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Evaluation of photosynthetic quantum yield is important for analyzing the phenotype of plants. Chlorophyll a fluorescence (ChlF) has been widely used to estimate plant photosynthesis and its regulatory mechanisms. The ratio of variable to maximum fluorescence, F v / F m , obtained from a ChlF induction curve, is commonly used to reflect the maximum photochemical quantum yield of photosystem II (PSII), but it is measured after a sample is dark-adapted for a long time, which limits its practical use. In this research, a least-squares support vector machine (LSSVM) model was developed to explore whether F v / F m can be determined from ChlF induction curves measured without dark adaptation. A total of 7,231 samples of 8 different experiments, under diverse conditions, were used to train the LSSVM model. Model evaluation with different samples showed excellent performance in determining F v / F m from ChlF signals without dark adaptation. Computation time for each test sample was less than 4 ms. Further, the prediction performance of test dataset was found to be very desirable: a high correlation coefficient (0.762 to 0.974); a low root mean squared error (0.005 to 0.021); and a residual prediction deviation of 1.254 to 4.933. These results clearly demonstrate that F v / F m , the widely used ChlF induction feature, can be determined from measurements without dark adaptation of samples. This will not only save experiment time but also make F v / F m useful in real-time and field applications. This work provides a high-throughput method to determine the important photosynthetic feature through ChlF for phenotyping plants.