Characterization and Phylogenetic Diversity of Carboxymethyl Cellulase Producing <Emphasis Type="Italic">Bacillus</Emphasis> Species from a Landfill Ecosystem

Total population of cellulose degrading bacteria was studied in a landfill ecosystem as a part of microbial diversity study. Samples were obtained from 3 and 5 feet depth of a local landfill being operated for past 10 years. Among many isolates, 22 bacterial strains were selected based on their capability to decompose carboxymethyl cellulose (CMC). These isolates were cultivated on agar medium with CMC as the carbon source. All isolates were Gram positive, endospore forming and alkalophilic bacteria with optimum growth pH 9–10. They were grouped based on the phenotypic and chemotaxonomic characters and representative strains of different groups along with high carboxymethyl cellulase (CMCase) producing strains were included for further characterization. Analysis of 16S rRNA gene indicated that these strains belong to different species of the genus Bacillus. Maximum CMCase activity of 4.8 U/ml at 50°C was obtained by strain LFC15. Results in the present study indicated the potential of waste land ecosystems such as landfill are potential source for isolation of industrially important microorganisms.     

Multi-stress resilience in plants recovering from submergence

Submergence limits plants' access to oxygen and light, causing massive changes in metabolism; after submergence, plants experience additional stresses, including reoxygenation, dehydration, photoinhibition and accelerated senescence. Plant responses to waterlogging and partial or complete submergence have been well studied, but our understanding of plant responses during post-submergence recovery remains limited. During post-submergence recovery, whether a plant can repair the damage caused by submergence and reoxygenation and re-activate key processes to continue to grow, determines whether the plant survives. Here, we summarize the challenges plants face when recovering from submergence, primarily focusing on studies of Arabidopsis thaliana and rice (Oryza sativa). We also highlight recent progress in elucidating the interplay among various regulatory pathways, compare post-hypoxia reoxygenation between plants and animals and provide new perspectives for future studies.     

Phenotypic variability in the Old World bollworm,Helicoverpa armigera (Hübner) populations in India

Helicoverpa armigera (Hübner) is one of the most prominent polyphagous species of the Heliothinae pest complex that inflicts severe damage to a wide range of crops in India. Knowledge regarding the population structure of the pest species, whether morphological or genetic, is considered as an essential tool in making effective management decisions. Thus here, we performed the phenotypic characterization of H. armigera populations collected from varied geographic locations across India. Studied populations differed significantly for several external morphometric traits studied at larval, pupal and adult stages. Significant differences were also observed with respect to the intensity of black pigmentation on larval body as well as adult eye and forewing colour patterns. Besides external phenotypic traits, the length of genital organs like aedeagus and valva in males, and bursa copulatrix and bursa seminalis in females also differed significantly amongst populations. The dendrogram based on selected traits showed clear cut differentiation of studied populations into two major groups, one including all the South Zone populations and the other having populations from North and Central Zones. Differences based on phenotyping in the present study indicate the possibility of the existence of different subspecies within the Indian populations of H. armigera.     

Ultrapotent miniproteins targeting the SARS-CoV-2 receptor-binding domain protect against infection and disease

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An inverse metabolic engineering approach for the design of an improved host platform for over-expression of recombinant proteins in Escherichia coli

ABSTRACT: BACKGROUND: A useful goal for metabolic engineering would be to generate non-growing but metabolically active quiescent cells which would divert the metabolic fluxes towards product formation rather than growth. However, for products like recombinant proteins which are intricately coupled to the growth process it is difficult to identify the genes that need to be knocked-out/knocked-in to get this desired phenotype. To circumvent this we adopted an inverse metabolic engineering strategy which would screen for the desired phenotype and thus help in the identification of genetic targets which need to be modified to get overproducers of recombinant protein. Such quiescent cells would obviate the need for high cell density cultures and increase the operational life span of bioprocesses. RESULTS: A novel strategy for generating a library consisting of randomly down regulated metabolic pathways in E. coli was designed by cloning small genomic DNA fragments in expression vectors. Some of these DNA fragments got inserted in the reverse orientation thereby generating anti-sense RNA upon induction. These anti-sense fragments would hybridize to the sense mRNA of specific genes leading to gene 'silencing'. This library was first screened for slow growth phenotype and subsequently for enhanced over-expression ability. Using Green Fluorescent Protein (GFP) as a reporter protein on second plasmid, we were able to identify metabolic blocks which led to significant increase in expression levels. Thus down-regulating the ribB gene (3, 4 dihydroxy-2-butanone-4-phosphate synthase) led to a 7 fold increase in specific product yields while down regulating the gene kdpD (histidine kinase) led to 3.2 fold increase in specific yields. CONCLUSION: We have designed a high throughput screening approach which is a useful tool in the repertoire of reverse metabolic engineering strategies for the generation of improved hosts for recombinant protein expression.     

Ecological variations in adult life table attributes of Aedes aegypti (L.) from the desert and coastal regions of India

The ecological variation in biological and adult life-table attributes of two populations of Aedes aegypti (Diptera: Culicidae) from the desert (Jodhpur) and coastal (Kolkata) regions of India are assessed to understand the reproductive and survival strategies. The results showed that females lived longer than males in both strains. The desert strain was more r-strategist because of its higher intrinsic rate of increase (r_m = 0.23), finite rate of increase (λ = 1.25), lower life expectancy of males (7.9 days) and females (14.4 days), mean generation time (T = 19.2 days) and doubling time (DT = 3.0 days). However, there was no difference in net reproductive rate (R₀) between the desert and coastal strains. The coastal strain showed a longer female life expectancy (22.0 days) than the desert strain (14.4 days). However, the fecundity (eggs/female/day) was lower in the coastal strain (11.4) than in the desert strain (15.1). Conclusively, the desert (Jodhpur) strain is adapted to a better r-strategy than the coastal (Kolkata) strain of Ae. aegypti, which might be helpful to flourish in harsh environmental conditions. This study may provide accurate predictions of Ae. aegypti population dynamics for vector management.