Phytoplankton species composition research is key to understanding phytoplankton ecological and biogeochemical functions. Hyperspectral optical sensor technology allows us to obtain detailed information about phytoplankton species composition. In the present study, a transfer learning method to inverse phytoplankton species composition using in situ hyperspectral remote sensing reflectance and hyperspectral satellite imagery was presented. By transferring the general knowledge learned from the first few layers of a deep neural network (DNN) trained by a general simulation dataset, and updating the last few layers with an in situ dataset, the requirement for large numbers of in situ samples for training the DNN to predict phytoplankton species composition in natural waters was lowered. This method was established from in situ datasets and validated with datasets collected in different ocean regions in China with considerable accuracy (R 2 = 0.88, mean absolute percentage error (MAPE) = 26.08%). Application of the method to Hyperspectral Imager for the Coastal Ocean (HICO) imagery showed that spatial distributions of dominant phytoplankton species and associated compositions could be derived. These results indicated the feasibility of species composition inversion from hyperspectral remote sensing, highlighting the advantages of transfer learning algorithms, which can bring broader application prospects for phytoplankton species composition and phytoplankton functional type research.focused on the identification of specific phytoplankton species or groups under blooming conditions using ocean color satellites with a moderate spectral resolution [14][15][16]. Furthermore, discrimination of phytoplankton phyla in blooming conditions has been achieved based on differences in optical signals among different phytoplankton populations [17]. However, these endeavors are constrained to a limited type of phytoplankton species or groups because of the limitations in band settings for multi-spectral remote sensing. Moreover, some studies have been focused on Case 1 water with relatively simple optical properties where optically active constituents co-vary with chlorophyll concentration [18,19]; however, it is more challenging to invert phytoplankton species composition from current ocean color remote sensing data with moderate spectral resolution in optically complex waters. Comparatively, hyperspectral remote sensing presents a promising tool to resolve the spectral variability of phytoplankton species [20].Several ocean color algorithms have also been proposed for phytoplankton information inversion in natural waters [21][22][23][24]. Among these efforts, a derivative spectroscopy/similarity index (SI) approach is the most common method for identifying dominant phytoplankton species or groups [25][26][27]. However, because SI-based approaches assign unknown spectra to the reference spectra that have the largest SI, only dominant species or groups can be identified, so it is difficult to determine phytoplankton species compo...