing will differentiate between phenylketonuria, tetrahydrobiopterin or DNAJC12 deficiency, with the latter characterised by a clinical spectrum ranging from mild autistic features or hyperactivity to severe intellectual disability, dystonia, and movement disorder, normal DHPR, reduced CSF HIAA and HVA. DNAJC12 deficiency should be considered early in the differential workup of hyperphenylalaninemia identified from newborn screening, with its genotyping performed once deficiencies of phenylalanine hydroxylase (PAH) and tetrahydrobiopterin (BH4) have been biochemically or genetically excluded.

Exogenous Auxin and Gibberellin on Fluoride Phytoremediation by Eichhornia crassipes.

High rates of fluorosis were reported worldwide as a result of human consumption of water with fluoride contents. Adjusting fluoride concentration in water as recommended by the World Health Organization (<1.5 mg L-1) is a concern and it needs to be conducted through inexpensive, but efficient techniques, such as phytoremediation. The application of phytohormones was investigated as a strategy to improve this process. Thus, the main goal of this research was to evaluate the effect of exogenous auxin and gibberellin on the tropical duckweed Eichhornia crassipes performance for fluoride phytoremediation. Definitive screening and central composite rotatable designs were used for experiments where fluoride concentration (5~15 mg L-1), phosphorus concentration (1~10 mg L-1), and pH (5~9) were assessed as well throughout 10 days. Fluoride contents were determined in solution and plant tissues by potentiometry. Higher concentrations of fluoride reflected on greater absorptions by plants, though in relative terms removal efficiencies were quite similar for all treatments (~60%). Auxin and acidic conditions favored fluoride removals per mass of plant. Fluoride accumulated mostly in leaves and auxin probably alleviated toxic effects on E. crassipes while gibberellin showed no effect. Therefore, E. crassipes could be employed as a fluoride accumulator plant for water treatment and exogenous auxin may be used to improve the process.

Isolation, Identification, and Characterization of Phosphate-Solubilizing Bacteria from Tunisian Soils.

Soil microorganisms play an important role in maintaining natural ecological balance through active participation in carbon, nitrogen, sulfur, and phosphorous cycles. Phosphate-solubilizing bacteria (PSB) are of high importance in the rhizosphere, enhancing the solubilization of inorganic phosphorus complexes into soluble forms available for plant nutrition. The investigation of this species of bacteria is of major interest in agriculture, as they can be used as biofertilizers for crops. In the present study, 28 isolates of PSB were obtained after the phosphate enrichment of soil samples from five Tunisian regions. Five PSB species were identified by 16S rRNA gene sequencing including Pseudomonas fluorescens, P. putida, and P. taiwanensis, Stenotrophomonas maltophilia, and Pantoea agglomerans. Solid and liquid Pikovskaya's (PVK) and National Botanical Research Institute's (NBRIP) media containing insoluble tricalcium phosphate were used for the evaluation of the phosphate solubilization ability of the bacterial isolates by two methods: visual evaluation of the solubilization zone around colonies (halo) and determination of solubilized phosphates in liquid medium by the colorimetric method of the vanado-molybdate yellow. Based on the results of the halo method, the isolate of each species that showed the higher phosphate solubilization index was selected for evaluation of phosphate solubilization by the colorimetric method. In the liquid media, the bacterial isolates showed phosphate solubilization ranging from 535.70 to 618.57 µg mL-1 in the NBRIP medium, and 374.20 to 544.28 µg mL-1 in the PVK medium, with the highest values produced by P. fluorescens. The best phosphate solubilization ability and higher reduction in broth pH, which indicates higher organic acid production, were achieved in NBRIP broth for most of the PSB. Strong correlations were observed between the average capability of PSB to solubilize phosphates and both the pH and total phosphorous content in the soil. The production of the hormone indole acetic acid (IAA), which can promote plant growth, was observed for all five PSB species. Among them, P. fluorescens obtained from the forest soil of northern Tunisia showed the highest production of IAA (50.4 ± 0.9 µg mL-1).

Non-Ionic Osmotic Stress Induces the Biosynthesis of Nodulation Factors and Affects Other Symbiotic Traits in Sinorhizobium fredii HH103.

(1) Background: Some rhizobia, such as Rhizobium tropici CIAT 899, activate nodulation genes when grown under osmotic stress. This work aims to determine whether this phenomenon also takes place in Sinorhizobium fredii HH103. (2) Methods: HH103 was grown with and without 400 mM mannitol. ß-galactosidase assays, nodulation factor extraction, purification and identification by mass spectrometry, transcriptomics by RNA sequencing, motility assays, analysis of acyl-homoserine lactones, and indole acetic acid quantification were performed. (3) Results: Non-ionic osmotic stress induced the production of nodulation factors. Forty-two different factors were detected, compared to 14 found in the absence of mannitol. Transcriptomics indicated that hundreds of genes were either activated or repressed upon non-ionic osmotic stress. The presence of 400 mM mannitol induced the production of indole acetic acid and acyl homoserine lactones, abolished swimming, and promoted surface motility. (4) Conclusions: In this work, we show that non-ionic stress in S. fredii HH103, caused by growth in the presence of 400 mM mannitol, provokes notable changes not only in gene expression but also in various bacterial traits, including the production of nodulation factors and other symbiotic signals.

Investigation of some endophytic fungi from five medicinal plants with growth promoting ability on maize (Zea mays L.).

AIMS: This study aimed to identify endophytic fungi from Anthemis altissima, Matricaria parthenium, Cichorium intybus, Achillea millefolium, and A. filipendulina with plant-promoting ability on the ZP684 maize hybrid-cultivar. METHODS AND RESULTS: Plants were collected from northeast-Iran and endophytic fungi were isolated and identified using partial large subunit nrDNA, internal transcribed spacer, translation elongation factor, and ß-tubulin genetic markers. Endophytic fungi that improved seed germination were studied under greenhouse conditions. Ninety-seven endophytic fungi were identified. Preussia africana, Bjerkandera adusta, Schizophyllum commune, Alternaria embellisia, Trichaptum biforme, Septoria malagutii, A. consortiale, Verticillium dahliae, Fusarium avenacearum, and Trametes versicolor significantly improved seed-germination. Alternaria consortiale produced the highest level of indole-3-acetic acid-like compounds and maize growth-promoting. CONCLUSIONS: Plant fungal colonization frequency increased with orthometric height. Sampling location Chahar Bagh at 2230 m contained the most endophytic fungi. Fusarium and Alternaria were the most frequently isolated endophytic genera. Therefore, medicinal plants are potential hosts for endophytic fungi that may be suitable biofertilizer agents in agriculture. SIGNIFICANCE AND IMPACT OF THE STUDY: This study helps to better understand the ecosystem functions by investigating of endophytic fungi distribution under different ecological conditions. Finding effective isolates among these microorganisms with a suitable plant-promoting ability on crops may help to reduce the use of chemical fertilizers in an agroecosystem.