Biological Potentials and Phytochemical Constituents of Raw and Roasted Nigella arvensis and Nigella sativa.

Nigella species are widely used to cure various ailments. Their health benefits, particularly from the seed oils, could be attributed to the presence of a variety of bioactive components. Roasting is a critical process that has historically been used to facilitate oil extraction and enhance flavor; it may also alter the chemical composition and biological properties of the Nigella seed. The aim of this study was to investigate the effect of the roasting process on the composition of the bioactive components and the biological activities of Nigella arvensis and Nigella sativa seed extracts. Our preliminary study showed that seeds roasted at 50 °C exhibited potent antimicrobial activities; therefore, this temperature was selected for roasting Nigella seeds. For extraction, raw and roasted seed samples were macerated in methanol. The antimicrobial activities against Streptococcus agalactiae, Streptococcus epidermidis, Streptococcus pyogenes, Candida albicans, Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, and Klebsiella oxytoca were determined by measuring the diameter of the zone of inhibition. The cell viability of extracts was tested in a colon carcinoma cell line, HCT-116, by using a microculture tetrazolium technique (MTT) assay. Amino acids were extracted and quantified using an automatic amino acid analyzer. Then, gas chromatography-mass spectrometry (GC-MS) analysis was performed to identify the chemical constituents and fatty acids. As a result, the extracts of raw and roasted seeds in both Nigella species showed strong inhibition against Klebsiella oxytoca, and the raw seed extract of N.arvensis demonstrated moderate inhibition against S. pyogenes. The findings of the MTT assay indicated that all the extracts significantly decreased cancer cell viability. Moreover, N. sativa species possessed higher contents of the measured amino acids, except tyrosine, cystine, and methionine. The GC-MS analysis of extracts showed the presence of 22 and 13 compounds in raw and roasted N. arvensis, respectively, and 9 and 11 compounds in raw and roasted N. sativa, respectively. However, heat treatment decreased the detectable components to 13 compounds in roasted N. arvensis and increased them in roasted N. sativa. These findings indicate that N. arvensis and N. sativa could be potential sources of anticancer and antimicrobials, where the bioactive compounds play a pivotal role as functional components.

Melatonin Ameliorates Thermotolerance in Soybean Seedling through Balancing Redox Homeostasis and Modulating Antioxidant Defense, Phytohormones and Polyamines Biosynthesis.

Global warming is impacting the growth and development of economically important but sensitive crops, such as soybean (Glycine max L.). Using pleiotropic signaling molecules, melatonin can relieve the negative effects of high temperature by enhancing plant growth and development as well as modulating the defense system against abiotic stresses. However, less is known about how melatonin regulates the phytohormones and polyamines during heat stress. Our results showed that high temperature significantly increased ROS and decreased photosynthesis efficiency in soybean plants. Conversely, pretreatment with melatonin increased plant growth and photosynthetic pigments (chl a and chl b) and reduced oxidative stress via scavenging hydrogen peroxide and superoxide and reducing the MDA and electrolyte leakage contents. The inherent stress defense responses were further strengthened by the enhanced activities of antioxidants and upregulation of the expression of ascorbate-glutathione cycle genes. Melatonin mitigates heat stress by increasing several biochemicals (phenolics, flavonoids, and proline), as well as the endogenous melatonin and polyamines (spermine, spermidine, and putrescine). Furthermore, the positive effects of melatonin treatment also correlated with a reduced abscisic acid content, down-regulation of the gmNCED3, and up-regulation of catabolic genes (CYP707A1 and CYP707A2) during heat stress. Contrarily, an increase in salicylic acid and up-regulated expression of the defense-related gene PAL2 were revealed. In addition, melatonin induced the expression of heat shock protein 90 (gmHsp90) and heat shock transcription factor (gmHsfA2), suggesting promotion of ROS detoxification via the hydrogen peroxide-mediated signaling pathway. In conclusion, exogenous melatonin improves the thermotolerance of soybean plants and enhances plant growth and development by activating antioxidant defense mechanisms, interacting with plant hormones, and reprogramming the biochemical metabolism.

Amelioration of heavy metal stress by endophytic Bacillus amyloliquefaciens RWL-1 in rice by regulating metabolic changes: potential for bacterial bioremediation.

