A simple high-throughput protocol for the extraction and quantification of inorganic phosphate in rice leaves

Premise Phosphorus (P) is an essential macronutrient that is often limited in agricultural systems. Determining inorganic phosphate (Pi) contents of plant tissues is crucial for evaluating plant P status. Here, we present a simple, high-throughput colorimetric microplate technique to measure Pi contents in rice (Oryza sativa) leaf tissues, based on the molybdenum blue reaction. Methods and Results We used a hole puncher to sample small equal areas of leaf tissue for Pi extraction. We removed the leaf grinding and weighing steps, which are time-consuming and normally required to release Pi from the tissues and to measure the biomass for data normalization, respectively. We showed that the punching method yielded comparable results to the conventional grinding method for two rice cultivars grown under various levels of P supply. Conclusions Compared with existing techniques, this protocol is more suited to an initial screening, enabling one researcher to determine the Pi contents of thousands of rice leaf samples within a few hours.