Fungal endophytes isolated from Withania somnifera Markedly exhibit antitumor activity against human glioblastoma.

In the current study, five fungal endophytes (WR-1, WR-2, WR-3, WR-4, WS-6) were isolated from the roots and stem of Withania somnifera (L.) Dunal, and tested fungal culture filtrates (FCF) against the brain tumour (glioblastoma) cell line (U-87). All FCFs showed anti-tumour activity with an IC50 value of 3.5, 6.0, 3.2, 6.0, and 0.95 µg/mL, respectively. HPLC fractionation of WS-6 showed the presence of 11 compounds (A1-A11), all with good anti-tumour activities (IC 50 3.5, 5.0, 5.0, 3.6, 2.5, 1.7, 2.3, 2.7, 0.9, 1.8 and 0.4 µg/mL, respectively). We report for the first time, the anti-tumour activity of 1-Docosene, Phenol, 2,4-bis(1,1-dimethylethyl), 1,2-Benzenedicarboxylic acid, diisooctyl ester, Hexadecane, 1,1-bis(dodecyloxy)-, Benzene, 1,1'-(2-methyl-2-(phenylthio)cyclopropylidene) bis-, Naphthalene, 1,2-dihydro-1-phenyl- (A1-A6), identified by GC-MS. The rest (A7-A11) had earlier reports of anticancer activities on cell lines other than U-87. We conclude that endophytic WS-6 (Aspergillus fumigatus) produces antitumor compounds that might be helpful in future drug development against brain tumours, especially glioblastoma.

Simultaneous Study of Analysis of Anti-inflammatory Potential of Dryopteris ramosa (C. Hope) C. Chr. using GC-Mass and Computational Modeling on the Xylene-induced Ear Oedema in Mouse Model.

INTRODUCTION: In the present study, we aimed to investigate the extraction and identification of the potential phytochemicals from the Methanolic Extract of Dryopteris ramosa (MEDR) using GC-MS profiling for validating the traditional uses of MEDR its efficacy in inflammations by using in-vitro, in-vivo and in silico approaches in anti-inflammatory models. METHODS: GC-MS analysis confirmed the presence of a total of 59 phytochemical compounds. The human red blood cells (HRBC) membrane stabilization assay and heat-induced hemolysis method were used as in-vitro anti-inflammatory activity of the extract. The in-vivo analysis was carried out through the Xylene-induced mice ear oedema method. It was found that MEDR at a concentration of 20 µg, 30 µg, and 40 µg showed 35.45%, 36.01%, and 36.33% protection to HRBC in a hypotonic solution, respectively. At the same time, standard Diclofenac at 30 µg showed 45.31% protection of HRBC in a hypotonic solution. RESULTS: The extract showed inhibition of 25.32%, 26.53%, and 33.31% cell membrane lysis at heating at 20 µg, 30 µg, and 40 µg, respectively. In comparison, standard Diclofenac at 30 µg showed 50.49% inhibition of denaturation to heat. Methanolic extract of the plant exhibited momentous inhibition in xylene-induced ear oedema in mice treated with 30 µg extract were 47.2%, 63.4%, and 78.8%, while inhibition in mice ear oedema treated with 60 µg extract was 34.7%, 43.05%, 63.21% and reduction in ear thickness of standard drug were 57.3%, 59.54%, 60.42% recorded at the duration of 1, 4 and 24 hours of inflammation. Molecular docking and simulations were performed to validate the anti-inflammatory role of the phytochemicals that revealed five potential phytochemicals i.e. Stigmasterol,22,23dihydro, Heptadecane,8methyl, Pimaricacid, Germacrene and 1,3Cyclohexadiene,_5(1,5dimethyl4hexenyl)-2methyl which revealed potential or significant inhibitory effects on cyclooxygenase-2 (COX-2), tumour necrosis factor (TNF-α), and interleukin (IL-6) in the docking analysis. CONCLUSION: The outcome of the study signifies that MEDR can offer a new prospect in the discovery of a harmonizing and alternative therapy for inflammatory disease conditions.

