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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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Heavy metal (HM) contamination in the soil and accumulation in plants poses a serious threat to crops and human health. HMs such as cadmium, mercury, lead and arsenic are widely acknowledged for their adverse effects on plants such as chlorosis, necrosis, low food quality and crop yields. Endophytic fungi secrete secondary metabolites and enhance the plant's ability to tolerate stressful conditions. However, the role of most fungal endophytes in their host plant growth or production of metabolites under HM stress conditions needs further understanding. In the present study, we studied the HM stress alleviation capability of the endophytic fungus, Paecilomyces lilacinus (MRF), isolated from the roots of Justicia adathoda. We studied two heavy metals, namely lead and cobalt. The culture filtrate (CF) of P. lilacinus revealed IAA (68.17 µg/mL), phenols (43.31 µg/mL), flavonoids (40.59 µg/mL), sugar content (97.83 µg/mL) and proline (17 µg/mL). Additionally, DPPH-free radical scavenging activity and the antibacterial potential against Salmonella typhi and Shigella sonnei of the CF demonstrated positive results. The gas chromatography mass spectrometry analysis of the CF manifested different constituents, including (1) Trichloromethane, (2) 3,7,11,15-Tetramethyl-2-hexadecen-1-ol, (3) phytol, (4) 1,2-Benzenedicarboxylic acid, (5) bis(2-ethylhexyl) ester, (6) squalene and (7) Cyclotrisiloxane Hexamethyl. Moreover, the plant growth-promoting activity of the P. lilacinus (MRF) strain revealed a robust increase in root and shoot growth, and the fresh and dry weight of S. lycoprsicum. Further, the IAA, phenols, flavonoids, sugar, proline, relative water content and protein contents also increased in the S. lycoprsicum inoculated with P. lilacinus as compared to the control plants. The present study revealed that the inoculation of P. lilacinus alleviates the damages of HM stress and improves the physicochemical characteristics of S. lycoprsicum. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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 enzymeproducing 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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Among the types of abiotic stresses, drought is one of the most serious growth limiting factors for crop plants. In recent years, various strategies have been employed to alleviate the adverse effects of drought stress in crops, but the use of plant growth-promoting rhizobacteria (PGPR) is among the standout techniques. Currently, we have isolated 55 strains of bacteria from the rhizosphere of Achyranthes aspera L. and Calotropis procera (Aiton). However, AGH3, AGH5, and AGH9 produced significant (p = 0.05) amounts of plant hormones and exhibited siderophore and phosphate solubilization activities. Bioassay experiments on Waito-C rice demonstrated an enhanced growth in the presence of the isolate AGH3. Moreover, the isolate AGH3 promoted the growth of Solynum lycopersicum L. under drought stress. The results revealed that AGH3-associated S. lycopersicum plants showed significantly (p = 0.05) reduced production of abscisic acid (ABA) and jasmonic acid (JA) as compared with the AGH3-free plants under polyethylene glycol (PEG) stress. In addition, high expressions of SlmiR 159 (from 6- to 10-fold), SlHsfA1a (from 1- to 4-fold), and SlHAKT1 (from 0.26- to 1-fold) genes were noticed in AGH3-associated S. lycopersicum plants under drought stress. From the results, it is concluded that rhizobacteria (AGH3) can be used as a pragmatic biofertilizer to ensure organic farming under normal as well as drought conditions. [ABSTRACT FROM AUTHOR]

