MYB Transcription Factor Family Research Articles

Transcriptome Identification of R2R3-MYB Gene Family Members in Pinus massoniana and PmMYB4 Response to Drought Stress

One of the largest families of transcription factors in plants, the MYB transcription factors family (Myeloblastosis, MYB TF), plays a vital role in regulating plant biochemical and physiological processes. The role of MYB TF in coping with stresses, such as drought, salt and cold, has been reported. Unfortunately, a comprehensive identification of R2R3-MYB TF in Masson pine (Pinus massoniana) has not been achieved. In this study, a total of 49 sequences were identified as R2R3-MYB TF. The structure, function and phylogenetic relationships of the conserved structural domains of Masson pine R2R3-MYB TF and Populus trichocarpa Torr. & A.Gray ex Hook. TFs were compared using bioinformatics tools. The results showed that Masson pine R2R3-MYB TF was divided into 24 groups, mainly located in the nucleus, and mostly lacking signal peptides and transmembrane structural domains with multiple phosphorylation sites. The drought stress-responsive R2R3-MYB gene, PmMYB4, was selected from the drought stress transcriptome based on analysis of the expression pattern and tissue specificity of PmMYB4 gene under abiotic stress using qPCR. The results showed that PmMYB4 can be involved in drought stress treatment through ABA signaling, as well as in multiple stress responses such as salt stress, and there were significant differences in the expression of PmMYB4 in the eight tissues. These results provide a reference scheme for the functional identification of R2R3-MYB transcription factors, which may be involved in plant responses to multiple stresses such as drought, and enrich our understanding of the functions of R2R3-MYB transcription factors in plants.

Genome-Wide Identification Analysis of the R2R3-MYB Transcription Factor Family in Cymbidium sinense for Insights into Drought Stress Responses

Cymbidium sinense represents a distinctive Orchidaceae plant that is more tolerant than other terrestrial orchids. Studies have shown that many members of the MYB transcription factor (TF) family, especially the R2R3-MYB subfamily, are responsive to drought stress. This study identified 103 CsMYBs; phylogenetic analysis classified these genes into 22 subgroups with Arabidopsis thaliana. Structural analysis showed that most CsMYB genes contained the same motifs, three exons and two introns, and showed a helix-turn-helix 3D structure in each R repeat. However, the members of subgroup 22 contained only one exon and no intron. Collinear analysis revealed that C. sinense had more orthologous R2R3-MYB genes with wheat than A. thaliana and rice. Ka/Ks ratios indicated that most CsMYB genes were under purifying negative selection pressure. Cis-acting elements analysis revealed that drought-related elements were mainly focused on subgroups 4, 8, 18, 20, 21, and 22, and Mol015419 (S20) contained the most. The transcriptome analysis results showed that expression patterns of most CsMYB genes were upregulated in leaves in response to slight drought stress and downregulated in roots. Among them, members in S8 and S20 significantly responded to drought stress in C. sinense. In addition, S14 and S17 also participated in these responses, and nine genes were selected for the real-time reverse transcription quantitative PCR (RT-qPCR) experiment. The results were roughly consistent with the transcriptome. Our results, thus, provide an important contribution to understanding the role of CsMYBs in stress-related metabolic processes.

GhMYB102 promotes drought resistance by regulating drought-responsive genes and ABA biosynthesis in cotton (Gossypium hirsutum L.)

The MYB transcription factor (TF) family is among the largest TF families and plays an important role in plant growth and stress response. However, few studies have investigated the role of the MYB gene in drought resistance in cotton. In this study, we analysed the drought transcriptomic data of cotton and identified that the GhMYB102 gene was significantly upregulated in upland cotton during the early stages of drought stress. Bioinformatics analysis showed that the amino acid sequence encoded by GhMYB102 contained two highly conserved MYB binding domains belonging to R2R3-MYB TFs. GhMYB102 was most closely related to AtMYB102. GhMYB102 is mainly expressed in roots and is induced by abiotic stresses and abscisic acid (ABA); it is localised in the nucleus and has transcriptional activation activity. Silencing of GhMYB102 decreased plant drought resistance. In addition, dual-luciferase assays and yeast single hybridisation analysis showed that GhMYB102 could directly bind the MYB motif elements in the promoter regions of GhNCED1 and GhZAT10. These results indicate that GhMYB102 plays a positive role in drought tolerance by regulating the expression of GhNCED1 and GhZAT10. Thus, GhMYB102 enhances drought resistance by participating in ABA biosynthesis or regulating the expression of drought-responsive genes.

