| Summary: | High-throughput and large-scale data are part of a new era of plant remote sensing science. Quantification of the yield, energetic content, and chlorophyll <i>a</i> fluorescence (ChlF) remains laborious and is of great interest to physiologists and photobiologists. We propose a new method that is efficient and applicable for estimating photosynthetic performance and photosystem status using remote sensing hyperspectroscopy with visible, near-infrared and shortwave spectroscopy (Vis-NIR-SWIR) based on rapid multivariate partial least squares regression (PLSR) as a tool to estimate biomass production, calorimetric energy content and chlorophyll <i>a</i> fluorescence parameters. The results showed the presence of typical inflections associated with chemical and structural components present in plants, enabling us to obtain PLSR models with R<sup>2</sup><sub>P</sub> and RPD<sub>P</sub> values greater than >0.82 and 3.33, respectively. The most important wavelengths were well distributed into 400 (violet), 440 (blue), 550 (green), 670 (red), 700–750 (red edge), 1330 (NIR), 1450 (SWIR), 1940 (SWIR) and 2200 (SWIR) nm operating ranges of the spectrum. Thus, we report a methodology to simultaneously determine fifteen attributes (i.e., yield (biomass), ΔH°area, ΔH°mass, Fv/Fm, Fv’/Fm’, ETR, NPQ, qP, qN, ΦPSII, P, D, SFI, PI<sub>(abs)</sub>, D.F.) with high accuracy and precision and with excellent predictive capacity for most of them. These results are promising for plant physiology studies and will provide a better understanding of photosystem dynamics in tobacco plants when a large number of samples must be evaluated within a short period and with remote acquisition data.
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