This study aimed to investigate the bioremediation efficiency of phytohormone-producing endophytic Bacillus amyloliquefaciens RWL-1 isolated from rice seeds. In this study, we tested RWL-1 against various heavy metals (Cu, Cr, Pb, and Cd). Among the tested heavy metals, RWL-1 showed the highest tolerance for Cu stress and we observed alterations in growth kinetics with various Cu concentrations (1, 2.5, and 5 mM). We confirmed the biosorption potential of RWL-1 by scanning electron microscopy coupled with energy-dispersive X-ray spectrometry showing that Cu ions were adsorbed on RWL-1 cell surfaces. We further tested RWL-1 for its plant growth promoting and stress reliance efficiency in response to a dose-dependent increase in soil Cu (1, 2.5, and 5 mM). The RWL-1 inoculation significantly increased seedling biomass and growth attributes compared with non-inoculated control seedlings with and without Cu stress. Moreover, RWL-1 inoculation significantly promoted a physiochemical response in seedlings with and without Cu stress by reducing Cu uptake, improving carbohydrate levels (glucose, sucrose, fructose, and raffinose), enhancing amino acids regulation, and augmenting antioxidant levels (POD, PPO, and GHS). Levels of stress-responsive phytohormones such as abscisic acid (ABA) and jasmonic acid were significantly reduced in RWL-1-inoculated seedlings as compared with non-inoculated control seedlings under normal condition and same levels of Cu stress. In conclusion, the inoculation of B. amyloliquefaciens RWL-1 can significantly improve plant growth in Cu-contaminated soil and reduce metal accumulation, thus making plants safer for consumption. This approach could be tremendously helpful for safe and sustainable agriculture in heavy metal-contaminated areas.

Bacillus amyloliquefaciens RWL-1 as a New Potential Strain for Augmenting Biochemical and Nutritional Composition of Fermented Soybean.

Soybean (Glycine max L.) is a good source of natural antioxidants and commonly consumed as fermented products such as cheonggukjang, miso, tempeh, and sufu in Asian countries. The aim of the current study was to examine the influence of novel endophytic bacterial strain, Bacillus amyloliquefaciens RWL-1 as a starter for soybean fermentation. During fermentation, the cooked soybeans were inoculated with different concentrations (1%, 3%, and 5%) of B. amyloliquefaciens RWL-1. The changes in 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, total phenolic contents, isoflavones (Daidzin, Genistin, Glycitin, Daidzein, Glycitein, and Genistein), amino acids (aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine, arginine, and proline) composition, and minerals (calcium, copper, iron, potassium, magnesium, manganese, sodium, nickel, lead, arsenic, and zinc) were investigated. The level of antioxidants, total phenolic contents, isoflavones, and total amino acids were higher in fermented soybean inoculated with 1% B. amyloliquefaciens RWL-1 after 60 h of fermentation as compared to control, 3% and 5% B. amyloliquefaciens RWL-1. Additionally, fermented soybean inoculated with 5% B. amyloliquefaciens RWL-1 showed the highest values for mineral contents. Changes in antioxidant activities and bioactive compounds depended on the concentration of the strain used for fermentation. From these results, we conclude that fermented soybean has strong antioxidant activity, probably due to its increased total phenolic contents and aglycone isoflavone that resulted from fermentation. Such natural antioxidants could be used in drug and food industries and can be considered to alleviate oxidative stress.

Indole-3-acetic-acid and ACC deaminase producing Leclercia adecarboxylata MO1 improves Solanum lycopersicum L. growth and salinity stress tolerance by endogenous secondary metabolites regulation.

