Aspergillus Terreus Strain Research Articles

><em>Invitro</em> Efficacy of Biosynthesized Silver Nanoparticles on <em>Trigonella foenum-graecum</em> L. Seed Germination and Growth

Nanoparticles have been suggested as a relatively new hope in agriculture area as a biofertilizer to make agriculture more sustainable. The purpose of this study was to compare effect of biologically synthesized silver nanoparticles on growth and development of plant during germination as well as seedling and vegetative period. Fertilizers are an important key factor for the production of any crop in both developed and developing countries. Nanoparticles as biofertilizer are a relatively new approach in agriculture area and it has had many promising results. It has potential contribution in slow release of fertilizers. In the present investigation, Silver nanoparticles were biologically synthesized using Aspergillus terreus and Aspergillus niger strains collected from culture center in aseptic conditions. It was observed that low concentrations of Ag nanoparticles being 20 ppm, 40 ppm and 60 ppm showed better growth than higher concentrations like 80 ppm and 100 ppm. The plants treated with low concentrations of Ag nanoparticle showed better growth than the control. So, to conclude there is significant difference in the growth of plants which had been treated with Ag nanoparticle and those without.

Lovastatin producing by wild strain of Aspergillus terreus isolated from Brazil

Lovastatin is a drug in the statin class which acts as a natural inhibitor of 3-hydroxy-3-methylglutaryl, a coenzyme reductase reported as being a potential therapeutic agent for several diseases: Alzheimer’s, multiple sclerosis, osteoporosis and due to its anti-cancer properties. Aspergillus terreus is known for producing a cholesterol reducing drug. This study sets out to evaluate the production of lovastatin by Brazilian wild strains of A. terreus isolated from a biological sample and natural sources. Carbon and nitrogen sources and the best physicochemical conditions using factorial design were also evaluated. The 37 fungal were grown to produce lovastatin by submerged fermentation. A. terreus URM5579 strain was the best lovastatin producer with a level of 13.96 mg/L. Soluble starch and soybean flour were found to be the most suitable substrates for producing lovastatin (41.23 mg/L) and biomass (6.1 mg/mL). The most favorable production conditions were found in run 16 with 60 g/L soluble starch, 15 g/L soybean flour, pH 7.5, 200 rpm and maintaining the solution at 32 °C for 7 days, which led to producing 100.86 mg/L of lovastatin and 17.68 mg/mL of biomass. Using natural strains and economically viable substrates helps to optimize the production of lovastatin and promote its use.

Determination of territtrem B in rice media with different strains of Aspergillus terreus by chromatography

Territrem B is a tremogenic mycotoxin produced by Aspergillus terreus. Animal studies have demonstrated that territrem B caused whole body tremor, salivation, convulsion and death. Territrem B is also a potent irreversible inhibitor of acetycholinesterase (AchE, E. C. 3.1.1.7). The aim of the present study is to assess the ability of strains of Aspergillus terreuss to produce territrem B in the rice media, which were obtained from Food Industry Research and Development Institute in Taiwan. In the 17 strains of A. terreus, only No. 32111 and 32664 strains produced territrem B-like blue fluorescence compound by TLC and HPLC analyses. Later on, this compound was identified as territrem B by mass spectroscopy. The amount of territrem B was determined by reverse phase HPLC on a C18 column with UV detection at 335 nm. The linear regression coefficient of the standard curve (Y=1428.5X+306.0) for territrem B at concentration ranging from 0 to 250 ng/L is 0.9998. In rice media, the recovery rates for territrem B at 50, 100 and 200 ng/L ranged from 91.5% to 95.2% with a coefficient of variation less than 2.1 %. The relative standard of territrem B ranged between 1.6-2.8% (intra-day) and 0.6-2.3% (interday). In rice culture of A. terreus 32111 and 32664 strains, the contents of territrem B were 0.20 and 0.31 mg/100 g rice, respectively. From the above results, it can be concluded that this HPLC method for determination of territrem B in rice media is suitable and only domestic strains can produce territrem B.

Draft Genome Sequence of the Aspergillus terreus High-Itaconic-Acid-Productivity Strain IFO6365.

Itaconic acid is an important organic acid used in the chemical industry. Aspergillus terreus strain IFO6365 is one of the highest-yielding itaconic acid-producing wild-type strains. Here, we report the draft genome sequence of IFO6365, enhancing the understanding of the role and biosynthesis of itaconic acid in this fungus.

