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.

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.

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.

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.

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.

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.

Exogenous melatonin induces drought stress tolerance by promoting plant growth and antioxidant defence system of soybean plants.

Melatonin is an indolamine bioactive molecule that regulates a wide range of physiological processes during plant growth and enhances abiotic stress tolerance. Here we examined the putative role of exogenous melatonin application (foliar or root zone) in improving drought stress tolerance in soybean seedlings. Pre-treatment of soybean seedlings with melatonin (50 and 100 µM) was found to significantly mitigate the negative effects of drought stress on plant growth-related parameters and chlorophyll content. The beneficial impacts against drought were more pronounced by melatonin application in the rhizosphere than in foliar treatments. The melatonin-induced enhanced tolerance could be attributed to improved photosynthetic activity, reduction of abscisic acid and drought-induced oxidative damage by lowering the accumulation of reactive oxygen species and malondialdehyde. Interestingly, the contents of jasmonic acid and salicylic acid were significantly higher following melatonin treatment in the root zone than in foliar treatment compared with the control. The activity of major antioxidant enzymes such as superoxide dismutase, catalase, polyphenol oxidase, peroxidase and ascorbate peroxidase was stimulated by melatonin application. In addition, melatonin counteracted the drought-induced increase in proline and sugar content. These findings revealed that modifying the endogenous plant hormone content and antioxidant enzymes by melatonin application improved drought tolerance in soybean seedlings. Our findings provide evidence for the stronger physiological role of melatonin in the root zone than in leaves, which may be useful in the large-scale field level application during drought.

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.

Evaluation potential of PGPR to protect tomato against Fusarium wilt and promote plant growth.

Soilborne fungal diseases are most common among vegetable crops and have major implications for crop yield and productivity. Eco-friendly sustainable agriculture practices that can overcome biotic and abiotic stresses are of prime importance. In this study, we evaluated the ability of plant growth-promoting rhizobacterium (PGPR) Bacillus aryabhattai strain SRB02 to control the effects of tomato wilt disease caused by Fusarium oxysporum f. sp. lycopersici (strain KACC40032) and promote plant growth. In vitro bioassays showed significant inhibition of fungal growth by SRB02. Inoculation of susceptible and tolerant tomato cultivars in the presence of SRB02 showed significant protection of the cultivar that was susceptible to infection and promotion of plant growth and biomass production in both of the cultivars. Further analysis of SRB02-treated plants revealed a significantly higher production of amino acids following infection by F. oxysporum. Analysis of plant defense hormones after inoculation by the pathogen revealed a significantly higher accumulation of salicylic acid (SA), with a concomitant reduction in jasmonic acid (JA). These results indicate that B. aryabhattai strain SRB02 reduces the effects of Fusarium wilt disease in tomato by modulating endogenous phytohormones and amino acid levels.

Extracellular vesicles in cancer diagnostics and therapeutics.

Cancer promotion, development, and malignant transformation is greatly influenced by cell-to-cell interactions in a complex tissue microenvironment. Cancer and stromal cells secrete soluble factors, as well as deport membrane-encapsulated structures, which actively contribute and mediate cell-to-cell interaction within a tumor microenvironment (TME). These membrane structures are recognized as extracellular vesicles (EVs), which include exosomes and microvesicles. They can carry and transport regulatory molecules such as oncogenic proteins, coding and non-coding RNAs, DNA, and lipids between neighboring cells and to distant sites. EVs mediate crucial pathophysiological effects such as the formation of premetastatic niches and the progression of malignancies. There is compelling evidence that cancer cells exhibit a significant amount of EVs, which can be released into the surrounding body fluids, compared with nonmalignant cells. EVs therefore have the potential to be used as disease indicator for the diagnosis and prognosis of cancers, as well as for facilitating research into the underlying mechanism and biomolecular basis of these diseases. Because of their ability to transport substances, followed by their distinct immunogenicity and biocompatibility, EVs have been used to carry therapeutically-active molecules such as RNAs, proteins, short and long peptides, and various forms of drugs. In this paper, we summarize new advancement in the biogenesis and physiological roles of EVs, and underpin their functional impacts in the process of cancer growth and metastasis. We further highlight the therapeutic roles of EVs in the treatment, prevention, and diagnosis of human malignancies.

