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Strawberry transgenic fruits ( Fragaria × ananassa cv. Elsanta) agroinfiltrated with FaMYB10 -pFRN construct. Control fruit, green– white fruit agroinfiltrated with the empty pFRN vector; FaMYB10 -RNAi fruits, green–white fruits agroinfiltrated with FaMYB10 -pFRN construct.
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This work characterized the role of the R2R3-MYB10 transcription factor (TF) in strawberry fruit ripening. The expression
of this TF takes place mainly in the fruit receptacle and is repressed by auxins and activated by abscisic acid (ABA), in
parallel to the ripening process. Anthocyanin was not produced when FaMYB10 expression was transiently sil...
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Citations
... MYB15 controls the CHS, DFR and ANS genes in Lilium regale (Yamagishi, 2016). MYB1 also suppresses the ANS gene in strawberry fruits (Medina-Puche et al. (2014); Kadomura-Ishikawa et al. (2015). Keeping in the view role of TFs in anthocyanin biosynthesis different groups have successfully done the metabolic engineering of R2R3-MYBs to improve anthocyanin content in fruits and vegetable crops (Niang and Kim 2018). ...
Anthocyanins are a class of natural water-soluble pigments found in plants, which are responsible for the varied colours of vegetative tissues and reproductive organs. The regulation of anthocyanin biosynthesis requires multiple factors, including enzymes, structural or regulatory genes, and transcriptional regulator proteins. While there has been notable advancement in comprehending anthocyanin biosynthesis, encompassing its structures and overarching regulation, there remains a gap in our understanding concerning the intricate interactions among various genes, molecular and biochemical mechanisms, and their feedback responses to anthocyanin regulatory networks. This article aims to provide an overview of the current understanding of the complex regulatory networks involved in the phenylpropanoid pathway, with a specific focus on anthocyanin biosynthesis.
... The R2R3 MYBs activators of anthocyanin and PA biosynthesis have been described in many fleshy fruitrelated species, including Malus × domestica (MdMYB10, MdMYB1, MdMYB3/MdMYB9, MdMYB11;Espley et al. 2007;Vimolmangkang et al. 2013;Hu et al. 2016), Prunus avium (PavMYB10.1; Jin et al. 2016), Actinidia chinensis (AcMYB10, AcMYB110; Peng et al. 2019), Vitis vinifera (VvMYBPAR; Koyama et al. 2014), Prunus persica (PpMYBPA1, PpMYB7; Ravaglia et al. 2013; Zhou et al. 2015), and Fragaria × ananassa (FaMYB10, FaMYB5, FaMYB9, FaMYB11;Schaart et al. 2013;Medina-Puche et al. 2014;Wang et al. 2020;Jiang et al. 2023). Nevertheless, the FaMYB1 TF was characterized as a suppressor of anthocyanin biosynthesis in F. × ananassa (Aharoni et al. 2001). ...
... Thus, the overexpression of FvMYB10 in Fragaria vesca fruit, produced greater red coloration and accumulation of anthocyanins both in pulp and peel, while silencing of FvMYB10 by RNAi produced white fruit (Lin- Wang et al. 2014). Transient suppression studies of FaMYB10 by RNAi in fruit receptacle showed that the anthocyanin accumulation decreased compared to control fruit (Medina-Puche et al. 2014). However, when the transient suppression of the repressor FaMYB1 was performed, the anthocyanin level was not affected, suggesting that silencing FaMYB1 did not generate an increase in the anthocyanin content (Medina-Puche et al. 2014). ...
... Transient suppression studies of FaMYB10 by RNAi in fruit receptacle showed that the anthocyanin accumulation decreased compared to control fruit (Medina-Puche et al. 2014). However, when the transient suppression of the repressor FaMYB1 was performed, the anthocyanin level was not affected, suggesting that silencing FaMYB1 did not generate an increase in the anthocyanin content (Medina-Puche et al. 2014). In contrast, Kadomura-Ishikawa et al. (2015) using RNAi for FaMYB1 silencing in F. × ananassa fruit observed a significant increase in the anthocyanin content with a concomitant reduction in the expression of both FaF3H and FaLAR, suggesting a redirection of the flavonoid precursors causing anthocyanins levels to increase at the expense of PAs. ...
