Biosynthesis of Silver Chloride Nanoparticles by Rhizospheric Bacteria and Their Antibacterial Activity against Phytopathogenic Bacterium Ralstonia solanacearum

Ralstonia solanacearum is the most destructive pathogen, causing bacterial wilt disease of eggplant. The present study aimed to develop green synthesis and characterization of silver chloride nanoparticles (AgCl-NPs) by using a native bacterial strain and subsequent evaluation of their antibacterial activity against R. solanacearum. Here, a total of 10 bacterial strains were selected for the biosynthesis of AgCl-NPs. Among them, the highest yield occurred in the synthesis of AgCl-NPs using a cell-free aqueous filtrate of strain IMA13. Ultrastructural observation revealed that the AgCl-NPs were spherical and oval with smooth surfaces and 5–35 nm sizes. XRD analysis studies revealed that these particles contained face-centered cubic crystallites of metallic Ag and AgCl. Moreover, FTIR analysis showed the presence of capping proteins, carbohydrates, lipids, and lipopeptide compounds and crystalline structure of AgCl-NPs. On the basis of phylogenetic analysis using a combination of six gene sequences (16S, gyrA, rpoB, purH, polC, and groEL), we identified strain IMA13 as Bacillus mojavensis. Three kinds of lipopeptide compounds, namely, bacillomycin D, iturin, and fengycin, forming cell-free supernatant produced by strain IAM13, were identified by MALDI-TOF mass spectrometry. Biogenic AgCl-NPs showed substantial antibacterial activity against R. solanacearum at a concentration of 20 µg/mL−1. Motility assays showed that the AgCl-NPs significantly inhibited the swarming and swimming motility (61.4 and 55.8%) against R. solanacearum. Moreover, SEM and TEM analysis showed that direct interaction of AgCl-NPs with bacterial cells caused rupture of cell wall and cytoplasmic membranes, as well as leakage of nucleic acid materials, which ultimately resulted in the death of R. solanacearum. Overall, these findings will help in developing a promising nanopesticide against phytopathogen plant disease management.

Ultrastructural Study of Platelet Behavior and Interrelationship in Sprouting and Intussusceptive Angiogenesis during Arterial Intimal Thickening Formation

Platelets in atherosclerosis, bypass stenosis, and restenosis have been extensively assessed. However, a sequential ultrastructural study of platelets in angiogenesis during the early phases of these lesions has received less attention. Our objective was the study of platelets in angiogenesis and vessel regression during intimal thickening (IT) formation, a precursor process of these occlusive vascular diseases. For this purpose, we used an experimental model of rat occluded arteries and procedures for ultrastructural observation. The results show (a) the absence of platelet adhesion in the de-endothelialized occluded arterial segment isolated from the circulation, (b) that intraarterial myriad platelets contributed from neovessels originated by sprouting angiogenesis from the periarterial microvasculature, (c) the association of platelets with blood components (fibrin, neutrophils, macrophages, and eosinophils) and non-polarized endothelial cells (ECs) forming aggregates (spheroids) in the arterial lumen, (d) the establishment of peg-and-socket junctions between platelets and polarized Ecs during intussusceptive angiogenesis originated from the EC aggregates, with the initial formation of IT, and (e) the aggregation of platelets in regressing neovessels (‘transitory paracrine organoid’) and IT increases. In conclusion, in sprouting and intussusceptive angiogenesis and vessel regression during IT formation, we contribute sequential ultrastructural findings on platelet behavior and relationships, which can be the basis for further studies using other procedures.

Developmental characteristics of sporogenous hyphae: a new observation between Brassica juncea var. tumida and Albugo candida

Brassica juncea var. tumida (tumorous stem mustard) is widely cultivated as a feature vegetable in Ningbo, Zhejiang Province, and is used as a raw material for pickles in China. White blister rust, caused by Albugo candida, has severe negative effects on tumorous stem mustard, resulting in significant economic losses. In this study, hypha colonization and extension, haustorial formation, and the development process of sporogenous hyphae and sporangia in A. candida-infected tissues were studied using light and transmission electron microscopy. Additionally, the complex and unique sporogenous hypha developmental process was revealed. During A. candida infection, cytological and ultrastructural characteristics were similar to those of cruciferous plants. During sporogenous hyphae development, an initially cylindrical sporogenous hypha with a thin inner wall at the apex developed from a cell with a thick inner wall. At the apical end of the sporogenous hypha, an electron-dense complex gathered, causing the disintegration of the inner and outer walls and the formation of new inner and outer walls. With the formation of a clavate sporogenous hypha, a deep constriction of the cell wall occurred at its subapex, in which a septum was produced and a primary sporangium was separated from the sporogenous hypha. Both layers of the sporogenous hypha wall took part in the formation of the sporangial wall but were not related the formation of the sporangial outer wall in a septum, providing direct evidence of the holoblastic formation of sporangia. Furthermore, ultrastructural observation demonstrated that the development of the primary sporangial walls within a septum occurred later than that of their lateral walls, including the degradation of constricted outer walls around the septum, formation of the inner wall and subsequent outer walls of a sporangium in the upper septum layers, confirming the earlier observation. Therefore, this study provided the model of sporogenous hyphae development and sporangial formation in chain.

