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Cloning, expression analysis and enzyme activity verification of dihydroflavonol 4-reductase from Cistanche tubulosa (Schenk) Wight flower
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Hai-ling QIU1, 2, Fang-ming WANG3, Bo-wen GAO4, Xin-yu MI1, 2, Ze-kun ZHANG1, 2, Yu DU1, 2, She-po SHI1, 2, Peng-fei TU3, *, Xiao-hui WANG1, 2, *
Acta Pharmaceutica Sinica | 2023, 58(4) : 1079 - 1089
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Acta Pharmaceutica Sinica | 2023, 58(4): 1079-1089
• Original Articles •
Cloning, expression analysis and enzyme activity verification of dihydroflavonol 4-reductase from Cistanche tubulosa (Schenk) Wight flower
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Hai-ling QIU1, 2, Fang-ming WANG3, Bo-wen GAO4, Xin-yu MI1, 2, Ze-kun ZHANG1, 2, Yu DU1, 2, She-po SHI1, 2, Peng-fei TU3, *, Xiao-hui WANG1, 2, *
Affiliations
  • 1. School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 102488, China
  • 2. Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China
  • 3. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Science, Peking University, Beijing 100191, China
  • 4. Baotou Medical College, Baotou 014060, China
Published: 2023-04-12 doi: 10.16438/j.0513-4870.2022-1008
Outline
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Dihydroflavonol 4-reductase (DFR) plays an essential role in the biosynthesis of anthocyanin and regulation of plant flower color. Based on the transcriptome data of Cistanche tubulosa (Schenk) Wight, a full-length cDNA sequence of CtDFR gene was cloned by reverse transcription-polymerase chain reaction (RT-PCR). CtDFR contains an open reading frame (ORF) of 1 263 bp which encodes 420 amino acids with a predicted molecular weight of 47.5 kDa. The sequence analysis showed that CtDFR contains a nicotinamide adenine dinucleotide phosphate (NADPH) binding domain and a specific substrate binding domain. The expression analysis indicated that CtDFR was highly expressed in red and purple flowers, and the relative expression levels were 4.04 and 19.37 times higher than those of white flowers, respectively. The recombinant CtDFR protein was expressed in E.coli BL21 (DE3) using vector pET-28a-CtDFR and was purified. In vitro enzyme activity analysis, CtDFR could reduce three types of dihydroflavonols including dihydrokaempferol, dihydroquercetin, and dihydromyricetin to leucopelargonidin, leucocyanidin and leucodelphinidin. Subcellular localization analysis showed that CtDFR was mainly localized in the cytoplasm. These results demonstrate that CtDFR plays an important role in regulation of flower color in C. tubulosa and make a valuable contribution for the further investigation on the regulation mechanism of C. tubulosa (Schenk) Wight flower color.

Cistanche tubulosa (Schenk) Wight  /  flower color  /  dihydroflavonol 4-reductase  /  expression analysis  /  enzyme activity analysis  /  subcellular localization
Hai-ling QIU, Fang-ming WANG, Bo-wen GAO, Xin-yu MI, Ze-kun ZHANG, Yu DU, She-po SHI, Peng-fei TU, Xiao-hui WANG. Cloning, expression analysis and enzyme activity verification of dihydroflavonol 4-reductase from Cistanche tubulosa (Schenk) Wight flower[J]. Acta Pharmaceutica Sinica, 2023 , 58 (4) : 1079 -1089 . DOI: 10.16438/j.0513-4870.2022-1008
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Article Info
doi: 10.16438/j.0513-4870.2022-1008
  • Receive Date：2022-08-23
  • Online Date：2025-11-21
  • Published：2023-04-12
Article Data
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History
  • Received：2022-08-23
  • Revised：2022-09-21
Funding
Affiliations
    1. School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 102488, China
    2. Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China
    3. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Science, Peking University, Beijing 100191, China
    4. Baotou Medical College, Baotou 014060, China
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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