Characterization and Pilot-Scale Application of the ZMS-2 Serine Protease with Novel Properties for the Eco-friendly Leather Processing.

Microbial alkaline proteases are dominating the global enzyme market with a share of over 65% due to their multifarious catalytic potentials. Hence, production of proteases with novel properties of commercial significance is highly desirable to meet the global enzyme demand. Here, we report the purification, characterization, and pilot-scale application of a serine protease from the desert soil bacterium Bacillus subtilis ZMS-2 with novel properties as dehairing agent in leather processing. The enzyme was purified 16.5-fold with a specific activity of 1543.5 U mg-1 and recovery percentage of 33.6% using ammonium sulfate precipitation, ion exchange, and gel filtration chromatography. The purified enzyme was characterized as a metal ion-, surfactant-, and denaturant-compatible alkaline serine protease having a molecular weight of 36.1 kDa with an optimum activity at pH 8.5 and 60 °C. The catalytic activity of the enzyme was enhanced by Zn+2 (204%), Ag+ (110%), H2O2 (123%), Triton X-100 (110%), iso-octane (109%), chloroform (110%), ethanol (105%), ethyl acetate (110%), and acetonitrile (128%). During pilot-scale applications, the optimum condition was found to be a combination of enzyme (1.5%, 460 U mL-1), sodium sulfide (2%), and calcium hydroxide (lime) (3%). Under this condition, the time required for complete dehairing was 90 min. Chemoenzymatically processed skins exhibited better physical properties than chemically processed skin, including tensile strength (16.35 ± 6.68 N/mm), ball burst (452.88 ± 6.06 N/mm), percent elongation (38.85 ± 1.06 N), tear strength (50.16 ± 4.42 N/mm), and softness (6.5 mm). Electron microscopy analysis of the treated skin showed complete removal of hairs with roots, confirming the keratin specificity of the enzyme. Moreover, the enzyme-assisted dehairing process reduced chemical oxygen demand (COD), biochemical oxygen demand (BOD), total dissolved solids (TDS), and total suspended solids (TSS) by 68, 77, 34, and 39%, respectively. Thus, the alkaline serine protease from B. subtilis ZMS-2 is a potential dehairing agent for the eco-friendly processing of animal skins on industrial scales.

Aspergillus welwitschiae BK Isolate Ameliorates the Physicochemical Characteristics and Mineral Profile of Maize under Salt Stress.

Abiotic stressors are global limiting constraints for plant growth and development. The most severe abiotic factor for plant growth suppression is salt. Among many field crops, maize is more vulnerable to salt, which inhibits the growth and development of plants and results in low productivity or even crop loss under extreme salinity. Consequently, comprehending the effects of salt stress on maize crop improvement, while retaining high productivity and applying mitigation strategies, is essential for achieving the long-term objective of sustainable food security. This study aimed to exploit the endophytic fungal microbe; Aspergillus welwitschiae BK isolate for the growth promotion of maize under severe salinity stress. Current findings showed that salt stress (200 mM) negatively affected chlorophyll a and b, total chlorophyll, and endogenous IAA, with enhanced values of chlorophyll a/b ratio, carotenoids, total protein, total sugars, total lipids, secondary metabolites (phenol, flavonoids, tannins), antioxidant enzyme activity (catalase, ascorbate peroxidase), proline content, and lipid peroxidation in maize plants. However, BK inoculation reversed the negative impact of salt stress by rebalancing the chlorophyll a/b ratio, carotenoids, total protein, total sugars, total lipids, secondary metabolites (phenol, flavonoids, tannins), antioxidant enzyme activity (catalase, ascorbate peroxidase), and proline content to optimal levels suitable for growth promotion and ameliorating salt stress in maize plants. Furthermore, maize plants inoculated with BK under salt stress had lower Na+, Cl- concentrations, lower Na+/K+ and Na+/Ca2+ ratios, and higher N, P, Ca2+, K+, and Mg2+ content than non-inoculated plants. The BK isolate improved the salt tolerance by modulating physiochemical attributes, and the root-to-shoot translocation of ions and mineral elements, thereby rebalancing the Na+/K+, Na+/Ca2+ ratio of maize plants under salt stress.