Ultrastructural Observation of Axon Complex of Epithelial Cells after Nerve Implantation into Denervated Fingers of Monkey

The recognition of visual shape in monkeys depends on a multi-layer pathway from primary visual cortex to lower temporal cortex. Visual stimulation is received by retina and then projected to the primary visual cortex VI region through lateral geniculate nucleus. There are a large number of neurons activated by linear stimulation such as short side and line segment. This paper mainly studies the ultrastructural observation of axon complex of epithelial cells after nerve implantation in monkey nerve loss fingers. Through the ultrastructural view of epithelial axon complex, we can master the changes of nerve regeneration function to skin cells and solve the problems caused by nerve defects, this paper mainly studies the method of nerve implantation, uses the neural interface model and the algorithm of nerve electrode, and compares the experiment with the monkey without nerve implantation, and then observe the synaptic ultrastructure under the electron microscope after the experiment to find that nerve implantation can promote the skin sensory organs. The results showed that the repair of sensory cells was faster than that of the skin sensory cells after nerve implantation in the ultrastructures of epithelial axon complex after nerve implantation in monkeys. 90% of the cells implanted with nerve were very fast to repair, which could provide useful information for the study of peripheral nerve regeneration in the nervous system. Nerve implantation regeneration has been a medical research the research focus of the topic,medical researchers hope to find an effective method of nerve implantation for skin cell repir.

Overexpression of Sesame Polyketide Synthase A Leads to Abnormal Pollen Development in Arabidopsis

Background: Sesame is a great reservoir of bioactive constituents and unique antioxidant components and is widely used for its nutritional and medicinal value. The expanding demands for sesame seeds are putting pressure on sesame breeders to develop reliable high-yielding varieties. Heterosis utilization is an efficient way to increase sesame yield. Polyketide synthases (PKSs) are critical enzymes in the biosynthesis of sporopollenin, a primary component of pollen exine. Their in planta functions are being investigated for application in crop breeding.Results: In this study, we cloned the sesame POLYKETIDE SYNTHASE A (SiPKSA) and examined its function in male sterility. SiPKSA was specifically expressed in sesame flower buds, and its expression was significantly higher in sterile sesame anthers than in fertile anthers at the tetrad and microspore development stage. Further overexpression of SiPKSA in Arabidopsis caused transgenic plants male sterile. Ultrastructural observation showed that the pollen grains of SiPKSA-overexpressing plants contained few cytoplasmic inclusions and exhibited an abnormal pollen wall structure, with a thicker exine layer compared with wild type. In agreement with it, the expression of a set of sporopollenin biosynthesis-related genes and the contents of fatty acids and phenolics were significantly altered in anthers of SiPKSA-overexpressing plants compared with wild type during anther development. Conclusion: These findings highlighted that overexpression of SiPKSA in Arabidopsis might cause excessive sporopollenin biosynthesis to influence pollen and pollen wall development, leading to male sterile, suggesting that its manipulation might improve hybrid breeding in sesame and other crop species.

Ultrastructural observation of Botrytis cinerea and physical changes in resistant and susceptible grapevines

The necrotrophic fungus Botrytis cinereais a major threat to grapevine cultivation worldwide. Here, a highly-resistant Chinese wild grapevine Vitis amurensis Rupr ‘Shuangyou’ (SY) and the susceptible V. vinifera ‘Red Globe’ (RG) were selected for study, and their pathogenic infection and biochemical responses to B. cinerea were evaluated. The results revealed more trichomes on and a thicker cuticle for leaves of SY than RG under scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Both SEM and TEM also showed that conidial germination, appressorium formation, and hyphal development of B. cinerea were delayed on the leaves of resistant SY. Fewer infected hyphae were also observed in leaves of resistant SY when compared with susceptible RG. The infected leaves of resistant SY harbored higher levels of cellulase and pectinase activity during the early infection stages of B. cinerea at 4 hours post-inoculation (hpi), and higher glucanase and chitinase activity were maintained in the inoculated leaves of SY from 4 hpi through 18 hpi. Lignin was deposited in the infected leaves of susceptible RG but not in resistant SY. Taken together, these results provide insights into the ultrastructural characterizations and physical changes in resistant and susceptible grapevines.

In vivo toxicity and antitumor activity of newly green synthesized reduced graphene oxide/silver nanocomposites

A novel biosynthesis of dual reduced graphene oxide/silver nanocomposites (rGO/AgNC) using the crude metabolite of Escherichia coli D8 (MF06257) strain and sunlight is introduced in this work. Physicochemical analysis of these rGO/AgNC revealed that they are sheet-like structures having spherically shaped silver nanoparticles (AgNPs) with an average particle size of 8 to 17 nm, and their absorption peak ranged from 350 to 450 nm. The biosynthesized rGO/AgNC were characterized by UV–vis and FT-IR spectra, X-ray diffraction, Zeta potential and transmission electron microscopy. After the injection of these nanocomposites to mice, their uptake by the kidney and liver has been proven by the ultrastructural observation and estimation of the hepatic and renal silver content. These nanocomposites caused a moderate toxicity for both organs. Changes in the liver and kidney functions and histopathological effects had been observed. The rGO/AgNC revealed a remarkable antitumor effect. They showed a dose-dependent cytotoxic effect on Ehrlich ascites carcinoma (EAC) cells in vitro. Treatment of mice bearing EAC tumors intraperitoneally with 10 mg/kg rGO/AgNC showed an antiproliferative effect on EAC cells, reduced ascites volume, and maintained mice survival. The results indicate that this green synergy of silver nanoparticles with reduced graphene oxide may have a promising potential in cancer therapy.