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Characterization of the enzyme activity in barley anthocyanin pathway and reaction of these pigments to electromagnetic irradiation

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Abstract

The enzymatic activities were indirectly investigated in the grain anthocyanin synthesis of blue barley by the HPLC data. These results were then evaluated together with absorbed light energy levels of the pigments. In addition, in two photochemical experiments, the change in aleurone pigmentation and the water activation was studied at the dormant grains, using focused sunlight and heat radiation. In the middle stage of the synthesis (at day after flowering, DAF 26), there was a significant cooling period in the weather. Differences of synthesis dynamics were found before and after the cooling period. The anthocyanin production after the cooling period (DAF 26–33) was more intense compared to the beginning of synthesis (17–22). In addition, a more intense degradation was detectable during the cooling period (22–26) than what was observed at the end of seed maturation (33–39). The most efficient light energy binders glucosides (delphinidin- and cyanidin-3-glu.) produced and degraded more dynamically than their more complex forms (malonylglucosides, rutinoside). Furthermore, among the pigments, the cyanidins are able to provide greater energy absorption. Differences in the synthesis dynamism of compounds indicate that individual enzymes and not a multienzyme complex operate in the last phase of the anthocyanin pathway, and by their operation, they can change the energy absorption level of aleurone. In irradiation of blue grains with focused sunlight (~ 400 to  ~ 2500 nm, 2–3 min), the aleurone anthocyanins facilitated the vitrified water activation. During intense heat irradiation (~ 8000 nm, 1 min), the laser light scattering associated with water content decreased more intensively within the blue grains compared to the white, indicating the IR absorption surplus for the pigments. Observation suggests that the blue pigments in aleurone can contribute to energy transfer in the direction of water, so they may have role in enhancing energy dissipation.

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Acknowledgements

I would like to express my gratitude to Dr. László Kursinszki for the help he provided in the HPLC experiment.

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  1. Eger, Hungary

    I. Diczházi

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  1. I. Diczházi
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Electronic supplementary material

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Figure S1. Average daily temperatures during the period of grain development. The boxed dates are times for the ears harvested (JPEG 75 kb)

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Figure S2. HPLC chromatogram of B-1133 blue barley anthocyanins. Peak numbers show the major anthocyanins: 1 delphinidin-3-O-glucoside, 2 delphinidin-3-O-rutinoside, 3 cyanidin-3-O-glucoside, 4 delphinidin-3-O-malonylglucoside, 5 cyanidin-3-O-malonylglucoside (JPEG 47 kb)

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Figure S3. One B-1133 barley grain is shown in dehulled state (a) and after metal plate insertion (b, *). Then, it is seen after focused solar light irradiation (c) followed by during and after acidic solution treatment (d 7 h, e 48 h). The yellow arrow and circle indicate the direction of the solar irradiation and the focal point (JPEG 138 kb)

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Figure S4. They are shown in the film the apparatus and setup for solar irradiation treatment of barley grain (AVI 55054 kb)

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Figure S5. Discolored B-1133 blue barley grains effect to focused solar light irradiation (a) and the red coloring on them in wet (b) and air-dried states (c) due to acidic solution treatment (JPEG 151 kb)

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Figure S6. Apparatus for detecting to laser light scattering change within the grains during heating. The names of devices are shown in the picture (JPEG 4627 kb)

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Figure S7. Laser light scattering change with inverted colors in case blue and white grains. In the snapshots of the films made during heating, the black–red–pink parts are the scattered, while the white–pale blue parts are nonscattered areas. The rules of ellipses drawing are written in Materials and Methods (JPEG 184 kb)

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Figure S8. The film presents the process of the green laser light scattering decrease when the blue barley grain was exposed to heat radiation (AVI 116462 kb)

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Figure S9. HPLC samples of the grain anthocyanins from B-1133 blue barley sorted by harvest dates (a 17, b 22, c 26, d 33, e 39 day after flowering) (JPEG 55 kb)

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Figure S10. Native, blue (a, b) and previously solar light-treated discolored (c) barley grains are shown in this picture after solar irradiation (a, b) and re-irradiation (c) when the light was focused in small focal points. Carbonization zones are marked with black circles, and the empty circle is belonging to dehydration zones (JPEG 149 kb)

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Supplementary material 12 (DOCX 21 kb)

Supplementary material 13 (DOCX 19 kb)

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Diczházi, I. Characterization of the enzyme activity in barley anthocyanin pathway and reaction of these pigments to electromagnetic irradiation. CEREAL RESEARCH COMMUNICATIONS 48, 317–324 (2020). https://doi.org/10.1007/s42976-020-00045-w

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