Purification and biochemical characterization of phytocystatin from Brassica alba

Phytocystatins belong to the family of cysteine proteinases inhibitors. They are ubiquitously found in plants and carry out various significant physiological functions. These plant derived inhibitors are gaining wide consideration as potential candidate in engineering transgenic crops and in drug designing. Hence it is crucial to identify these inhibitors from various plant sources. In the present study a phytocystatin has been isolated and purified by a simple two‐step procedure using ammonium sulfate saturation and gel filtration chromatography on Sephacryl S‐100HR from Brassica alba seeds (yellow mustard seeds).The protein was purified to homogeneity with 60.3% yield and 180‐fold of purification. The molecular mass of the mustard seed cystatin was estimated to be nearly 26 000 Da by sodium dodecyl sulfate polyacrylamide gel electrophoresis as well as by gel filtration chromatography. The stokes radius and diffusion coefficient of the mustard cystatin were found to be 23A° and 9.4 × 10^?7 cm²s^?1 respectively. The isolated phytocystatin was found to be stable in the pH range of 6–8 and is thermostable up to 60 °C. Kinetic analysis revealed that the phytocystatin exhibited non‐competitive type of inhibition and inhibited papain more efficiently (K_i = 3 × 10^?7 M) than ficin (K_i = 6.6 × 10^?7 M) and bromelain (Ki = 7.7 × 10^?7 M respectively). CD spectral analysis shows that it possesses 17.11% alpha helical content. Copyright © 2016 John Wiley & Sons, Ltd.     

Partial substitution of organic nitrogen with synthetic nitrogen enhances rice yield,grain starch metabolism and related genes expression under the dual cropping system

Improving grain filling in the presernt farming systems is crucial where grain filling is a concern due to the extreme use of chemical fertilizers (CF). A field experiment was conducted at the experimental station of Guangxi University, China in 2019 to test the hypothesis that cattle manure (CM) and poultry manure (PM) combined with CF could improve rice grain filling rate, yield, biochemical and qualitative attributes. A total of six treatments, i.e., no fertilizer (T₁), 100% CF (T₂), 60% CM + 40% CF (T₃), 30% CM + 70% CF (T₄), 60% PM + 40% CF (T₅), and 30% PM + 70% CF (T₆) were used in this study. Results showed that the combined treatment T₆increased starch metabolizing enzymes activity (SMEs), such as ADP-glucose phosphorylase (ADGPase) by 8 and 12%, soluble starch synthase (SSS) by 7 and 10%, granule bound starch synthesis (GBSS) by 7 and 9%, and starch branching enzyme (SBE) by 14 and 21% in the early and late seasons, respectively, compared with T₂. Similarly, higher rice grain yield, grain filling rate, starch, and amylose content were also recorded in combined treatments. In terms of seasons, higher activity of SMEs , grain starch, and amylose content was noted in the late-season compared to the early season. The increment in these traits was mainly attributed to a lower temperature in the late season during the grain filling period. Furthermore, our results suggested that an increment in starch accumulation and grain filling rate were mainly associated with the enhanced sink capacity by regulating key enzyme activities involved in Suc-to-starch conversion. In-addition, RT-qPCR analysis showed higher expression levels of AGPS2b, SSS1, GBSS1, and GBSE11b genes, which resultantly increased the activities of SMEs during the grain filling period under combined treatments. Linear regression analysis revealed that the activity of ADGPase, SSS, GBSS, and SBE were highly positively correlated with starch and amylose accumulation. Thus, we concluded that a combination of 30% N from PM or CM with 70% N from CF is a promising option in terms of improving rice grain yield and quality. Our study provides a sustainable fertilizer management strategy to enhance rice grain yield and quality at the lowest environmental cost.     

Smash ridge tillage strongly influence soil functionality,physiology and rice yield

The practice of smash-ridging on dry land crop cultivation has shown much promise. However, the mechanism how does soil functionality and root traits can affect rice yield under smash ridge tillage with reduced nitrogen fertilization have not yet been explored. To fill this knowledge gap, we used three tillage methods—smash-ridging 40 cm (S40), smash-ridging 20 cm (S20), and traditional turn-over plowing 20 cm (T)—and two rice varieties (hybrid rice and conventional rice) and measured soil quality, root traits, rice yield and their correlation analysis at different growth stages. Soil physical and chemical properties were significantly improved by smash-ridging, including improvements in root morphological and physiological traits during three growth stages compared with T. S40 had the highest leaf area index (LAI), plant height (PH), and biomass accumulation (BA). Increment in biomass and panicle number (PN) resulted in higher grain yield (GY) of 6.9–9.4% compared with T. Correlation analysis revealed that root total absorption area (RTAA), root active absorption area (RAA), and root area ratio (RAR) were strongly correlated with soil quality. Root injury flow (RIF) and root biomass accumulation (RBA) were strongly correlated with LAI and above-ground plant biomass accumulation (AGBA). Conclusively, S40 is a promising option for improving soil quality, root traits, and consequently GY.     

Thermodynamic, spectroscopic studies and catechol oxidase activity of copper (II) complexes with amphiphilic d-galacturonic acid derived ligands

The interactions of three amphiphilic glycoligands derived from d-galacturonic acid (L¹, L² and L³) with copper (II) ions were investigated in aqueous solution and/or in aqueous-methanol media. The combination of potentiometry, UV-Vis spectrophotometry and Electron Paramagnetic Resonance (EPR) was used to determine the formation constants of the complexes and their relative structures in solution. The complexation sites were identified using electronic absorption and EPR spectroscopies. Copper complexes were obtained as square planar or square pyramidal mononuclear or dinuclear species. Solid compounds were synthesized and tested as catalysts in the autooxidation of catechols in methanol and in aqueous micellar media. Mononuclear species were found to be catalytically active in both media, whereas dinuclear ones do not show any significant catecholase activity.