Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in Mulberry

Mulberry (<i>Morus alba</i> L.) is a significant economic tree species in China. The lignin component serves as a critical limiting factor that impacts both the forage quality and the conversion efficiency of mulberry biomass into biofuel. Cinnamoyl CoA reductase (CCR; EC 1.21.1.44) and...

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Main Authors: Shuai Huang, Xiaoru Kang, Rumeng Fu, Longyan Zheng, Peijun Li, Fengjuan Tang, Nan Chao, Li Liu
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/13/24/3512
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author Shuai Huang
Xiaoru Kang
Rumeng Fu
Longyan Zheng
Peijun Li
Fengjuan Tang
Nan Chao
Li Liu
author_facet Shuai Huang
Xiaoru Kang
Rumeng Fu
Longyan Zheng
Peijun Li
Fengjuan Tang
Nan Chao
Li Liu
author_sort Shuai Huang
collection DOAJ
description Mulberry (<i>Morus alba</i> L.) is a significant economic tree species in China. The lignin component serves as a critical limiting factor that impacts both the forage quality and the conversion efficiency of mulberry biomass into biofuel. Cinnamoyl CoA reductase (CCR; EC 1.21.1.44) and cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.95) are the key enzymes that catalyze the final two reductive steps in the biosynthesis of monolignols. In this study, we conducted a comprehensive functional analysis to validate the predominant <i>CCR</i> genes involved in monolignol biosynthesis. In this study, we initially validated the predominant <i>CCR</i> genes implicated in monolignol biosynthesis through an extensive functional analysis. Phylogenetic analysis, tissue-specific expression profiling and enzymatic assays indicated that <i>MaCCR1</i> is the authentic CCR involved in lignin biosynthesis. Furthermore, the expression level of <i>MaCCR1</i> exhibited a significant positive correlation with lignin content, and the down-regulation of <i>MaCCR1</i> via virus-induced gene silencing resulted in altered lignin content in mulberry. The down-regulation of <i>MaCCR1</i> and <i>MaCAD3/4,</i> both individually and concurrently, exhibited markedly different effects on lignin content and mulberry growth. Specifically, the simultaneous down-regulation of <i>MaCCR1</i> and <i>MaCAD3/4</i> significantly altered lignin content in mulberry, resulting in dwarfism of the plants. Conversely, the down-regulation of <i>MaCAD3/4</i> alone not only decreased lignin content but also led to an increase in biomass. These findings offer compelling evidence elucidating the roles of <i>MaCCRs</i> in mulberry and identify specific target genes, thereby providing a crucial foundation for the genetic modification of lignin biosynthesis.
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spelling doaj-art-b25d2b4b4ab9488ea5167a918214c0ec2024-12-27T14:47:47ZengMDPI AGPlants2223-77472024-12-011324351210.3390/plants13243512Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in MulberryShuai Huang0Xiaoru Kang1Rumeng Fu2Longyan Zheng3Peijun Li4Fengjuan Tang5Nan Chao6Li Liu7Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaJiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaJiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaJiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaJiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaJiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaJiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaJiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, ChinaMulberry (<i>Morus alba</i> L.) is a significant economic tree species in China. The lignin component serves as a critical limiting factor that impacts both the forage quality and the conversion efficiency of mulberry biomass into biofuel. Cinnamoyl CoA reductase (CCR; EC 1.21.1.44) and cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.95) are the key enzymes that catalyze the final two reductive steps in the biosynthesis of monolignols. In this study, we conducted a comprehensive functional analysis to validate the predominant <i>CCR</i> genes involved in monolignol biosynthesis. In this study, we initially validated the predominant <i>CCR</i> genes implicated in monolignol biosynthesis through an extensive functional analysis. Phylogenetic analysis, tissue-specific expression profiling and enzymatic assays indicated that <i>MaCCR1</i> is the authentic CCR involved in lignin biosynthesis. Furthermore, the expression level of <i>MaCCR1</i> exhibited a significant positive correlation with lignin content, and the down-regulation of <i>MaCCR1</i> via virus-induced gene silencing resulted in altered lignin content in mulberry. The down-regulation of <i>MaCCR1</i> and <i>MaCAD3/4,</i> both individually and concurrently, exhibited markedly different effects on lignin content and mulberry growth. Specifically, the simultaneous down-regulation of <i>MaCCR1</i> and <i>MaCAD3/4</i> significantly altered lignin content in mulberry, resulting in dwarfism of the plants. Conversely, the down-regulation of <i>MaCAD3/4</i> alone not only decreased lignin content but also led to an increase in biomass. These findings offer compelling evidence elucidating the roles of <i>MaCCRs</i> in mulberry and identify specific target genes, thereby providing a crucial foundation for the genetic modification of lignin biosynthesis.https://www.mdpi.com/2223-7747/13/24/3512cinnamyl alcohol dehydrogenasecinnamoyl CoA reductase<i>Morus</i>ligninenzymatic assayVIGS
spellingShingle Shuai Huang
Xiaoru Kang
Rumeng Fu
Longyan Zheng
Peijun Li
Fengjuan Tang
Nan Chao
Li Liu
Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in Mulberry
Plants
cinnamyl alcohol dehydrogenase
cinnamoyl CoA reductase
<i>Morus</i>
lignin
enzymatic assay
VIGS
title Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in Mulberry
title_full Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in Mulberry
title_fullStr Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in Mulberry
title_full_unstemmed Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in Mulberry
title_short Simultaneous Down-Regulation of Dominant Cinnamoyl CoA Reductase and Cinnamyl Alcohol Dehydrogenase Dramatically Altered Lignin Content in Mulberry
title_sort simultaneous down regulation of dominant cinnamoyl coa reductase and cinnamyl alcohol dehydrogenase dramatically altered lignin content in mulberry
topic cinnamyl alcohol dehydrogenase
cinnamoyl CoA reductase
<i>Morus</i>
lignin
enzymatic assay
VIGS
url https://www.mdpi.com/2223-7747/13/24/3512
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