MYB transcription factors (TFs) are involved in the regulation of flavonoid biosynthesis in both Chilean white strawberry (Fragaria chiloensis) and commercial strawberry (Fragaria × ananassa). There is evidence that MYB1 TF repress anthocyanin biosynthesis, but would also be participating in the regulation of the proanthocyanidin (PA) polymer synthesis, either in the activation or repression of the expression of PA-related genes. We investigated the role of the MYB1 TF in the regulation of the PA-associated structural gene leucoanthocyanidin reductase (LAR). The promoter regions of LAR were isolated from F. chiloensis and F. × ananassa genomes and named as pFcLAR and pFaLAR, respectively. Promoter analysis revealed several putative binding sites for MYBs and bHLH TFs. Then, dual luciferase assays were conducted to investigate whether MYB1 can act as a transactivator of LAR promoters in both species. In the long version of LAR promoters of both species (1381 bp upstream of ATG), the addition of MYB1 to the MYB10-bHLH3 complex repressed the activation. However, when using the medium version of the pFcLAR (815 bp upstream of ATG), MYB1 alone significantly activated the promoter, and no repression was observed when MYB1 was added to the MYB10-bHLH3 complex in the short pFa/FcLAR promoters (316 bp upstream of ATG). We propose that the FcMYB1 TF could regulate PA biosynthesis, and that its role, as either a repressor or activator, is dependent of other partners and the DNA sequence context of the promoter regions.
... TFs are typically composed of multiple functionally independent binding domains, which categorize them into different families based on these domains [2,3]. The v-MYB avivan myoblastsis virus oncogene homolog (MYB) TFs family is the largest TFs family in plants working in plant growth and development, secondary metabolism and resistance to biotic and abiotic stress [4][5][6][7]. The first MYB was identified in the avian myoblast virus, and the first plant MYB was isolated from Zea mays [8,9]. ...
... ODORANT1 (ODO1) is the first R2R3 type MYB TFs discovered in Petunia [42], ODO1, BENZENOID II (EOBII), EOBI and LATE ELONGATED HYPOCOTYL (LHY) regulate the odor production of flowers by activating odor related genes in P. hybrida [43,44]. FaMYB10 and FaEOBII in Fragaria × ananassa participate in the regulation of eugenol [6,45]. Overexpression of SmMYB39 alters the synthesis of rosmarinic acid by inhibiting C4H and tyrosine aminotransferase genes, reducing the content of 4-coumaric acid, rosmarinic acid, and other substances. ...
The v-MYB avivan myoblastsis virus oncogene homolog (MYB) family is the largest gene family of the transcription factor in plants, involved in plant growth and development, secondary metabolism and resistance to biotic/abiotic stress. Antirrhinum majus (snapdragon) is an ideal material for studying ornamental traits. Nevertheless, there has been no systematic investigation into the AmMYB family of snapdragons. In this study, we identified a total of 162 members of the AmMYB gene family in snapdragons. Gene structure analysis showed that the AmMYB family within the same subgroup had a similar structure and motifs. Analysis of gene duplication events revealed that the amplification of the AmMYB family was driven by whole-genome duplication (WGD) and dispersed duplication. The analysis of cis-acting elements in the promoter region of AmMYB genes reveals a collaborative involvement of light-responsive growth and development elements, stress resistance elements, and hormone-responsive elements jointly participating in the regulation of the AmMYB gene. Collinearity analysis demonstrates significant functional distinctions between AmMYB and monocotyledonous plants. The classification of AmMYB members results in 3 main subgroups with 36 smaller subgroups. All AmMYB genes are distributed across all eight chromosomes, with no apparent correlation between subfamily distribution and chromosome length. Through phylogenetic analysis and RNA-seq analysis, we have identified 9 R2R3-MYB genes that potentially play a role in the regulation of floral volatile organic compounds (FVOCs) biosynthesis. Their expression patterns were verified by qRT-PCR experiments. This study establishes a robust foundation for further investigations into the functionality of AmMYB genes and their molecular mechanisms underlying FVOC biosynthesis in snapdragons.
