Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniata
Chalcone isomerase (CHI), catalyzing isomerization of chalcones, is a crucial enzyme in flavonoid biosynthesis. Three CHI genes were isolated from Clivia miniata and designated as CmCHI1, CmCHI2 and CmCHI3, respectively. Multiple sequence alignments and phylogenetic analysis showed that CmCHI1 and C...
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Maximum Academic Press
2021-01-01
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| Series: | Ornamental Plant Research |
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| Online Access: | https://www.maxapress.com/article/doi/10.48130/OPR-2021-0002 |
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| author | Yue Liu Xinxin Xue Chunli Zhao Jia Zhang Meng Liu Xiangyu Li Yueqing Li Xiang Gao |
| author_facet | Yue Liu Xinxin Xue Chunli Zhao Jia Zhang Meng Liu Xiangyu Li Yueqing Li Xiang Gao |
| author_sort | Yue Liu |
| collection | DOAJ |
| description | Chalcone isomerase (CHI), catalyzing isomerization of chalcones, is a crucial enzyme in flavonoid biosynthesis. Three CHI genes were isolated from Clivia miniata and designated as CmCHI1, CmCHI2 and CmCHI3, respectively. Multiple sequence alignments and phylogenetic analysis showed that CmCHI1 and CmCHI2 were members of type I CHI proteins, whereas CmCHI3 belonged to type IV CHI proteins. Subcellular localization analysis found that all three CmCHIs had diffused distribution in the cytoplasm similar to green fluorescent protein (GFP). Anthocyanin biosynthesis and gene expression analysis demonstrated that CmCHIs were highly expressed in anthocyanin accumulated tissues. To further functionally characterize the role of CmCHIs, an in vitro enzymatic activity assay was carried out using the purified recombinant proteins. Results showed that CmCHI1 and CmCHI2 could completely convert the substrate naringenin chalcone (NC) into the product naringenin (NA), whereas CmCHI3 seemed nonfunctional as no increment of NA was detected. Further genetic transformation of Arabidopsis tt5-1 mutant validated that CmCHI1 and CmCHI2 rather than CmCHI3 could complement the chi deficient phenotypes. In summary, CmCHI1 and CmCHI2 are the real active CHI genes in Clivia miniata. The results not only broaden our knowledge on flavonoid biosynthesis in C. miniata but also lay a new foundation for further flavonoid modification in C. miniata. |
| first_indexed | 2024-03-07T20:10:18Z |
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| id | doaj.art-d34c4340bd22464bad43a968bb3a4270 |
| institution | Directory Open Access Journal |
| issn | 2769-2094 |
| language | English |
| last_indexed | 2024-03-07T20:10:18Z |
| publishDate | 2021-01-01 |
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| series | Ornamental Plant Research |
| spelling | doaj.art-d34c4340bd22464bad43a968bb3a42702024-02-28T01:52:27ZengMaximum Academic PressOrnamental Plant Research2769-20942021-01-011111010.48130/OPR-2021-0002OPR-2021-0002Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniataYue Liu0Xinxin Xue1Chunli Zhao2Jia Zhang3Meng Liu4Xiangyu Li5Yueqing Li6Xiang Gao7College of Horticulture, Jilin Agricultural University, Changchun 130118, ChinaDepartment of Botany, University of British Columbia, 2329 West Mall Vancouver, BC Canada V6T 1Z4College of Horticulture, Jilin Agricultural University, Changchun 130118, ChinaKey Laboratory of Molecular Epigenetics of MOE and Institute of Genetics & Cytology, Northeast Normal University, Changchun 130024, ChinaKey Laboratory of Molecular Epigenetics of MOE and Institute of Genetics & Cytology, Northeast Normal University, Changchun 130024, ChinaKey Laboratory of Molecular Epigenetics of MOE and Institute of Genetics & Cytology, Northeast Normal University, Changchun 130024, ChinaKey Laboratory of Molecular Epigenetics of MOE and Institute of Genetics & Cytology, Northeast Normal University, Changchun 130024, ChinaKey Laboratory of Molecular Epigenetics of MOE and Institute of Genetics & Cytology, Northeast Normal University, Changchun 130024, ChinaChalcone isomerase (CHI), catalyzing isomerization of chalcones, is a crucial enzyme in flavonoid biosynthesis. Three CHI genes were isolated from Clivia miniata and designated as CmCHI1, CmCHI2 and CmCHI3, respectively. Multiple sequence alignments and phylogenetic analysis showed that CmCHI1 and CmCHI2 were members of type I CHI proteins, whereas CmCHI3 belonged to type IV CHI proteins. Subcellular localization analysis found that all three CmCHIs had diffused distribution in the cytoplasm similar to green fluorescent protein (GFP). Anthocyanin biosynthesis and gene expression analysis demonstrated that CmCHIs were highly expressed in anthocyanin accumulated tissues. To further functionally characterize the role of CmCHIs, an in vitro enzymatic activity assay was carried out using the purified recombinant proteins. Results showed that CmCHI1 and CmCHI2 could completely convert the substrate naringenin chalcone (NC) into the product naringenin (NA), whereas CmCHI3 seemed nonfunctional as no increment of NA was detected. Further genetic transformation of Arabidopsis tt5-1 mutant validated that CmCHI1 and CmCHI2 rather than CmCHI3 could complement the chi deficient phenotypes. In summary, CmCHI1 and CmCHI2 are the real active CHI genes in Clivia miniata. The results not only broaden our knowledge on flavonoid biosynthesis in C. miniata but also lay a new foundation for further flavonoid modification in C. miniata.https://www.maxapress.com/article/doi/10.48130/OPR-2021-0002anthocyaninenzymatic assayflavonoidplant transformationtransient protoplast transfection assay |
| spellingShingle | Yue Liu Xinxin Xue Chunli Zhao Jia Zhang Meng Liu Xiangyu Li Yueqing Li Xiang Gao Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniata Ornamental Plant Research anthocyanin enzymatic assay flavonoid plant transformation transient protoplast transfection assay |
| title | Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniata |
| title_full | Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniata |
| title_fullStr | Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniata |
| title_full_unstemmed | Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniata |
| title_short | Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in Clivia miniata |
| title_sort | cloning and functional characterization of chalcone isomerase genes involved in anthocyanin biosynthesis in clivia miniata |
| topic | anthocyanin enzymatic assay flavonoid plant transformation transient protoplast transfection assay |
| url | https://www.maxapress.com/article/doi/10.48130/OPR-2021-0002 |
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