Nanocurcumin: A Double-Edged Sword for Microcancers.

There is compelling evidence that drug molecules isolated from natural sources are hindered by low systemic bioavailability, poor absorption, and rapid elimination from the human body. Novel approaches are urgently needed that could enhance the retention time as well as the efficacy of natural products in the body. Among the various adopted approaches to meet this ever-increasing demand, nanoformulations show the most fascinating way of improving the bioavailability of dietary phytochemicals through modifying their pharmacokinetics and pharmacodynamics. Curcumin, a yellowish pigment isolated from dried ground rhizomes of turmeric, exhibits tremendous pharmacological effects, including anticancer activities. Several in vitro and in vivo studies have shown that curcumin mediates anticancer effects through the modulation (upregulation and/or downregulations) of several intracellular signaling pathways both at protein and mRNA levels. Scientists have introduced multiple modern techniques and novel dosage forms for enhancing the delivery, bioavailability, and efficacy of curcumin in the treatment of various malignancies. These novel dosage forms include nanoparticles, liposomes, micelles, phospholipids, and curcumin-encapsulated polymer nanoparticles. Nanocurcumin has shown improved anticancer effects compared to conventional curcumin formulations. This review discusses the underlying molecular mechanism of various nanoformulations of curcumin for the treatment of different cancers. We hope that this study will make a road map for preclinical and clinical investigations of cancer and recommend nano curcumin as a drug of choice for cancer therapy.

Inoculation with Indole-3-Acetic Acid-Producing Rhizospheric Rhodobacter sphaeroides KE149 Augments Growth of Adzuki Bean Plants Under Water Stress.

The use of plant growth-promoting rhizobacteria is economically viable and environmentally safe for mitigating various plant stresses. Abiotic stresses such as flood and drought are a serious threat to present day agriculture. In the present study, the indole-3-acetic acid-producing rhizobacterium R. sphaeroides KE149 was selected, and its effect on the growth of adzuki bean plants was investigated under flood stress (FS) and drought stress (DS). IAA quantification of bacterial pure culture revealed that KE149 produced significant amount of IAA. KE149 inoculation significantly decreased stress-responsive endogenous abscisic acid and jasmonic acid and increased salicylic acid in plants under DS and FS. KE149 inoculation also increased proline under DS and methionine under FS. Moreover, KE149 inoculation significantly increased the calcium (Ca), magnesium (Mg), and potassium (K) content and lowered the sodium (Na) content in the plant shoot under stress. KE149-treated plants had significantly higher root length, shoot length, stem diameter, biomass, and chlorophyll content under both normal and stressed conditions. These results suggest that KE149 could be an efficient biofertilizer for mitigating water stress.

Effect of Ammonia and Indole-3-acetic Acid Producing Endophytic Klebsiella pneumoniae YNA12 as a Bio-Herbicide for Weed Inhibition: Special Reference with Evening Primroses.

Information on the use of endophytic bacteria as a bio-herbicide for the management of weed control in agricultural fields is limited. The current study aimed to isolate endophytic bacteria from evening primroses and to screen them for their bio-herbicidal activity. Two isolated endophytic bacteria (Pantoea dispersa YNA11 and Klebsiella pneumoniae YNA12) were initially screened for citrate utilization and for indole-3-acetic acid (IAA) and catalase production. The preliminary biochemical assessment showed YNA12 as a positive strain. Ammonia, catalase, and IAA in its culture filtrate were quantified. Gas Chromatography/Mass Spectroscopy- Selective Ion Monitoring (GC/MS-SIM) analysis revealed the production of IAA by YNA12 in a time-dependent manner. YNA12 also exhibited significant ammonia-producing potential and catalase activity against hydrogen peroxide. The YNA12 culture filtrate significantly inhibited the germination rate of evening primrose seeds, resulting in a marked reduction in seedling length and biomass compared with those of the control seeds. Moreover, the culture filtrate of YNA12 significantly accelerated the endogenous abscisic acid (ABA) production and catalase activity of evening primrose seedlings. Macronutrient regulation was adversely affected in the seedlings exposed to the culture filtrate of YNA12, leading to inhibition of seed germination. The current results suggest that endophytic YNA12 may be used as a potent bio-herbicidal agent for controlling weed growth and development.

