We have characterized a new commercial chlorophyll (Chl) and flavonoid (Flav) meter called Dualex 4 Scientific (Dx4). We compared this device to two other Chl meters, the SPAD-502 and the CCM-200. In addition, Dx4 was compared to the leaf-clip Dualex 3 that measures only epidermal Flav. Dx4 is factory-calibrated to provide a linear response to increasing leaf Chl content in units of µg cm–2, as opposed to both SPAD-502 and CCM-200 that have a non-linear response to leaf Chl content. Our comparative calibration by Chl extraction confirmed these responses. It seems that the linear response of Dx4 derives from the use of 710 nm as the sampling wavelength for transmittance. The major advantage of Dx4 is its simultaneous assessment of Chl and Flav on the same leaf spot. This allows the generation of the nitrogen balance index (NBI) used for crop surveys and nitrogen nutrition management. The Dx4 leaf clip, that incorporates a GPS receiver, can be useful for non-destructive estimation of leaf Chl and Flav contents for ecophysiological research and ground truthing of remote sensing of vegetation. In this work, we also propose a consensus equation for the transformation of SPAD units into leaf Chl content, for general use.

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A new, commercial, fluorescence-based optical sensor for plant constituent assessment was recently introduced. This sensor, called the Multiplex® (FORCE-A, Orsay, France), was used to monitor grape maturation by specifically monitoring anthocyanin accumulation. We derived the empirical anthocyanin content calibration curves for Champagne red grape cultivars, and we also propose a general model for the influence of the proportion of red berries, skin anthocyanin content and berry size on Multiplex® indices. The Multiplex® was used on both berry samples in the laboratory and on intact clusters in the vineyard. We found that the inverted and log-transformed far-red fluorescence signal called the FERARI index, although sensitive to sample size and distance, is potentially the most widely applicable. The more robust indices, based on chlorophyll fluorescence excitation ratios, showed three ranges of dependence on anthocyanin content. We found that up to 0.16 mg cm−2, equivalent to approximately 0.6 mg g−1, all indices increase with accumulation of skin anthocyanin content. Excitation ratio-based indices decrease with anthocyanin accumulation beyond 0.27 mg cm−2. We showed that the Multiplex® can be advantageously used in vineyards on intact clusters for the non-destructive assessment of anthocyanin content of vine blocks and can now be tested on other fruits and vegetables based on the same model.

In this study a method was designed to assess non-destructively the type of UV-screening compounds present in the leaf epidermis. The method is based on the recording and calculation of the ratio of UV-excitation spectra of chlorophyll fluorescence (FER) from the adaxial and abaxial sides of bifacial leaves, or from older and younger segments of monocotyledonous leaves. The logarithm of this ratio (logFER) matched the absorption spectrum of the UVabsorbers present in the leaf, as confirmed by its overlap with the absorption spectrum of the methanolic extract of the leaf or of the isolated epidermis. By using the logFER approach, it was possible to demonstrate that the concentration but not the classes of compounds present in the epidermis that are responsible for UV-screening is affected by the side and the age of the leaves. In contrast, measurements from the leaves of seven dicots and one monocot indicated large difference in the classes of these compounds between species. Finally, it was shown that the logFER in the UV is independent of the emission wavelength, and that the method can be used for quantitative measurements. This method expands to the spectral domain the use of ChlF for the estimation of the leaf epidermal transmittance.

Dualex (dual excitation) is a field-portable instrument, hereby described, for the assessment of polyphenolic compounds in leaves from the measurement of UV absorbance of the leaf epidermis by double excitation of chlorophyll fluorescence. The instrument takes advantage of a feedback loop that equalizes the fluorescence level induced by a reference red light to the UV-light-induced fluorescence level. This allows quick measurement from attached leaves even under field conditions. The use of light-emitting diodes and of a leaf-clip configuration makes Dualex a user-friendly instrument with potential applications in ecophysiological research, light climate analysis, agriculture, forestry, horticulture, pest management, selection of medicinal plants, and wherever accumulation of leaf polyphenolics is involved in plant responses to the environment.

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