MYB Transcription Factor Family in Pearl Millet: Genome-Wide Identification, Evolutionary Progression and Expression Analysis under Abiotic Stress and Phytohormone Treatments.

Transcription factors (TFs) are the regulatory proteins that act as molecular switches in controlling stress-responsive gene expression. Among them, the MYB transcription factor family is one of the largest TF family in plants, playing a significant role in plant growth, development, phytohormone signaling and stress-responsive processes. Pearl millet (Pennisetum glaucum L.) is one of the most important C4 crop plants of the arid and semi-arid regions of Africa and Southeast Asia for sustaining food and fodder production. To explore the evolutionary mechanism and functional diversity of the MYB family in pearl millet, we conducted a comprehensive genome-wide survey and identified 279 MYB TFs (PgMYB) in pearl millet, distributed unevenly across seven chromosomes of pearl millet. A phylogenetic analysis of the identified PgMYBs classified them into 18 subgroups, and members of the same group showed a similar gene structure and conserved motif/s pattern. Further, duplication events were identified in pearl millet that indicated towards evolutionary progression and expansion of the MYB family. Transcriptome data and relative expression analysis by qRT-PCR identified differentially expressed candidate PgMYBs (PgMYB2, PgMYB9, PgMYB88 and PgMYB151) under dehydration, salinity, heat stress and phytohormone (ABA, SA and MeJA) treatment. Taken together, this study provides valuable information for a prospective functional characterization of the MYB family members of pearl millet and their application in the genetic improvement of crop plants.

Genome-wide identification and expression pattern analysis of the SABATH gene family in Neolamarckia cadamba.

Plant SABATH methyltransferases are a class of enzymes that catalyze the transfer of the methyl group from S-adenosyl-L-methionine (SAM) to the carboxyl group or the nitrogen group of the substrate to form small molecule methyl esters or N-methylated compounds, which are involved in various secondary metabolite biosynthesis and have important impacts on plant growth, development, and defense reactions. We previously reported the monoterpenoid indole alkaloids (MIAs) cadambine biosynthetic pathway in Neolamarckia cadamba, a woody tree species that provides an important traditional medicine widely used to treat diseases such as diabetes, leprosy, and cancer in Southeast Asia. However, the functions of NcSABATHs in cadambine biosynthesis remain unclear. In this study, 23 NcSABATHs were identified and found to be distributed on 12 of the total 22 chromosomes. Gene structure, conserved motifs, and phylogenetic analysis showed that NcSABATHs could be divided into three groups. According to cis-element analysis, the NcSABATH promoters contained a large number of elements involved in light, plant hormone, and environmental stress responses, as well as binding sites for the BBR-BPC, DOF, and MYB transcription factor families. Based on RNA-seq data and qRT-PCR analysis, the NcSABATH genes exhibited diverse tissue expression patterns. Furthermore, NcSABATH7/22, which clustered with LAMT in the same clade, were both up-regulated under MeJA treatment. The correlation analysis between gene expression and cadambine content showed that NcSABATH7 potentially participated in cadambine biosynthesis. Taken together, our study not only enhanced our understanding of SABATH in N. cadamba but also identified potential candidate genes involved in cadambine biosynthesis.

Deciphering aroma formation during flowering in nectar tree (Tilia amurensis): insights from integrated metabolome and transcriptome analysis.