Silicon and Gibberellins: Synergistic Function in Harnessing ABA Signaling and Heat Stress Tolerance in Date Palm (Phoenix dactylifera L.).

Date palm is one of the most economically vital fruit crops in North African and Middle East countries, including Oman. A controlled experiment was conducted to investigate the integrative effects of silicon (Si) and gibberellic acid (GA3) on date palm growth and heat stress. The exogenous application of Si and GA3 significantly promoted plant growth attributes under heat stress (44 ± 1 °C). The hormonal modulation (abscisic acid [ABA] and salicylic acid [SA]), antioxidant accumulation, and the expression of abiotic stress-related genes were evaluated. Interestingly, heat-induced oxidative stress was markedly reduced by the integrative effects of Si and GA3 when compared to their sole application, with significant reductions in superoxide anions and lipid peroxidation. The reduction of oxidative stress was attributed to the enhancement of polyphenol oxidase, catalase, peroxidase, and ascorbate peroxidase activities as well as the upregulation of their synthesis related genes expression viz. GPX2, CAT, Cyt-Cu/Zn SOD, and glyceraldehyde3-phosphate dehydrogenase gene (GAPDH). The results showed the activation of heat shock factor related genes (especially HsfA3) during exogenous Si and GA3 as compared to the control. Furthermore, the transcript accumulation of ABA signaling-related genes (PYL4, PYL8, and PYR1) were significantly reduced with the combined treatment of Si and GA3, leading to reduced production of ABA and, subsequently, SA antagonism via its increased accumulation. These findings suggest that the combined application of Si and GA3 facilitate plant growth and metabolic regulation, impart tolerance against stress, and offers novel stress alleviating strategies for a green revolution in sustainable food security.

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.

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.

Phytohormones enabled endophytic Penicillium funiculosum LHL06 protects Glycine max L. from synergistic toxicity of heavy metals by hormonal and stress-responsive proteins modulation.

This study investigates the stress-mitigating effects of endophytic Penicillium funiculosum LHL06 on soybean roots via modulation of physio-biochemical, molecular, and proteomic responses to combined heavy metal (Ni, Cu, Pb, Cr, and Al) toxicity. Preliminary screening revealed that LHL06 can tolerate and remediate combined heavy metal contamination in its media and upregulate gibberellins (GA1, GA3, GA4, GA7 and GA9) and indole-3-acetic acid (IAA) production. Inoculation of LHL06 resulted in marked reduction of metals uptake in roots and shoots by downregulating heavy metal ATPase genes (GmHMA13, GmHMA14, GmHMA19) and GmMATE1 compared to non-inoculated plants; in turn, this decreased abscisic acid and jasmonic acid levels. Moreover, triggering of free amino acid metabolism in LHL06-inoculated roots significantly upregulated expression of stress-related proteins (glutathione S-transferase L3, isoflavone reductase-like, chalcone isomerase A, NAD(P)H dehydrogenase (quinone), FQR1-like 1 isoform X2, and Peroxidase 3) to combat metals toxicity. Compared to non-inoculated-plants, LHL06-inoculated-plants exhibited higher antioxidant activity and transcript accumulation of glutathione S-transferase (GmGST8 and GmGST3), G6PDH, and GmSOD1[Cu-Zn], which decreased metal-induced reactive oxygen species. Therefore, LHL06-inoculation remediate combined metal contamination in soil, activate signaling network of stress-responsive hormones and antioxidant systems for promoting growth and tolerance, and reduce metal-accumulation, thereby making plants safer for consumption.

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.