Eos kiss digital x7i camera

Fluorescence was visually evaluated by comparing the photographs taken by a Canon EOS Kiss Digital X7i camera (Canon, Tokyo, Japan) under UV light excitation (30 W LED, light intensity of 150 μmol m⁻² s⁻¹, peak wavelength 365 nm; NS365-FLB-30WR, Nitride semiconductors Co., Ltd, Tokushima, Japan) or blue light excitation (60 W LED, light intensity of 300 μmol m⁻² s⁻¹, peak wavelength 470 nm; VBL-SL150BB, Valore, Kyoto, Japan). Two excitation filters were used: the UV LED light was covered with a UL360 filter (OMG Co. Ltd, Osaka, Japan) to eliminate visible light, and the blue LED light was covered with a SV0490 filter (Asahi Spectra, Tokyo, Japan) to eliminate long-wavelength light. A yellowish sharp cut filter (SC-52, Fujifilm, Tokyo, Japan) was used as an emission filter for blue light excitation. Both LED light sources were placed approximately 300 mm apart from the uppermost petals of each plant to obtain images under the same conditions (f-ratio, ISO speed, shutter speed), as listed in Supplementary Table S2.

For protein extraction from the leaves and petals of each cultivar, we used the 8-min Plant Tissue Total Protein Extraction Kit (101Bio, Palo Alto, CA, USA) with native lysis buffer, following the manufacturer’s instructions. Protein concentration of each extract was determined using a BCA protein assay kit (Pierce Biotechnology, Rockland, IL, USA). Fluorescence excitation and emission of 10 mg mL⁻¹ (50 μL) of each protein extract were scanned with a M1000 Pro microplate reader (TECAN, Männedorf, Switzerland). For native-PAGE analysis, 2 µg of purified polyhistidine-tagged proteins and 80 µg of each crude protein extract were loaded on 10% polyacrylamide gels (c-PAGEL C10L; ATTO, Tokyo, Japan) and photograph was taken by using a Canon EOS Kiss Digital X7i camera under UV LED lights as described above. The image acquisition condition was listed in Supplementary Table S2.