Anthocyanin Production Using Rough Bluegrass Treated with High-Intensity Light

in HortScience

Anthocyanins are plant pigments that are in demand for medicinal and industrial uses. However, anthocyanin production is limited due to the harvest potential of the species currently used as anthocyanin sources. Rough bluegrass (Poa trivialis L.) is a perennial turfgrass known for accumulating anthocyanins, and may have the potential to serve as a source of anthocyanins through artificial light treatments. The objectives of this research were to determine optimal light conditions that favor anthocyanin synthesis in rough bluegrass, and to determine the suitability of rough bluegrass as a source of anthocyanins. When exposed to high-intensity white light, rough bluegrass increased anthocyanin content by 100-fold on average, and anthocyanin contents greater than 0.2% of dry tissue weight were observed in some samples. Blue light, at intensities between 150 and 250 μmol·m−2·s−1, was the only wavelength that increased anthocyanin content. However, when red light was applied with blue light at 30% or 50% of the total light intensity, anthocyanin content was increased compared with blue light alone. Further experiments demonstrated that these results may be potentially due to a combination of photosynthetic and photoreceptor-mediated regulation. Rough bluegrass is an attractive anthocyanin production system, since leaf tissue can be harvested while preserving meristematic tissues that allow new leaves to rapidly grow; thereby allowing multiple harvests in a single growing season and greater anthocyanin yields.

Contributor Notes

This article is a part of a thesis by senior author in partial fulfillment of the requirements for the PhD degree at The Ohio State University.

Salaries and research support were provided in part by State and Federal funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University. Journal Article HCS-16-10.

We thank the James B. and Harriet Beard Graduate Fund. We also thank J.R. Simplot Company for seed used in these experiments.

Corresponding author. E-mail: petrella.21@osu.edu.

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    High-performance liquid chromatography chromatograms of rough bluegrass (RB); cyanidin-3-glucoside; Cy3g (1), and cyanidin-3-malyonlyglucoside; Cy3mg (2). (A) RB anthocyanins; (B) saponified RB anthocyanins; and (C) purple corn (PCorn) anthocyanins. Sample saponification was used to confirm the presence of cyanidin-3-malyonylglucoside, and retention times were compared with PCorn as a standard. Ultraviolet-vis spectral profiles were also used to confirm anthocyanin identity (DG). Spectral profiles of cyanidin-3-glucoside from (D) RB and (E) PCorn exhibited the same profile and maxima (279 and 517 nm). Spectral profiles of cyanidin-3-malonylglucoside from (F) RB and (G) PCorn exhibited the same profile and maxima (279 and 519 nm). Anthocyanins were identified using mass spectrometry data (Table 2) along with retention times and ultraviolet-vis absorption spectral profiles.

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    (A) Chlorophyll a and b content, and (B) chlorophyll a:b ratios of dark- and light-grown rough bluegrass (RB) seedlings. Relative anthocyanin concentration of (C) light-grown and (D) dark-grown RB seedlings treated with a white light (control), blue, far-red, or red light-emitting diode light (200 µmol·m−2·s−1). Relative anthocyanin concentration = (Abs530 − 0.25 Abs657)/g fresh weight. Error bars represent treatment se, and letters represent mean separation using Tukey’s honest significant difference (n = 3). Different letters indicate a significant statistical difference (P = 0.05).

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    Relative anthocyanin concentration of light-grown rough bluegrass seedlings treated with blue light-emitting diode light of increasing intensity. Plants were treated using the following light intensities: 25, 50, 100, 150, 200, 250, 300, or 350 µmol·m−2·s−1. Relative anthocyanin concentration = (Abs530 − 0.25 Abs657)/g fresh weight. Error bars represent treatment se, and letters represent mean separation using Tukey’s honest significant difference (n = 3). Different letters indicate a significant statistical difference (P = 0.05).

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    Relative anthocyanin concentration of (A) light-grown seedlings and (B) mature (3-month old) rough bluegrass plants. Plants were treated with 200 µmol·m−2·s−1 of light-emitting diode light consisting of the following blue/red distributions: 0% blue (100% red), 30% blue (70% red), 50% blue (50% red), 70% blue (30% red), or 100% blue (0% red). Relative anthocyanin concentration = (Abs530 − 0.25 Abs657)/g fresh weight. Error bars represent treatment se, and letters represent mean separation using Tukey’s honest significant difference (n = 3). Different letters indicate a significant statistical difference (P = 0.05).

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    Relative anthocyanin concentration of light-grown (A) and dark-grown (B) rough bluegrass seedlings treated with or without 2.8 mg·L−1 amicarbazone (AMI), and exposed to the following LED light treatments: 170 µmol·m−2·s−1 blue light plus 0, 10, 25, 50, or 100 µmol·m−2·s−1 or red light. Relative anthocyanin concentration = (Abs530 − 0.25 Abs657)/g fresh weight. Error bars represent treatment se, and letters represent mean separation using Tukey’s honest significant difference (n = 3). Different letters indicate a significant statistical difference (P = 0.05).

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    Relative anthocyanin concentration of (A) light-grown and (B) dark-grown rough bluegrass seedlings treated with or without 2.8 mg·L−1 amicarbazone (AMI), and exposed to the following light-emitting diode light treatments: 170 µmol·m2·s−1 red light plus 0, 10, 25, 50, or 100 µmol·m2·s−1 or blue light. Relative anthocyanin concentration = (Abs530 − 0.25 Abs657)/g fresh weight. Error bars represent treatment se, and letters represent mean separation using Tukey’s honest significant difference (n = 3). Different letters indicate a significant statistical difference (P = 0.05).

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