Gas chromatography-mass spectrometry associated global analysis of rice root exudates under aseptical conditions(Plant Nutrition)

Components of rice root exudates were surveyed by gas chromatography-mass spectrometry (GC-MS) and the effect of phosphate (P) starvation was investigated. Rice seedlings were aseptically grown in controlled environments under P-supplied (+P) or P-deficient (-P) conditions. Root exudates were collected from the culture solution 4 and 8 days after treatment (D4 and D8, respectively), which contain the first and second 4-day intervals. The collected solution was lyophilized and then eluted with methanol. In the present study, primary metabolites, such as sugars, amino acids, organic acids and fatty acids, were mainly targeted. After derivatization with methoxyamine hydrochloride and N-methyl-N-trifluoroacetamide, the samples were analyzed by GC-MS. Sugars and various amino acids (such as aspartic acid and L-isoleucine) were mainly detected and the components changed with the growth stage rather than the P content in the culture solution. The relative amount of sugars (such as glucose and fructose) and amino acids (such as aspartic acid and L-isoleucine) decreased in the D8 samples. Of the 78 detected compounds, orthogonal partial least-squares discrimination analysis (O-PLS-DA) showed that four compounds were related to the P content of the nutrient solution and 58 were related to the sampling date (days after treatment). The results suggest that the physiological change during seedling development was unexpectedly large and that the response to the environment was rather small.