Tilia amurensis is a significant ornamental and economically-important tree species, known for its fragrant flowers, which are a source of high-quality honey production. However, the regulatory mechanisms involved in aroma formation during flower development in T. amurensis remains limited. The current study revealed the detection of plant hormones at every assessed stage of flower development. Among them, auxin and brassinosteroid contents significantly increased at stage 3, potentially regulating crucial functions during T. amurensis flower development. Moreover, the study examined the levels and change patterns of secondary metabolites and employed a combination of transcriptomics and metabolomics to comprehensively assess essential structural genes implicated in the biosynthesis pathways of terpenoid and phenylpropanoid. A comprehensive set of 89,526 differentially expressed genes (DEGs) was uncovered, including candidate structural genes ACAT, HDS, TPS, 4CL, CAD, and CCOAMT, which are specifically involved in the biosynthesis of terpenoids and phenylpropanoids. Maslinic acid, 2α,3α-dihydroxyursolic acid, and betulinic acid were accumulated in the terpenoid biosynthesis pathway. In contrast, metabolites with differential accumulation, such as phenylalanine, coniferyl alcohol, and cinnamic acid, were specifically enriched in the phenylpropanoid biosynthesis pathway. The C2H2, MYB, and NAC transcription factor families are crucially associated with the terpenoid and phenylpropanoid biosynthesis pathways. Two transcription factors, C2H2-17 and MYB-24, exhibited strong co-expression with structural genes in two networks, and were identified as central regulatory factors. These findings establish a solid groundwork for elucidating the generation of floral fragrance and provide comprehensive genetic and metabolic information for further studies on T. amurensis.

Transcriptome-wide analysis reveals core transcriptional regulators associated with culm development and variation in Dendrocalamus sinicus, the strongest woody bamboo in the world

Transcription factors (TFs) play indispensable roles in plant development and stress responses. As the largest woody bamboo species in the world, Dendrocalamus sinicus is endemic to Yunnan Province, China, and possesses two natural variants characterized by culm shape, namely straight or bent culms. Understanding the transcriptional regulation network of D. sinicus provides a unique opportunity to clarify the growth and development characteristics of woody bamboos. In this study, 10,236 TF transcripts belonging to 57 families were identified from transcriptome data of two variants at different developmental stages, from which we constructed a transcriptional regulatory network and unigene-coding protein-TFs interactive network of culm development for this attractive species. Gene function enrichment analysis revealed that hormone signaling and MAPK signaling pathways were two most enriched pathways in TF-regulated network. Based on PPI analysis, 50 genes interacting with nine TFs were screened as the core regulation components related to culm development. Of them, 18 synergistic genes of seven TFs, including nuclear cap-binding protein subunit 1, transcription factor GTE9-like, and ATP-dependent DNA helicase DDX11 isoform X1, involved in culm-shape variation. Most of these genes would interact with MYB, C3H, and ARF transcription factors. Six members with two each from ARF, C3H, and MYB transcription factor families and six key interacting genes (IAA3, IAA19, leucine-tRNA ligase, nuclear cap-binding protein subunit 1, elongation factor 2, and coiled-coil domain-containing protein 94) cooperate with these transcription factors were differentially expressed at development stage of young culms, and were validated by quantitative PCR. Our results represent a crucial step towards understanding the regulatory mechanisms of TFs involved in culm development and variation of D. sinicus.

DNA-binding domain in MYB transcription factor family in date palm

Date palm (Phoenix dactylifera L.) is one of the most important economic plants which belongs to the Arecaceae family in arid and semi-arid areas. The genetic map of the date palm genome is incomplete, which hindered the expansion of molecular studies. Bioinformatics is a way that the researcher compares different plant species to find similarities and differences between them. MYB is a transcription factor family that has well-known functions in many plants, especially Arabidopsis. DNA-binding domains are pieces of peptides (a group of amino acids) that are folded independently and are highly attracted to DNA. The study aims to identify the DNA-binding domains for the MYB family in Date Palm to diagnose drought-related genes in the date palm genome through bioinformatics. The main approach followed in the research is to use a set of approved sites to align members of MYB proteins in Arabidopsis with the date palm genome to determine the most closely related MYB proteins in date palms. The results indicated a group of date palm proteins, which considered candidate proteins or genes to have an effective role in resisting or reducing the negative effects of water stress on date palms. Also, numbers of palm proteins have the same alignment effect and DNA-binding domains, which means that they may be close to each other in function and structure.

