Isolation and characterization of a newly discovered plant growth-promoting endophytic fungal strain from the genus Talaromyces.

In the Kandi zone of Punjab, India, root and rhizospheric soil samples were collected from the local vegetation near the Shivalik mountain foothills. Fifteen fungal colonies exhibiting distinct cultural morphology on Potato Dextrose Agar (PDA) plates were selected for plant-microbe interaction studies. Among these, the isolate HNB9 was identified as a nonpathogenic root colonizer. Morphological and molecular analyses confirmed HNB9 as Talaromyces albobiverticillius, characterized by the secretion of a red pigment as a secondary metabolite. Plants colonized with T. albobiverticillius HNB9 exhibited enhanced growth, manifesting in increased shoot and root length compared to untreated controls. This study unveiled the first evidence that a species from the Talaromyces genus, specifically T. albobiverticillius, possesses dual capabilities of root colonization and plant growth promotion. Moreover, HNB9 demonstrated the production of plant growth-regulating compounds like Indole Acetic Acid (IAA) and proficient solubilization of crucial nutrients (Phosphorous, Zinc, and Silica) through plate culture methods. This finding represents a significant contribution to the understanding of root-colonizing fungi with plant growth-promoting attributes, challenging the existing knowledge gap within the Talaromyces genus.

Biocontrol potential of mycogenic copper oxide nanoparticles against Alternaria brassicae.

The biological synthesis of nanoparticles using fungal cultures is a promising and novel tool in nano-biotechnology. The potential culture of Trichoderma asperellum (T. asperellum) has been used to synthesize copper oxide nanoparticles (CuO NPs) in the current study. The necrotrophic infection in Brassica species is caused due to a foliar pathogen Alternaria brassicae (A. brassicae). Mycogenic copper oxide nanoparticles (M-CuO NPs) were characterized by spectroscopic and microscopic techniques such as UV-visible spectrophotometry (UV-vis), transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The antifungal potential of CuO NPs was studied against A. brassicae. M-CuO NPs exhibited a surface plasmon resonance (SPR) at 303 nm, and XRD confirmed the crystalline phase of NPs. FTIR spectra confirmed the stretching of amide bonds, and the carbonyl bond indicated the presence of enzymes in T. asperellum filtrate. SEM and TEM confirmed the spherical shape of M-CuO NPs with an average size of 22 nm. Significant antifungal potential of M-CuO NPs was recorded, as it inhibited the growth of A. brassicae up to 92.9% and 80.3% in supplemented media with C-CuO NPs at 200 ppm dose. Mancozeb and propiconazole inhibited the radial growth up to 38.7% and 44.2%. SEM confirmed the morphological changes in hyphae and affected the sporulation pattern. TEM revealed hardly recognizable organelles, abnormal cytoplasmic distribution, and increased vacuolization, and light microscopy confirmed the conidia with reduced diameter and fewer septa after treatment with both types of NPs. Thus, M-CuO NPs served as a promising alternative to fungicides.

Phytofabricated zinc oxide nanoparticles as a nanofungicide for management of Alternaria blight of Brassica.

Plant pathogens resistant to the commercially available fungicides and bactericides even at higher concentrations are the biggest challenge for the farmers to control the losses due to plant diseases. The antibacterial and antifungal potential of nanomaterials makes them a suitable candidate for the control of plant diseases. Thus, the present study reports the phytofabricated zinc oxide nanoparticles (ZnO Np's) using aqueous plant leaf extract of Terminalia bellerica (Baheda). Characterization of ZnO nanoparticles was done by ultraviolet-visible (UV-Vis) studies, X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infra-red (FT-IR) analysis, and transmission electron microscopy (TEM). The presence of pure hexagonal wurtzite crystalline structure of ZnO nanoparticles was confirmed by XRD analysis. The TEM images revealed the spherical to hexagonal shaped ZnO nanoparticles with sizes ranging from 20 to 30 nm. The stabilization of synthesized ZnO nanoparticles through the interactions of terpenoids, steroids, phenylpropanoids, flavonoids, phenolic acids, and enzymes present in the leaf extract was suggested by FTIR analysis. The mechanism of the formation of ZnO nanoparticles using Terminalia bellerica (Baheda) (Tb-ZnO Np's) as a bioactive compound is proposed. These phytofabricated ZnO nanoparticles (Tb-ZnO Np's) have shown significant antifungal potential against Alternaria brassicae the causal agent of Alternaria blight disease/leaf spot disease in Brassica species. The microscopic results confirm the changes in mycelium morphology and reduction in the number of spore germination at 0.2 mg/mL concentration Tb-ZnO Np's.

