Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> Cotton

The red plant phenotype of <i>R1</i> cotton is a genetic marker produced by light-induced anthocyanin accumulation. <i>GhPAP1D</i> controls this trait. There are two 228 bp tandem repeats upstream of <i>GhPAP1D</i> in <i>R1</i> cotton. In this study, G...

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Main Authors: Shuyan Li, Kunpeng Zhang, Chenxi Fu, Chaofeng Wu, Dongyun Zuo, Hailiang Cheng, Limin Lv, Haiyan Zhao, Jianshe Wang, Cuicui Wu, Xiaoyu Guo, Guoli Song
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Language:English
Published: MDPI AG 2025-08-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/15/2367
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author Shuyan Li
Kunpeng Zhang
Chenxi Fu
Chaofeng Wu
Dongyun Zuo
Hailiang Cheng
Limin Lv
Haiyan Zhao
Jianshe Wang
Cuicui Wu
Xiaoyu Guo
Guoli Song
author_facet Shuyan Li
Kunpeng Zhang
Chenxi Fu
Chaofeng Wu
Dongyun Zuo
Hailiang Cheng
Limin Lv
Haiyan Zhao
Jianshe Wang
Cuicui Wu
Xiaoyu Guo
Guoli Song
author_sort Shuyan Li
collection DOAJ
description The red plant phenotype of <i>R1</i> cotton is a genetic marker produced by light-induced anthocyanin accumulation. <i>GhPAP1D</i> controls this trait. There are two 228 bp tandem repeats upstream of <i>GhPAP1D</i> in <i>R1</i> cotton. In this study, GUS staining assays in transgenic <i>Arabidopsis thaliana</i> (L.) Heynh. demonstrated that tandem repeats in the <i>GhPAP1D</i> promoter-enhanced transcriptional activity. <i>GhPAP1D</i> is a homolog of <i>A. thaliana AtPAP1</i>. <i>AtPAP1’s</i> expression is regulated by photomorphogenesis-related transcription factors such as AtHY5 and AtBBXs. We identified the homologs of <i>A. thaliana AtHY5</i>, <i>AtBBX21</i>, and <i>AtBBX24</i> in <i>R1</i> cotton, designated as <i>GhHY5</i>, <i>GhBBX21</i>, and <i>GhBBX24</i>, respectively. Y1H assays confirmed that GhHY5, GhBBX21, and GhBBX24 each bound to the <i>GhPAP1D</i> promoter. Dual-luciferase reporter assays revealed that GhHY5 weakly activated the promoter activity of <i>GhPAP1D</i>. Heterologous expression assays in <i>A. thaliana</i> indicated that <i>GhBBX21</i> promoted anthocyanin accumulation, whereas <i>GhBBX24</i> had the opposite effect. Dual-luciferase assays showed GhBBX21 activated <i>GhPAP1D</i> transcription, while GhBBX24 repressed it. Further study indicated that GhHY5 did not enhance GhBBX21-mediated transcriptional activation of <i>GhPAP1D</i> but alleviates GhBBX24-induced repression. Together, our results demonstrate that GhBBX21 and GhBBX24 antagonistically regulate anthocyanin accumulation in <i>R</i>1 cotton under GhHY5 mediation, providing insights into light-responsive anthocyanin biosynthesis in cotton.