Efficacy of Fungi in the Decolorization and Detoxification of Remazol Brilliant Blue Dye in Aquatic Environments.

Industrial effluents result in water pollution and affect the biological activity of aquatic and terrestrial life. In this study, efficient fungal strains were isolated from the aquatic environment and identified as Aspergillus fumigatus (SN8c) and Aspergillus terreus (SN40b). The isolates were selected based on their potential to efficiently decolorize and detoxify Remazol brilliant blue (RBB) dye, which is extensively used in different industries. Initially, a total of 70 different fungal isolates were screened. Among these, 19 isolates demonstrated dye decolorization capabilities, and SN8c and SN40b revealed the highest decolorization capabilities in liquid medium. The maximum estimated decolorization for SN8c was 91.3% and for SN40b, 84.5% at 40 mg/L of RBB dye in the presence of glucose (1 gm/L), after 5 days of incubation at different levels of pH, temperature, nutrient source, and concentration. RBB dye decolorization using SN8c and SN40b isolates was at a maximum of 99% at pH 3-5, whereas minimum decolorization was recorded as 71.29% and 73.4% SN8c, respectively, at pH 11. The maximum decolorization of the dye was 93% and 90.9% in a defined glucose concentration of 1 gm/L, and a 63.01% decrease was recorded in the decolorization activity at a low level of glucose concentration (0.2 gm/L). Finally, the decolorization and degradation were detected using UV spectrometry and HPLC. Toxicity tests of pure dye and treated dye samples were checked against the seed germination of different plants and the larvae mortality of Artemia salina. This study revealed that indigenous aquatic fungal flora can recover contaminated sites and support aquatic and terrestrial life.

The Recent Advances in the Utility of Microbial Lipases: A Review.

Lipases are versatile biocatalysts and are used in different bioconversion reactions. Microbial lipases are currently attracting a great amount of attention due to the rapid advancement of enzyme technology and its practical application in a variety of industrial processes. The current review provides updated information on the different sources of microbial lipases, such as fungi, bacteria, and yeast, their classical and modern purification techniques, including precipitation and chromatographic separation, the immunopurification technique, the reversed micellar system, aqueous two-phase system (ATPS), aqueous two-phase flotation (ATPF), and the use of microbial lipases in different industries, e.g., the food, textile, leather, cosmetics, paper, and detergent industries. Furthermore, the article provides a critical analysis of lipase-producing microbes, distinguished from the previously published reviews, and illustrates the use of lipases in biosensors, biodiesel production, and tea processing, and their role in bioremediation and racemization.

Potent Bioactivity of Endophytic Fungi Isolated from Moringa oleifera Leaves.

Plant species are known to harbor large number of endophytes, which stays in plant tissues as symbionts. These endophytes secrete large array of bioactive compounds that have potency against certain diseases with no side effects. We have collected leaf samples of the Moringa oleifera plant from the Pakistan Forest Institute, Khyber Pakhtunkhwa, Pakistan for the isolation of beneficial endophytes. The strains isolated from the leaves of M. oleifera were coded with MOL and tested for antimicrobial, antifungal, germicidal, phytotoxic, insecticidal, cytotoxic, and anti-inflammatory activities. The isolates, MOL1, MOL16, MOL19, and MOL21, possessed antibacterial activity against Staphylococcus aureus, whereas MOL7 inhibited 55% of the growth of Escherichia coli. MOL3 inhibited the growth of E. coli, S. aureus, and Pseudomonas aeruginosa. The strains, MOL1 and MOL7, showed antifungal activity against Candida albicans and Saccharomyces cerevisiae, while the strains, MOL11 and MOL17, showed activity against Verticillium chlamydosporium. The isolates, MOL3, MOL7, MOL9, MOL15, MOL17, MOL18, and MOL19, inhibited the growth of Lemna minor (duckweed) at 100 µg/ml. MOL2 exhibited strong activity in the brine shrimp assay, while MOL1, MOL2, MOL5, MOL6, MOL12, MOL17, MOL19, and MOL20 showed insecticidal, and MOL3 demonstrated larvicidal and antileishmanial activity. The isolated potent endophytes were identified as Aspergillus, Penicillium, Fusarium, Tricoderma, Rhizoctonia, Mucor, Alternaria, Pestalotiopsis, Acremonium, and Cladosporium through morphological and microscopic characteristics of the colonies.