Rapid industrialization and global warming have threatened the plants with multiple abiotic stresses, such as heavy metals and drought stress. For crop cultivation, the conventional approach of cleaning the soils by excavation is very costly and not feasible for large scale. Establishing toxin-free and droughtresistant crops is a major challenge in the environment under natural and anthropogenic pressure. In the past decades, copper contamination of agricultural land has become an emerging concern. For dry land reclamation, several new strategies, including bioremediation (phytoremediation and microbial remediation), have been used. Owing to the potential of Cu hyperaccumulators, the current project aims to enhance the drought tolerance and the phytoremediation potential of Solanum lycopersicum L. with the inoculation of copper and 12% polyethylene glycol (PEG)-induced drought stress-tolerant endophytic fungus Porostereum spadiceum AGH786 under the combined stress of copper heavy metal and PEG-induced drought stress. When S. lycopersicum L. was watered with individual stress of copper (Cu) concentration (400 ppm) in the form of copper sulfate (CuSO4.5H2O), 12% PEG-induced drought stress and the combined stress of both negatively affected the growth attributes, hormonal, metabolic, and antioxidant potential, compared with control. However, the multistress-resistant AGH786 endophytic fungus ameliorated the multistress tolerance response in S. lycopersicum L. by positively affecting the growth attributes, hormonal, metabolic, and antioxidant potential, and by restricting the root-to-shoot translocation of Cu and inducing its sequestration in the root tissues of affected plants. AGH786-associated plants exhibited a reduction in the severity of copper (Cu) and drought stress, with higher levels of SlCOPT (Cu transporters) and SlMT (metallothionine) gene expressions in root and shoot tissues, indicating that AGH786 contributed to resistance to copper metal toxicity and drought stress in the host S. lycopersicum L. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Fungal endophytes are not widely known for their role in bioactive metabolite production and salinity stress alleviation in different crop plants. Presently, we investigated the salt stress (NaCl, KCl, and H2SO4) mitigation capabilities of fungal endophyte Candida membranifaciens (FH15) isolated from Euphorbia milii L. The pure culture filtrate (CF) of C. membranifaciens revealed siderophore production and solubilization of phosphate, with high levels of indoleacetic acid (IAA: 35.8µg/mL), phenolics (70 µg/mL), and flavonoids (50 µg/mL) by using a UV spectrophotometer. The LC/MS analysis of the CF showed different phenols and flavonoids that were identified as Salicylic acid, Baicalein, Aconitic acid, Feruloylquinic acid, Coniferyl aldehyde hexoside, Pentose, Chlorogenic acid, Myricetin, Propoxyphene, and Amino-flunitrazepam. Inoculation of maize seedlings with C. membranifaciens significantly (p = 0.05) enhanced the fresh and dry biomass, carotenoid, and chlorophyll contents under 100 mM salt stress conditions. Similarly, the catalase, peroxidase activity, phenols, proline flavonoids and relative water contents (RWC) of the maize plants were enhanced. More interestingly, the inoculation of C. membranifaciens on maize revealed a higher endogenous IAA level as compared to non-inoculated control plants. Endophyte C. membranifaciens inoculation on maize seedlings under salt stress revealed a 20.87% and 16.60% increase in fresh and dry biomass, as well as significantly enhanced root shoot length and allied growth attributes, in addition to an alleviation of the adverse effects of salinity stress. Conclusively, endophytic C. membranifaciens significantly enhanced the growth attributes of maize and mitigated the adverse effects of salinity stress. Such endophytic fungal strain could be used for further field trails to enhance agricultural productivity and facilitate sustainable agricultural practices. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) and chromium (Cr) in agricultural soils is becoming a serious threat in tomato. The complete removal of these heavy metals (HMs) from contaminated soil is challenging. However, the use of plant endophytes might be beneficial. The pragmatic role of endophytic fungi Aspergillus flavus (Ch-01) in tomato under Cd and Cr stress was, therefore, investigated. A. flavus enhanced growth attributes in tomato seedlings under Cd and Cr stress. Besides, it remediated Cd and Cr toxicity by reducing their translocation from the roots to the upper part of the tomato plants. In fact, A. flavus associated plants developed tolerance against Cd and Cr toxicity via the expression of SlGSH1 and SlPCS1 genes. Both genes helped in metal chelation and mitigation of Cd and Cr toxicity. From the results, it is concluded that A. flavus is a plant growth promoting and HMs alleviating endophyte, hence it might be used as a biofertilizer. [ABSTRACT FROM AUTHOR]

