Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) Hara
The synthesis of secondary metabolites in plants often includes glycosylation modifications. Often, the final step of constructing plant secondary metabolites is completed by glycosylation transferases, which are also involved in many cell processes. In this study, a UDP-glycosyltransferase gene (&l...
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MDPI AG
2022-08-01
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| author | Conglong Lian Jinxu Lan Bao Zhang Hao Yang Kaihua Guo Jingjing Li Suiqing Chen |
| author_facet | Conglong Lian Jinxu Lan Bao Zhang Hao Yang Kaihua Guo Jingjing Li Suiqing Chen |
| author_sort | Conglong Lian |
| collection | DOAJ |
| description | The synthesis of secondary metabolites in plants often includes glycosylation modifications. Often, the final step of constructing plant secondary metabolites is completed by glycosylation transferases, which are also involved in many cell processes. In this study, a UDP-glycosyltransferase gene (<i>UGT</i>) was amplified from <i>Isodon rubescens</i> (Hemsl.) Hara with RT-PCR and named <i>IrUGT86A1-like</i> (GenBank: MZ913258). Here, we found that <i>IrUGT86A1-like</i> gene is 1450 bp in length and encodes for 479 amino acids. Bioinformatics analysis revealed that <i>IrUGT86A1-like</i> is a stable and hydrophilic protein, located in the cytoplasm with a transmembrane domain. Phylogenetic analysis showed that IrUGT86A1-like protein has the closest genetic relationship with the UDP-glycosyltransferase 86A1-like protein (XP_042054241.1) of <i>Salvia splendens</i>. RT-qPCR analysis demonstrated that the expression of <i>IrUGT86A1-like</i> gene varied in different tissues; leaves had the highest expression followed by flowers, stems, and roots had the lowest expression. This expression trend is similar to the distribution of oridonin content in different tissues of <i>I. rubescens</i>. Additionally, <i>IrUGT86A1-like</i> gene was found to be positively enhanced by NaCl and MeJA treatment, and in contrast was down-regulated by ABA treatment. Finally, the prokaryotic expression vector pEASY<sup>®</sup>-Blunt E1-IrUGT86A1 was successfully used to express about 53 KD of IrUGT86A1-like protein. This research builds a foundation for further investigation on the function of this gene in the synthesis and modification of secondary metabolites. |
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| spelling | doaj.art-4a3150402607470780ee98852ecd671d2023-11-23T17:22:22ZengMDPI AGLife2075-17292022-08-01129133410.3390/life12091334Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) HaraConglong Lian0Jinxu Lan1Bao Zhang2Hao Yang3Kaihua Guo4Jingjing Li5Suiqing Chen6School of Pharmacy, Henan Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry, Henan University of Chinese Medicine, Boxue Road, Jinshui District, Zhengzhou 450046, ChinaSchool of Pharmacy, Henan Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry, Henan University of Chinese Medicine, Boxue Road, Jinshui District, Zhengzhou 450046, ChinaSchool of Pharmacy, Henan Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry, Henan University of Chinese Medicine, Boxue Road, Jinshui District, Zhengzhou 450046, ChinaSchool of Pharmacy, Henan Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry, Henan University of Chinese Medicine, Boxue Road, Jinshui District, Zhengzhou 450046, ChinaSchool of Pharmacy, Henan Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry, Henan University of Chinese Medicine, Boxue Road, Jinshui District, Zhengzhou 450046, ChinaSchool of Pharmacy, Henan Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry, Henan University of Chinese Medicine, Boxue Road, Jinshui District, Zhengzhou 450046, ChinaSchool of Pharmacy, Henan Key Laboratory of Chinese Medicine Resources and Chinese Medicine Chemistry, Henan University of Chinese Medicine, Boxue Road, Jinshui District, Zhengzhou 450046, ChinaThe synthesis of secondary metabolites in plants often includes glycosylation modifications. Often, the final step of constructing plant secondary metabolites is completed by glycosylation transferases, which are also involved in many cell processes. In this study, a UDP-glycosyltransferase gene (<i>UGT</i>) was amplified from <i>Isodon rubescens</i> (Hemsl.) Hara with RT-PCR and named <i>IrUGT86A1-like</i> (GenBank: MZ913258). Here, we found that <i>IrUGT86A1-like</i> gene is 1450 bp in length and encodes for 479 amino acids. Bioinformatics analysis revealed that <i>IrUGT86A1-like</i> is a stable and hydrophilic protein, located in the cytoplasm with a transmembrane domain. Phylogenetic analysis showed that IrUGT86A1-like protein has the closest genetic relationship with the UDP-glycosyltransferase 86A1-like protein (XP_042054241.1) of <i>Salvia splendens</i>. RT-qPCR analysis demonstrated that the expression of <i>IrUGT86A1-like</i> gene varied in different tissues; leaves had the highest expression followed by flowers, stems, and roots had the lowest expression. This expression trend is similar to the distribution of oridonin content in different tissues of <i>I. rubescens</i>. Additionally, <i>IrUGT86A1-like</i> gene was found to be positively enhanced by NaCl and MeJA treatment, and in contrast was down-regulated by ABA treatment. Finally, the prokaryotic expression vector pEASY<sup>®</sup>-Blunt E1-IrUGT86A1 was successfully used to express about 53 KD of IrUGT86A1-like protein. This research builds a foundation for further investigation on the function of this gene in the synthesis and modification of secondary metabolites.https://www.mdpi.com/2075-1729/12/9/1334<i>Isodon rubescens</i>UDP-glycosyltransferaseRT-qPCRprokaryotic expression |
| spellingShingle | Conglong Lian Jinxu Lan Bao Zhang Hao Yang Kaihua Guo Jingjing Li Suiqing Chen Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) Hara Life <i>Isodon rubescens</i> UDP-glycosyltransferase RT-qPCR prokaryotic expression |
| title | Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) Hara |
| title_full | Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) Hara |
| title_fullStr | Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) Hara |
| title_full_unstemmed | Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) Hara |
| title_short | Molecular Cloning and Functional Analysis of <i>IrUGT86A1-like</i> Gene in Medicinal Plant <i>Isodon rubescens</i> (Hemsl.) Hara |
| title_sort | molecular cloning and functional analysis of i irugt86a1 like i gene in medicinal plant i isodon rubescens i hemsl hara |
| topic | <i>Isodon rubescens</i> UDP-glycosyltransferase RT-qPCR prokaryotic expression |
| url | https://www.mdpi.com/2075-1729/12/9/1334 |
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