... Insights are provided by analyzes of phylogenetic relationships, which associate the TFs FvMYB11, 3,9,21,22,41,45,77,75,64,105 with the accumulation of proanthocyanidins and flavonoids (Xu et al. 2021). The most elucidated MYBs are FaMYB10 and FaMYB1, which act as positive and negative regulators, respectively, in the biosynthesis of flavonoids and anthocyanins (Medina-Puche et al. 2014;Wei et al. 2018). ...
... AtTCP3 forms protein complexes with another TF, R2R3-MYB (Li and Zachgo 2013). The R2R3 MYB FaMYB10 serves as a signal in ABA-mediated anthocyanin synthesis during ripening (Medina-Puche et al. 2014). However, the role of TCPs identified in Fragaria 9 ananassa still need to be validated by functional analysis and protein-protein interactions in order to define strategies to increase the biosynthesis of phenylpropanoids in strawberry. ...
The biosynthesis of phenylpropanoids is regulated by a complex molecular, biochemical and physiological network. Modulation of phenylpropanoid metabolism occurs by endogenous (transcription and post-transcription factors, homeostasis modulators, hormones) and extrinsic (biotic and abiotic agents) signals, both during the growth and development of strawberries. In the context of endogenous modulation, during the transition from maturation to ripening of strawberries, the most significant alteration in the synthesis and accumulation of phenylpropanoids occurs, and phytohormones are intensely involved in regulating this event. The phytohormone abscisic acid (ABA) is highlighted, which plays a central role in strawberry ripening and seems necessary for anthocyanin synthesis. In this review, we report the main mechanisms involved before and during the biosynthesis of phenylpropanoid compounds in strawberry ripening, focusing on anthocyanin synthesis.
... In apples, of the allelic homologues, fruit flesh and foliage anthocyanin accumulation have been demonstrated to be regulated by MdMYB10, whereas MdMYB1 and MdMYBA are expressed in the fruit skin [17][18][19]. Some MYBs have also been characterized in strawberries, including FaMYB10 [20], FaMYB123 [21], and an anthocyanin biosynthesis inhibitor FaMYB1 [22]. Among them, MYB10 codes for a dominant regulatory transcription factor are involved in the anthocyanin accumulation of strawberry fruits. ...
Anthocyanins widely accumulate in the vegetative and reproductive tissues of strawberries and play an important role in stress resistance and fruit quality. Compared with other fruits, little is known about the molecular mechanisms regulating anthocyanin accumulation in strawberry vegetative tissues. In this study, we revealed an R2R3–MYB transcription factor, FaMYB10-like (FaMYB10L), which positively regulated anthocyanin accumulation and was induced by light in the petiole and runner of cultivated strawberry. FaMYB10L is a homologue of FveMYB10-like and a nuclear localization protein. Transient overexpression of FaMYB10L in a white fruit strawberry variety (myb10 mutant) rescued fruit pigmentation, and further qR–PCR analysis revealed that FaMYB10L upregulated the expression levels of anthocyanin biosynthesis-related genes and transport gene. A dual luciferase assay showed that FaMYB10L could activate the anthocyanin transport gene FaRAP. Anthocyanin accumulation was observed in FaMYB10L-overexpressing strawberry calli, and light treatment enhanced anthocyanin accumulation. Furthermore, transcriptomic profiling indicated that the DEGs involved in the flavonoid biosynthesis pathway and induced by light were enriched in FaMYB10L-overexpressing strawberry calli. In addition, yeast two-hybrid assays and luciferase complementation assays indicated that FaMYB10L could interact with bHLH3. These findings enriched the light-involved regulatory network of anthocyanin metabolism in cultivated strawberries.
... Flavonoids are the most common polyphenol compounds such as anthocyanins, PAs, and flavonols present in strawberry fruits ). Overexpression of FaMYB10 in Fragaria ananassa or FvMYB10 in F. vesca accelerated anthocyanin formation, whereas knockdown of FaMYB10/FvMYB10 resulted in white fruit (Lin- Medina-Puche et al. 2014;Li et al. 2020). These findings confirm the role of FvMYB10 in the regulation of anthocyanin biosynthesis in strawberry fruit . ...
