Enzymes and Phytohormones from Micromonospora

Actinobacteria produce diverse and huge amounts of enzymes that are widely used in different industrial purposes. Specific properties of enzymes allow to run the reactions under milder conditions with improved yield and reduced wastes. Further redesign for natural enzymes is very essential because they are often not suitable for biocatalytic processes. Recently, new microbial natural and creating enzymes are discovered synchronous with the late advanced technologies of genomics, metagenomics, proteomics, efficient expression systems and emerging recombinant DNA. The ongoing development of enzyme biotechnology aids in the improvement of the industrial biocatalysis field. Thermophilic actinobacteria produce thermostable enzymes that are widely used in industrial processes. In contrast, psychrophilic actinobacteria grow well at low temperatures and subsequently their enzymes are more effective at low temperatures. The advanced DNA sequencing technique allows determining and identifying the sequences and functions of all the genes that synthesize proteins that are widely use in the industry. Recombinant strains can be obtained by using certain biotechnological tools to potentially increase enzymes production on a large scale. The ongoing development in this field will lead to the improvement of different industrial purposes such as food, chemicals, textiles, leather, pharmaceuticals, and so on

Plant Pathogens

Plants cover the most area of the earth’s living environment as trees, grasses, flowers, and so on. Plants play different important roles in the environment such as ecosystem balance and food supplement for animals and humans. Moreover, wild or cultivated plants are considered the powerful biofertilizers for the soil, where the plant debris after death and degradation provides the soil with sufficient organic matters. Accordingly, plant care is a great duty and hard mission, which must be constantly improved. The study of plant pathogens belongs to the branch of biology known as plant pathology. The latter is also concerned to overcome the plant diseases arising from the biotic and/or abiotic origin. Biotic (infectious) diseases are developed owing to microbial infection, while abiotic (noninfectious) diseases are developed due to environmental factors. In this chapter, we are concerned with plant pathogens or phytopathogenic microbes such as bacteria, viruses, fungi, mollicutes, and so on

Exosome/Liposome-like Nanoparticles: New Carriers for CRISPR Genome Editing in Plants

Rapid developments in the field of plant genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) systems necessitate more detailed consideration of the delivery of the CRISPR system into plants. Successful and safe editing of plant genomes is partly based on efficient delivery of the CRISPR system. Along with the use of plasmids and viral vectors as cargo material for genome editing, non-viral vectors have also been considered for delivery purposes. These non-viral vectors can be made of a variety of materials, including inorganic nanoparticles, carbon nanotubes, liposomes, and protein- and peptide-based nanoparticles, as well as nanoscale polymeric materials. They have a decreased immune response, an advantage over viral vectors, and offer additional flexibility in their design, allowing them to be functionalized and targeted to specific sites in a biological system with low cytotoxicity. This review is dedicated to describing the delivery methods of CRISPR system into plants with emphasis on the use of non-viral vectors

Polianthes tuberosa-Mediated Silver Nanoparticles from Flower Extractand Assessment of Their Antibacterial and Anticancer Potential: An In Vitro Approach

Plant-mediated metallic nanoparticles have beenreported for a diversified range of applications in biological sciences. In the present study, we propose the Polianthes tuberosa flower as a reducing and stabilizing agent for the synthesis of silver nanoparticles (PTAgNPs). The PTAgNPs were exclusively characterized using UV–Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), atomic force microscopy, zeta potential, and transmission electron microscopy (TEM) studies. In a biological assay, we investigated the antibacterial and anticancer activity of silver nanoparticles in the A431 cell line. The PTAgNPs demonstrated a dose-dependent activity in E. coli and S. aureus, suggesting the bactericidal nature of AgNPs. The PTAgNPs exhibited dose-dependent toxicity in the A431 cell line, with an IC50 of 54.56 µg/mL arresting cell growth at the S phase, as revealed by flow cytometry analysis. The COMET assay revealed 39.9% and 18.15 severities of DNA damage and tail length in the treated cell line, respectively. Fluorescence staining studies indicate that PTAgNPs cause reactive oxygen species (ROS) and trigger apoptosis. This research demonstrates that synthesized silver nanoparticles have a significant effect on inhibiting the growth of melanoma cells and other forms of skin cancer. The results show that these particles can cause apoptosis or cell death in malignant tumor cells. This suggests that they could be used to treat skin cancers without harming normal tissues