Two R2R3-MYB transcription factors, CsMYB33 and CsMYB78 are involved in the regulation of anthocyanin biosynthesis in Cannabis sativa L.

Cannabis sativa L. is an economically important plant which is renowned for its wide range of uses. Anthocyanins , a class of flavonoids , impart pigmentation to the plant parts and play important roles in plant environment interactions. However, to the best of our knowledge, the molecular basis of anthocyanin biosynthesis in C. sativa remains elusive. Since MYB family of transcription factors plays a vital role in regulating anthocyanin biosynthesis, herein, we identified 99 R2R3-MYB genes in the C. sativa genome. Based on the phylogeny as well as the expression profiling of genes in two accessions of C. sativa differing in the content of anthocyanin pigmentation in sugar leaves, two putative regulators of anthocyanin biosynthesis, named as CsMYB33 and CsMYB78 were identified. Transient overexpression of CsMYB33 and CsMYB78 activate anthocyanin biosynthesis in the leaves of Nicotiana benthamiana . As compared to CsMYB78, CsMYB33 was found to be stronger in terms of activation of anthocyanin biosynthesis. Constitutive expression of CsMYB33 also resulted in the enhanced anthocyanin pigmentation in the leaves and sepals of transgenic lines of Nicotiana tabacum . Taken together, the present study provides valuable insights into the regulatory genes of anthocyanin biosynthesis in C. sativa , which will be useful for its biotechnological improvement. • A total of 99 R2R3-MYB family of transcription factor genes were identified in Cannabis sa tiva L. genome. • The phylogeny, gene structure and gene expression analyses revealed putative function of some of the R2R3-CsMYB genes. • CsMYB33 and CsMYB78 are the two key transcriptional activators of anthocyanin biosynthesis in C. sativa . • In comparison of CsMYB78, CsMYB33 was found to be more effective in activation of anthocyanin biosynthesis.

SmMYB4 Is a R2R3-MYB Transcriptional Repressor Regulating the Biosynthesis of Phenolic Acids and Tanshinones in Salvia miltiorrhiza.

Salvia miltiorrhiza Bunge is one of the most famous traditional Chinese medicinal plants. The two most important classes of pharmaceutically relevant compounds in S. miltiorrhiza are phenolic acids and tanshinones. The MYB family of transcription factors may efficiently regulate the secondary metabolism in plants. In this study, a subgroup 4 R2R3MYB transcription factor gene, SmMYB4, was isolated from S. miltiorrhiza and functionally characterized using overexpression and a RNAi-mediated silencing. We achieved a total of six overexpressions and eight RNAi transgenic lines from the Agrobacterium leaf disc method. The content of the total phenolics, rosmarinic acid, and salvianolic acid B markedly decreased in the SmMYB4-overexpressing lines but increased in the SmMYB4-RNAi lines. The content of the total tanshinones, cryptotanshinone, and tanshinone IIA decreased in the SmMYB4-overexpressing transgenic lines but increased in the SmMYB4-RNAi lines. A gene expression analysis demonstrated that SmMYB4 negatively regulated the transcription of the critical enzyme genes involved in the phenolic acid and tanshinone biosynthesis. The genetic control of this transcriptional repressor may be used to improve the content of these bioactive compounds in the cultivated S. miltiorrhiza.

