A new tool for direct non-invasive evaluation of chlorophyll a content from diffuse reflectance measurements.

Chlorophyll is a key biochemical component that is responsible for photosynthesis and is an indicator of plant health. The effect of stressors can be determined by measuring the amount of chlorophyll a, which is the most abundant chlorophyll, in vegetation in general. Nowadays, invasive methods and vegetation indices are used for establishing chlorophyll amount or an approximation to this value, respectively. This paper demonstrates that H-point curve isolation method (HPCIM) is useful for isolating the signal of chlorophyll a from non-invasive diffuse reflectance measurements of leaves. Spinach plants have been chosen as an example. For applying the HPCIM only the registers of both, a standard and the sample are needed. The results obtained by HPCIM and the invasive method were statistically similar for spinach leaves: 144±6mg/m (n=5) and 155±40mg/m (n=5), respectively. However, more precise values were achieved with the HPCIM, which also involved minimal experimental effort. The HPCIM method was applied to spinach plants stressed by the action of several pesticides and water scarcity, showing a decrease of chlorophyll a content with time, which is related with a loss of health. The results obtained were compared with those achieved by two different reflectance vegetation indices (Macc01 and NDVI). Although NDVI and HPCIM gave similar footprints for the plants tested, vegetation indices fail in the estimation of real content of the chlorophyll a. The HPCIM could contribute to improve the knowledge of the chlorophyll a content of vegetation like health indicator, by applying it to a much employed non-invasive technique such as diffuse reflectance, which can be used in place or in remote sensing mode.