Plants can be severely affected by heat stress due to rapid environmental changes. Use of endophytic fungi is a new tool to protect crops from the environmental stresses. Here we claim a potent endophyte isolated from Sonchus asper L. The aim was to explore the stress adaptive mechanism of sunflower and soybean mediated by Aspergillus niger (SonchL-7) under high temperature. Inoculation with A. niger boosted plant height, biomass and chlorophyl contents, while significantly curtailed the concentration of lipid peroxidation and reactive oxygen species (ROS) under thermal stress at 40°C. Moreover, the ROS-scavenging activities, like ascorbic acid oxidase (AAO), catalase (CAT), glutathione reductase (GR), superoxide dismutase (SOD) and peroxidase (POD) were augmented. Also, proline and phenolics were enhanced in the tested crops, while ABA concentration was significantly reduced. These positive results suggested that A. niger can be used as a heat-stress ameliorative tool for crops in the future. [ABSTRACT FROM AUTHOR]

High temperature is one of the leading threats to the plants that severely affects crop quality as well as quantity. Endophytic fungi might be a new tool to safeguard crops against the perilous effects of global warming. In this context, we isolated a thermal stress mitigating endophytic fungus from the fern Dryopteris flix L. The phylogenetic study and 18S rRNA sequence similarity confrmed the potential strain as Aspergillus violaceofuscus. The culture fltrate of A. violaceofuscus exhibited higher concentration of secondary metabolites that enhanced the total chlorophyll content, plant height and biomass of sunfower and soybean seedlings under heat stress. Conversely, the A. violaceofuscus associated plants achieved low levels of reactive oxygen species, abscisic acid, catalase, ascorbic acid oxidase, proline and an overall improved the nutritional value. The current study suggests that A. violaceofuscus can be used as heat stress adaptive tool in crops to achieve sustainable agriculture. [ABSTRACT FROM AUTHOR]

A total of 149,551 cases of diarrheal infections, 142,739 case of skin infections, 132,485 cases of acute respiratory illness and 49,420 probable malaria cases were recently reported in flood-affected areas of Pakistan (Ochani et al., [3]). Currently, Pakistan is experiencing monsoon rainfall that is three times greater than usual, according to the World Weather Attribution (WWA, [6]). Authorities should take infection-control measures based on local weather conditions, extent of flooding and the ease of transmission of infection. [Extracted from the article]

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. [ABSTRACT FROM AUTHOR]

Aim: To investigate the HCV 3a core sequence variation and amino acid substitutions of patients from Lahore, Pakistan. Materials & methods: Blood samples from HCV positive patients (n = 232) were collected for viral genotypes. Moreover, the nucleotide sequencing was performed for core gene of 20 samples. Results: Viral genotyping showed that 69.82% (n = 162) belonged to 3a genotype, 9.05% (1a; n = 21), 2.15% (3b; n = 5) and 18.98% were untypable (n = 44). Phylogenetic analyses suggest majority of our isolates clustered with previously reported reference isolates from Pakistan. The remaining isolates clustered with HCV-core sequences reported from Vietnam, Japan, Thailand, Iran, USA, Bangladesh, Malaysia and Morocco. Conclusion: We report HCV-core substitutions (G60E, R70Q, C91A, A94Q and Q63E/D) that could be associated with treatment response in Pakistani patients. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Plant growth and gibberellins (GAs) biosynthesis are two separate but linked processes, involving many genes but fewer have been reported for their role in plant growth and development. Due to little information on the genes involved in such processes, the known plant growth promoting and GAs producing fungal endophytePenicillium citrinumKACC43900 was used as potential tool to obtain a blueprint of the putative growth promoting and GAs synthesizing proteins. For proteomic analysis, the seedlings ofGlehnia littoraliswere treated with culture filtrate ofP. citrinumKACC43900, which revealed significant differences between 2 dimensional gel electrophoresis profile of the crude protein extracts of the treated and control samples. Matrix-assisted laser desorption/ionization analysis of the 56 selected spots led to the identification of 41 proteins. A significant number (31.5%) of these highly expressed proteins were associated with plant growth regulation, including beta-expansin EXPB4, ent-kaur 16-ene synthase, gibberellin 3-oxidase, and cytochrome P450 family proteins. Proteins involved in regulating energy metabolism and intracellular redox conditions, such as glyceraldehyde-3-phosphate dehydrogenase and ribulose-1,5-bisphospahate carboxylase/oxygenase, were also expressed. It was concluded that culture filtrate ofP. citrinumKACC43900 altered the gene expression pattern of hostG. littoralis. Current study highlighted the importance of proteomics as a starting tool for any post-genomic research. [ABSTRACT FROM AUTHOR]