Transcriptome analysis of MYB transcription factors family and PgMYB genes involved in salt stress resistance in Panax ginseng

BackgroundAs the king of all herbs, the medicinal value of ginseng is self-evident. The perennial nature of ginseng causes its quality to be influenced by various factors, one of which is the soil environment. During plant growth and development, MYB transcription factors play an important role in responding to abiotic stresses and regulating the synthesis of secondary metabolites. However, there are relatively few reports on the MYB transcription factor family in Panax ginseng.ResultsThis study identified 420 PgMYB transcripts under 117 genes ID in the Jilin ginseng transcriptome database. Phylogenetic analysis showed that PgMYB transcripts in Jilin ginseng were classified into 19 functional subclasses. The GO annotation result indicated that the functional differentiation of PgMYB transcripts was annotated to 11 functional nodes at GO Level 2 in ginseng. Expression pattern analysis of PgMYB transcripts based on the expression data (TPM) that PgMYB transcripts were revealed spatiotemporally specific in expression patterns. We performed a weighted network co-expression network analysis on the expression of PgMYB transcripts from different samples. The co-expression network containing 51 PgMYB transcripts was formed under a soft threshold of 0.85, revealing the reciprocal relationship of PgMYB in ginseng. Treatment of adventitious roots of ginseng with different concentrations of NaCl revealed four up-regulated expression of PgMYB transcripts that can candidate genes for salt resistance studies in ginseng.ConclusionsThe present findings provide data resources for the subsequent study of the functions of MYB transcription factor family members in ginseng, and provide an experimental basis for the anti-salt functions of MYB transcription factors in Panax ginseng.

Integrated metabolomics and transcriptomics insights on flavonoid biosynthesis of a medicinal functional forage, Agriophyllum squarrosum (L.), based on a common garden trial covering six ecotypes.

Agriophyllum squarrosum (L.) Moq., well known as sandrice, is an important wild forage in sandy areas and a promising edible and medicinal resource plant with great domestication potential. Previous studies showed flavonoids are one of the most abundant medicinal ingredients in sandrice, whereby isorhamnetin and isorhamnetin-3-glycoside were the top two flavonols with multiple health benefits. However, the molecular regulatory mechanisms of flavonoids in sandrice remain largely unclear. Based on a common garden trial, in this study, an integrated transcriptomic and flavonoids-targeted metabolomic analysis was performed on the vegetative and reproductive periods of six sandrice ecotypes, whose original habitats covered a variety of environmental factor gradients. Multiple linear stepwise regression analysis unveiled that flavonoid accumulation in sandrice was positively correlated with temperature and UVB and negatively affected by precipitation and sunshine duration, respectively. Weighted co-expression network analysis (WGCNA) indicated the bHLH and MYB transcription factor (TF) families might play key roles in sandrice flavonoid biosynthesis regulation. A total of 22,778 differentially expressed genes (DEGs) were identified between ecotype DL and ecotype AEX, the two extremes in most environmental factors, whereby 85 DEGs could be related to known flavonoid biosynthesis pathway. A sandrice flavonoid biosynthesis network embracing the detected 23 flavonoids in this research was constructed. Gene families Plant flavonoid O-methyltransferase (AsPFOMT) and UDP-glucuronosyltransferase (AsUGT78D2) were identified and characterized on the transcriptional level and believed to be synthases of isorhamnetin and isorhamnetin-3-glycoside in sandrice, respectively. A trade-off between biosynthesis of rutin and isorhamnetin was found in the DL ecotype, which might be due to the metabolic flux redirection when facing environmental changes. This research provides valuable information for understanding flavonoid biosynthesis in sandrice at the molecular level and laid the foundation for precise development and utilization of this functional resource forage.

Overexpression of a Fragaria vesca MYB Transcription Factor Gene (FvMYB82) Increases Salt and Cold Tolerance in Arabidopsis thaliana.