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spelling doaj-art-4e50bc8b39cd4508a7e71f7f9e5fd7f02025-08-20T03:36:22ZengMDPI AGPlants2223-77472025-08-011415236710.3390/plants14152367Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> CottonShuyan Li0Kunpeng Zhang1Chenxi Fu2Chaofeng Wu3Dongyun Zuo4Hailiang Cheng5Limin Lv6Haiyan Zhao7Jianshe Wang8Cuicui Wu9Xiaoyu Guo10Guoli Song11Anyang Institute of Technology, Anyang 455000, ChinaAnyang Institute of Technology, Anyang 455000, ChinaAnyang Institute of Technology, Anyang 455000, ChinaAnyang Institute of Technology, Anyang 455000, ChinaInstitute of Cotton Research of Chinese Academy of Agricultural Science, Anyang 455000, ChinaInstitute of Cotton Research of Chinese Academy of Agricultural Science, Anyang 455000, ChinaInstitute of Cotton Research of Chinese Academy of Agricultural Science, Anyang 455000, ChinaAnyang Institute of Technology, Anyang 455000, ChinaAnyang Institute of Technology, Anyang 455000, ChinaCotton Research Institute, Shanxi Agricultural University, Yuncheng 044000, ChinaAnyang Institute of Technology, Anyang 455000, ChinaInstitute of Cotton Research of Chinese Academy of Agricultural Science, Anyang 455000, ChinaThe red plant phenotype of <i>R1</i> cotton is a genetic marker produced by light-induced anthocyanin accumulation. <i>GhPAP1D</i> controls this trait. There are two 228 bp tandem repeats upstream of <i>GhPAP1D</i> in <i>R1</i> cotton. In this study, GUS staining assays in transgenic <i>Arabidopsis thaliana</i> (L.) Heynh. demonstrated that tandem repeats in the <i>GhPAP1D</i> promoter-enhanced transcriptional activity. <i>GhPAP1D</i> is a homolog of <i>A. thaliana AtPAP1</i>. <i>AtPAP1’s</i> expression is regulated by photomorphogenesis-related transcription factors such as AtHY5 and AtBBXs. We identified the homologs of <i>A. thaliana AtHY5</i>, <i>AtBBX21</i>, and <i>AtBBX24</i> in <i>R1</i> cotton, designated as <i>GhHY5</i>, <i>GhBBX21</i>, and <i>GhBBX24</i>, respectively. Y1H assays confirmed that GhHY5, GhBBX21, and GhBBX24 each bound to the <i>GhPAP1D</i> promoter. Dual-luciferase reporter assays revealed that GhHY5 weakly activated the promoter activity of <i>GhPAP1D</i>. Heterologous expression assays in <i>A. thaliana</i> indicated that <i>GhBBX21</i> promoted anthocyanin accumulation, whereas <i>GhBBX24</i> had the opposite effect. Dual-luciferase assays showed GhBBX21 activated <i>GhPAP1D</i> transcription, while GhBBX24 repressed it. Further study indicated that GhHY5 did not enhance GhBBX21-mediated transcriptional activation of <i>GhPAP1D</i> but alleviates GhBBX24-induced repression. Together, our results demonstrate that GhBBX21 and GhBBX24 antagonistically regulate anthocyanin accumulation in <i>R</i>1 cotton under GhHY5 mediation, providing insights into light-responsive anthocyanin biosynthesis in cotton.https://www.mdpi.com/2223-7747/14/15/2367anthocyanin biosynthesisBBX proteinscotton
spellingShingle Shuyan Li
Kunpeng Zhang
Chenxi Fu
Chaofeng Wu
Dongyun Zuo
Hailiang Cheng
Limin Lv
Haiyan Zhao
Jianshe Wang
Cuicui Wu
Xiaoyu Guo
Guoli Song
Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> Cotton
Plants
anthocyanin biosynthesis
BBX proteins
cotton
title Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> Cotton
title_full Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> Cotton
title_fullStr Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> Cotton
title_full_unstemmed Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> Cotton
title_short Two B-Box Proteins, GhBBX21 and GhBBX24, Antagonistically Modulate Anthocyanin Biosynthesis in <i>R1</i> Cotton
title_sort two b box proteins ghbbx21 and ghbbx24 antagonistically modulate anthocyanin biosynthesis in i r1 i cotton
topic anthocyanin biosynthesis
BBX proteins
cotton
url https://www.mdpi.com/2223-7747/14/15/2367
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