The color that results from anthocyanin accumulation has a direct effect on the dietary and market values of fresh fruit. Fruit color pigments such as anthocyanins are produced via the flavonoid pathway, where the biosynthetic genes are coordinatedly regulated by the myeloblastosis, basic helix–loop–helix, and WD40 a beta-transducin repeat, which is a short structural motif consisting of about 40 amino acids that commonly terminating in a tryptophan–aspartic acid (W–D) dipeptide “MYB-bHLH-WD40/WDR” (MBW) regulatory complex. This article discusses the representatives of this regulatory complex, such as bHLH, MYB, WD40, and their regulatory role in fruit coloration. Further, the joint role of MYBs, bHLH TFs, and structural genes has been described in this article. This article will help plant breeders in the future to breed colorful, anthocyanin, and nutrient-rich fruit-bearing plant species.
... For example, in strawberry fruit, drought conditions increased ABA, which was correlated with elevated anthocyanin concentration (as well as AsA) without a reduction in fruit yield. A previous study in strawberry clearly demonstrated how the presence of ABA regulated the expression of the anthocyanin-regulating MYB TF, FaMYB10, which, in turn, elevated the anthocyanin biosynthetic genes and fruit colour(Medina-Puche et al., 2013). In apple, an alternative molecular model of drought-induced anthocyanin production has been proposed, whereby the ethylene response factor ERF38 partners with the homologous MYB1 to drive the anthocyanin biosynthetic pathway genes(An et al., 2020c). ...
For many fruit crops, the colour of the fruit outwardly defines its eating quality. Fruit pigments provide reproductive advantage for the plant as well as providing protection against unfavourable environmental conditions and pathogens. For consumers these colours are considered attractive and provide many of the dietary benefits derived from fruits. In the majority of species, the main pigments are either carotenoids and/or anthocyanins. They are produced in the fruit as part of the ripening process, orchestrated by phytohormones and an ensuing transcriptional cascade, culminating in pigment biosynthesis. Whilst this is a controlled developmental process, the production of pigments is also attuned to environmental conditions such as light quantity and quality, availability of water and ambient temperature. If these factors intensify to stress levels, fruit tissues respond by increasing (or ceasing) pigment production. In many cases, if the stress is not severe, this can have a positive outcome for fruit quality. Here, we focus on the principal environmental factors (light, temperature and water) that can influence fruit colour.
... Peach PpMYB10.1 and PpMYB10.3 interacted with PpbHLH proteins to activate the expression of CHS, F3 H and UFGT, thereby promoting the synthesis of anthocyanin [20]. In strawberry, FaMYB5/FaMYB10-FaEGL3 (bHLH)-FaLWD1/FaLWD1like (WD40) formed an 'MBW' complex to positively regulate anthocyanin synthesis in strawberry fruit [21,22], FaMYB10 promotes anthocyanin synthesis in strawberry, and FaMYB10 mutation is the main cause of color difference between yellow woodland strawberry 'Yellow wonder' and red woodland strawberry 'Ruegen' [23,24]. The interaction between FaMYB123 and FabHLH3 in strawberry regulates anthocyanin synthesis by regulating late biosynthetic genes (LBGs) [25]. ...
MYB and BBX transcription factors play important roles in flavonoid biosynthesis. Here, we obtained transgenic woodland strawberry with stable overexpression of FaMYB5, demonstrating that FaMYB5 can increase anthocyanin and proanthocyanidin content in roots, stems and leaves of woodland strawberry. In addition, bimolecular fluorescence complementation assays and yeast two-hybridization demonstrated that the N-terminal (1-99aa) of FaBBX24 interacts with FaMYB5. Transient co-expression of FaBBX24 and FaMYB5 in cultivated strawberry 'Xiaobai' showed that co-expression strongly promoted the expression of F3'H, 4CL-2, TT12, AHA10 and ANR and then increased the content of anthocyanin and proanthocyanidin in strawberry fruits. We also determined that FaBBX24 is also a positive regulator of anthocyanin and proanthocyanidin biosynthesis in strawberry. The results reveal a novel mechanism by which the FaMYB5-FaBBX24 module collaboratively regulates anthocyanin and proanthocyanidin in strawberry fruit.