BACKGROUND: The utilization of plant growth-promoting microbes is an environment friendly strategy to counteract stressful condition and encourage plants tolerance. In this regards, the current study was designed to isolate ACC deaminase and indole-3-acetic acid (IAA) producing halotolerant bacteria to promote tomato (Solanum lycopersicum L.) growth and tolerance against salinity stress. RESULTS: The selected bacterial isolate MO1 was identified as Leclercia adecarboxylata and IAA quantification results revealed that MO1 produced significant amount of IAA (9.815 ± 0.6293 µg mL- 1). The MO1 showed the presence of ACC (1-Aminocyclopropane-1-Carboxylate) deaminase responsible acdS gene and tolerance against salinity stress. A plant microbe interaction experiment using tomato (Solanum lycopersicum L.) with glycine betaine (GB) as a positive control was carried out to investigate the positive role MO1 in improving plant growth and stress tolerance. The results indicated that MO1 inoculation and GB application significantly increased growth attributes under normal as well as saline condition (120 mM NaCl). The MO1 inoculation and GB treatment approach conferred good protection against salinity stress by significantly improving glucose by 17.57 and 18.76%, sucrose by 34.2 and 12.49%, fructose by 19.9 and 10.9%, citric acid by 47.48 and 34.57%, malic acid by 52.19 and 28.38%, serine by 43.78 and 69.42%, glycine by 14.48 and 22.76%, methionine by 100 and 124.99%, threonine by 70 and 63.08%, and proline by 36.92 and 48.38%, respectively, while under normal conditions MO1 inoculation and GB treatment also enhanced glucose by 19.83 and 13.19%, sucrose by 23.43 and 15.75%, fructose by 15.79 and 8.18%, citric acid by 43.26 and 33.14%, malic acid by 36.18 and 14.48%, serine by 46.5 and 48.55%, glycine by 19.85 and 29.77%, methionine by 22.22 and 38.89%, threonine by 21.95 and 17.07%, and proline by 29.61 and 34.68% compared to levels in non-treated plants, respectively. In addition, the endogenous abscisic acid (ABA) level was noticeably lower in MO1-inoculated (30.28 and 30.04%) and GB-treated plants (45 and 35.35%) compared to their corresponding control plants under normal condition as well as salinity stress, respectively. CONCLUSION: The current findings suggest that the IAA- and ACC-deaminase-producing abilities MO1 can improve plants tolerance to salinity stress.

Metabolic and proteomic alteration in phytohormone-producing endophytic Bacillus amyloliquefaciens RWL-1 during methanol utilization.

INTRODUCTION: Methanol utilization by bacteria is important for various industrial processes. Methylotrophic bacteria are taxonomically diverse and some species promote plant growth and induce stress tolerance. However, methylotrophic potential of bacterial endophytes is poorly understood. OBJECTIVE: The current study aimed to evaluate the metabolomic and proteomic changes in endophytic Bacillus amyloliquefaciens RWL-1 caused by its methanol utilization and the resultant influence on its phytohormone production. METHODS: B. amyloliquefaciens RWL-1 was grown in LB medium with different concentrations [0 (control), 0.5, 1, 1.5, 2, 2.5, 3, 3.5, and 4%) of methanol to examine its methylotrophic potential. SDS-PAGE analysis was carried out for bacterial protein confirmation. Moreover, the phytohormones (indole 3 acetic acid (IAA), gibberellins (GAs), abscisic acid (ABA)) produced by RWL-1 in methanol supplemented medium were quantified by GC-MS/SIM (6890N Network GC system, and 5973 Network Mass Selective Detector; Agilent Technologies, Santa Clara, CA, USA), while the antioxidants were estimated spectrophotometrically (T60 UV-VIS spectrophotometer, Leicester, UK). The amino acid quantification was carried out by amino acid analyzer (HITACHI L-8900, Japan). Furthermore, Nano-liquid chromatography (LC)-MS/MS analysis was performed with an Agilent system (Wilmington, DE, USA) for proteomic analysis while mascot algorithm (Matrix science, USA) was used to identify peptide sequences present in the protein sequence database. RESULTS: RWL-1 showed significant growth in media supplemented with 2 and 3.5% methanol, when compared with other concentrations. Mass spectroscopy analysis revealed that RWL-1 utilizes methanol efficiently as a carbon source. In the presence of methanol, RWL-1 produced significantly higher levels of IAA but lower levels of ABA, when compared with the control. Further, enzymatic antioxidants and functional amino acids were significantly up-regulated, with predominant expression of glutamic acid and alanine. Nano-liquid chromatography, quadrupole time-of-flight analysis, and quantitative analysis of methanol-treated bacterial cells showed expression of eight different types of proteins, including detoxification proteins, unrecognized and unclassified enzymes with antioxidant properties, proteases, metabolism enzymes, ribosomal proteins, antioxidant proteins, chaperones, and heat shock proteins. CONCLUSION: Results demonstrate that RWL-1 can significantly enhance its growth by utilizing methanol, and could produce phytohormones when growing in methanol-supplemented media, with increased expression of specific proteins and different biochemicals. These results will be useful in devising strategies for utilizing methylotrophic bacterial endophytes as alternative promoters of plant growth. Understanding RWL-1 ability to utilize methanol. The survival and phytohormones production by Bacillus amyloliquefaciens RWL-1 in methanol supplemented media whistle inducing metabolic and proteomic changes.