Aspergillus violaceofuscus alleviates cadmium and chromium stress in Okra through biochemical modulation.

Endophytic fungi from the Chilli were used to help okra plants exposed to cadmium (Cd) or chromium (Cr) stress. Initially, the strain Ch06 produced higher amounts of indole acetic acid (IAA) (230.5 µg/mL), sugar (130.7 µg/mL), proteins (128.2 µg/mL), phenolics (525.6 µg/mL) and flavonoids (98.4 µg/mL) in Czapek broth supplemented with Cd or Cr. The production of IAA and other metabolites in such a higher concentration suggested that Ch06 might improve plant growth under heavy metal stress. For this reason, an experiment was designed, in which biomass of Ch06 (at 2g/100g of sand) were applied to the okra plants exposed to Cd or Cr stress (at 100 or 500 µg/g). The results exhibited that Ch06 improved the total chlorophyll (36.4±0.2 SPAD), shoot length (22.6±0.2 cm), root length (9.1±0.6 cm), fresh weight (5±0.6 g), dry weight (1.25±0.01 g), sugars (151.6 µg/g), proteins (114.8 µg/g), proline (6.7 µg/g), flavonoids (37.9 µg/g), phenolics (70.7 µg/g), IAA (106.7 µg/g), catalase (0.75 enzyme units/g tissue) and ascorbic acid oxidaze (2.2 enzyme units/g tissue) of the associated okra plants. Similar observations have been recorded in Ch06 associated okra plants under Cd and Cr stress. Also, Ch06 association reduced translocation of Cd (35% and 45%) and Cr (47% and 53%) to the upper parts of the okra plants and thus reduced their toxicity. The internal transcribed spacer (ITS) region amplification of 18S rDNA (ribosomal deoxyribo nucleic acid) exhibited that the potent strain Ch06 was Aspergillus violaceofuscus. The results implied that A. violaceofuscus has the ability to promote host species growth exposed to Cd and Cr. Moreover, it helped the host plants to recover in Cd and Cr polluted soils, hence can be used as biofertilizer.

A Review on the Role of Endophytes and Plant Growth Promoting Rhizobacteria in Mitigating Heat Stress in Plants.

Among abiotic stresses, heat stress is described as one of the major limiting factors of crop growth worldwide, as high temperatures elicit a series of physiological, molecular, and biochemical cascade events that ultimately result in reduced crop yield. There is growing interest among researchers in the use of beneficial microorganisms. Intricate and highly complex interactions between plants and microbes result in the alleviation of heat stress. Plant-microbe interactions are mediated by the production of phytohormones, siderophores, gene expression, osmolytes, and volatile compounds in plants. Their interaction improves antioxidant activity and accumulation of compatible osmolytes such as proline, glycine betaine, soluble sugar, and trehalose, and enriches the nutrient status of stressed plants. Therefore, this review aims to discuss the heat response of plants and to understand the mechanisms of microbe-mediated stress alleviation on a physio-molecular basis. This review indicates that microbes have a great potential to enhance the protection of plants from heat stress and enhance plant growth and yield. Owing to the metabolic diversity of microorganisms, they can be useful in mitigating heat stress in crop plants. In this regard, microorganisms do not present new threats to ecological systems. Overall, it is expected that continued research on microbe-mediated heat stress tolerance in plants will enable this technology to be used as an ecofriendly tool for sustainable agronomy.

Porostereum spadiceum-AGH786 Regulates the Growth and Metabolites Production in Triticum aestivum L. Under Salt Stress.

