Flavonoids are important secondary metabolites in strawberry as they fulfill a wide variety of physiological functions. In addition, they are beneficial for human health. Previous studies have shown for selected enzymes from the flavonoid pathway that flavonoid biosynthesis shows two peaks during fruit development. We provide optimized protocols for the determination of the activities of the key flavonoid enzymes: phenylalanine ammonia lyase, chalcone synthase/chalcone isomerase, flavanone 3-hydroxylase, dihydroflavonol 4-reductase, flavonol synthase, flavonoid 3-O-glucosyltransferase, and flavonoid 7-O-glucosyltransferase. Using these protocols we were able to demonstrate two distinct activity peaks during fruit ripening at early and late developmental stages for all enzymes with the exception of flavonol synthase. The first activity peak corresponds to the formation of flavanols, while the second peak is clearly related to anthocyanin and flavonol accumulation. The results indicate that flavonoid 3-O-glucosyltransferase activity is not essential for redirection from flavanol to anthocyanin formation in strawberry.

Prohexadione-Ca is a structural mimic of 2-oxoglutarate, and according to this property, it is able to inhibit dioxygenase enzymes, which require 2-oxoglutarate as a cosubstrate. Such enzymes are involved in flavonoid biosynthesis; therefore, prohexadione-Ca treatment leads to alterations in the flavonoid metabolism in grapevine tissues. Because of the fact that phenolic compounds often are responsible for enhanced plant resistance, modification of phenylpropanoid metabolism using elicitation can be considered as a new potential strategy in plant protection. The phenolic compounds were analyzed by high-performance liquid chromatography combined with chemical reaction detection. Tissue treatment induced the accumulation of unusual flavonoids, which were identified as derivatives of pentahydroxyflavanone, eriodictyol, and luteoliflavan. Concentrations of constitutive flavonoids were also affected by the bioregulator treatment. The alterations of the flavonoid profiles are discussed with respect to substrate preferences of relevant enzymes.

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Laser-induced fluorescence spectroscopy (LIFS) was nondestructively applied on strawberries (EX = 337 nm, EM = 400-820 nm) to test the feasibility of quantitatively determining native phenolic compounds in strawberries. Eighteen phenolic compounds were identified in fruit skin by UV and MS spectroscopy and quantitatively determined by use of rp-HPLC for separation and diode-array or chemical reaction detection. Partial least-squares calibration models were built for single phenolic compounds by means of nondestructively recorded fluorescence spectra in the blue-green wavelength range using different data preprocessing methods. The direct orthogonal signal correction resulted in r (2) = 0.99 and rmsep < 8% for p-coumaroyl-glucose, and r (2) = 0.99 and rmsep < 24% for cinnamoyl-glucose. In comparison, the correction of the fluorescence spectral data with simultaneously recorded reflectance spectra did not further improve the calibration models. Results show the potential of LIFS for a rapid and nondestructive assessment of contents of p-coumaroyl-glucose and cinnamoyl-glucose in strawberry fruits.