Exo-ethylene application mitigates waterlogging stress in soybean (Glycine max L.).

BACKGROUND: Waterlogging (WL) is a key factor hindering soybean crop productivity worldwide. Plants utilize various hormones to avoid various stress conditions, including WL stress; however, the physiological mechanisms are still not fully understood. RESULTS: To identify physiological mechanisms during WL stress, different phytohormones, such as ethephon (ETP; donor source of ethylene), abscisic acid, gibberellins, indole-3-acetic acid, kinetin, jasmonic acid, and salicylic acid were exogenously applied to soybean plants. Through this experiment, we confirmed the beneficial effects of ETP treatment. Thus, we selected ETP as a candidate hormone to mitigate WL. Further mechanistic investigation of the role of ETP in waterlogging tolerance was carried out. Results showed that ETP application mitigated WL stress, significantly improved the photosynthesis pigment, and increased the contents of endogenous GAs compared to those in untreated plants. The amino acid contents during WL stress were significantly activated by EPT treatments. The amino acid contents were significantly higher in the 100 µM ETP-treated soybean plants than in the control. ETP application induced adventitious root initiation, increased root surface area, and significantly increased the expressions of glutathione transferases and relative glutathione activity compared to those of non-ETP-treated plants. ETP-treated soybeans produced a higher up-regulation of protein content and glutathione S-transferase (GSTs) than did soybeans under the WL only treatment. CONCLUSIONS: In conclusion, the current results suggest that ETP application enabled various biochemical and transcriptional modulations. In particular, ETP application could stimulate the higher expression of GST3 and GST8. Thus, increased GST3 and GST8 induced 1) increased GSH activity, 2) decreased reactive oxygen species (ROS), 3) mitigation of cell damage in photosynthetic apparatus, and 4) improved phenotype consecutively.

Endophytic fungus Paecilomyces formosus LHL10 produces sester-terpenoid YW3548 and cyclic peptide that inhibit urease and α-glucosidase enzyme activities.

Endophytic fungi have been used to obtain novel bioactive secondary metabolites with potential applications in medical and agricultural sectors, which can also act as lead targets for pharmaceutical and medicinal potential. In the present study, the endophytic fungus Paecilomyces formosus LHL10 isolated from the root of cucumber plant was tested for its enzyme inhibitory potential. The ethyl acetate (EtOAc) extract of LHL10 was screened for its inhibitory effect on acetylcholinesterase (AChE), α-glucosidase, urease, and anti-lipid peroxidation. The findings suggest that the EtOAc extract from LHL10 possesses significant inhibitory potential against urease and α-glucosidase. The EtOAc extract was thus, subjected to advanced column chromatographic techniques for the isolation of pure compounds. The structure elucidation was carried out through spectroscopic analysis and comparison with literature data, and these compounds were confirmed as known a sester-terpenoid (1) and a known cyclic peptide (2). The enzyme inhibition bioassay indicated that Compounds 1 and 2 exhibited remarkable inhibitory rate against α-glucosidase and urease, with an IC50 value of 61.80 ± 5.7, 75.68 ± 6.2 and 74.25 ± 4.3, 190.5 ± 10.31 µg/g, respectively. Thus, the current study concludes the enzyme inhibitory potential of endophyte LHL10 and provides the basis for further investigations of bioactive compounds, which could be used as potent drugs for enzyme inhibition.

Mechanisms of Cr(VI) resistance by endophytic Sphingomonas sp. LK11 and its Cr(VI) phytotoxic mitigating effects in soybean (Glycine max L.).