Isolation and Characterization a Novel Catabolic Gene Cluster Involved in Chlorobenzene Degradation in Haloalkaliphilic Alcanivorax sp. HA03

Chlorobenzene (CB) poses a serious risk to human health and the environment, and because of its low degradation rate by microorganisms, it persists in the environment. Some bacterial strains can use CB as growth substrates and their degradative pathways have evolved; very little is known about these pathways and the enzymes for CB degradation in high pH and salinity environments. Alcanivorax sp. HA03 was isolated from the extremely saline and alkaline site. HA03 has the capability to degrade benzene, toluene and chlorobenzene (CB). CB catabolic genes were isolated from HA03, which have a complete gene cluster comprising α and β subunits, ferredoxin and ferredoxin reductase (CBA1A2A3A4), as well as one gene-encoding enzyme for chlorocatechol 1,2-dioxygenase (CC12DOs). Based on the deduced amino acid sequence homology, the gene cluster was thought to be responsible for the upper and lower catabolic pathways of CB degradation. The CBA1A2A3A4 genes probably encoding a chlorobenzene dioxygenase was confirmed by expression during the growth on CB by RT-PCR. Heterologous expression revealed that CBA1A2A3A4 exhibited activity for CB transformation into 3-chlorocatechol, while CC12DOs catalyze 3-chlorocatechol, transforming it into 2-chloromucounate. SDS-PAGE analysis indicated that the sizes of CbA1 and (CC12DOs) gene products were 51.8, 27.5 kDa, respectively. Thus, Alcanivorax sp. HA03 constitutes the first bacterial strain described in the metabolic pathway of CB degradation under high pH and salinity conditions. This finding may have obvious potential for the bioremediation of CB in both highly saline and alkaline contaminated sites

A Brief History of Endophyte Detection Techniques in Grasses

Endophytes are the plant mutualists that live asymptomatically inside plant tissue and are found in nearly whole plant kingdom. Endophytic fungi receive shelter and nutrition from host plants and in return provide great advantages to the host. Grasses are a useful forage species and are of great agricultural and socio-economic value. The presence of endophytes in these grasses provide protection, persistence and improved yield against herbivores, insects, pathogens, drought and several other biotic and abiotic stresses. This review summarizes traditional and modern molecular techniques to identify endophytes from turf and forage grasses. Traditional approaches include direct observation, staining, laser micro dissection and pressure catapulting and cultivation-dependent methods that provide a morphological identification of endophytic mycobiota in grass tissues. Earlier studies on endophytes using these methods resulted in several technical implications which molecular approaches are able to solve now-a-days. Molecular approaches include DNA extraction, PCR based DNA Fingerprinting techniques, Denaturing Gradient Gel Electrophoresis, Sanger sequencing, Pyrosequencing, Immunoblot assay, Biosensors, DNA Barcoding and Molecular Phylogenetics etc. A comparison of these detection techniques will facilitate other researchers as well to develop new ways for the detection of endophytes that will contribute to the improvement of grassland in future

Application of Nanocomposites from Bees Products and Nano-Selenium in Edible Coating for Catfish Fillets Biopreservation