The MYB transcription factor (TF) family is one of the largest transcription families in plants, which is widely involved in the responses to different abiotic stresses, such as salt, cold, and drought. In the present study, a new MYB TF gene was cloned from Fragaria vesca (a diploid strawberry) and named FvMYB82. The open reading frame (ORF) of FvMYB82 was found to be 960 bp, encoding 319 amino acids. Sequence alignment results and predictions of the protein structure indicated that the FvMYB82 contained the conserved R2R3-MYB domain. Subcellular localization analysis showed that FvMYB82 was localized onto the nucleus. Furthermore, the qPCR showed that the expression level of FvMYB82 was higher in new leaves and roots than in mature leaves and stems. When dealing with different stresses, the expression level of FvMYB82 in F. vesca seedlings changed markedly, especially for salt and cold stress. When FvMYB82 was introduced into Arabidopsis thaliana, the tolerances to salt and cold stress of FvMYB82-OE A. thaliana were greatly improved. When dealt with salt and cold treatments, compared with wild-type and unloaded line (UL) A. thaliana, the transgenic lines had higher contents of proline and chlorophyll, as well as higher activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). However, the transgenic A. thaliana had lower level of malondialdehyde (MDA) and electrolytic leakage (EL) than wild-type and UL A. thaliana under salt and cold stress. Meanwhile, FvMYB82 can also regulate the expression of downstream genes associated with salt stress (AtSnRK2.4, AtSnRK2.6, AtKUP6, and AtNCED3) and cold stress (AtCBF1, AtCBF2, AtCOR15a, and AtCOR78). Therefore, these results indicated that FvMYB82 probably plays an important role in the response to salt and cold stresses in A. thaliana by regulating downstream related genes.

Analysis of Floral Organ Development and Sex Determination in Schisandra chinensis by Scanning Electron Microscopy and RNA-Sequencing.

S. chinensis is a typical monoecious plant, and the number and development of female flowers determines the yield of S. chinensis. Due to a lack of genetic information, the molecular mechanism of sex differentiation in S. chinensis remains unclear. In this study, the combination of scanning electron microscopy (SEM) and RNA sequencing (RNA-seq) was used to understand the way of sex differentiation of S. chinensis and to mine the related genes of sex determination. The result shows the development of male and female S. chinensis flowers was completed at the same time, the unisexual S. chinensis flowers did not undergo a transition stage between sexes, and sex may have been determined at an early stage in flower development. The results of the gene function analysis of the plant hormone signaling pathway and sucrose metabolism pathway suggest that auxin and JA could be the key hormones for sex differentiation in S. chinensis, and sucrose may promote pollen maturation at the later stage of male flower development. Two AGAMOUS (GAG) genes, 10 AGAMOUS-like MADS-box (AGLs) genes, and the MYB, NAC, WRKY, bHLH, and Trihelix transcription factor families may play important roles in sex determination in S. chinensis. Taken together, the present findings provide valuable genetic information on flower development and sex determination in S. chinensis.

IbMYB308, a Sweet Potato R2R3-MYB Gene, Improves Salt Stress Tolerance in Transgenic Tobacco

The MYB (v-myb avian myeloblastosis viral oncogene homolog) transcription factor family plays an important role in plant growth, development, and response to biotic and abiotic stresses. However, the gene functions of MYB transcription factors in sweet potato (Ipomoea batatas (L.) Lam) have not been elucidated. In this study, an MYB transcription factor gene, IbMYB308, was identified and isolated from sweet potato. Multiple sequence alignment showed that IbMYB308 is a typical R2R3-MYB transcription factor. Further, quantitative real-time PCR (qRT-PCR) analysis revealed that IbMYB308 was expressed in root, stem, and, especially, leaf tissues. Moreover, it showed that IbMYB308 had a tissue-specific profile. The experiment also showed that the expression of IbMYB308 was induced by different abiotic stresses (20% PEG-6000, 200 mM NaCl, and 20% H2O2). After a 200 mM NaCl treatment, the expression of several stress-related genes (SOD, POD, APX, and P5CS) was upregulation in transgenic plants, and the CAT activity, POD activity, proline content, and protein content in transgenic tobacco had increased, while MDA content had decreased. In conclusion, this study demonstrated that IbMYB308 could improve salt stress tolerance in transgenic tobacco. These findings lay a foundation for future studies on the R2R3-MYB gene family of sweet potato and suggest that IbMYB308 could potentially be used as an important positive factor in transgenic plant breeding to improve salt stress tolerance in sweet potato plants.

Anthocyanin Accumulation and Molecular Analysis of Correlated Genes by Metabolomics and Transcriptomics in Sister Line Apple Cultivars.