Chromium Cr(VI) is highly toxic and leads to impaired phenotypic plasticity of economically important crops. The current study assessed an endophytic-bacteria assisted metal bio-remediation strategy to understand stress-alleviating mechanisms in Glycine max L (soybean) plants inoculated with Sphingomonas sp. LK11 under severe Cr(VI) toxicity. The screening analysis showed that high Cr concentrations (5.0 mM) slightly suppressed LK11 growth and metal uptake by LK11 cells, while significantly enhancing indole-3-acetic acid (IAA) production. Endophytic LK11 significantly upregulated its antioxidant system compared to control by enhancing reduced glutathione (GSH), catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) activities to counteract Cr-induced oxidative stress. Cr toxicity induced cell morphological alteration, as shown by SEM-EDX analysis and triggered significant lipid peroxidation. The interaction between LK11 and soybean in Cr-contaminated soil significantly increased plant growth attributes and down-regulated the synthesis of endogenous defense-related phytohormones, salicylic acid and abscisic acid, by 20% and 37%, respectively, and reduced Cr translocation to the roots, shoot, and leaves. Additionally, Cr-induced oxidative stress was significantly reduced in LK11-inoculated soybean, regulating metal responsive reduced GSH and enzymatic antioxidant CAT. Current findings indicate that LK11 may be a suitable candidate for the bioremediation of Cr-contaminated soil and stimulation of host physiological homeostasis.

Characterization of New Bioactive Enzyme Inhibitors from Endophytic Bacillus amyloliquefaciens RWL-1.

Endophytic bacteria are known to produce a wide array of bioactive secondary metabolites with beneficial effects on human health. In the current study, a novel endophytic bacterial strain, Bacillus amyloliquefaciens RWL-1, was isolated from the seeds of Oryza sativa. Initially, the crude extract of RWL-1 was assessed for potential biological effects of enzyme inhibition and cytotoxicity and was found to exhibit a broad spectrum inhibition for α-glucosidase (37 ± 0.09%) and urease (49.4 ± 0.53%). The screening results were followed by bioassay-guided isolation of secondary metabolite(s) from RWL-1. Extensive chromatographic and spectrophotometry analyses revealed the presence of compound 1 (S)-2-hydroxy-N-((S)-1-((S)-8-hydroxy-1-oxoisochroman-3-yl)-3-methylbutyl)-2-((S)-5-oxo-2,5-dihydrofuran-2-yl)acetamide. Further bioassays of compound 1 showed significant inhibition of α-glucosidase (52.98 ± 0.8%) and urease (51.27 ± 1.0%), compared with positive control values of 79.14 ± 1.9% and 88.24 ± 2.2%, and negative controls (0.08 ± 0.1% and 0.05 ± 0.01%), respectively. The current study suggests that bacterial endophytes are a rich source of novel bioactive compounds with high therapeutic value.

Comparison of antioxidants potential, metabolites, and nutritional profiles of Korean fermented soybean (Cheonggukjang) with Bacillus subtilis KCTC 13241.

This study was carried out to determine the effect of different concentrations of Bacillus subtilis (0, 1, 3, 5, and 7%) on the antioxidant potential and biochemical constituents of traditional Korean fermented soybean, Cheonggukjang (CKJ). The antioxidant capacity was studied using the reducing power, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) assays and the total phenolic contents (TPC) were measured using the Folin-Ciocalteu method. CKJ prepared using 1% B. subtilis revealed the highest TPC (5.99 mg/g), total amino acids (7.43 mg/g), DPPH (94.24%), and ABTS (86.03%) radical-scavenging activity and had the highest value of palmitic acid (11.65%), stearic acid (2.87%), and linolenic acid (11.76%). Results showed that the calcium, iron, sodium, and zinc contents increased in the CKJ prepared using 7% B. subtilis from 1481.38 to 1667.32, 41.38 to 317.00, 48.01 to 310.07, and 32.82 to 37.18 mg/kg respectively. In conclusion, the present results indicate that the fermentation of soybean with B. subtilis (KCTC 13241) significantly augments the nutritional and antioxidant potential of CKJ and it can be recommended as a health-promoting food source.

Effects of Bacterial Fermentation on the Biochemical Constituents and Antioxidant Potential of Fermented and Unfermented Soybeans Using Probiotic Bacillus subtilis (KCTC 13241).

Fermented soybeans, cheonggukjang (CKJ), are considered to be more wholesome than soybeans in Korea. To select the best soybean cultivar for making functional CKJ, a comparison was made between the biological activities of four soybean cultivars in their unfermented soybean (UFS) and CKJ states. Changes in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays, superoxide dismutase (SOD)-like activity, total phenolic compounds, total amino acids, and isoflavones were investigated. The levels of DPPH, ABTS, SOD-like activity, and total phenolic compounds increased in CKJ among all cultivars. The isoflavone aglycone and total amino acids showed the highest amount in CKJ prepared from soybean cultivar Aga 3. These results suggest that the improved antioxidant activity of CKJ in all cultivars might occur because of the higher levels of aglycones and total phenolic compounds achieved during fermentation. Moreover, CKJ prepared from soybean cultivar Aga 3 showed higher antioxidant activity than the other cultivars and so can be considered for the commercial production of functional foods in the future.