Production, purification and applications of raw starch degrading and calcium-independent α-amylase from soil rich in extremophile.

Calcium independent, raw starch hydrolyzing, acidic α-amylase (66 kDa) was synthesized by Bacillus subtilis S113 that is an aerobic, rod-shaped and Gram +ve bacteria. Purification of the enzyme was performed by HiTrap Capto Q (Ion-exchange chromatography; 19.28 fold; 22.41% yield). The purified enzyme was found stable at broad acidic pH (4-6.5) and high-temperature range (40-80 °C), that fulfilled the necessary criteria and laid the foundation to be utilized in starch saccharification industry. Kinetic studies of the enzyme revealed that Km and Vmax of the enzyme was 0.22% and 357.14 U/mg respectively. Scanning electron microscopy studies showed that the enzyme was capable of completely hydrolyzing raw wheat and potato starch, further confirming its role in the starch industry. It was found that only 7.93% of the activity was loss at 4 °C when kept for one year.

Piriformospora indica promotes the growth of the in-vitro-raised Cymbidium aloifolium plantlet and their acclimatization.

Cymbidium aloifolium is known for its ornamental and medicinal values. It has been listed as threatened orchid species. In this study, in vitro propagated C. aloifolium plantlets were interacted with the Piriformospora indica. The growth assay was performed for 45 days; the plant growth pattern such as number and length of roots and shoots were measured. Microscopic study of the root section stained by trypan blue was done to detect the peloton formation. The methanol extracts of the fungal colonized plant as well as uncolonized (control) plant were prepared and various metabolites were identified by gas chromatography mass spectroscopy. Acclimatization was done in a substrate composition of coco peat: gravel: charcoal in ratio 2:2:1. P. indica-colonized plantlet showed the highest growth with the formation of clamdospore in the root section. The growth regulator such as auxin, ascorbic acid, andrographolide, hexadecanoic acid, and DL-proline were identified. After three months of field transfer, plantlet colonized by P. indica survived and remained healthy as compared to uncolonized control plantlet.

Dual symbiosis between Piriformospora indica and Azotobacter chroococcum enhances the artemisinin content in Artemisia annua L.

At present, Artemisia annua L. is the major source of artemisinin production. To control the outbreaks of malaria, artemisinin combination therapies (ACTs) are recommended, and hence an ample amount of artemisinin is required for ACTs manufacture to save millions of lives. The low yield of this antimalarial drug in A. annua L. plants (0.01-1.1%) ensues its short supply and high cost, thus making it a topic of scrutiny worldwide. In this study, the effects of root endophyte, Piriformospora indica strain DSM 11827 and nitrogen fixing bacterium, Azotobacter chroococcum strain W-5, either singly and/or in combination for artemisinin production in A. annua L. plants have been studied under poly house conditions. The plant growth was monitored by measuring parameters like height of plant, total dry weight and leaf yield with an increase of 63.51, 52.61 and 79.70% respectively, for treatment with dual biological consortium, as compared to that of control plants. This significant improvement in biomass was associated with higher total chlorophyll content (59.29%) and enhanced nutrition (especially nitrogen and phosphorus, 55.75 and 86.21% respectively). The concentration of artemisinin along with expression patterns of artemisinin biosynthesis genes were appreciably higher in dual treatment, which showed positive correlation. The study suggested the potential use of the consortium P. indica strain DSM 11827 and A. chroococcum strain W-5 in A. annua L. plants for increased overall productivity and sustainable agriculture.

