A biomolecular isolation framework for eco-systems biology

Mixed microbial communities are complex, dynamic and heterogeneous. It is therefore essential that biomolecular fractions obtained for high-throughput omic analyses are representative of single samples to facilitate meaningful data integration, analysis and modeling. We have developed a new methodological framework for the reproducible isolation of high-quality genomic DNA, large and small RNA, proteins, and polar and non-polar metabolites from single unique mixed microbial community samples. The methodology is based around reproducible cryogenic sample preservation and cell lysis. Metabolites are extracted first using organic solvents, followed by the sequential isolation of nucleic acids and proteins using chromatographic spin columns. The methodology was validated by comparison to traditional dedicated and simultaneous biomolecular isolation methods. To prove the broad applicability of the methodology, we applied it to microbial consortia of biotechnological, environmental and biomedical research interest. The developed methodological framework lays the foundation for standardized molecular eco-systematic studies on a range of different microbial communities in the future.     

Field tests with insecticides and insect growth regulators to control insect pests of cowpea and its effects on certain beneficial insects

A small‐plot tests were conducted on cowpea, Vigna ungiculata (L.) Walp, to determine the effectiveness of 14 selected insecticides representing four insecticide classes (organophosphates, carbamates, pyrethroids, and insect growth regulators, IGRs), and four insecticide/IGR mixtures on cowpea insect pests and its effects on certain beneficial insects. By day 3 after treatement, the insecticides phenthoate, isoxathion, cyanophos, carbaryl, and cypermethrin used at sub‐recommended rates reduced the leafhopper, Empoasca spp., populations by >83% than those in the control. On that day, all insecticide treatments significantly reduced the cowpea aphid, Aphis craccivora, numbers below that of the control. The prothiofos, isoxathion, pirimicarb, and fenpropathrin treatments provided continuing control to both leafhoppers and aphids through day 21 after spraying. It seems that most insecticide treatments were not effective in whitefly, Bemisia tabaci, control. The best control of the whitefly immatures was obtained by day 3 after spraying in plots received thiodicarb (76%) and fenpropathrin (60%). All selected insecticides and rates used had very low residual effect against B. tabaci immatures. By day 3 posttreatment, the carbaryl (2.02 kg/ha) caused completely protection for cowpea pods frMw blue pea butterfly, Lampides boeticus, larvae. The IGRs, flufenoxuron and Dowco‐439, applied alone and those applied in mixtures with insecticides, carbaryl/teflubenzuron, chlorpyrifos/XRD‐473, and methomyl/ flufenoxuron, exhibited satisfactory control (>81%) to butterfly larvae on day 7 posttreatment. All treatments did not exhibit a considerable residual activity against L. boeticus larvae through 2–3 weeks followed spray. Most insecticides applied at the higher rates used significantly reduced the numbers of limabean pod borer, Etiella zinckenella, larvae found in cowpea pods collected on day 7 after application. The IGRs, flufenoxuron and Dowco‐439, applied alone, at 0.119 kg/ha, or in mixtures, methomyl/flufenoxuron (0.167 kg/ha) and chlorpyrifos/Dowco‐439 (0.911 kg/ha) caused >73% control of E. zinckenella larvae through day 21 posttreatment. The best control of cowpea weevil, Callosobru‐chus spp., adults (>83%), on day 3 after spraying, was obtained in treatments of cyanophos (1.19 kg/ha), prothiofos (1.43 kg/ha), flufenoxuron (0.119 kg/ha), and chlorpyrifos/Dowco‐439 (0.911 kg/ha). The IGR flufenoxuron applied alone or in mixture, methomyl/flufenoxuron (0.164 kg/ ha) exhibited satisfactory residual activity against Callosobruchus adults through day 21 posttreatment. Percentage seed damage by the larvae of cowpea weevils was significantly lower in most treatments than that of control. Populations of insect predators found in all treated plots were extremely reduced than those found in untreated plots. By day 21 after spraying, the IGRs flufenoxuron, XRD‐473, and Dowco‐439, applied at the low rate of 0.071 kg/ha, seemed to be less effect against insect predators than other insecticides used.     