Indoleacetic acid production and plant growth promoting potential of bacterial endophytes isolated from rice (Oryza sativa L.) seeds.

Bacterial endophytes from the phyllosphere and rhizosphere have been used to produce bioactive metabolites and to promote plant growth. However, little is known about the endophytes residing in seeds. This study aimed to isolate and identify seed-borne bacterial endophytes from rice and elucidate their potential for phytohormone production and growth enhancement. The isolated endophytes included Micrococcus yunnanensis RWL-2, Micrococcus luteus RWL-3, Enterobacter soli RWL-4, Leclercia adecarboxylata RWL-5, Pantoea dispersa RWL-6, and Staphylococcus epidermidis RWL-7, which were identified using 16S rRNA sequencing and phylogenetic analysis. These strains were analyzed for indoleacetic acid (IAA) production by using GC-MS and IAA was found in the range of 11.50 ± 0.77 µg ml-1 to 38.80 ± 1.35 µg ml-1. We also assessed the strains for plant growth promoting potential because these isolates were able to produce IAA in pure culture. Most of the growth attributes of rice plants (shoot and root length, fresh and dry biomass, and chlorophyll content) were significantly increased by bacterial endophytes compared to the controls. These results show that IAA producing bacterial endophytes can improve hostplant growth traits and can be used as bio-fertilizers.

Biochemical Constituents and in Vitro Antioxidant and Anticholinesterase Potential of Seeds from Native Korean Persimmon Genotypes.

In the current study, the functional and biochemical potential of the seeds of four persimmon cultivars (PC1, PC2, PC3 and PC4) and their role against oxidative stress and acetylcholinesterase (AChE) inhibition were evaluated. In terms of biochemical compositions, free amino acids, fatty acids and organic acids analysis was performed. The free amino acids ranged from 2617.31 (PC2) to 3773.01 µg∙g(-1) dry weight (PC4). Oleic acid and linoleic acid were the principal fatty acids, which were significantly higher in PC4 and PC1, respectively. PC4 presented the highest amount of organic acid content (4212 mg∙kg(-1)), whereas PC2 presented the lowest (2498 mg∙kg(-1)). PC2 contained higher total phenolic content and flavonoid content, whereas PC3 had the lowest amount as compared to other cultivars. The in vitro DPPH, ABTS and superoxide anion radicals scavenging activity increased in a dose-dependent manner, whereas PC2 showed significantly higher scavenging activities as compared to PC1, PC2 and PC4 types. In the case of AChE inhibition, PC4 showed a moderate activity (67.34% ± 1.8%). In conclusion, the current findings reveal that the studied persimmon seeds cultivars are a source of bioactive natural antioxidants and AChE inhibitors. Such natural products could be employed in pharmaceutical and food industries, whilst can also be considered for the treatment of neurodegenerative diseases such as Alzheimer's.

Plant growth-promoting potential of endophytic fungi isolated from Solanum nigrum leaves.

Fungal endophytes have been characterized as producers of phytohormones and potent promoters of plant growth. In this study, two fungal endophytes, Fusarium tricinctum RSF-4L and Alternaria alternata RSF-6L, were isolated from the leaves of Solanum nigrum. Culture filtrates (CFs) from each isolate were initially screened for indole compounds, and assayed for their ability to promote the growth of Dongjin rice plants. Nearly all plant growth attributes examined (i.e., chlorophyll content, root-shoot length, and biomass production) were significantly enhanced upon treatment with fungal CFs. Subsequently, gas chromatography/mass spectrometry analyses were utilized to confirm the presence of phytohormones in the CF of each fungal endophytic isolate. These analyses revealed that RSF-4L and RSF-6L produced 54 and 30 µg/mL indole acetic acid, respectively, within their respective cultures. These findings suggest that the endophytes isolated in this study synthesize bioactive compounds that could play important roles in promoting plant growth.

