Rapid and accurate crop chlorophyll content estimation is crucial for guiding field management and improving crop yields. This study explored the potential for potato chlorophyll content estimation based on unmanned aerial vehicle (UAV) multispectral imagery. To search the optimal estimation method, three parts of research were conducted as following. First, a combination of support vector machines (SVM) and a gaussian mixture model (GMM) thresholding method was proposed to estimate fractional vegetation cover (FVC) during the potato growing period, and the proposed method produced efficient estimates of FVC; among all the selected vegetation indices (VIs), the soil adjusted vegetation index (SAVI) had the highest accuracy. Second, the recursive feature elimination (RFE) algorithm was utilized to screen the VIs and texture features derived from multispectral images: three Vis, including modified simple ratio (MSR), ratio vegetation index (RVI) and normalized difference vegetation index (NDVI); three texture features, including correlation in the NIR band (corr-NIR), correlation in the red-edge band (corr-Red-edge) and homogeneity in the NIR band (hom-NIR), showed higher contribution to chlorophyll content estimation. Finally, a stacking model was constructed with K-Nearest Neighbor (KNN), a light gradient boosting machine (light-GBM), SVM algorithm as the base model and linear fitting as the metamodel, and four machine learning algorithms (SVM, KNN, light-GBM and stacking) were used to build the chlorophyll content estimation model suitable for different growing seasons. The results were: (1) The performance of the estimation model could be improved based on both VIs and texture features over using single-type features, and the stacking algorithm yielded the highest estimation accuracy with an R2 value of 0.694 and an RMSE value of 0.553; (2) When FVC was added, the estimation model accuracy was further improved, and the stacking algorithm also produced the highest estimation accuracy with R2 value of 0.739, RMSE value of 0.511 (3) When comparing modeling algorithms, stacking algorithms had greater advantages in the estimation chlorophyll content with potato plants than using single machine learning algorithms. This study indicates that taking into account the combination of VIs reflecting spectral characteristics, texture features reflecting spatial information and the FVC reflecting canopy structure properties can accomplish higher chlorophyll content estimation accuracy, and the stacking algorithm can integrate the advantages of a single machine learning model, with great potential for estimation of potato chlorophyll content.