... In recent years, a number of transcription factors (TFs) have been identified and functionally characterized as regulators of fruit ripening or quality formation in strawberry (Sanchez-Gomez et al. 2022). For example, FaMYB10 and FaMYB1, members of the R2R3-type MYB TF family, regulate the biosynthesis of phenylpropanoids, flavonoids, and anthocyanins in strawberry fruit (Aharoni et al. 2001;Lin-Wang et al. 2010;Medina-Puche et al. 2014). The MADS-box proteins FaMADS9 and FaSHP (SHATTERPROOF) are associated with strawberry development and ripening (Seymour et al. 2011;Daminato et al. 2013), while the MYB and DNA binding with One Finger (DOF)-like TFs FaEOBII (Emission of Benzenoid II) and FaDOF2 can promote the biosynthesis of eugenol, a volatile phenylpropanoid in ripe strawberry (Koeduka et al. 2006;Molina-Hidalgo et al. 2017). ...
The NAC transcription factor RIF (Ripening Inducing Factor) was previously reported to be necessary for the ripening of octoploid strawberry (Fragaria × ananassa) fruit, but the mechanistic basis of RIF-mediated transcriptional regulation and how RIF activity is modulated remains elusive. Here we show that FvRIF in diploid strawberry, Fragaria vesca, is a key regulator in the control of fruit ripening, and that knockout mutations of FvRIF result in a complete block of fruit ripening. DNA affinity purification sequencing coupled with transcriptome deep sequencing suggests that 2080 genes are direct targets of FvRIF-mediated regulation, including those related to various aspects of fruit ripening. We provide evidence that FvRIF modulates anthocyanin biosynthesis and fruit softening by directly regulating the related core genes. Moreover, we demonstrate that FvRIF interacts with and serves as a substrate of mitogen-activated protein kinase 6 (FvMAPK6), which regulates the transcriptional activation function of FvRIF by phosphorylating FvRIF at Thr-310. Our findings uncover the FvRIF-mediated transcriptional regulatory network in controlling strawberry fruit ripening and highlight the physiological significance of phosphorylation modification on FvRIF activity in ripening.
... Overexpression of FaMYB10 in octoploid strawberry or FvMYB10 in woodland strawberry substantially promoted anthocyanin accumulation including in leaves, petioles, and fruits; the flesh color of woodland strawberries changed from white to red (Lin-Wang et al. 2010. However, silencing or mutation of FaMYB10/FvMYB10 reduced anthocyanin accumulation, resulting in white or lightly colored fruits (Lin-Wang et al. 2014;Medina-Puche et al. 2014). In addition, genome analysis of several woodland strawberry accessions with white fruit showed that the white fruit color was due to natural mutations in the MYB10 gene (Hawkins et al. 2016;Castillejo et al. 2020;Qiao Qin et al. 2021). ...
Fruit color is a very important external commodity factor for consumers. Compared to the most typical red octoploid strawberry (Fragaria × ananassa), the pink strawberry often sells for a more expensive price and has a higher economic benefit due to its outstanding color. However, few studies have examined the molecular basis of pink colored strawberry fruit. Through an EMS mutagenesis of woodland strawberry (Fragaria vesca), we identified a mutant with pink fruits and green petioles. BSA-Seq analysis and gene function verification confirmed that the responsible mutation resides in a gene encoding flavanone-3-hydroxylase (F3H) in the anthocyanin synthesis pathway. This nonsynonymous mutation results in an arginine to histidine change at position 130 of F3H. Molecular docking experiments showed that the arginine to histidine mutation results in a reduction of intermolecular force-hydrogen bonding between the F3H protein and its substrates. Enzymatic experiments showed a greatly reduced ability of the mutated F3H protein to catalyze the conversion of the substrates, and hence a blockage of the anthocyanin synthesis pathway. The discovery of a key residue in the F3H gene controlling anthocyanin synthesis provides a clear target of modification for the molecular breeding of strawberry varieties with pink-colored fruits, which may be of great commercial value.