Synergistic anti-candidal activity and mode of action of Mentha piperita essential oil and its major components.

CONTEXT: Mentha piperita L. (Lamiaceae) has been used in folk medicine since antiquity. Its essential oil (mint EO) and major bioactive components have antimicrobial properties but their mechanism of action is still not clear. OBJECTIVE: The present work aims to elucidate M. piperita's anti-Candida activity and mode of action. MATERIALS AND METHODS: Chemical constituents of mint EO were identified by GC-MS by injecting 0.1 ml sample in a splitless mode. MIC was determined by the broth dilution method. Synergy with fluconazole (FLC) was evaluated by checkerboard assay and FICI. Mid log phase cells harvested from YPD media were used for proton extrusion measurement and the rate of glucose-induced H(+) efflux gives PM-ATPase activity. Cell membrane integrity was estimated by total ergosterol content and scanning microscopy at respective MIC and sub-MIC values. In vitro hemolytic activity was performed to rule out possible cytotoxicity of the test compounds. RESULTS: The MIC value of mint EO, carvone, menthol, and menthone was 225, 248, 500, and 4200 µg/ml, respectively. At their respective MICs, these compounds showed 47, 42, 35, and 29% decrease in PM-ATPase activity besides showing synergy with FLC. In case of FLC-resistant strains, the decrease in H(+) efflux was by 52, 48, 32, and 30%, a trend similar to the susceptible cases. Exposed Candida cells showed a 100% decrease in the ergosterol content, cell membrane breakage, and alterations in morphology. DISCUSSION AND CONCLUSION: Our studies suggest that mint EO and its lead compounds exert antifungal activity by reducing ergosterol levels, inhibiting PM-ATPase leading to intracellular acidification, and ultimately cell death. Our results suggest that mint EO and its constituents are potential antifungal agents and need to be further investigated.

Scalable, sustainable cost-effective surgical care: a model for safety and quality in the developing world, part I: challenge and commitment.

BACKGROUND: With an estimated backlog of 4,000,000 patients worldwide, cleft lip and cleft palate remain a stark example of the global burden of surgical disease. The need for a new paradigm in global surgery has been increasingly recognized by governments, funding agencies, and professionals to exponentially expand care while emphasizing safety and quality. This three-part article examines the evolution of the Operation Smile Guwahati Comprehensive Cleft Care Center (GCCCC) as an innovative model for sustainable cleft care in the developing world. METHODS: The GCCCC is the result of a unique public-private partnership between government, charity, and private enterprise. In 2009, Operation Smile, the Government of Assam, the National Rural Health Mission, and the Tata Group joined together to work towards the common goal of creating a center of excellence in cleft care for the region. RESULTS: This partnership combined expertise in medical care and training, organizational structure and management, local health care infrastructure, and finance. A state-of-the-art surgical facility was constructed in Guwahati, Assam which includes a modern integrated operating suite with an open layout, advanced surgical equipment, sophisticated anesthesia and monitoring capabilities, central medical gases, and sterilization facilities. CONCLUSION: The combination of established leaders and dreamers from different arenas combined to create a synergy of ambitions, resources, and compassion that became the backbone of success in Guwahati.

Scalable, sustainable cost-effective surgical care: a model for safety and quality in the developing world, part II: program development and quality care.

