Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to Acetylshikonin
The BAHD acyltransferase family is a unique class of plant proteins that acylates plant metabolites and participates in plant secondary metabolic processes. However, the BAHD members in <i>Lithospermum erythrorhizon</i> remain unknown and uncharacterized. Although the heterologously expr...
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2022-11-01
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| Series: | Life |
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| author | Xuan Wang Zhuoyu He Huan Yang Cong He Changyi Wang Aliya Fazal Xiaohui Lai Liangjie Yang Zhongling Wen Minkai Yang Shenglin Ma Wencai Jie Jinfeng Cai Tongming Yin Bao Liu Yonghua Yang Jinliang Qi |
| author_facet | Xuan Wang Zhuoyu He Huan Yang Cong He Changyi Wang Aliya Fazal Xiaohui Lai Liangjie Yang Zhongling Wen Minkai Yang Shenglin Ma Wencai Jie Jinfeng Cai Tongming Yin Bao Liu Yonghua Yang Jinliang Qi |
| author_sort | Xuan Wang |
| collection | DOAJ |
| description | The BAHD acyltransferase family is a unique class of plant proteins that acylates plant metabolites and participates in plant secondary metabolic processes. However, the BAHD members in <i>Lithospermum erythrorhizon</i> remain unknown and uncharacterized. Although the heterologously expressed <i>L. erythrorhizon</i> BAHD family member LeSAT1 in <i>Escherichia coli</i> has been shown to catalyze the conversion of shikonin to acetylshikonin in vitro, its in vivo role remains unknown. In this study, the characterization, evolution, expression patterns, and gene function of <i>LeBAHDs</i> in <i>L. erythrorhizon</i> were explored by bioinformatics and transgenic analysis. We totally identified 73 <i>LeBAHDs</i> in the reference genome of <i>L. erythrorhizon</i>. All <i>LeBAHDs</i> were phylogenetically classified into five clades likely to perform different functions, and were mainly expanded by dispersed and WGD/segmental duplication. The in vivo functional investigation of the key member <i>LeBAHD1/LeSAT1</i> revealed that overexpression of <i>LeBAHD1</i> in hairy roots significantly increased the content of acetylshikonin as well as the conversion rate of shikonin to acetylshikonin, whereas the CRISPR/Cas9-based knockout of <i>LeBAHD1</i> in hairy roots displayed the opposite trend. Our results not only confirm the in vivo function of <i>LeBAHD1/LeSAT1</i> in the biosynthesis of acetylshikonin, but also provide new insights for the biosynthetic pathway of shikonin and its derivatives. |
| first_indexed | 2024-03-09T18:54:44Z |
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| institution | Directory Open Access Journal |
| issn | 2075-1729 |
| language | English |
| last_indexed | 2024-03-09T18:54:44Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
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| series | Life |
| spelling | doaj.art-5899ef78bfac4b8094fdffcf1c4265872023-11-24T05:30:46ZengMDPI AGLife2075-17292022-11-011211177510.3390/life12111775Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to AcetylshikoninXuan Wang0Zhuoyu He1Huan Yang2Cong He3Changyi Wang4Aliya Fazal5Xiaohui Lai6Liangjie Yang7Zhongling Wen8Minkai Yang9Shenglin Ma10Wencai Jie11Jinfeng Cai12Tongming Yin13Bao Liu14Yonghua Yang15Jinliang Qi16State Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaYili Key Laboratory of Applied Research and Development on Active Ingredients of Chinese Herbal Medicine, Yili National Agricultural Science and Technology Park at Xinjiang, Yili 835600, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaCo-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, ChinaCo-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, ChinaKey Laboratory of Molecular Epigenetics of the Ministry of Education (MOE), Northeast Normal University, Changchun 130024, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaState Key Laboratory of Pharmaceutical Biotechnology, Institute for Plant Molecular Biology, School of Life Sciences, Nanjing University, Nanjing 210023, ChinaThe BAHD acyltransferase family is a unique class of plant proteins that acylates plant metabolites and participates in plant secondary metabolic processes. However, the BAHD members in <i>Lithospermum erythrorhizon</i> remain unknown and uncharacterized. Although the heterologously expressed <i>L. erythrorhizon</i> BAHD family member LeSAT1 in <i>Escherichia coli</i> has been shown to catalyze the conversion of shikonin to acetylshikonin in vitro, its in vivo role remains unknown. In this study, the characterization, evolution, expression patterns, and gene function of <i>LeBAHDs</i> in <i>L. erythrorhizon</i> were explored by bioinformatics and transgenic analysis. We totally identified 73 <i>LeBAHDs</i> in the reference genome of <i>L. erythrorhizon</i>. All <i>LeBAHDs</i> were phylogenetically classified into five clades likely to perform different functions, and were mainly expanded by dispersed and WGD/segmental duplication. The in vivo functional investigation of the key member <i>LeBAHD1/LeSAT1</i> revealed that overexpression of <i>LeBAHD1</i> in hairy roots significantly increased the content of acetylshikonin as well as the conversion rate of shikonin to acetylshikonin, whereas the CRISPR/Cas9-based knockout of <i>LeBAHD1</i> in hairy roots displayed the opposite trend. Our results not only confirm the in vivo function of <i>LeBAHD1/LeSAT1</i> in the biosynthesis of acetylshikonin, but also provide new insights for the biosynthetic pathway of shikonin and its derivatives.https://www.mdpi.com/2075-1729/12/11/1775<i>Lithospermum erythrorhizon</i>acetylshikoninshikoninBAHDLeSAT1hairy root |
| spellingShingle | Xuan Wang Zhuoyu He Huan Yang Cong He Changyi Wang Aliya Fazal Xiaohui Lai Liangjie Yang Zhongling Wen Minkai Yang Shenglin Ma Wencai Jie Jinfeng Cai Tongming Yin Bao Liu Yonghua Yang Jinliang Qi Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to Acetylshikonin Life <i>Lithospermum erythrorhizon</i> acetylshikonin shikonin BAHD LeSAT1 hairy root |
| title | Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to Acetylshikonin |
| title_full | Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to Acetylshikonin |
| title_fullStr | Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to Acetylshikonin |
| title_full_unstemmed | Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to Acetylshikonin |
| title_short | Genome-Wide Identification of <i>LeBAHDs</i> in <i>Lithospermum erythrorhizon</i> and In Vivo Transgenic Studies Confirm the Critical Roles of <i>LeBAHD1/LeSAT1</i> in the Conversion of Shikonin to Acetylshikonin |
| title_sort | genome wide identification of i lebahds i in i lithospermum erythrorhizon i and in vivo transgenic studies confirm the critical roles of i lebahd1 lesat1 i in the conversion of shikonin to acetylshikonin |
| topic | <i>Lithospermum erythrorhizon</i> acetylshikonin shikonin BAHD LeSAT1 hairy root |
| url | https://www.mdpi.com/2075-1729/12/11/1775 |
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