Consortia of cyanobacteria/microalgae and bacteria in desert soils: an underexplored microbiota

Applied Microbiology and Biotechnology - Desert ecosystem is generally considered as a lifeless habitat with extreme environmental conditions although it is colonized by extremophilic...     

Potassium release characteristics, potassium balance, and fingermillet (Eleusine coracana G.) yield sustainability in a 27- year long experiment on an Alfisol in the semi-arid tropical India

Aims

In Alfisols, potassium (K) deficiency limits productivity, as these soils are poor in K-bearing minerals such as mica. As nutrient management practices greatly influence K nutrition of crops especially in the longer term, we evaluated the effects of 27 (1978–2004) years of cropping fingermillet (Eleusine coracana G.) under different manure and mineral fertilizer treatments on K release, balance and yield sustainability on K deficient Alfisols in the semi-arid tropical region of southern India.

Methods

Fingermillet (variety: PR-202) was grown each year under rainfed conditions with 5 different nutrient management treatments: control (no amendment), 10 Mg ha^?1 farm yard manure (FYM), 10 Mg ha^?1 FYM +50 % NPK, 10 Mg ha^?1 FYM +100 % NPK and 100 % NPK. Potassium release characteristics in the soil profile were determined using 1 N boiling HNO₃ (strong extracting solution), 0.01 M HCl (medium extracting solution) and 0.01 M CaCl₂ (mild extracting solution).

Results

Continuous cropping of Alfisols for 27 years resulted in a decrease in K supplying capacity due to soil K depletion through crop K uptake. In soils without K addition, inherent soil supply could not meet the K requirement of fingermillet; thus, a negative K balance following 27 years of cropping affected K nutrition of the crop in all the treatments. As a result, the highest sustainable yield index (SYI) was observed using an integrated nutrient supply (combined application of nutrients from organic and inorganic sources), and the lowest index was obtained without K additions.

Conclusion

For balanced nutrient management in cereal production systems, K nutrition needs urgent attention in the K deficient Alfisol region of southern India. Addition of any amount of organic manures available at field level offers an alternative strategy for maintaining soil K fertility to improve and sustain crop productivity.     

Structural investigation of zymogenic and activated forms of human blood coagulation factor VIII: a computational molecular dynamics study

Background

Human blood coagulation factor VIII (fVIII) is a large plasma glycoprotein with sequential domain arrangement in the order A1-a1-A2-a2-B-a3-A3-C1-C2. The A1, A2 and A3 domains are interconnected by long linker peptides (a1, a2 and a3) that possess the activation sites. Proteolysis of fVIII zymogen by thrombin or factor Xa results in the generation of the activated form (fVIIIa) which serves as a critical co-factor for factor IXa (fIXa) enzyme in the intrinsic coagulation pathway.

Results

In our efforts to elucidate the structural differences between fVIII and fVIIIa, we developed the solution structural models of both forms, starting from an incomplete 3.7 Å X-ray crystal structure of fVIII zymogen, using explicit solvent MD simulations. The full assembly of B-domainless single-chain fVIII was built between the A1-A2 (Ala1-Arg740) and A3-C1-C2 (Ser1669-Tyr2332) domains. The structural dynamics of fVIII and fVIIIa, simulated for over 70 ns of time scale, enabled us to evaluate the integral motions of the multi-domain assembly of the co-factor and the possible coordination pattern of the functionally important calcium and copper ion binding in the protein.

Conclusions

MD simulations predicted that the acidic linker peptide (a1) between the A1 and A2 domains is largely flexible and appears to mask the exposure of putative fIXa enzyme binding loop (Tyr555-Asp569) region in the A2 domain. The simulation of fVIIIa, generated from the zymogen structure, predicted that the linker peptide (a1) undergoes significant conformational reorganization upon activation by relocating completely to the A1-domain. The conformational transition led to the exposure of the Tyr555-Asp569 loop and the surrounding region in the A2 domain. While the proposed linker peptide conformation is predictive in nature and warrants further experimental validation, the observed conformational differences between the zymogen and activated forms may explain and support the large body of experimental data that implicated the critical importance of the cleavage of the peptide bond between the Arg372 and Ser373 residues for the full co-factor activity of fVIII.     

