Measurement of Total Phosphorus and Polyphosphate in Chlamydomonas reinhardtii

Yujie Yang; Suna Ren; Xianqing Jia; Houqing Zeng; Long Wang; Yiyong Zhu; Keke Yi

doi:10.21769/BioProtoc.4692

Measurement of Total Phosphorus and Polyphosphate in Chlamydomonas reinhardtii

Bio Protoc. 2023 Jun 5;13(11):e4692. doi: 10.21769/BioProtoc.4692.

Authors

Yujie Yang¹, Suna Ren^{1

2}, Xianqing Jia¹, Houqing Zeng³, Long Wang¹, Yiyong Zhu⁴, Keke Yi^{1

5}

Affiliations

¹ State Key Laboratory of Efficient Utilization of Arid and Semi-arid Arable Land in Northern China, Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
² College of Resources and Environment, Anhui Agricultural University, Hefei 230036, China.
³ College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 311121, China.
⁴ Jiangsu Provincial Key Lab for Organic Solid Waste Utilization, National Engineering Research Center for Organic-based Fertilizers, Jiangsu Collaborative Innovation Center for Solid Organic Waste Resource Utilization, Nanjing Agricultural University, Nanjing, China.
⁵ KeKe Yi's Lab https://yilab.life.

Abstract

Phosphorus is an essential nutrient for plants. Green algae usually store excess P as polyphosphate (polyP) in the vacuoles. PolyP, a linear chain of three to hundreds of phosphate residues linked by phosphoanhydride bonds, is important for cell growth. Based on the previous method of polyP purification with silica gel columns (Werner et al., 2005; Canadell et al., 2016) in yeast cells, we developed a protocol to purify and determine the total P and polyP in Chlamydomonas reinhardtii by a quick, simplified, and quantitative method. We use hydrochloric acid or nitric acid to digest polyP or total P in dried cells and analyze P content using the malachite green colorimetric method. This method may be applied to other microalgae.

Keywords: Chlamydomonas; Green algae; Plant nutrient; Polyphosphate; Total phosphorus; Vacuolar phosphate.