BACKGROUND: The Guwahati Comprehensive Cleft Care Center (GCCCC) is committed to free medical and surgical care to patients afflicted with facial deformities in Assam, India. A needs-based approach was utilized to assemble numerous teams, processes of care, and systems aimed at providing world-class care to the most needy of patients, and to assist them with breaking through the barriers that prohibit them from obtaining services. METHODS: A team of international professionals from various disciplines served in Guwahati full time to implement and oversee patient care and training of local counterparts. Recruitment of local professionals in all disciplines began early in the scheme of the program and led to gradual expansion of all medical teams. Emphasis was placed on achieving optimal outcome for each patient treated, as opposed to treating the maximum number of patients. RESULTS: The center is open year round to offer full-time services and follow-up care. Along with surgery, GCCCC provides speech therapy, child life counseling, dental care, otolaryngology, orthodontics, and nutrition services for the cleft patients under one roof. Local medical providers participated in a model of graded responsibility commiserate with individualized skill and progress, and gradually assumed all leadership positions and now account for 92% of the workforce. Institutional infrastructure improvements positioned and empowered teams of skilled local providers while implementing systemized perioperative processes. CONCLUSION: This needs-based approach to program development in Guwahati was successful in optimization of quality and safety in all clinical divisions.

Scalable, sustainable cost-effective surgical care: a model for safety and quality in the developing world, part III: impact and sustainability.

BACKGROUND: The Guwahati Comprehensive Cleft Care Center (GCCCC) utilizes a high-volume, subspecialized institution to provide safe, quality, and comprehensive and cost-effective surgical care to a highly vulnerable patient population. METHODS: The GCCCC utilized a diagonal model of surgical care delivery, with vertical inputs of mission-based care transitioning to investments in infrastructure and human capital to create a sustainable, local care delivery system. Over the first 2.5 years of service (May 2011-November 2013), the GCCCC made significant advances in numerous areas. Progress was meticulously documented to evaluate performance and provide transparency to stakeholders including donors, government officials, medical oversight bodies, employees, and patients. RESULTS: During this time period, the GCCCC provided free operations to 7,034 patients, with improved safety, outcomes, and multidisciplinary services while dramatically decreasing costs and increasing investments in the local community. The center has become a regional referral cleft center, and governments of surrounding states have contracted the GCCCC to provide care for their citizens with cleft lip and cleft palate. Additional regional and global impact is anticipated through continued investments into education and training, comprehensive services, and research and outcomes. CONCLUSION: The success of this public private partnership demonstrates the value of this model of surgical care in the developing world, and offers a blueprint for reproduction. The GCCCC experience has been consistent with previous studies demonstrating a positive volume-outcomes relationship, and provides evidence for the value of the specialty hospital model for surgical delivery in the developing world.

In vitro plant development and root colonization of Coleus forskohlii by Piriformospora indica.

The present study was conducted for optimization of in vitro substrates under aseptic conditions for interaction of Piriformospora indica with the medicinal plant Coleus forskohlii. It aims to test the effects of different substrates on P. indica colonization as well as growth parameters of the in vitro raised C. forskohlii. Interaction of in vitro C. forskohlii with root endophyte P. indica under aseptic condition resulted in increase in growth parameters in fungus colonized plants. It was observed that P. indica promoted the plant's growth in all irrespective of substrates used for co-culture study. The growth was found inferior in liquid compared to semisolid medium as well as there was problem of hyperhydricity in liquid medium. P. indica treated in vitro plantlets were better adapted for establishment under green house compared to the non treated plants due to fungal intervention.

Root endophyte Piriformospora indica DSM 11827 alters plant morphology, enhances biomass and antioxidant activity of medicinal plant Bacopa monniera.

Unorganized collections and over exploitation of naturally occurring medicinal plant Bacopa monniera is leading to rapid depletion of germplasm and is posing a great threat to its survival in natural habitats. The species has already been listed in the list of highly threatened plants of India. This calls for micropropagation based multiplication of potential accessions and understanding of their mycorrhizal associations for obtaining plants with enhanced secondary metabolite contents. The co-cultivation of B. monniera with axenically cultivated root endophyte Piriformospora indica resulted in growth promotion, increase in bacoside content, antioxidant activity and nuclear hypertrophy of this medicinal plant.