Anticancer activity of gelatin-asparaginase nanobiocomposite against cervical and brain cancer cell lines

l-Asparaginase acts as a therapeutic agent for cancer. The free l-asparaginase has innumerable side effects on the human body. The source of l-asparaginase also plays a major role in the effectiveness of the therapeutic agent. In the present research work, l-asparaginase was produced from a fungal strain Aspergillus terreus. The free enzyme was then immobilised onto a nanobiocomposite consisting of Gelatin and β-cyclodextrin. Both Gelatin and β-Cyclodextrin are biodegradable and are non-toxic. The nanobiocomposite was subjected to characterization studies like the UV–Vis spectra analysis which confirms the presence of the nanobiocomposite. The FTIR analysis of the nanobiocomposite confirmed the functional group present in the nanobiocomposite. The NMR analysis confirmed the chemical shift of the nanobiocomposite. The AFM analysis was used to study surface morphology of the nanobiocomposite. The anticancer activity for the brain cancer cell line and cervical cancer cell line was found as 42.13% at 500 μg/ml and 48.60% at 62.5 μg/ml respectively.

Bioprocess development for production of a process-apt xylanase with multifaceted application potential for a range of industrial processes

Discovery of new/novel enzymes with process-apt features has been a continuous practice. A wide range pH-stable and thermostable xylanase was reported hitherto from a newly isolated fungal strain Aspergillus terreus S9, from mushroom compost. This is the first ever report where-in a pH-stable and thermostable xylanase is being reported from a strain of A. terreus. Solid state fermentation was executed on agro-industrial residues for xylanase production. Maximum xylanase production was supported by wheat bran (40 U/ml), however, substantial xylanase yield was observed on other residues like corn cob powder (38.4 U/ml), cotton cake (38 U/ml), malt (37.6 U/ml) and almond hulls (35.2 U/ml). Almond hulls are being reported for the first time as the substrate for xylanase production. Plackett–Burman design was used to earmark the most effective process variables (wheat bran, medium pH, and incubation time), and the same were optimized by design of experiments (DoE) based on response surface methodology to achieve a yield enhancement of 1.82-fold. A. terreus S9 xylanase exhibited an excellent and brisk clarification of mosambi and orange juices. DoE based optimization of process parameters (enzyme dose, treatment time and incubation temperature) resulted in 1.23-fold and 1.28-fold enhanced orange and mosambi juice clarification, respectively. Furthermore, admirable dough raising capacity and pulp brightening ability of A. terreus S9 xylanase shows its promising potential for bread making, and eco-friendly biobleaching processes, respectively. The xylanase must be in-depth investigated to decipher the molecular mechanisms of diverse applications.

Tryptoquivalines W and X, two new compounds from a Hawaiian fungal strain and their biological activities

Two new compounds tryptoquivalines W (1) and X (2) were isolated from a Hawaiian soil fungal strain Aspergillus terreus FS107. The soil sample was collected on the top of Mauna Kea, the tallest mountain in Hawaii. The structures of compounds 1 and 2 were determined on the basis of MS spectroscopic and NMR analysis, and NMR calculation. The absolute configuration (AC) was determined by ECD calculations. Compounds 4 and 5 showed inhibition against NF-κB with IC50 values of 3.45 and 6.76 μM, respectively.

Structural insights into the catalytic mechanism of lovastatin hydrolase

Structural insights into the catalytic mechanism of lovastatin hydrolase

SCREENING OF FILAMENTOUS FUNGI FROM THE ATLANTIC FOREST BIOME PRODUCING ENZYMES OF THE PECTINOLYTIC COMPLEX

Fungi are the major producers of enzymes such as cellulases, pectinases and xylanases, so these microorganisms were selected in the present work to analyze the ideal conditions to improve the production of the activity of the pectinase enzyme. A total of 36 fungi previously collected from the Atlantic Forest of Parana were cultivated in a test tube containing 10 to 15 mL of BDA medium for 7 days at 40 °C. Among all analyzed, the filamentous fungus Aspergillus terreus strain PA3A5T was the best producer of pectinases, whose greatest enzymatic activity occurred at a temperature of 60 °C and at pH 5. The ideal culture medium for inducing production Enzyme was the defined medium Vogel supplemented with passion fruit peel at 1% (w/v) as a carbon source and incubated for 96 hours. To our knowledge, this is the first time that these data are reported in the literature.

