The release of root exudates (REs) provides an important source of soil organic carbon. This work revealed the molecular composition of REs of different plant species including alfalfa (Medicago satioa L.), bean (Phaseolus oulgaris L.), barley (Hordeum oulgare L.), maize (Zea mays), wheat (Triticum aestioum L.), ryegrass (Lolium perenne L.) and pumpkin (Cucurbita maxima) using electrospray ionization coupled with Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR MS). The combination of positive ion mode (+ESI) and negative ion mode (-ESI) increased the number of the molecules detected by ESI FT-ICR MS, and a total of 8758 molecules were identified across all the samples. In detail, lipids and proteins and unsaturated hydrocarbons were more easily detected in +ESI mode, while aromatic compounds with high O/C were readily ionized in -ESI mode, and only 38% of the total assigned formulas were shared by -ESI and +ESI modes. Multivariate statistical analysis of the formulas indicated that the close related plants species secreted REs with similar molecular components. Moreover, the unsaturation degree and nitrogen content were the two key parameters able to distinguish the similarities and differences of molecular components of REs between plant species. The results provided a feasible analysis method for characterization of the molecular components of REs and for the first time characterized the molecular components of REs of a variety of plant species using ESI FT-ICR MS. (C) 2020 The Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V.