Quantitative Detection of Inositol Hexakisphosphate (InsP6) in Crop Plants Using Polyacrylamide Gel Electrophoresis (PAGE)

Inositol phosphates are essential for cell development and signaling in all living organisms. Inositol hexakisphosphate (InsP6) is the most abundant phosphoinositol in both plants and animals. While the concentration of inorganic phosphorous (Pi) is often limited in soil, some plants overcome this limitation by creating a phosphate reservoir that serves as a source of Pi during phosphate deficiency. Although this strategy benefits plant development and signaling under adverse environmental conditions, excessive accumulation of Pi in crop plants has raised serious concerns about its toxicity and ill effects on human health. Consumption of crop plants with high InsP6 content or food products made from these crops is found to reduce nutrient intake significantly by way of chelating essential metal cations in human and livestock fed by such plants. Therefore, it is necessary to determine InsP6 contents in crop plants. Several methods have been developed for the screening and detection of InsP6 in plants. These detection methods however, are complex, labor-intensive, and often provide inaccurate results. We have developed a fast, reliable, and cost-effective method for the detection and quantification of InsP6 in plants using polyacrylamide gel electrophoresis (PAGE) with potential applications in industry, quality control labs, and research projects.

A novel microcosm to identify inherently competitive microorganisms with the ability to mineralize phytate in solum

Fertilizer phosphorus(P)is a finite resource,necessitating the development of innovative solutions for P fertilizer efficiency in agricultural systems.Myo-inositol hexakisphosphate(phytate)constitutes the majority of identified organic P in many soil types and is poorly available to plants.Incorporating phytase-producing biofertilizers into soil presents a viable and environmentally acceptable way of utilizing P from phytate,while reducing the need for mineral P application.A deeper understanding of the microbial ecology in relation to degradation of phytate under natural soil conditions is however needed to obtain successful biofertilizer candidates able to compete in complex soil environments.Here we present the development of a microcosm for studying microbial communities able to colonize and utilize Ca-phytate hotspots in solum.Our results provide evidence that the recruited microbial population mineralizes Ca-phytate.Furthermore,quantification of bacterial genes associated with organic P cycling in alkaline soils indicated that the phosphatases PhoX and PhoD may play a larger role in phytate mineralization in soil than previously recognized.Amplicon sequencing and BioLog®catabolism studies show that hotspots containing Ca-phytate,recruited a different set of microorganisms when compared to those containing an addition of C source alone,with the genus Streptomyces specifically enriched.We propose that Streptomyces represents an hitherto unexplored resource as P biofertilizer with competitive advantage for utilizing CaPhy in an inherently competitive soil environment.We further conclude that the use of our newly designed microcosm presents an innovative approach for isolating soil microorganisms with the potential to degrade precipitated phytate in solum.