Quercetin Accumulation Research Articles

Nutrient depletion as a key factor for manipulating gene expression and product formation in different branches of the flavonoid pathway

The content of flavonoids increases in response to nitrogen and phosphorus depletion in plants. Manipulation of these macronutrients may therefore be used to control the levels of desirable compounds and improve plant quality. Key enzymes in the shikimate pathway, which feeds precursors into the flavonoid pathway, are regulated post-translationally by feedback from aromatic amino acids, and possibly by redox control through photosynthesis. Use of microarrays for global transcript analysis in Arabidopsis has revealed that transcript levels are less influenced by mineral nutrients in the shikimate pathway compared with the flavonoid pathway. The responses in the shikimate pathway appear complex, whereas in the flavonoid pathway, a single gene often responds similarly to mineral depletion, high light intensity and sucrose. MYB [production of anthocyanin pigment 1 (PAP1)/production of anthocyanin pigment 2 (PAP2)] and bHLH [GLABRA3 (GL3)] transcription factors are important for the nutrient depletion response. PAP1/2 stimulate gross activation of the flavonoid pathway, and different investigations support merging signal transduction chains for various abiotic treatments on PAP1/2. Flavonol synthase is not part of the PAP1/2 regulon, and expression is mainly enhanced by high light intensity and sucrose, not mineral depletion. Nevertheless, both cyanidin and flavonol derivatives increase in response to nitrogen depletion. Kaempferols are the dominating flavonols in Arabidopsis leaves under normal cultivation conditions, but quercetin accumulation can be triggered by nitrogen depletion in combination with other abiotic factors.

Responses of Nine Trifolium repens L. Populations to Ultraviolet-B Radiation: Differential Flavonol Glycoside Accumulation and Biomass Production

This study aimed to quantify and identify flavonoids involved in the response of nine populations of white clover (Trifolium repens L.) to ultraviolet-B radiation (UV-B). Plants were grown for 12 weeks in controlled environment rooms with or without supplemental UV-B radiation of 13.3kJ m−2d−1. Methanol–water extractable flavonoids were quantified using high performance liquid chromatography (HPLC). Two major peaks showed significant enhancement in the HPLC chromatogram in response to supplemental UV-B. The structures of the compounds responsible were identified by1H and13C nuclear magnetic resonance (NMR) spectroscopy to be the flavonols quercetin-3-O-β- D -xylopyranosyl-(1→2)-β- D -galactopyranoside and kaempferol-3-O-β- D -xylopyranosyl-(1→2)-β- D -galactopyranoside. With supplemental UV-B, quercetin glycoside levels increased on average by 200% while the kaempferol glycoside response was much smaller. Significant differences in flavonol accumulation were found among T. repens populations, both constitutively and in response to UV-B. Stress-adapted populations displayed particularly high flavonol levels under UV-B. There was an inverse correlation between plant productivity and quercetin accumulation. Furthermore, higher quercetin accumulation under UV-B was correlated with tolerance against UV-B-induced growth reduction. In conclusion, within-species comparisons in T. repens lend support to a distinct role for ortho -dihydroxylated flavonoids in the adaptation to UV-B stress and suggest particular advantages in this UV-B-induced biochemical adaptation for populations characterized by low habitat and plant productivity.

The role of low temperatures in the development of the red blush on apple fruit (`Granny Smith')

Combinations of low and high temperatures with light were applied to detached `Granny Smith' apples to assess their role in the reported effect of cold nights and warm days on blush formation in apple skin. Of the temperature combinations tried, 4°C for 16 h in the dark followed by UVB–Visible light at 20°C for 75 h was the most effective in increasing anthocyanin concentrations in green fruit skin. UVB–Visible light was essential for anthocyanin accumulation, regardless of the temperature combination. UVB–Visible light appeared to induce quercetin accumulation without an effect of temperature. It was concluded that cool nights followed by warm days when UVB and light is incident on the fruit skin will bring about blush formation in apple fruit.

Time courses for phytochrome-induced enzyme levels in phenylpropanoid metabolism (phenylalanine ammonia-lyase, naringenin-chalcone synthase) compared with time courses for phytochrome-mediated end-product accumulation (anthocyanin, quercetin)

Time course for changes in the levels of enzymes characteristic of general phenylpropanoid metabolism (phenylalanine ammonia-lyase, PAL; EC 4.3.1.5) and of the flavonoid-glycoside branch pathway (naringenin-chalcone synthase, CHS; EC 2.3.1.74) were measured in the cotyledons of mustard (Sinapis alba L.) seedlings and compared with the rates of accumulation of related end products (anthocyanin and quercetin). Induction of enzyme levels and of end-product accumulation was carried out with red and far-red (FR) light, operating via phytochrome. The data are compatible with the concept that the phytochrome-mediated appearance of enzymes such as PAL and CHS is indeed a prerequisite for the appearance of anthocyanins and flavonols. However, there is no close correlation between enzyme levels and the rates of synthesis of end products which could justify the identification of specific rate-limiting enzymes. Rather, the data indicate that there is a second phytochrome-dependent step, beyond enzyme induction, where the actual rate of flavonoid accumulation is determined. Anthocyanin and quercetin accumulation respond differently to light. However, the relative action of continuous FR, red light pulses and 'stored phytochrome signal' is the same in both cases. This indicates that the mode of operation of phytochrome is the same in both cases. The two syntheses differ only in the degree of responsiveness towards phytochrome. The time course for changes in CHS levels in continuous FR, i.e. under conditions of phytochrome photosteady state, is similar to the time course for PAL levels whereas the time courses in darkness, following transfer from FR to darkness, are totally different. In the case of CHS, a transient rise is observed whereas, with PAL, an instantaneous drop in enzyme level occurs after transfer from FR to darkness. It is concluded that the 'stored phytochrome signal' operates in darkness in the case of CHS but not in the case of PAL.

Phytochrome-induced flavonoid biosynthesis in mustard (Sinapis alba L.) cotyledons. Enzymic control and differential regulation of anthocyanin and quercetin formation

Phytochrome-induced increases in enzyme activities for phenylalanine ammonia-lyase (EC 4.3.1.5) and chalcone isomerase (EC 5.5.1.6), and in amounts of the related end products, anthocyanin and the flavonol, quercetin, were measured in cotyledons of mustard (Sinapis alba L.). There was no correlation between the activities of these enzymes and the rate of anthocyanin accumulation; however, some correlation was found with the quercetin accumulation rate. Since anthocyanin and flavonol accumulation is spatially separated in mustard (flavonols in the upper epidermis, anthocyanin in the lower epidermis), it was possible to measure anthocyanin-associated phenylalanine ammonia-lyase independently. This activity correlated well with the accumulation rate for anthocyanin during the first few hours after induction. The phytochrome effect on anthocyanin formation differed from that on quercetin formation: anthocyanin was strongly induced by continuous far-red light and by both continuous red light and red light pulses, whereas quercetin was only effectively induced by continuous far-red light.