Red coloration in apples, an important quality trait, is primarily attributed to the accumulation of anthocyanins. Centuries of breeding have produced a wide variety of apples with different levels of anthocyanins in response to genetic and environmental stimuli. The Huashuo apple shows a much darker red color than its sister line, Huarui. Thirteen different anthocyanins were detected in Huashuo and Huarui apples, of which ten were significantly more abundant in Huashuo apples, confirming that the color difference is indeed attributed to high anthocyanins accumulation rather than the types of anthocyanins. In particular, the contents of cyanidin 3-O-galactoside levels were highest among anthocyanins in both cultivars, reaching >5000 μg·g−1 at the last color transition stage in Huashuo apples, while only >3000 μg·g−1 in Huarui apples. Moreover, the expression of most structural genes, especially DFR, CHI, and 4CL associated with anthocyanin synthesis, were higher in Huashuo apples than in Huarui apples. Combined transcriptomics, metabolomics, and qRT-PCR analysis revealed that six transcription factors from the MYB and bZIP transcription factor families likely play key roles in the dark coloring of Huashuo apples. These results provide deeper insights into apple coloring and suggest a series of candidate genes for breeding anthocyanin-rich cultivars.

Coexpression Network Analysis Based Characterisation of the R2R3-MYB Family Genes in Tolerant Poplar Infected with Melampsora larici-populina

R2R3-MYB protein is the most abundant class of MYB transcription factor family in plants. The transcript profiles of two E4 races of Melampsora larici-populina-tolerant poplars and an intolerant poplar were investigated to characterise the role of the R2R3-MYB family genes in the poplar–E4 interaction. In this study, 217 R2R3-MYBs were identified, and 83 R2R3-MYB genes were assigned to 22 different coexpression modules by weighted gene coexpression network analysis. Most R2R3-MYB genes were unchanged in the early period of E4 infection in both tolerant and intolerant poplars. However, there were obvious increases in differentially expressed R2R3-MYB genes in tolerant poplars at 2 and 4 dpi when defence responses occurred, suggesting that differently expressed R2R3-MYB genes at these time points may play an important role in poplar resistance to E4 infection. In total, 34 R2R3-MYB genes showed differential expression at 2 and 4 dpi between tolerant and intolerant poplars. Among them, 16 differentially expressed R2R3-MYB genes were related to 43 defence-related genes that had significant differences between tolerant and intolerant poplars. There might be coregulatory relationships between R2R3-MYBs and other TFs during poplar–E4 interaction. Some differentially expressed R2R3-MYB genes were related to genes involved in flavonoid biosynthesis and IAA or free SA signal transduction and might help activate defence response during poplar–E4 interaction. MYB194 could be an important node in the convergence of IAA and SA signalling.

The clock component OsLUX regulates rice heading through recruiting OsELF3-1 and OsELF4s to repress Hd1 and Ghd7

IntroductionCircadian clocks coordinate internal physiology and external environmental factors to regulate cereals flowering, which is critical for reproductive growth and optimal yield determination. ObjectivesIn this study, we aimed to confirm the role of OsLUX in flowering time regulation in rice. Further research illustrates how the OsELF4s–OsELF3-1–OsLUX complex directly regulates flowering-related genes to mediate rice heading. MethodsWe identified a circadian gene OsLUX by the MutMap method. The transcription levels of flowering-related genes were evaluated in WT and oslux mutants. OsLUX forms OsEC (OsELF4s–OsELF3-1–OsLUX) complex were supported by yeast two-hybrid, pull down, BiFC, and luciferase complementation assays (LCA). The EMSA, Chip-qPCR, luciferase luminescence images, and relative LUC activity assays were performed to examine the targeted regulation of flowering genes by the OsEC (OsELF4s–OsELF3-1–OsLUX) complex. ResultsThe circadian gene OsLUX encodes an MYB family transcription factor that functions as a vital circadian clock regulator and controls rice heading. Defect in OsLUX causes an extremely late heading phenotype under natural long-day and short-day conditions, and the function was further confirmed through genetic complementation, overexpression, and CRISPR/Cas9 knockout. OsLUX forms the OsEC (OsELF4s–OsELF3-1–OsLUX) complex by recruiting OsELF3-1 and OsELF4s, which were required to regulate rice heading. OsELF3-1 contributes to the translocation of OsLUX to the nucleus, and a compromised flowering phenotype results upon mutation of any component of the OsEC complex. The OsEC complex directly represses Hd1 and Ghd7 expression via binding to their promoter's LBS (LUX binding site) element. ConclusionOur findings show that the circadian gene OsLUX regulates rice heading by directly regulating rhythm oscillation and core flowering-time-related genes. We uncovered a mechanism by which the OsEC target suppresses the expression of Hd1 and Ghd7 directly to modulate photoperiodic flowering in rice. The OsEC (OsELF4s–OsELF3-1–OsLUX)–Hd1/Ghd7 regulatory module provides the genetic targets for crop improvement.