An ethnobotanical research project was carried out in order to investigate the floral diversity of Utror and Gabral, two twin valleys situated in the remote north western part of District Swat, Pakistan. The area exhibit unique and diverse flora, as it is located in the Hindu-Kush mountain region, adjacent to Himalayas and Karakorams. We observed that twin valleys contain 9 species of Bryophytes belonging to 5 families, 34 species of Pteridophytes belonging to 6 families, 10 plant species of Gymnosperms belonging to 3 families and 610 plant species of Angiosperms belonging to 93 families (104 species of Monocot belonging to 13 families and 506 plant species of Dicot belonging to 80 families). [ABSTRACT FROM AUTHOR]

Crop productivity continues to decline due to a wide array of biotic and abiotic stresses. Salinity is one of the worst abiotic stresses, as it causes huge losses to crop yield each year. Kinetin (Kn) has been reported as plant growth regulator since long, but its role in improving plant growth and food quality under saline conditions through mediating phytohormonal cross-talk is poorly studied. Current study was designed to evaluate the impact of exogenously applied Kn on growth, isoflovones and endogenous phytohormones of soybean grown under NaCl induced salt stress. Soybean plants were grown in perlite (semi hydroponic), and under controlled green-house conditions. Elevated levels of exogenous Kn significantly mitigated the adverse effect of NaCl and rescued plant growth attributes, i.e., plant height, fresh and dry biomass of soybean plants grown in all treatments. Higher diadzen, glycitin, and genistin contents were observed in plants treated with elevated Kn in the presence or absence of NaCl induce salt stress. The gibberellins (GAs) biosynthesis pathway was up-regulated by Kn as the bioactive GA1 and GA4 contents were significantly higher in Kn treated plants, as compared to control, while GAs level reduced in NaCl treated plants. Contrary to GAs, the abscisic acid contents declined with Kn but promoted in NaCl stressed soybean plants. The endogenous jasmonic acid and salicylic acid contents of soybean enhanced with elevated Kn application, but they showed an antagonistic response under salt stress. Current study supports the active role of Kn to ameliorate the adverse effects of salt stress on the growth and food quality of soybean. The favorable role of Kn toward soybean growth under salt stress may be attributed to its potential to modulate crosstalk between the various phytohormones involved in soybean growth and its resistance to salinity stress. [ABSTRACT FROM AUTHOR]

Theclinical outcomes of patients infected with hepatitis C virus (HCV) range fromacute resolving hepatitis to chronic liver diseases such as liver cirrhosis or hepatocellular carcinoma. Identification of the infecting virus genotype is indispensable for the exploration of many aspects of HCV infection, including epidemiology, pathogenesis, and response to antiviral therapy. 1419 individuals were screened for anti-HCV in this study, of which 166 (11.7%) were found reactive by ICT (Immunochromatographic test). These 166 anti-HCV positive and 26 normal individuals were further analyzed. RNA was extracted from serum and reverse-transcribed to cDNAand the core region of HCVgenomewas targeted and amplified bymultiplex PCR. HCVRNAwas detected in 121 individuals, of which 87 were male and 34 were female. Genotype 3a was the most prevalent among all the genotypes observed followed by 3b. Genotypes 1a, 2a, and 2b were found in 10.89%, 13.22%, and 6.61% patients, respectively. 25.41% of the HCV RNA positive samples were not typed. 6.05% of patients were found having mixed genotypes. These findings will not only help the physicians to prescribe more appropriate treatment for the HCV infection but will also draw the attention of health-related policy makers to devise strategies to curb the disease more effectively. [ABSTRACT FROM AUTHOR]