Efficacy of endophytic SB10 and glycine betaine duo in alleviating phytotoxic impact of combined heat and salinity in Glycine max L. via regulation of redox homeostasis and physiological and molecular responses.

Adverse environmental stresses occurring simultaneously exhibit a lethal effect on crop productivity at the global level. Here, we investigated the individual and synergistic effects of endophytic T. virens SB10 and glycine betaine (GB) on the physiological and biochemical responses of Glycine max L. to alleviate the devastating effects of combined heat and salinity (HS) stress. Screening against HS stress tolerance showed that SB10 has significant tolerance against heat stress and produces hormones such as gibberellins and indole-3-acetic acid upon GB amendment of the growth medium under HS stress. Moreover, the current findings illustrated that the synergistic application of SB10 and GB was effective in alleviating the negative effects of HS stress on plant growth and physiology. The findings revealed that SB10 + GB led to a reduction in proline accumulation and Na+ uptake. It also maintained a high K+/Na + ratio by regulating GmHKT1 and GmSOS1 expression and enhanced macronutrient uptake (N, Ca, K) in plants. In turn, plants exhibited a higher growth rate and gaseous exchange attributes coupled with the upregulation of APX, SOD, POD, and GSH antioxidant activities and transcript accumulation of GmSOD1 and GmAPX1 to overcome HS-induced oxidative damage. Furthermore, SB10 + GB downregulated DREB2, DREB1B, and GmNCED3 expression and resulted in the reduced accumulation of endogenous ABA while enhancing endogenous SA accumulation via upregulation of PAL genes. In addition, enhanced accumulation of bioactive gibberellins (GA1, GA3, GA4, and GA7) was detected under HS stress in the SB10 + GB treatment group. Moreover, SB10 + GB also significantly regulated GmHsp90A2 and GmHsfA2 expression in tolerance against HS stress. The combination of SB10 and GB was shown to be an effective and alternative approach for growing G. max at high temperature coupled with saline conditions for sustainable agriculture.

Modulation of salt stress through application of citrate capped silver nanoparticles and indole acetic acid in maize.

The present study was conducted to determine the effect of indole acetic acid (IAA) and Citrate Capped Silver Nanoparticles (Cit-AgNPs) on various attributes of maize under induced salinity stress. Seeds of the said variety were collected from Cereal Crop Research Institute (CCRI) Pirsabaq, Nowshera, sterilized and sown in earthen pots filled with 2 kg silt and soil (1:2) in triplicates in the green house of the Botany Department, University of Peshawar. Nanoparticles were analyzed by scanning electron microscopy (SEM), Energy Dispersive X-Ray Spectroscopy (EDX), Thermo-gravimetric analysis (TGA) and Differential thermal analysis (DTA). Results of SEM revealed spherical morphology of Cit-AgNPs while EDX showed various elemental composition. TGA showed dominant weight loss up to 300 °C while the DTA showed major exothermic peaks at 420 °C. High Salinity concentration (80 mM) imposed significant detrimental impacts by reducing the agronomic attributes, photosynthetic pigments, osmolytes and antioxidant enzymes, which was remarkably ameliorated by the foliar application of Cit-AgNPs and IAA. Agronomic attributes including leaf, root and shoot fresh and dry weight was improved by 52-74%, 43-69% and 36-79% in individual as well as combined treatments of IAA and NPs. Photosynthetic pigments were amplified by 35-63%, total osmolytes were augmented by 39-68% and antioxidant enzymes including SOD and POD were boosted by 42-57% and 37-62% respectively, in combined as well as individual application. Conclusively, Cit-AgNPs are considered as salt mitigating entities that enhance the tolerance level of crop plants along with IAA, which may be beneficial for the plants growing in saline stressed environment.

Anti-inflammatory and analgesic potential of leaf extract of Justicia adhatoda L. (Acanthaceae) in Carrageenan and Formalin-induced models by targeting oxidative stress.

