Myeloblastosis Genes Research Articles

MYB transcription factors, their regulation and interactions with non-coding RNAs during drought stress in Brassica juncea

BackgroundBrassica juncea(L.) Czern is an important oilseed crop affected by various abiotic stresses like drought, heat, and salt. These stresses have detrimental effects on the crop’s overall growth, development and yield. Various Transcription factors (TFs) are involved in regulation of plant stress response by modulating expression of stress-responsive genes. The myeloblastosis (MYB) TFs is one of the largest families of TFs associated with various developmental and biological processes such as plant growth, secondary metabolism, stress response etc. However, MYB TFs and their regulation by non-coding RNAs (ncRNAs) in response to stress have not been studied in B. juncea. Thus, we performed a detailed study on the MYB TF family and their interactions with miRNAs and Long non coding RNAs.ResultsComputational investigation of genome and proteome data presented a comprehensive picture of the MYB genes and their protein architecture, including intron-exon organisation, conserved motif analysis, R2R3 MYB DNA-binding domains analysis, sub-cellular localization, protein-protein interaction and chromosomal locations. Phylogenetically, BjuMYBs were further classified into different subclades on the basis of topology and classification in Arabidopsis. A total of 751 MYBs were identified in B. juncea corresponding to 297 1R-BjuMYBs, 440 R2R3-BjuMYBs, 12 3R-BjuMYBs, and 2 4R-BjuMYBs types. We validated the transcriptional profiles of nine selected BjuMYBs under drought stress through RT-qPCR. Promoter analysis indicated the presence of drought-responsive cis-regulatory components. Additionally, the miRNA-MYB TF interactions was also studied, and most of the microRNAs (miRNAs) that target BjuMYBs were involved in abiotic stress response and developmental processes. Regulatory network analysis and expression patterns of lncRNA-miRNA-MYB indicated that selected long non-coding RNAs (lncRNAs) acted as strong endogenous target mimics (eTMs) of the miRNAs regulated expression of BjuMYBs under drought stress.ConclusionsThe present study has established preliminary groundwork of MYB TFs and their interaction with ncRNAs in B. juncea and it will help in developing drought- tolerant Brassica crops.

Genome-wide analysis of the MYB gene family and functional analysis of BhMYB79 in wax gourd

Wax gourd (Benincasa hispida) is an important cucurbit crop with economic and medicinal value. The myeloblastosis (MYB) gene family is one of the largest gene families in plants and regulates various biological processes, whereas the MYB gene family has not been systematically studied in wax gourd. In this study, we performed genome-wide identification of the MYB gene family in wax gourd and analyzed their phylogenetic relationship, MYB DNA-binding domain (MYB DBD), gene structure, protein motif, synteny, duplication mode and expression pattern. As a result, a total of 215 BhMYB genes (BhMYBs) were identified, belonging to four subfamilies: 1R-, 2R-, 3R- and 4R-MYB subfamilies. Genes of 1R-MYB subfamily and 2R-MYB subfamily were subdivided into different subgroups respectively. The analysis of MYB DBD, gene structure and protein motif showed that the most genes in the same subgroup have similar characteristics and the 2R-MYB genes were more conserved than the 1R-MYB genes. Interestingly, the long terminal retrotransposons (LTR-RTs) were found in the long introns of several BhMYBs. The results of synteny analysis showed that there were more syntenic gene pairs between wax gourd and other cucurbit crops, while the least number of syntenic gene pairs existed between wax gourd and rice. Gene duplication was the main reason for the expansion of the MYB gene family in wax gourd, with the transposed duplication (TRD) mode contributing more. All duplication BhMYB genes were under purifying selection pressure. Further expression analysis showed that many BhMYBs exhibited obvious tissue-specific expression and several BhMYBs were significantly induced by one or more abiotic stresses. BhMYB79 was particularly expressed in roots and significantly induced by salt, drought, cold and heat stresses, overexpression of which led to reduced tolerance to salt stress in Arabidopsis. In conclusion, our results provide a systematic analysis of wax gourd MYB gene family and facilitate the biological role study of BhMYB79 during wax gourd salt stress response process.

Molecular Characterization of MYB Transcription Factors in Camellia chekiangoleosa Reveals That CcMYB33 Is an Important Regulator Involved in Multiple Developmental Processes

Camellia chekiangoleosa is an economically important woody plant from the Genus Camellia in Theaceae, and its seed kernels are rich in edible oils of high health value. Yet, little is known about the molecular regulation of growth and development in C. chekiangoleosa. In this study, we characterized the MYB (Myeloblastosis) gene family that was widely involved in plant development and stress responses, and identified 235 members from the C. chekiangoleosa genome. Based on transcriptomic analysis of multiple tissues, we obtained tissue-specific expression profiles of the MYB genes. We found that 37 MYB genes were highly expressed during seed development, and among them, CcMYB33 (GAMYB) was specifically expressed in the seed coat, suggesting that it may be an important regulator. We cloned full-length sequences of the CcMYB33 gene and further analyzed its sequence characteristics and expression pattern. Our results indicated that CcMYB33 is an R2R3-type MYB transcription factor that is closely related to GAMYB genes of Arabidopsis thaliana. We showed that ectopic expression of CcMYB33 in Arabidopsis lines caused pleiotropical developmental defects, including abnormal leaves, fused stamen, and early flowering, among other things. This work identified important MYB regulators in the regulation of development and growth in C. chekiangoleosa, providing support for further molecular and genetic studies.

Genome-Wide Identification and Characterization of MYB Gene Family and Analysis of Its Sex-Biased Expression Pattern in Spinacia oleracea L.