The role of the most fungal endophytes in the host plant growth and production of metabolites under stress conditions is still unknown. Fungal endophytes occur in almost all plants to benefit the host plants exposed to biotic and abiotic stress. In the present work, we investigated salt (NaCl) stress alleviation capability of a fungal endophyte (Porostereum spadiceum-AGH786). The culture filtrate (CF: 1.5 mL.) of P. spadiceum-AGH786 contained IAA (158 µg/ml), SA (29.3 µg/ml), proline (114.6 µg/ml), phenols (167.4 µg/ml), lipids (71.4 µg/ml), sugar (133.2 µg/ml), flavonoids (105.04 µg/ml). Smaller amounts of organic acids, such as butyric acid (5.8 µg/ml), formic acid (2.34 µg/ml), succinic acid (2.02 µg/ml), and quinic acid (2.25 µg/ml) were also found in CF of P. spadiceum-AGH786. Similarly, the CF displayed antioxidant activity in 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. Moreover, wheat plants colonized by P. spadiceum-AGH786 showed significantly (P = 0.05) higher polyphenol oxidases activity (2.2 mg/g DW) under normal conditions as compared to the NaCl-treated plants. We also observed that P. spadiceum-AGH786 improved biomass (0.30 g) of wheat plants subjected to 140 mM NaCl stress. The results conclude that the wheat plant colonization by P. spadiceum-AGH786 greatly improved the plant growth under 70 mM and 140 mM NaCl stress. Thus, the biomass of the P. Spadiceum-AGH786 can be used in saline soil to help the host plants.

ACC deaminase-producing endophytic fungal consortia promotes drought stress tolerance in M.oleifera by mitigating ethylene and H2O2.

Introduction: Drought has become more prevalent due to dramatic climate change worldwide. Consequently, the most compatible fungal communities collaborate to boost plant development and ecophysiological responses under environmental constraints. However, little is known about the specific interactions between non-host plants and endophytic fungal symbionts that produce growth-promoting and stress-alleviating hormones during water deficits. Methods: The current research was rationalized and aimed at exploring the influence of the newly isolated, drought-resistant, ACC deaminase enzyme-producing endophytic fungi Trichoderma gamsii (TP), Fusarium proliferatum (TR), and its consortium (TP+TR) from a xerophytic plant Carthamus oxycantha L. on Moringa oleifera L. grown under water deficit induced by PEG-8000 (8% osmoticum solution). Results: The current findings revealed that the co-inoculation promoted a significant enhancement in growth traits such as dry weight (217%), fresh weight (123%), root length (65%), shoot length (53%), carotenoids (87%), and chlorophyll content (76%) in comparison to control plants under water deficit. Total soluble sugars (0.56%), proteins (132%), lipids (43%), flavonoids (52%), phenols (34%), proline (55%), GA3 (86%), IAA (35%), AsA (170%), SA (87%), were also induced, while H2O2 (-45%), ABA (-60%) and ACC level (-77%) was decreased by co-inoculation of TP and TR in M. oleifera plants, compared with the non-inoculated plants under water deficit. The co-inoculum (TP+TR) also induced the antioxidant potential and enzyme activities POX (325%), CAT activity (166%), and AsA (21%), along with a lesser decrease (-2%) in water potential in M. oleifera plants with co-inoculation under water deficit compared with non-inoculated control. The molecular analysis for gene expression unraveled the reduced expression of ethylene biosynthesis and signaling-related genes up to an optimal level, with an induction of antioxidant enzymatic genes by endophytic co-inoculation in M. oleifera plants under water deficit, suggesting their role in drought stress tolerance as an essential regulatory function. Conclusion: The finding may alert scientists to consider the impacts of optimal reduction of ethylene and induction of antioxidant potential on drought stress tolerance in M. oleifera. Hence, the present study supports the use of compatible endophytic fungi to build a bipartite mutualistic symbiosis in M. oleifera non-host plants to mitigate the negative impacts of water scarcity in arid regions throughout the world.

Taxonomic investigation of selected rust fungi using scanning electron microscopy from Khyber Pakhtunkhwa, Pakistan.