Draft Genome Sequence of Aspergillus terreus High-Itaconic-Acid-Productivity Mutant TN-484.

Itaconic acid is an important organic acid used in the chemical industry. Aspergillus terreus strain TN-484 is a high-itaconic-acid-productivity mutant derived from strain IFO6365. Here, we report the draft genome sequence of strain TN-484, advancing the understanding of the biosynthesis of itaconic acid in filamentous fungi.

Identification and inoculation of fungal strains from Cedrus deodara rhizosphere involve in growth and alleviation of high nitrogen stress.

Cedrus deodara is economically and ethnobotanically an important forest tree and is shown to be at decline in Northern areas of Pakistan in recent years mainly due to high concentration of Nitrogen in forests. Ectomycorrhizal (ECM) association forming fungi enables the forest trees to develop optimally by absorbing water from the rhizosphere through their absorptive hyphae and by making available the nutrients by mobilization of N and P from the organic substrates. This study was conducted to identify the ECM strains from C. deodara rhizosphere and to analyse the impact of high N load on the C. deodara seedlings to establish N critical load value for coniferous forests of Pakistan. Six new fungal strains were identified from the rhizosphere of C. deodara and were registered at GenBank (NCBI) as Emmia latemarginata strain ACE1, Aspergillus terreus strain ACE2, Purpureocillium lilacinum strain ACE3, Talaromyces pinophilus strain ACE4, A. fumigatus strain ACE5 and T. pinophilus strain ACE6 with accession numbers MH145426, MH145427, MH145428, MH145429, MH145430 and MH547115. Four out of six isolated strains were inoculated with seedlings of C. deodara singly and in consortium (CN) in combination with nitrogen load of 0 (C), 25 (T1), 50 (T2), 100 kg N ha−1 yr−1 (T3). Agronomic, physiological and gene expression studies for ExpansinA4 (EXPA4) and Cystatins (Cys) were made to analyse the impact of fungal strains in relation to high N stress. This study suggests a positive impact of T1 (25 kg N ha−1 yr−1) Nitrogen load and a negative impact of T3 (100 kg N ha−1 yr−1) on growth parameters and expression patterns of EXPA4 and Cys genes. Peroxidase (POX) activity decreased in the order ACE5 > ACE2 > C > ACE3 > ACE1 > CN. However, the results of Superoxide dismutase (SOD) showed decreasing trend in the order ACE5 > C > CN > ACE1 > ACE2 > ACE3. Strain ACE3 was shown to have a positive impact on the seedlings in terms of growth, physiology and expression of genes. Present study suggests that newly identified fungal strains showing positive impact on the growth and physiology of C. deodara could be used for the propagation of this economically important plant in Pakistan after pathogenicity test.

Evaluating aeration and stirring effects to improve itaconic acid production from glucose using Aspergillus terreus

The effects of the bioreactor conditions, in particular the mode and intensity of aeration and mixing were studied on itaconic acid (IA) fermentation efficiency by Aspergillus terreus strain from glucose substrate. IA was produced in batch system by systematically varying the oxygen content of the aeration gas (from 21 to 31.5 vol% O2) and the stirring rate (from 150 to 600 rpm). The data were analyzed kinetically to characterize the behavior of the process, and besides, the performances were evaluated comparatively with the literature. It turned out that the operation of the bioreactor with either the higher inlet O2 concentration (31.5 vol% O2) or faster stirring (600 rpm) could enhance biological IA generation the most, resulting in yield and volumetric productivity of 0.31 g IA/g glucose and 0.32 g IA/g glucose and 3.15 g IA/L day and 4.26 g IA/L day, respectively. Overall, the significance of fermentation settings was shown in this work regarding IA production catalyzed by A. terreus and notable advances could be realized by adjusting the aeration and stirring towards an optimal combination.Graphic abstract

Large-Scale Production of Bioactive Terrein by Aspergillus terreus Strain S020 Isolated from the Saudi Coast of the Red Sea.