Cyclin-dependent kinase-mediated phosphorylation and the negative regulatory domain of transcription factor B-Myb modulate its DNA binding

B-Myb is a highly conserved member of the vertebrate Myb family of transcription factors that plays a critical role in cell-cycle progression and proliferation. Myb proteins activate Myb-dependent promoters by interacting specifically with Myb-binding site (MBS) sequences using their DNA-binding domain (DBD). Transactivation of MBS promoters by B-Myb is repressed by its negative regulatory domain (NRD), and phosphorylation of the NRD by Cdk2-CyclinA relieves the repression to activate B-Myb–dependent promoters. However, the structural mechanisms underlying autoinhibition and activation of B-Myb–mediated transcription have been poorly characterized. Here, we determined that a region in the B-Myb NRD (residues 510–600) directly associates with the DBD and inhibits binding of the DBD to the MBS DNA sequence. We demonstrate using biophysical assays that phosphorylation of the NRD at T515, T518, and T520 is sufficient to disrupt the interaction between the NRD and the DBD, which results in increased affinity for MBS DNA and increased B-Myb–dependent promoter activation in cell assays. Our biochemical characterization of B-Myb autoregulation and the activating effects of phosphorylation provide insight into how B-Myb functions as a site-specific transcription factor.

Transcriptomic and metabolomic analyses reveal that exogenous strigolactones alleviate the response of melon root to cadmium stress

Strigolactones (SLs) are classified into plant hormones, playing a key role as a mediator of plant growth in response to several abiotic stresses. Cadmium (Cd), a common heavy metal and soil pollutant, can suppress plant growth and development. In this work, we explored the effects of exogenous SLs on root formation in response to Cd stress using melon seeds subjected to seven germination treatments: CK (control), Cd (300 μmol·L-1 CdCl2), and SL1–SL5 (CdCl2-stressed seeds pretreated with 0.1, 0.5, 1, 2, and 3 μmol·L-1 GR24 solutions). The results indicated that SLs increased the antioxidant enzyme activities and root vigor and decreased the malondialdehyde (MDA) contents in the roots of Cd-stressed melon seedlings. Then we used transcriptomic and metabolomic analyses to explore the mechanisms by which exogenous SLs protect against Cd stress. There were 242 significant differentially expressed genes (DEGs) (78 upregulated, 164 downregulated) and 247 significantly differentially expressed metabolites (DEMs) (222 upregulated, 25 downregulated) between the Cd and SL3 treatments. SLs altered the expression of genes related to redox formation processes, including peroxidase (POD), lipoxygenase (LOX), glutamate dehydrogenase (GDH), and glutathione S-transferase (GST). In addition, we found that SLs regulated the expression of the MYB, AP2/ERF, bHLH, and WRKY transcription factor families. The combined transcriptomic and metabolomic analyses revealed that the DEGs and DEMs involved in Cd stress alleviation were mainly related to the gene expression of jasmonic acid (JA) and flavonoid biosynthesis. SLs might induce LOX-related genes to regulate JA biosynthesis. Moreover, SLs might promote flavonoid biosynthesis by regulating eleven flavonoid-related genes and eight metabolites. The results provide a new perspective for studying the adaptation of plants to Cd stress.