Rusts comprises the largest natural group of plant pathogens including approximately 8% of all described Fungi. Rust fungi are extremely plant pathogens responsible for great losses to agriculture productivity. Rust species belong to several genera among which more than half are Puccinia species. In Pakistan, rust causes severe damage to agriculture crops. Current study was carried out to identify and characterize different rust species common in the research area through microscopy and Scanning electron microscopy (SEM) in Khyber Pakhtunkhwa, Pakistan. Morpho-anatomical investigation of each collected rust species was carried out using different standard protocols. The dimensions of spores were measured and snapped under a stereomicroscope. SEM was used to examine the shape, size, and ornamentation of the spores of each rust fungus. Results revealed documentation of seven rust fungi, that is, Melampsora euphorbiae, Phragmidium barclayi, Puccinia nepalensis, P. exhausta, P. menthae, Uromyces capitatus, and Uromyces decorates belong to four different genera, were recorded. SEM revealed that spermogonia and Aecia were missing in most of the rust fungus studied. Uredinia was found in a scattered, irregular, lengthy, and epidermis-enclosed form. Urediniospores were found to be ovulating, elongated, echinulate, globose to sub-globose, ellipsoid to ovoid, and globose to sub-globose. Telia was found as sub-epidermal, amphigenous, dispersed, minute, and spherical cells. Teliospores ranged in form from cylindrical to oblong. The germ pores were detected in both apical (top cell) and basal (bottom cell) idiosyncratic and pedicel-attached cells. The techniques used in the current investigation will aid mycologists in rust identification and microscopic characterization.

Industrial polluted soil borne fungi decolorize the recalcitrant azo dyes Synozol red HF-6BN and Synozol black B.

Today's world needs to control the industrial pollution through smarter ways. Presently, we observed the capacity of soil borne fungi to digest Synozol Red HF-6BN and Synozol Black B. Initially, 86 fungal strains were isolated from soil samples randomly collected from industrial sites. Among these, 31 isolates were capable of dye decolorization on solid media, with SN12f and SN13a isolates showed the highest decolorization capacity. The dye decolorization by both strains was higher (80-95%), when incubated for 120 h under optimized conditions of pH, concentration, nutrient source and temperature. The dye (Synozol red HF-6BN and Synozol black B) decolorization by SN12f isolate was maximum (˃90%) at pH7, whereas the SN13a decolorized 90% of Synozol red HF-6BN and 89% of Synozol black B at pH3. The SN13a and SN12f isolates at 40 mg/L showed de-colorization of 94.71%, 81.4% (for Synozol red HF-6BN) and 90.5%, 84.4% (Synozol black B), respectively. Our isolates also mitigated the toxic effect of azo dyes on the growth of phosphate solubilizing soil bacteria. In fact, the untreated effluent showed toxic effects on the growth of beneficial bacterial by developing zone of inhibition (16.5 mm around Aeromonas spp., 14.5 mm around Sallmonella while 14.25 mm around Citrobacter spp). However, the fungal treated dye was unable to develop zone of inhibition. Laccase activity was positive for both of fungal isolates after incubation on Bassnell Hass Medium (0.0733 U/mL for SN12f and 0.0439 U/mL SN13a). Using molecular approaches (ITS region), SN12f was identified as Aspergillus nidulans, while SN13a as Aspergillus fumigatus. The current study showed that local fungal flora can reclaim the contaminated soils and support the agro-friendly micro-flora.

A promising growth promoting Meyerozyma caribbica from Solanum xanthocarpum alleviated stress in maize plants.