The diversity of symbiotic fungi derived from two marine sponges and sediment collected off Obhur, Jeddah (Saudi Arabia), was investigated in the current study. A total of 23 isolates were purified using a culture-dependent approach. Using the morphological properties combined with internal transcribed spacer-rDNA (ITS-rDNA) sequences, 23 fungal strains (in the majority Penicillium and Aspergillus) were identified from these samples. The biological screening (cytotoxic and antimicrobial activities) of small-scale cultures of these fungi yielded several target fungal strains which produced bioactive secondary metabolites. Amongst these isolates, the crude extract of Aspergillus terreus strain S020, which was cultured in fermentation static broth, 21 L, for 40 days at room temperature on potato dextrose broth, displayed strong antimicrobial activities against Pseudomonas aeruginosa and Staphylococcus aureus and significant antiproliferative effects on human carcinoma cells. Chromatographic separation of the crude extract by silica gel column chromatography indicated that the S020 isolate could produce a series of chemical compounds. Among these, pure crystalline terrein was separated with a high yield of 537.26 ± 23.42 g/kg extract, which represents the highest fermentation production of terrein to date. Its chemical structure was elucidated on the basis of high-resolution electrospray ionization mass spectrometry (HRESIMS) or high-resolution mass spectrometry (HRMS), 1D, and 2D NMR spectroscopic analyses and by comparison with reported data. The compound showed strong cytotoxic activity against colorectal carcinoma cells (HCT-116) and hepatocellular carcinoma cells (HepG2), with IC50 values of 12.13 and 22.53 µM, respectively. Our study highlights the potential of A. terreus strain S020 for the industrial production of bioactive terrein on a large scale and the importance of future investigations of these strains to identify the bioactive leads in these fungal extracts.

Molecular Identification, Antifungal Susceptibility Testing, and Mechanisms of Azole Resistance in Aspergillus Species Received within a Surveillance Program on Antifungal Resistance in Spain.

Antifungal resistance is one of the major causes of the increasing mortality rates for fungal infections, especially for those caused by Aspergillus spp. A surveillance program was established in 2014 in the Spanish National Center for Microbiology for tracking resistance in the most prevalent Aspergillus species. A total of 273 samples were included in the study and were initially classified as susceptible or resistant according to EUCAST breakpoints. Several Aspergillus cryptic species were found within the molecularly identified isolates. Cyp51 mutations were characterized for Aspergillus fumigatus, Aspergillus terreus, and Aspergillus flavussensu stricto strains that were classified as resistant. Three A. fumigatus sensu stricto strains carried the TR34/L98H resistance mechanism, while two harbored G54R substitution and one harbored the TR46/Y121F/T289A mechanism. Seventeen strains had no mutations in cyp51A, with ten of them resistant only to isavuconazole. Three A. terreussensu stricto strains harbored D344N substitution in cyp51A, one of them combined with M217I, and another carried an A249G novel mutation. Itraconazole-resistant A. flavussensu stricto strains harbored P220L and H349R alterations in cyp51A and cyp51C, respectively, that need further investigation on their implication in azole resistance.

Nano-Biotechnology: Developing Nano-silver coated cotton fabrics by means of biosynthesis of silver nanoparticles using Aspergillus terrues strain (MTCC9618) against staphylococcus aureus

Biosynthesis of silver nanoparticles, especially fungal mediated method is given attention in the development of new drugs for resistance pathogens, molecular diagnosis, drug delivery therapy and in catalytic sensor due to its cost effective, none toxicity and eco-friendly. The present study focused on the fungal mediated biosynthesis of silver nanoparticles using Aspergillus terrues strain (MTCC 9618). The synthesized nanoparticles was monitored by spectrophotometer SEM, XRD and its band gap was determined by Tuac equation. After while the fungal crude cells was exposed to 5mM silver nitrate the reduction reaction was recorded according to red shift colorchange. Based on ultra violet spectrophotometeran absorbance was recorded in a distinct pick around 430nm - 450nm and also the band gab was determined using Tuac equationsuch that 2.08eV, 2.02eV, 2.0eV and 1.96eV at about 10min, 30min, 12h and 24h respectively. The AgNPs coated cotton fabrics was developed through direct exposed to extracellular metabolites and 100ppm colloidal solution of AgNPs. The antimicrobial efficacy of the synthesized AgNPs coted cotton fabrics against gram positive Hospital staphylococcus aureus pathogenstrains was conducted by disk diffusion assay. In which the antimicrobial efficacy of coated cotton-Ag against human pathogens was proofed how the staphylococcus aureus had susceptible too and (16mm) zone of inhibition was recorded. Based on disk diffusion assay at 10ug/mL minimum inhibition concentration (MIC)10.5 mm inhibition zone was noted consequently, this study accomplished that Aspergillus terreus strain mediated biosynthesis of silver nanoparticles is cost effective, time saving, eco-friendly and small spherical (<10nm) had produced against to Physio-chemical means. The bio-synthesized silver nanoparticles cotton fabrics publicized that a higher efficacy of antimicrobial activity against staphylococcus aureus and the result was considerable suggested in widely range used in textile and pharmaceutical industries to enrich durability, strength, quality of products against a clinical pathogens application as well bad odor and spoilage of dusts from fabrics.