An endophytic Pseudonocardia species induces the production of artemisinin in Artemisia annua.

Endophytic actinobacteria colonize internal tissues of their host plants and are considered as a rich and reliable source of diverse species and functional microorganisms. In this study, endophytic actinobacterial strain YIM 63111 was isolated from surface-sterilized tissue of the medicinal plant Artemisia annua. We identified strain YIM 63111 as a member of the genus Pseudonocardia. A. annua seedlings grown under both sterile and greenhouse conditions were inoculated with strain YIM 63111. The growth of A. annua seedlings was strongly reduced when YIM 63111 was inoculated at higher concentrations under sterile conditions. However, no growth inhibition was observed when A. annua was grown under greenhouse conditions. Using an enhanced green fluorescent protein (EGFP) expressing YIM 63111 strain, we also observed the endophytic colonization of A. annua seedling using confocal laser-scanning microscopy. The transcription levels of the key genes involved in artemisinin biosynthesis were investigated using real time RT-PCR, revealing that cytochrome P450 monooxygenase (CYP71AV1) and cytochrome P450 oxidoreductase (CPR) expression were up-regulated in A. annua upon inoculation with strain YIM 63111 under certain conditions. The up-regulation of these genes was associated with the increased accumulation of artemisinin. These results suggest that endophytic actinobacteria effectively stimulate certain plant defense responses. Our data also demonstrate the use of Pseudonocardia sp. strain YIM 63111 as a promising means to enhance artemisinin production in plants.

The root endophyte fungus Piriformospora indica leads to early flowering, higher biomass and altered secondary metabolites of the medicinal plant, Coleus forskohlii.

This study was undertaken to investigate the influence of plant probiotic fungus Piriformospora indica on the medicinal plant C. forskohlii. Interaction of the C. forskohlii with the root endophyte P. indica under field conditions, results in an overall increase in aerial biomass, chlorophyll contents and phosphorus acquisition. The fungus also promoted inflorescence development, consequently the amount of p-cymene in the inflorescence increased. Growth of the root thickness was reduced in P. indica treated plants as they became fibrous, but developed more lateral roots. Because of the smaller root biomass, the content of forskolin was decreased. The symbiotic interaction of C. forskohlii with P. indica under field conditions promoted biomass production of the aerial parts of the plant including flower development. The plant aerial parts are important source of metabolites for medicinal application. Therefore we suggest that the use of the root endophyte fungus P. indica in sustainable agriculture will enhance the medicinally important chemical production.

Arbuscular mycorrhiza-like biotechnological potential of Piriformospora indica, which promotes the growth of Adhatoda vasica Nees

We report the growth promoting potential of Piriformospora indica, which is a newly discovered arbuscular mycorrhiza-like fungus. It is a facultative symbiont and unlike arbuscular mycorrhizal fungi, it can be cultured in vitro. Adhatoda vasica is a medicinal plant. Rapid proliferation of roots was recorded in A. vasica withan important root colonization estimated to 95 percent after 6 months. P. indica improved growth of A. vasica. This association forms a new host-symbiont combination.

Positive growth responses of the medicinal plants Spilanthes calva and Withania somnifera to inoculation by Piriformospora indica in a field trial.

The medicinal plants Spilanthes calva and Withania somnifera were inoculated with Piriformospora indica, a plant growth-promoting root endophyte, in nurseries and subsequently transferred to the field. A significant increase in growth and yield of both plant species was recorded relative to uninoculated controls. Shoot and root length, biomass, basal stem, leaf area, overall size, number of inflorescences and flowers and seed production were all enhanced in the presence of the fungus. Net primary productivity was also higher than in control plants. The results clearly indicate the commercial potential of P. indica for large-scale cultivation of S. calva and W. somnifera.