Fungal endophytes are known to secrete a large array of secondary metabolites (phenols, flavonoids, indole acetic acid (IAA) etc.) that facilitate crops under stress conditions. Considering this, a potent plant growth promoting endophyte (SXSp1) from the spines and leaves of Solanum xanthocarpum L. has been isolated. The isolated strain ably secreted high quantities of indole-3-acetic acid, phenols and flavonoids. Also, it exhibited phosphate solubilization, siderophore and had 2,2 diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. The SXSp1 also resisted the salinity stress up to 150 mM. LC/MS analysis of SXSp1 culture filtrate (CF) revealed the presence of p-hydroxyl benzoic acid, diadzein, genistien, myricetin and caffeoyl-d-glucose. Moreover, the inoculation of maize plants with SXSp1 significantly (P=0.05) promoted the chlorophyll and carotenoid contents, root and shoot lengths, plant fresh and dry weights, catalase and peroxidase activities, proline, phenolics, flavonoids and relative water contents (RWCs) under salinity. More interestingly, SXSp1-associated plants showed lower endogenous abscisic acid (ABA) and higher endogenous IAA contents that helped the plants to resist salinity stress up to 100 mM. After sequencing, internal transcribed spacer (ITS) regions (ITS1 and ITS4) and phylogenetic analysis, the SXSp1 was identified as Meyerozyma caribbica.

Gibberellin application ameliorates the adverse impact of short-term flooding on Glycine max L.

Flooding is an abiotic stress that creates hypoxic conditions triggered by redox potential leading to restricted growth and grain yield in plants. In the current study, we have investigated the effect of exogenous gibberellins (GA4+7) on soybean under flooding stress. A regulatory role of GAs on biochemical changes in soybean plants [including chlorophyll contents, endogenous bioactive GA1 and GA4, endogenous jasmonic acid (JA) and abscisic acid (ABA)] has been elucidated after 3 and 6 h of flooding stress. The modulation of stress-related bio-chemicals and their genetic determinants [for instance, ABA (Timing of CAB expression1-TOC1, ABA-receptor-ABAR) and NO (S-nitrosoglutathione reductase-GSNOR1, NO overproducer1-NOX, and nitrite reductase-NR)] in response to short-term flooding stress were also explored. The current study showed that exogenous GAs rescued chlorophyll contents, enhanced endogenous bioactive GA1 and GA4 levels, endogenous jasmonic acid (JA) and checked the rate of ABA biosynthesis under short-term flooding. The exo-GAs induced the glutathione activity and reduced the resulting superoxide anion contents during short-term flooding in Pungsannamul soybean. Exo-GAs also triggered the endogenous S-nitrosothiols (precursor for increased NO production) that have been decreased over the time. Moreover, the exo-GAs could impinge a variety of biochemical and transcriptional programs that are ameliorative to plant growth during short-term flooding stress. The presence of GA1 and GA4 also confirms the presence of both C13-hydroxylation pathway and non-C13-hydroxylation pathway in soybean, respectively.

Anthracene biodegradation capacity of newly isolated rhizospheric bacteria Bacillus cereus S13.

Biodegradation of hazardous pollutants is of immense importance for maintaining a clean environment. However, the concentration of such contaminants/pollutants can be minimized with the help of microorganisms that has the ability to degrade the toxic pollutants into non-toxic metabolites. In the current study, 23 bacterial isolates were purified from the rhizospheric soil of Sysimbrium irio, growing as a wild plant in the vicinity of gas filling stations in Peshawar city. The isolated strains were initially screened on solid nutrient agar and further purified by culturing it on anthracene amended mineral media (PNR). The bacterial growth and anthracene disappearance were observed by calculating optical density (OD). The isolates showed a concentration-dependent growth on anthracene amended PNR media at 30°C and pH7. Also, an increase in bacterial OD from 0.351 to 1.80 with increased shaking speed was noticed. On the contrary, alternate carbon sources (glucose, fructose, sucrose) or nitrogen sources (KNO3, NaNO3, NH4NO3 and CaNO3) posed inhibitory effect on bacterial growth during anthracene degradation. The recorded efficiency of anthracene degradation by the selected bacterial isolate (1.4×1023 CFUmL-1 and 1.80 OD) was 82.29%, after 120 h of incubation. The anthracene was degraded to 9, 10, dihydroxy-anthracene and anthraquinone, detected through GC-MS. The efficient bacterial isolate was identified as S13, a new strain of Bacillus cereus, using 16S rRNA analysis, showing 98% homology. The isolated bacterial strain S13 may be used as a potential tool for bioremediation of toxic hydrocarbons and to keep the environment free from PAH pollutants.