Biomass-Derived Production of Itaconic Acid as a Building Block in Specialty Polymers.

Biomass, the only source of renewable organic carbon on Earth, offers an efficient substrate for bio-based organic acid production as an alternative to the leading petrochemical industry based on non-renewable resources. Itaconic acid (IA) is one of the most important organic acids that can be obtained from lignocellulose biomass. IA, a 5-C dicarboxylic acid, is a promising platform chemical with extensive applications; therefore, it is included in the top 12 building block chemicals by the US Department of Energy. Biotechnologically, IA production can take place through fermentation with fungi like Aspergillus terreus and Ustilago maydis strains or with metabolically engineered bacteria like Escherichia coli and Corynebacterium glutamicum. Bio-based IA represents a feasible substitute for petrochemically produced acrylic acid, paints, varnishes, biodegradable polymers, and other different organic compounds. IA and its derivatives, due to their trifunctional structure, support the synthesis of a wide range of innovative polymers through crosslinking, with applications in special hydrogels for water decontamination, targeted drug delivery (especially in cancer treatment), smart nanohydrogels in food applications, coatings, and elastomers. The present review summarizes the latest research regarding major IA production pathways, metabolic engineering procedures, and the synthesis and applications of novel polymeric materials.

Itaconic acid fermentation using activated charcoal-treated corn stover hydrolysate and process evaluation based on Aspen plus model

Itaconic acid production using lignocellulose materials is a promising approach to replace sugar substrates such as glucose that are expensive. However, the complicated detoxification of the hydrolysate is pre-requisite to remove the lignocellulose-derived inhibitors to enable high organic acid fermentation. In this study, the hydrolysate prepared using dry acid pretreated and biodetoxified corn stover was tested for itaconic acid production by the fungal strain Aspergillus terreus M69. Corn stover hydrolysate containing 0.85 g/L acetic acid severely inhibited the organic acid production, but a treatment on the hydrolysate with activated charcoal to remove partial acetic acid helped to produce 33.6 g/L itaconic acid at a yield of 0.56 g/g. Most acetic acid released during enzymatic hydrolysis other than pretreatment was responsible for the inhibition of itaconic acid production. The technol-economic analysis showed that the minimum itaconic acid selling price was $1.647 per kg, which was lower than its market price. This study demonstrates the great potential of itaconic acid production using lignocellulose.

Potential of fungi isolated from the dumping sites mangrove rhizosphere soil to degrade polythene

Polythene is the most widely used plastic around the globe. Among the total plastic waste generated, polythene contributes the maximum share (64%). Various strategies/methods are being utilized to deal with the increasing rate of plastic waste, but among all the methods, bioremediation is regarded as the ecofriendly and widely accepted method. In the current investigation, we have attempted to discover the elite polythene deteriorating fungi (isolated from the rhizosphere soil of Avicennia marina). From 12 different eco-geographical locations along the West Coast of India, total 109 fungal isolates were recorded. The polythene deteriorating fungi were screened at varied pH (3.5, 7 and 9.5) based on changes in weight and tensile strength of the treated polythene at ambient temperature with continuous shaking for 60 days. BAYF5 isolate (pH 7) results in maximum reduction in weight (58.51 ± 8.14) whereas PNPF15 (pH 3.5) recorded highest reduction in tensile strength (94.44 ± 2.40). Surprisingly, we have also reported weight gain, with highest percent weight gain (28.41 ± 6.99) with MANGF13 at pH 9.5. To test the reproducibility of the results, the elite polythene degrading fungal isolates based on weight loss and reduction in tensile strength were only used for repetition experiment and the results based on the reduction in tensile strength were found only reproducible. Polythene biodegradation was further confirmed using Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR) analysis. The most efficient polythene deteriorating fungal isolates were identified as Aspergillus terreus strain MANGF1/WL and Aspergillus sydowii strain PNPF15/TS using both morphological keys and molecular tools.