Bee products, e.g., chitosan and propolis (Pro), have extraordinary importance in many disciplines including food biopreservation. Fish meat is highly susceptible to vast spoilage, especially catfish (Clarias gariepinus) products. The current work involved the extraction of bees’ chitosan nanoparticles (BCht), Pro, Pro-mediated SeNPs and their composites, to evaluate them as potential antimicrobial and preservative nano-compounds, for the preservation of catfish fillets and augment their quality. BCht was extracted from bees (Apis mellifera) corpses and had a 151.9 nm mean particle diameter. The Pro was used for biosynthesis of SeNPs, which had 11.2 nm mean diameters. The entire compounds/composites exhibited powerful antibacterial acts against Escherichia coli, Staphylococcus aureus and Salmonella typhimurium, where S aureus had the uppermost resistance. BCht/Pro/SeNPs were the most forceful toward all bacterial strains. The constructed edible coatings (ECs) from produced compounds/composites (BCht, Pro, Pro/SeNPs, Pro/BCht and BCht/Pro/SeNPs) had elevated efficiency for preserving catfish fillets during cold storages for 7 days. The microbiological (total counts, psychrophilic bacteria, yeast and molds), spoilage chemical parameters (TVB-N, TBARS) and sensorial attributes (appearance, odor, color, overall quality) of ECs-treated fillets indicated the nanocomposite’s efficiency for protecting the fish from microbial growth, the progress of chemical spoilage indicators and maintaining the sensorial quality of treated stored fillets. The most effective nanocomposite for maintaining the entire fillet’s quality was the BCht/Pro/SeNP. The based ECs on BNCt, Pro/SeNPs and their nanocomposites could be endorsed for prospective employment in the biopreservation of various seafoods

Green Synthesis of Gold Nanoparticles Using <i>Polianthes tuberosa</i> L. Floral Extract

The developments of green-based metallic nanoparticles (gold) are gaining tremendous interest, having potential applications in health care and diagnosis. Therefore, in the present study, Polianthes tuberosa flower filtered extract was used as a reducing and stabilizing agent to synthesize gold nanoparticles (PtubAuNPs). The PtubAuNPs were extensively characterized by UV–visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, and X-ray diffraction. The antibacterial activity of PtubAuNPs was determined by the agar well diffusion method; the PtubAuNPs performed extreme antagonistic activity against the tested pathogens. Furthermore, the cytotoxicity of the PtubAuNPs was evaluated in MCF 7 cells by MTT assay. The PtubAuNPs induced toxicity in MCF 7 cells with the least concentration of 100 µg/mL in a dose-dependent method by inducing apoptosis. Overall, the study manifested that PtubAuNPs are a potent nanomaterial that can be employed as an antimicrobial and anticancer agent

Nettle-Leaf Extract Derived ZnO/CuO Nanoparticle-Biopolymer-Based Antioxidant and Antimicrobial Nanocomposite Packaging Films and Their Impact on Extending the Post-Harvest Shelf Life of Guava Fruit

Green synthesized metal oxide nanoparticles (NPs) have prominent applications in antimicrobial packaging systems. Here we have attempted for the fabrication of chitosan-based nanocomposite film containing Urtica dioica leaf extract derived copper oxide (CuO) and zinc oxide (ZnO) NPs for shelf-life extension of the packaged guava fruits. Electron microscopy and spectroscopy analysis of the CuO and ZnO NPs exhibited nano-scale size, spherical morphologies, and negative ζ-potential values. The NPs possessed appreciable antioxidant and antimicrobial activity (AMA) in order of CuO NPs > ZnO NPs > nettle extract. Therefore, this work establishes for the first time the successful synthesis of CuO NPs and compares its antimicrobial and antioxidant properties with ZnO NPs. On incorporation in chitosan, the polymer nanocomposite films were developed by solvent casting technique. The developed films were transparent, had low antioxidant but substantial AMA. The NP supplementation improved the film characteristics as evident from the decrease in moisture content, water holding capacity, and solubility of the films. The nanocomposite films improved the quality attributes and shelf life of guava fruits by one week on packaging and storage compared to unpackaged control fruits. Therefore, this study demonstrates the higher antimicrobial potential of the nettle leaf extract derived CuO/ZnO NPs for development of antimicrobial nanocomposite films as a promising packaging solution for enhancing the shelf life of various perishable fruits