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3 protocols using α carotene

1

Carotenoid Extraction and Quantification

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α-carotene and β-carotene standards were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Lutein standard was purchased from CaroteNature GmbH (Ostermundigen, Switzerland). Lycopene standard was purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). Special grade n-Hexane, acetone, ethanol, toluene, and pyrogallol were purchased from Wako Pure Chemical Industries, Ltd. HPLC grade methyl tert-butyl ether, acetonitrile, and methanol were purchased from Wako Pure Chemical Industries, Ltd. Cellulose was purchased from Amano Enzyme Inc. (Nagoya, Japan). Pectinase was purchased from Yakult Pharmaceutical Industry Co., Ltd. (Tokyo, Japan). Citric acid was purchased from Showa Kako Co., Ltd., (Osaka, Japan). Pectin was purchased from Unitec Foods Co., Ltd., (Tokyo, Japan).
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2

Carotenoid Extraction and Quantification from Flowering Petals

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Carotenoid were extracted from fresh petals at the flowering stage and detected following the methods of Cao et al.51 (link). Carotenoid analysis was performed using LC-2010AHT HPLC (Shimadzu, Kyoto, Japan) with C30 column (YMC, Kyoto, Japan). Carotenoids were identified by the typical retention time of the standard compounds, including violaxanthin (Sigma-Aldrich, Saint Louis, America), lutein (Solarbio, Beijing, China), α-carotene and β-carotene (Wako, Osaka, Japan). The identification of prolycopene was performed based on reported the typical retention time and relative order of carotenoid compound peaks22 (link),43 (link),51 (link). Carotenoid content was quantified according to Morris’ method52 (link). The total carotenoid content was the sum of all the detected carotenoid compound contents. Three biological replicates were used for all analyses and the calculation of means and standard deviations were conducted. The significant difference between 92S105 and 15S1040 was analyzed by t-test.
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3

Carotenoid Extraction and Analysis

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2.1 Chemicals Lutein, zeaxanthin, and β-cryptoxanthin were purchased from Extrasynthese (Genay, France) . α-Carotene and methyl tert-butyl ether (MTBE) were obtained from Wako Pure Chemical Industries (Osaka, Japan) . β-Carotene and lycopene were obtained from Sigma-Aldrich (St. Louis, MO, USA) . All other reagents were of the highest grade available.
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