| Rice is not only one of the most important staple food crops but also one of the essential monocot model organisms, the study on rice has long been one of the most popular field in biology. As the rapid development of “omics” technologies, metabolomics has been widely used in the field of plant biology in various plant species, including rice. Extraction method plays an important role in metabolic profiling, which influences significantly the final output of the metabolomics analysis. In this study, we intended to evaluate the effects of different extraction methods on rice seed metabolic profiling and to propose selection strategy of extraction methods for various purposes. To achieve this aim, we chose three different extracting solutions, which are commonly used in different plant metabolomics studies, to perform the extraction step of metabolomics analysis in rice mature seeds of two cultivars. The tested three extraction solutions included:(1) water: acetonitrile: isopropanol= 2:3:3(v: v: v);(2) water: methanol= 2:8(v:v);(3) methanol: chloroform: water=4:5:3(v:v:v), the two rice cultivars were Yangdao 6 and Wuyungeng 7, and the metabolomics platform was ultra high performance liquid chromatography and mass spectrometry/mass spectrometry(UHPLC-MS/MS)(Agilent).In total, we have identified 165 known metabolites in rice seeds using UHPLC-MS/MS. The statistic analysis data showed that different extraction methods affect not only the detection coverage as well as the yield of metabolite, but also the metabolic variation between the two tested rice seeds. Our data showed that the third extraction method had the best detection coverage and yield but at the cost of a double workload in the same time, while the second method could be quite effective for a high throughput metabolomics study on rice seeds. In summary, this study provided a scientific base for the selection of suitable sample extraction methods in rice seed metabolomics study for different purposes. |
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