Endophytic Fungus Aspergillus japonicus Mediates Host Plant Growth under Normal and Heat Stress Conditions.

We have isolated an endophytic fungus with heat stress alleviation potential from wild plant Euphorbia indica L. The phylogenetic analysis and 18S rDNA sequence homology revealed that the designated isolate was Aspergillus japonicus EuR-26. Analysis of A. japonicus culture filtrate displayed higher concentrations of salicylic acid (SA), indoleacetic acid (IAA), flavonoids, and phenolics. Furthermore, A. japonicus association with soybean and sunflower had improved plant biomass and other growth features under high temperature stress (40°C) in comparison to endophyte-free plants. In fact, endophytic association mitigated heat stress by negotiating the activities of abscisic acid, catalase, and ascorbic acid oxidase in both soybean and sunflower. The nutritional quality (phenolic, flavonoids, soluble sugars, proteins, and lipids) of the A. japonicus-associated seedlings has also improved under heat stress in comparison to endophyte-free plants. From the results, it is concluded that A. japonicus can modulate host plants growth under heat stress and can be used as thermal stress alleviator in arid and semiarid regions of the globe (where mean summer temperature exceeds 40°C) to sustain agriculture.

Effect of Methanolic Extract of Dandelion Roots on Cancer Cell Lines and AMP-Activated Protein Kinase Pathway.

Ethnomedicinal knowledge of plant-derived bioactives could help us in discovering new therapeutic compounds of great potential. Certainly, dandelion has been used in traditional ethno-medicinal systems (i.e., Chinese, Arabian, Indian, and Native American) to treat different types of cancer. Though, dandelion is highly vigorous, but the potential mode of action is still unclear. In the current study, the antiproliferative activity of methanolic extracts of dandelion root (MEDr) on cell viability of HepG2, MCF7, HCT116, and normal Hs27 was investigated. It was observed that MEDr (500 µg/mL) drastically decreased the growth of HepG2 cell line, while the effect on MCF7 and HCT116 cell lines was less pronounced and no effect has been observed in Hs27 cell lines. The MEDr also enhanced the phosphorylation level of AMPK of HepG2 cells, which considered crucial in cancer treatment and other metabolic diseases. The AMPK activation by MEDr noticed in the current study has never been reported previously. The results regarding the number of apoptotic cells (HepG2 cells) were in line with the cell viability test. The current observations clearly demonstrated the potency of MEDr against liver cancer with validation that dandelion could control AMPK and thus cancer in the treated cell lines.

Enzyme inhibitory metabolites from endophytic Penicillium citrinum isolated from Boswellia sacra.

Fungal endophytes establish an important niche within the host plant through the secretion of chemical constituents. Isolation of bioactive metabolites could be a vital source for inhibiting the function of enzymes such as α-glucosidase and urease. The present study aimed to elucidate the potential of endophytes associated with Boswellia sacra through bioassay-guided isolation and identification of secondary metabolites with enzyme inhibitory ability. Endophytic fungal strains viz. Penicillium citrinum, P. spinulosum, Fusarium oxysporum, Alternaria alternata and Aspergillus caespitosus were identified through genomic DNA extraction, PCR amplification, sequencing and phylogenetic analysis. The enzymes inhibition analysis of the ethyl acetate extract from pure cultures suggested that P. citrinum possess significantly higher enzyme inhibitory activities compared to other strains. The active strain was subjected to chromatographic isolation and nuclear magnetic resonance methods to identify bioactive compounds. The bioactive extracts resulted in the isolation of 11-oxoursonic acid benzyl ester (1), n-nonane (2), 3-decene-1-ol (3), 2-Hydroxyphenyl acetic acid (4), and Glochidacuminosides A (5). Among pure compound, 11-oxoursonic acid benzyl ester (1) showed significantly higher enzyme inhibition activity compared to other metabolites. Our results suggest that the endophytic microorganism associated with the arid-land tree can offer a rich source of biologically active chemical constituents that could help discover lead drugs for enzyme inhibition.