A first genetic linkage map of mulberry (Morus spp.) using RAPD, ISSR, and SSR markers and pseudotestcross mapping strategy

To lay the foundation for molecular breeding efforts, the first genetic linkage map of mulberry (2n=2x=28) was constructed with 50 F1 full-sib progeny using randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), and simple sequence repeat (SSR) markers and two-way pseudotestcross mapping strategy. We selected 100 RAPD, 42 ISSR, and 9 SSR primers that amplified 517 markers, of which 188 (36.36%) showed a test-cross configuration, corresponding to the heterozygous condition in one parent and null in the other. Two separate female and male maps were constructed using 94 each of female- and male-specific testcross markers, containing 12 female linkage groups and 14 male linkage groups. At a minimum logarithm of the odds (LOD) score threshold of 6.0 and at a maximum map distance of 20 cM, the female map covered a 1,196.6-cM distance, with an average distance of 15.75 cM and maximum map distance of 37.9 cM between two loci; the male-specific map covered a 1,351.7-cM distance, with an average distance of 18.78 cM and a maximum map distance between two loci is of 34.7 cM. The markers distributed randomly in all linkage groups without any clustering. All 12 linkage groups in the female-specific map consisted of 4–10 loci ranging in length from 0 to 140.4 cM, and in the male-specific map, the 13 largest linkage groups (except linkage group 12, which contained three loci) consisted of 4–12 loci, ranging in length from 53.9 to 145.9 cM and accounting for 97.22% of the total map distance. When mapping, progeny pass through their juvenile phase and assume their adult characters, mapping morphological markers and identification of quantitative trait loci for adaptive traits will be the primary target. In that sense, our map provides reference information for future molecular breeding work on Morus and its relatives.     

Membranes of five-fold alamethicin-resistant<Emphasis Type="Italic">Staphylococcus aureus,Enterococcus faecalis</Emphasis> and<Emphasis Type="Italic">Bacillus cereus</Emphasis> show decreased interactions with alamethicin due to changes in membrane fluidity and surface charge

The aim of this study was to understand development of resistance to alamethicin (a model barrel stave pore forming antimicrobial peptide) by investigating changes in phospholipid profile, fatty acid side chain analysis and extent of alamethicin insertion in biomimetic membrane prepared form wild type strains and five folds alamethicin resistant variants ofStaphylococcus aureus NCDC 110,Enterococcus faecalis NCDC 114 andBacillus cereus NCDC 66. The wild type strains NCDC 110, 114, 66, were sensitive to alamethicin with IC₅₀ 5.5, 3.25 and 2.0 μg/ml respectively. Wild type strains were cultured in the presence of alamethicin to select resistant variants with IC₅₀ 29.0, 17.0 and 9.5 μg/ml respectively. The phospholipid profile analysis revealed increase in amino-group containing phospholipids to amino-group lacking phospholipids ratio between wild-type and resistant variant inS. aureus and B. cereus but decreased inE. faecalis. Predominant fatty acids in all strains were composed of even number of carbons. Linoleic acid was detected only in resistant strain ofB. cereus. As indicated by saturated-to-unsaturated fatty acids ratio, the membrane fromS. aureus andE. faecalis became more rigid, whereas, inB. cereus it became more fluid. Using a colorimetricin vitro assay, a decrease in alamethicin insertion in the biomimetic membrane could be observed upon acquisition of resistance. The membranes of five-fold alamethicin-resistantS. aureus, E. faecalis andB. cereus revealed changes in membrane fluidity and surface charge upon acquisition of resistance to alamethicin.     

ARF6 activated by the LHCG receptor through the cytohesin family of guanine nucleotide exchange factors mediates the receptor internalization and signaling

The luteinizing hormone chorionic gonadotropin receptor (LHCGR) is a G(s)-coupled GPCR that is essential for the maturation and function of the ovary and testis. LHCGR is internalized following its activation, which regulates the biological responsiveness of the receptor. Previous studies indicated that ADP-ribosylation factor (ARF)6 and its GTP-exchange factor (GEF) cytohesin 2 regulate LHCGR internalization in follicular membranes. However, the mechanisms by which ARF6 and cytohesin 2 regulate LHCGR internalization remain incompletely understood. Here we investigated the role of the ARF6 signaling pathway in the internalization of heterologously expressed human LHCGR (HLHCGR) in intact cells using a combination of pharmacological inhibitors, siRNA and the expression of mutant proteins. We found that human CG (HCG)-induced HLHCGR internalization, cAMP accumulation and ARF6 activation were inhibited by Gallein (βγ inhibitor), Wortmannin (PI 3-kinase inhibitor), SecinH3 (cytohesin ARF GEF inhibitor), QS11 (an ARF GAP inhibitor), an ARF6 inhibitory peptide and ARF6 siRNA. However, Dynasore (dynamin inhibitor), the dominant negative mutants of NM23-H1 (dynamin activator) and clathrin, and PBP10 (PtdIns 4,5-P2-binding peptide) inhibited agonist-induced HLHCGR and cAMP accumulation but not ARF6 activation. These results indicate that heterotrimeric G-protein, phosphatidylinositol (PI) 3-kinase (PI3K), cytohesin ARF GEF and ARF GAP function upstream of ARF6 whereas dynamin and clathrin act downstream of ARF6 in the regulation of HCG-induced HLHCGR internalization and signaling. In conclusion, we have identified the components and molecular details of the ARF6 signaling pathway required for agonist-induced HLHCGR internalization.