Maofu Li, Yuan Yang, Hua Wang, Pei Sun, Shuting Zhou, Yanhui Kang, Xiangyi Sun, Min Jin, Wanmei Jin
{"title":"The Mutations in <i>RcMYB114</i> Affect Anthocyanin Glycoside Accumulation in Rose.","authors":"Maofu Li, Yuan Yang, Hua Wang, Pei Sun, Shuting Zhou, Yanhui Kang, Xiangyi Sun, Min Jin, Wanmei Jin","doi":"10.3390/biology14030258","DOIUrl":null,"url":null,"abstract":"<p><p>In plants, the R2R3-MYB transcription factors are one of the largest MYB gene families. These MYB transcription factors are very important for regulating plant growth and development. RcMYB114, RcbHLH, and RcWD40 promote anthocyanin accumulation by forming the MBW (MYB-bHLH-WD40) complex and determine the rose flower's color. <i>RcMYB114</i> genomic sequences differ between the red petal and white varieties. Two non-synonymous substitutions were found in the open reading frame. It leads to a change in amino acids. Here, the anthocyanin content showed that there was no anthocyanin in white petals, while the anthocyanin content in red petals increased firstly at stage 2, decreased slightly at stage 4, and then increased again at stage 5. The spatiotemporal expression pattern analysis showed that <i>RcMYB114</i> was not expressed in all petals and tissues of white petals at different flower development stages. In red petal varieties, <i>RcMYB114</i> was highly expressed in petals, followed by styles, and not expressed in stems, young leaves, and stage 1 of flower development. However, <i>RcMYB114</i> has the highest expression level at the blooming stage. The <i>RcMYB114</i> sequence contains 9 SNPs in the coding region, 7 of which were synonymous substitutions that had no effect on the translation product and 2 of which were non-synonymous substitutions that resulted in amino acid alteration at positions 116 and 195, respectively. The <i>RcMYB114</i> gene in red rose was named <i>RcMYB114a</i>, and in white rose was <i>RcMYB114b</i>. <i>RcMYB114c</i> was mutated into leucine via artificial mutation; it was valine at position 116 of <i>RcMYB114a</i>, and Glycine mutated into Arginine at position 195 of <i>RcMYB114a</i> was <i>RcMYB114d</i>. <i>RcMYB114b</i> was the double mutation at positions 116 and 195 of <i>RcMYB114a</i>. The results of yeast two-hybrid experiments showed that RcMYB114a and its missense mutations RcMYB114b, RcMYB114c, and RcMYB114d could both interact with RcbHLH and RcWD40 to form the MYB-bHLH-WD40 complex. A transient transformation experiment in tobacco confirmed that <i>RcMYB114a</i> and its missense mutations <i>RcMYB114b</i>, <i>RcMYB114c</i>, and <i>RcMYB114d</i> could significantly promote the high expression of related structural genes in tobacco, together with the <i>RcbHLH</i> gene, which led to the accumulation of anthocyanins and produced the red color of the leaves. The <i>RcMYB114a</i> gene and its missense mutations <i>RcMYB114b</i>, <i>RcMYB114c</i>, and <i>RcMYB114d</i> interacted with the <i>RcbHLH</i> gene and significantly regulated the accumulation of anthocyanins. The two non-synonymous mutations of <i>RcMYB114</i> do not affect the function of the gene itself, but the content of the anthocyanins accumulated was different. This study should provide clues and references for further research on the molecular mechanism underlying the determination of rose petal color.</p>","PeriodicalId":48624,"journal":{"name":"Biology-Basel","volume":"14 3","pages":""},"PeriodicalIF":3.6000,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11939692/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biology-Basel","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3390/biology14030258","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
In plants, the R2R3-MYB transcription factors are one of the largest MYB gene families. These MYB transcription factors are very important for regulating plant growth and development. RcMYB114, RcbHLH, and RcWD40 promote anthocyanin accumulation by forming the MBW (MYB-bHLH-WD40) complex and determine the rose flower's color. RcMYB114 genomic sequences differ between the red petal and white varieties. Two non-synonymous substitutions were found in the open reading frame. It leads to a change in amino acids. Here, the anthocyanin content showed that there was no anthocyanin in white petals, while the anthocyanin content in red petals increased firstly at stage 2, decreased slightly at stage 4, and then increased again at stage 5. The spatiotemporal expression pattern analysis showed that RcMYB114 was not expressed in all petals and tissues of white petals at different flower development stages. In red petal varieties, RcMYB114 was highly expressed in petals, followed by styles, and not expressed in stems, young leaves, and stage 1 of flower development. However, RcMYB114 has the highest expression level at the blooming stage. The RcMYB114 sequence contains 9 SNPs in the coding region, 7 of which were synonymous substitutions that had no effect on the translation product and 2 of which were non-synonymous substitutions that resulted in amino acid alteration at positions 116 and 195, respectively. The RcMYB114 gene in red rose was named RcMYB114a, and in white rose was RcMYB114b. RcMYB114c was mutated into leucine via artificial mutation; it was valine at position 116 of RcMYB114a, and Glycine mutated into Arginine at position 195 of RcMYB114a was RcMYB114d. RcMYB114b was the double mutation at positions 116 and 195 of RcMYB114a. The results of yeast two-hybrid experiments showed that RcMYB114a and its missense mutations RcMYB114b, RcMYB114c, and RcMYB114d could both interact with RcbHLH and RcWD40 to form the MYB-bHLH-WD40 complex. A transient transformation experiment in tobacco confirmed that RcMYB114a and its missense mutations RcMYB114b, RcMYB114c, and RcMYB114d could significantly promote the high expression of related structural genes in tobacco, together with the RcbHLH gene, which led to the accumulation of anthocyanins and produced the red color of the leaves. The RcMYB114a gene and its missense mutations RcMYB114b, RcMYB114c, and RcMYB114d interacted with the RcbHLH gene and significantly regulated the accumulation of anthocyanins. The two non-synonymous mutations of RcMYB114 do not affect the function of the gene itself, but the content of the anthocyanins accumulated was different. This study should provide clues and references for further research on the molecular mechanism underlying the determination of rose petal color.
期刊介绍:
Biology (ISSN 2079-7737) is an international, peer-reviewed, quick-refereeing open access journal of Biological Science published by MDPI online. It publishes reviews, research papers and communications in all areas of biology and at the interface of related disciplines. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files regarding the full details of the experimental procedure, if unable to be published in a normal way, can be deposited as supplementary material.