Plants are regarded as a valuable and inexpensive source of new drug development, and a variety of plant compounds are now being used in clinical trials to treat a variety of ailments. The goal of this work was to characterize and evaluate the anti-inflammatory and antioxidant effects of Justicia adhatoda L. leaf extract (Acanthaceae). The presence of alkaloids, saponins, tannins, phytosterols, phenols, and proteins in the leaf extract of J. adhatoda was determined using phytochemical screening. While the identification of different compounds in the leaf extract was carried out by HPLC analysis. Similarly, the anti-inflammatory potential of the leaf extract was assessed in Carrageenan and Formalin-induced inflammatory mice models. The phytochemical analysis of the leaf extract indicated a positive test for alkaloids, saponins, tannins, phytosterols, phenols, proteins, and amino acids, while the negative test for carbohydrates, and glycosides, flavonoids, and diterpenes. Moreover, among the detected compounds, gallic acid was found in the highest concentration with a 45.42% composition. The leaf extract showed the highest antibacterial activity against E. coli, while the lowest activity against Listeria was observed. The leaf extract of J. adhatoda revealed promising anti-inflammatory, analgesic, and antioxidants activities both in vitro and in vivo. Similarly, the detected compounds portrayed variable pharmacokinetic as well as binding affinities with the target proteins. In conclusion, the leaf extract exhibited significant antioxidants and antibacterial activities using in vitro assays. Similarly, the extract also revealed promising anti-inflammatory activities in vivo while exhibiting variable Pharmacokinetics and binding affinities towards protein target using computational tools.

Influence of Scion/Rootstock reciprocal effects on post-harvest and metabolomics regulation in stored peaches.

Peach is an important stone fruit crop cultivated at commercial scale in Pakistan. While, appropriate selection of rootstock has significant impact on the quality of peach fruit. Therefore, in the current study the influence of three rootstocks viz. 'GF-677', 'Peshawar Local' and 'Swat Local' were evaluated on the quality of 'Flordaking' peaches following cold storage during two consecutive years. The fruit from these rootstocks were kept at 0 °C for five weeks were studied for various fruit physical (weight loss, colour, firmness) and biochemical (pH, soluble solids content (SSC), titratable acidity (TA), SSC:TA ratio, fruit juice pH, sugars (total, reducing and non-reducing sugars), ascorbic acid (vitamin C) and free radicals scavenging activities) were evaluated. During both years, fruit harvested form trees grafted on 'GF-677' exhibited reduced fruit weight loss, changes in Chroma (C*) and highest fruit firmness, Lightness (L*), ascorbic acid contents and radical scavenging activities as compared to fruit harvested from tree grafted on other rootstocks. In conclusion, the post-harvest quality of scion 'Flordaking' peach fruit was significantly influenced and best quality can be obtained when it is grafted on 'GF-677' rootstock.

Synergistic interaction of fungal endophytes, Paecilomyces formosus LHL10 and Penicillium funiculosum LHL06, in alleviating multi-metal toxicity stress in Glycine max L.

Heavy metal accumulation in crop grains due to hazardous metal contamination is considered a great concern. However, phytobeneficial fungi are reported to have important abilities for the biosafety of crops grown in contaminated soil. Therefore, the current study was undertaken to explore the mutualistic association of plant growth-promoting endophytic fungi in reducing heavy metal concentration in the seeds of soybean plants subsequently grown in contaminated soil, without comprising seed quality and biochemical profile. The results revealed that endophytic Paecilomyces formosus LHL10 and Penicillium funiculosum LHL06 synergistically produced higher amounts of GAs and IAA in a co-cultured medium. Moreover, the co-inoculation of LHL06 and LHL10 to soybean plants grown under multi-metal toxic conditions significantly mitigated the adverse effects of heavy metal toxicity and increased the seed production (number of pods per plants, number of seeds per pod, and 100 seed weight) of soybean plants grown under control and multi-metal toxic conditions. Moreover, the levels of carbohydrates (glucose, sucrose, and fructose), minerals (iron, calcium, magnesium, and potassium), amino acids (serine, glutamic acids, glycine, methionine, lysine, arginine, and proline), and antioxidants (superoxide dismutase, catalase, and peroxidase) were significantly enhanced in sole and co-inoculated plants under control and stress conditions. Whereas organic acids (citric acid, tartaric acid, malic acid, and succinic acid), lipid peroxidation (MDA) products, multi-metal accumulation (nickel, cadmium, copper, lead, chromium, and aluminum), and stress-responsive endogenous abscisic acid levels were significantly decreased in seeds of soybean plants grown under control and multi-metal toxic conditions upon LHL06 and LHL10 sole and co-inoculation. The current results suggested the positive biochemical regulation in seeds for improving the nutritional status and making it safe for human consumption.