Diversity of Sertoli cells in the testis of Saduria entomon L. (Crustacea,Isopoda)

Summary

The present paper is the first to give a comprehensive and detailed characterization of Sertoli cells in the isopod, Saduria entomon, based on transmission electron microscopy. Two types of Sertoli cells, A and B, were distinguished which clearly differ in their location in the wall of the testicular tubule, and in their morphology, ultrastructure, and function. Their occurrence is closely connected with the characteristic arrangement of germ cells inside the tubule. Sertoli A cells occupy only the part of the tubule containing spermatogonia and primary spermatocytes and they are associated with these cells by means of numerous ramified processes running in many directions. They are irregular in shape, but their shape and the ultrastructure are stable during maturation of the germ cells. Sertoli B cells, which compose most of the testicular tubule wall, form a columnar epithelium. They send long processes into the lumen of the tubule by means of which they make contact with maturing spermatids. The cytoarchitecture of the processes is highly variable and reflects their role in spermiogenesis and the formation of sperm bundles. After spermiation, when the apical part of the Sertoli cells has become flattened, they phagocytoze the residual cytoplasmic masses of spermatids, which undergo degradation in heterophagic vacuoles. Simultaneously, numerous autophagic vesicles appear.     

The Significance of Leaf Area in Evapotranspiration

Evapotranspiration rates increased with leaf area and fell withits decline at the time of flowering of cotton plants (Gossypiumbarbadense L.) and this relation was maintained while the rateof pan evaporation was steadily decreasing. A similar relationshipwas established for the hyacinth bean (Dolichos lablab L.) butevapotranspiration dropped at the onset of flowering, at thetime when leaf area was still increasing and the evaporatingpotential of the atmosphere began to rise. Evapotranspirationper unit area of leaf surface per day declined progressivelywith age for both species.     

Mycorrhiza and Phosphate Solubilizing Bacteria: Potential Bioagents for Sustainable Phosphorus Management in Agriculture

Phosphorus (P) is a critical nutrient that plays an essential role in improving soil fertility for optimum plant growth and productivity. It is one of the most deficient macro-nutrients in agricultural soils after nitrogen and is considered inadequate for plant growth and production. To P availability in soils, the farmers are applying huge amounts of synthetic P fertilizers that adversely affect the wider environment, groundwater, soil fertility and microbial population. Many beneficial microbes are known to release and supply soluble P for improving growth and yield of a variety of plants in a sustainable manner in P deficient soils. Thus, inoculation of these microbes, including arbuscular mycorrhizal fungi (AMF) and phosphate solubilizing bacteria (PSB) to soil to enhance crop production without harming the environment, is an alternative approach to chemical fertilizers. The combined role of AMF and PSB in P solubilization is not well understood and the application and mode of action of these microbial groups are often naive due to variation in the environment. Therefore, the current review article would develop a better understanding of the interactive role and mechanisms of AMF and PSB in improving P availability from both organic and inorganic sources in a sustainable crop production system. Finally, the current review would loop out further avenues for researchers interested to commercially produce effective AMF and PSB-based biofertilizers for sustainable management of phosphorus over a wide range of agricultural crops worldwide.     

Investigation of polyacrylamide hydrogel-based molecularly imprinted polymers using protein gel electrophoresis

In conjunction with polyacrylamide gel electrophoresis (PAGE), molecular imprinting methods have been applied to produce a multilayer mini-slab in order to evaluate how selectively and specifically a hydrogel-based molecularly imprinted polymer (MIP) binds bovine haemoglobin (BHb, ~64.5 kDa). A three-layer mini-slab comprising an upper and lower layer and a MIP, or a non-imprinted control polymer dispersion middle layer has been investigated. The discriminating MIP layer, also based on polyacrylamide, was able to specifically bind BHb molecules in preference to a protein similar in molecular weight such as bovine serum albumin (BSA, ~66 kDa). Protein staining allowed us to visualise the protein retention strength of the MIP layer under the influence of an electric field. This method could be applied to other proteins with implications in effective protein capture, disease diagnostics, and protein analysis.