The CRISPR/Cas9 System for Targeted Genome Engineering in Free-Living Fungi: Advances and Opportunities for Lichenized Fungi

Studies using whole genome sequencing, computational and gene expression, targeted genome engineering techniques for generating site-specific sequence alterations through non-homologous end joining (NHEJ) by genomic double-strand break (DSB) repair pathway with high precision, resulting in gene inactivation have elucidated the complexity of gene expression, and metabolic pathways in fungi. These tools and the data generated are crucial for precise generation of fungal products such as enzymes, secondary metabolites, antibiotics etc. Artificially engineered molecular scissors, zinc finger nucleases (ZFNs), Transcriptional activator-like effector nucleases (TALENs; that use protein motifs for DNA sequence recognition in the genome) and CRISPR associated protein 9 (Cas9;CRISPR/Cas9) system (RNA-DNA recognition) are being used in achieving targeted genome modifications for modifying traits in free-living fungal systems. Here, we discuss the recent research breakthroughs and developments which utilize CRISPR/Cas9 in the metabolic engineering of free-living fungi for the biosynthesis of secondary metabolites, enzyme production, antibiotics and to develop resistance against post-harvest browning of edible mushrooms and fungal pathogenesis. We also discuss the potential and advantages of using targeted genome engineering in lichenized fungal (mycobiont) cultures to enhance their growth and secondary metabolite production in vitro can be complemented by other molecular approaches

Evaluation of natural products for the management of damping-off of tomato incited by Pythium aphanidermatum (Edson) fitz

The effect of various natural products from plants and animals were evaluated for the presence of antifungal activity against Pythium aphanidermatum causing damping-off of tomato. Among the plant species tested, bulbs of A. sativum (10%) and Allium cepa var. aggregatum (20%), leaves of Lawsonia inermis, Piper betle (20% each), Eucalyptus globulus and Vitex negundo (40% each) exhibited complete inhibition of the mycelial growth of P. aphanidermatum. Out of 4 animal excreta screened, Pig dung (40%) extract totally inhibited the mycelial growth of P. aphanidermatum. Garlic bulb, eucalyptus leaf and hen litter extract were found to retain the fungitoxicity when extracted at 80 °C for 10 minutes. A. sativum bulb followed by E. globulus leaf extracted in acetone was found to be significantly superior over other natural products. Among the natural products tested, the minimum mycelial dry weight of P. aphanidermatum was obtained with garlic bulb followed by eucalyptus leaf extract. The plant products combined together retained strong inhibitory effect in solid and liquid media against P. aphanidermatum. Upon mixing, the more loss of toxicity was observed when botanicals were mixed with animal excreta, particularly with pig dung. Among the natural products tested, the water extracts of A. sativum plus E. globulus and A. sativum plus L. inermis combinations recorded the maximum percentage of seed germination, growth and vigour of tomato seedlings, respectively. Also, when the various natural products tested by seed treatment, soil drenching and seed treatment plus soil drenching, garlic plus eucalyptus treatment recorded maximum seedling emergence and lesser damping off incidence in all three methods of application. Among the three methods, seed treatment plus soil drenching (St+Sd) was found to be superior than other two methods

Influence of Ni and Sn Perovskite NiSn(OH)₆ Nanoparticles on Energy Storage Applications

New NiSn(OH)6 hexahydroxide nanoparticles were synthesised through a co-precipitation method using various concentrations of Ni2+ and Sn4+ ions (e.g., 1:0, 0:1, 1:2, 1:1, and 2:1; namely, N, S, NS-3, NS-2, and NS-1) with an ammonia solution. The perovskite NiSn(OH)6 was confirmed from powder X-ray diffraction and molecule interactions due to different binding environments of Ni, Sn, O, and water molecules observed from an FT-IR analysis. An electronic transition was detected from tin (Sn 3d) and nickel (Ni 2p) to oxygen (O 2p) from UV-Vis/IR spectroscopy. Photo luminescence spectroscopy (PL) identified that the emission observed at 400–800 nm in the visible region was caused by oxygen vacancies due to various oxidation states of Ni and Sn metals. A spherical nanoparticle morphology was observed from FE-SEM; this was due to the combination of Ni2+ and Sn4+ increasing the size and porosity of the nanoparticle. The elemental (Ni and Sn) distribution and binding energy of the nanoparticle were confirmed by EDAX and XPS analyses. Among the prepared various nanoparticles, NS-2 showed a maximum specific capacitance of 607 Fg−1 at 1 Ag−1 and 56% capacitance retention (338 Fg−1 and 5 Ag−1), even when increasing the current density five times, and excellent cycle stability due to combining Ni2+ with Sn4+, which improved the ionic and electrical conductivity. EIS provided evidence for NS-2’s low charge transfer resistance compared with other prepared samples. Moreover, the NS-2//AC (activated carbon) asymmetric supercapacitor exhibited the highest energy density and high-power density along with excellent cycle stability, making it the ideal material for real-time applications

Effect of Yttrium doping on antibacterial and antioxidant property of LaTiO3

Abstract The advancement of multidrug-resistant bacterial strains and their adverse effects is one of the most significant global health issues. The perovskite nanomaterial with combined antioxidant and antibacterial activities in one molecule has the potential for improved therapeutic solutions. In this work, Yttrium-doped Lanthanum Titanate (LaTi1 −x Y x O3, where x = 0, 0.05, and 0.1) was synthesized using auto combustion technique. Excellent crystalline structure with a tetragonal system is revealed by X-ray diffraction analysis (XRD). UV–Visible diffuse reflectance spectroscopy (UV–Vis DRS), Fourier transform infrared (FTIR), and photoluminescence (PL) were used to study its optical characteristics. The field emission scanning electron microscope (FE-SEM) shows rod-like pellet-shaped Yttrium-doped nanostructures, and the elements present were confirmed with the Energy Dispersive X-Ray Analysis (EDAX). Various concentrations of the synthesized materials were tested for antibacterial activity against Gram-positive (Staphylococcus aureus 902) and Gram-negative (E. coli 443) strains using the agar-well diffusion method with gentamicin antibiotic as a positive control. High antibacterial activity of 87.1% and 83.3% was shown by 10% Yttrium-doped LaTiO3 (LY(0.1)TO) at 500 mg/mL against both positive and negative stains, respectively. Moreover, the antioxidant properties of synthesized materials were assessed with IC50 values of 352.33 µg/mL, 458.055 µg/mL, and 440.163 µg/mL for samples LaTi1 − x Y x O3, where x = 0, 0.05, and 0.1 respectively. The antibacterial and antioxidant capabilities of the proposed samples illustrate their applicability in various biomedical applications