Screening and optimizing fermentation production of l-asparaginase by Aspergillus terreus strain S-18 isolated from the Brazilian Caatinga Biome.

The aim of this study was to find new eukaryotic sources of the l-asparaginase (l-ASNase), since the prokaryotic sources of the enzyme are well-reported as causing allergic hypersensitivity reactions in a significant number of patients. This report describes screening for l-ASNase production by filamentous fungi isolated from the Brazilian Caatinga, and the optimization of fermentation parameters to increase fungal growth and improve yield in the production of l-ASNase. Thirty-two filamentous fungi were investigated in this study. When Aspergillus terreus strain S-18 was cultured in a proline-enriched medium, intracellular l-ASNase was expressed in concurrence with reduced l-glutaminase (l-GLUase) and protease activities. Fermentation conditions were then optimized in a 5-l bioreactor system to produce a maximum volumetric yield of 108U total of l-ASNase activity. The work reported here represents the first attempt to produce l-ASNase by filamentous fungi isolated from Brazil and offers a promising alternative eukaryotic source for l-ASNase production. In order to minimize the side effects caused by bacterial l-ASNase, the search of eukaryotic micro-organism for l-ASNase was carried out in fungi. This study demonstrates the diversity of filamentous fungi isolated from the Brazilian Caatinga Biome and the importance of knowledge of the microbial metabolism to obtain high concentrations of biotechnological products.

Compatibility of Inherent Fungal Endophytes of Withania somnifera with Trichoderma viride and its Impact on Plant Growth and Withanolide Content

Withania somnifera (Ashwagandha), also known as Indian ginseng, is an important ancient medicinal plant, used in the Indian traditional systems of medicine. In view of increasing demand for roots of Ashwagandha, the present study was undertaken to investigate the compatibility of inherent fungal endophytes along with the biocontrol agent, Trichoderma viride, for enhancing W. somnifera plant growth and root secondary metabolites (withaferin A). It has frequently been emphasized by the World Health Organization the use of healthy roots of Ashwagandha for therapeutic applications. To maintain quality of W. somnifera roots, an option could be eco-friendly management of root-knot diseases and co-inoculation of native endophytes along with T. viride. The in vitro antagonistic activity of T. viride (TV) against the W. somnifera pathogens, Alternaria alternata and Sclerotium rolfsii, showed 64.3% and 69.5% growth inhibition, respectively. Here, we investigated the compatibility of TV along with the native endophytic fungi Aspergillus terreus strain 2aWF (2aWF), Penicillium oxalicum strain 5aWF (5aWF), and Sarocladium kiliense strain 10aWF (10aWF) for the cultivation of W. somnifera. The co-inoculation of TV and native endophytic fungi resulted in increased shoot, root weight, and plant height to 65–150%, 35–74.5%, and 15–35%, respectively, compared to untreated plants. Withanolide A content in leaves of TV-treated plants increased significantly by 260%, whereas in co-inoculation treatments, it was enhanced up to 109–242%. However, no considerable change was noticed with withaferin A content in leaves, except the 2aWF + TV treatment significantly increased by 27%. In contrast, withanolide A content in roots was not affected by TV alone but co-inoculation with endophyte treatments significantly increased its content (19–73%). TV alone had increased chlorophyll a by 23%; however, in combination treatments, it increased up to 115–164% compared to control. Besides secondary metabolites in roots and leaves, co-inoculation of TV and native endophytes modulated the expression of the withanolide biosynthetic pathway genes HMGR, DXR, FPPS, SQS, SQE, CAS, SMT1, STE1, and CYP710A1 compared to control treatments. Apart from withanolide biosynthetic pathway genes, co-inoculation of TV also ameliorated the host-resistant-related gene NPR1 which was upregulated by ninefold in the TV treatment and 3- to 7-fold in the combination treatment. Overall, our results show that co-inoculation of TV along with inherent endophytes of W. somnifera enhanced plant growth and withanolides accumulation.