Recently, plant growth-promoting endophytic fungi with gibberellins-producing ability were isolated from the roots of coastal sand dune plants. Carex kobomugi Ohwi in coastal sand dunes was treated with the supernatant solution of Penicillium citrinum KACC43900 isolated from Ixeris repens (L.) A. Gray. After treatment, the length of leaf blades, photosynthesis, chlorophyll fluorescence, and soil respiration were measured in the field. The contents of chlorophyll and carotenoid were determined with leaf blades selected for measurement of chlorophyll fluorescence. Fungal culture filtrate (FCF) increased leaf blade length, the contents of chlorophyll a (Chl a) and chlorophyll b (Chl b), total chlorophyll (total Chl [a + b]), and carotenoid in leaf blades of C. kobomugi. FCF also increased net photosynthetic rate (A), transpiration rate (E), carboxylation efficiency (A/Ci), and water-use efficiency (A/E). However, the ratio of variable fluorescence to maximal fluorescence (Fv/Fm) did not show significant difference between control and FCF-treated plants. Soil respiration rates were higher in the site treated with FCF than the control. Although growth response and physiological characteristics of C. kobomugi were different according to the treatment methods of FCF, injection or spray treatment, plant growth of C. kobomugi was increased with FCF treatment. [ABSTRACT FROM AUTHOR]

The beneficial effects of plant growth promoting fungi (PGPF) on plant growth and development are well documented. However, limited information is available on gibberellin (GA) production capacity of PGPF of endophytic origin. In current study, 11 fungal endophytes were isolated from cucumber roots and then screened on Waito-C rice, in order to identify plant growth promoting fungal strains. The fungal isolate GAH7 provided the maximum shoot length (11.3 cm) in comparison to control treatment (7.8 cm). In a separate experiment, bioassay of GAH7 significantly promoted growth attributes of cucumber. The GAH7 culture filtrate (CF) was found to contain physiologically active gibberellins in higher concentrations (GA1, 0.81 ng/ml; GA3, 4.34 ng/ml and GA4, 9.31 ng/ml) in conjunction with physiologically inactive GA9 (0.74 ng/ml), GA15 (0.97 ng/ml), GA19 (1.67 ng/ml) and GA20 (0.46 ng/ml). Isolate GAH7 produced higher amounts of GA3, GA4, GA9 and GA19 than wild type Fusarium fujikuroi, which was used as control for GA production. Gibberellins were analyzed through gas chromatograph/mass spectrometer (GC/MS) with selected ion monitoring (SIM). The fungal isolate GAH7 was later identified as a new strain of Phoma on the basis of sequence homology (99%) and phylogenetic analysis of 18S rDNA sequence. [ABSTRACT FROM AUTHOR]

Plant growth-promoting endophytic fungi with gibberellin-producing ability were isolated from the roots of Carex kobomugi Ohwi, a common sand-dune plant, and bioassayed for plant growth-promotion. A new strain, Arthrinium phaeospermum KACC43901, promoted growth of waito-c rice and Atriplex gemelinii. Analysis of its culture filtrate showed the presence of bioactive GA1 (0.5 ng/ml), GA3 (8.8 ng/ml), GA4 (4.7 ng/ml) and GA7 (2.2 ng/ml) along with physiologically inactive GA5 (0.4 ng/ml), GA9 (0.6 ng/ml), GA12 (0.4 ng/ml), GA15 (0.4 ng/ml), GA19 (0.9 ng/ml) and GA24 (1.8 ng/ml). The fungal isolate was identified through sequence homology and phylogenetic analysis of 18S rDNA (internal transcribed region). [ABSTRACT FROM AUTHOR]

Plants are susceptible to various environmental constrains, including heat stress due to their sessile nature. Endophytic fungi can be used as a novel technique to protect crop plants against the injurious effects of thermal stress. Endophytic fungi were isolated from Adiantum capillus-veneris L. and tested against heat stress in Glycine max L. and Helianthus annuus L. The results exhibited increased levels of the plant's chlorophyll, height and biomass in Aspergillus foetidus (AdR-13) inoculated host crop species. Conversely, a significant decrease in lipid peroxidation and reactive oxygen species (ROS) production was noted in A. foetidus-associated host crop species. Likewise, the amounts of ROS-degrading antioxidants (glutathione reductase (GR), peroxidase (POD), ascorbic acid oxidase (AAO), superoxide dismutase (SOD), catalase (CAT)) as well as phenolics were increased, while the amounts of proline and abscisic acid (ABA) were decreased in fungal-associated test crops. Total lipids, proteins and sugars were noted to be high in A. foetidus-associated test crops. From the results, we concluded that A. foetidus have a role in heat stress mitigation that might help to sustain the production of important crops in the future. [ABSTRACT FROM AUTHOR]