The members of the myeloblastosis (MYB) family of transcription factors (TFs) participate in a variety of biological regulatory processes in plants, such as circadian rhythm, metabolism, and flower development. However, the characterization of MYB genes across the genomes of spinach Spinacia oleracea L. has not been reported. Here, we identified 140 MYB genes in spinach and described their characteristics using bioinformatics approaches. Among the MYB genes, 54 were 1R-MYB, 80 were 2R-MYB, 5 were 3R-MYB, and 1 was 4R-MYB. Almost all MYB genes were located in the 0-30 Mb region of autosomes; however, the 20 MYB genes were enriched at both ends of the sex chromosome (chromosome 4). Based on phylogeny, conserved motifs, and the structure of genes, 2R-MYB exhibited higher conservation relative to 1R-MYB genes. Tandem duplication and collinearity of spinach MYB genes drive their evolution, enabling the functional diversification of spinach genes. Subcellular localization prediction indicated that spinach MYB genes were mainly located in the nucleus. Cis-acting element analysis confirmed that MYB genes were involved in various processes of spinach growth and development, such as circadian rhythm, cell differentiation, and reproduction through hormone synthesis. Furthermore, through the transcriptome data analysis of male and female flower organs at five different periods, ten candidate genes showed biased expression in spinach males, suggesting that these genes might be related to the development of spinach anthers. Collectively, this study provides useful information for further investigating the function of MYB TFs and novel insights into the regulation of sex determination in spinach.

Identification of MYB gene family and functional analysis of GhMYB4 in cotton (Gossypium spp.).

Myeloblastosis (MYB) transcription factors (TFs) form a large gene family involved in a variety of biological processes in plants. Little is known about their roles in the development of cotton pigment glands. In this study, 646 MYB members were identified in Gossypium hirsutum genome and phylogenetic classification was analyzed. Evolution analysis revealed assymetric evolution of GhMYBs during polyploidization and sequence divergence of MYBs in G. hirustum was preferentially happend in D sub-genome. WGCNA (weighted gene co-expression network analysis) showed that four modules had potential relationship with gland development or gossypol biosynthesis in cotton. Eight differentially expressed GhMYB genes were identified by screening transcriptome data of three pairs of glanded and glandless cotton lines. Of these, four were selected as candidate genes for cotton pigment gland formation or gossypol biosynthesis by qRT-PCR assay. Silencing of GH_A11G1361 (GhMYB4) downregulated expression of multiple genes in gossypol biosynthesis pathway, indicating it could be involved in gossypol biosynthesis. The potential protein interaction network suggests that several MYBs may have indirect interaction with GhMYC2-like, a key regulator of pigment gland formation. Our study was the systematic analysis of MYB genes in cotton pigment gland development, providing candidate genes for further study on the roles of cotton MYB genes in pigment gland formation, gossypol biosynthesis and future crop plant improvement.

Genome-wide identification and expression analysis of anthocyanin synthesis-related R2R3-MYB gene family in purple passion fruit (Passiflora edulis)

The myeloblastosis (MYB) transcription factors (TFs) participate in a variety of physiological and biochemical processes, especially in the anthocyanin biosynthesis in plants. However, the comprehensive functional study of MYB TFs, especially in regulating anthocyanin accumulation in passion fruit (Passiflora edulis) remains limited. To gain a better understanding of the evolutionary history of this family, we conducted a genome-wide analysis of MYB TFs in passion fruit, including phylogenetic analysis, gene structure, conserved motifs, chromosomal locations, and collinearity analysis. A total of 254 MYB genes were identified in the passion fruit genome, comprising 128 1R-MYBs, 119 R2R3-MYBs, 5 3R-MYBs and 2 4R-MYBs. Based on the classification of Arabidopsis thaliana MYB (AtMYB) genes, the passion fruit R2R3-MYB (PeMYB) genes were further divided into 37 subgroups, which were dispersed in 9 chromosomes. The analyses of gene structure and protein conserved motif analyses further supports the clustering results of the phylogenetic tree. RT-qPCR validation revealed that the relative expression of PeMYB67 and PeMYB86 increased during fruit development, while PeMYB95 exhibited no significant changes. The expression levels of other eight MYB genes involving PeMYB4, PeMYB13, PeMYB32, PeMYB40, PeMYB78, PeMYB115, PeMYB25 and PeMYB118 manifested declining regulation alongside fruit ripening. They were either positively or negatively correlated with the accumulation of anthocyanins during fruit growth. Overall, this study provides a comprehensive overview of the passion fruit MYB superfamily genes and lays a foundation for future cloning and functional analysis of PeMYB genes related to anthocyanin accumulation in passion fruit.

Full-length fruit transcriptomes of southern highbush (Vaccinium sp.) and rabbiteye (V. virgatum Ait.) blueberry

BackgroundBlueberries (Vaccinium sp.) are native to North America and breeding efforts to improve blueberry fruit quality are focused on improving traits such as increased firmness, enhanced flavor and greater shelf-life. Such efforts require additional genomic resources, especially in southern highbush and rabbiteye blueberries.ResultsWe generated the first full-length fruit transcriptome for the southern highbush and rabbiteye blueberry using the cultivars, Suziblue and Powderblue, respectively. The transcriptome was generated using the Pacific Biosciences single-molecule long-read isoform sequencing platform with cDNA pooled from seven stages during fruit development and postharvest storage. Raw reads were processed through the Isoseq pipeline and full-length transcripts were mapped to the ‘Draper’ genome with unmapped reads collapsed using Cogent. Finally, we identified 16,299 and 15,882 non-redundant transcripts in ‘Suziblue’ and ‘Powderblue’ respectively by combining the reads mapped to Northern Highbush blueberry ‘Draper’ genome and Cogent analysis. In both cultivars, > 80% of sequences were longer than 1,000 nt, with the median transcript length around 1,700 nt. Functionally annotated transcripts using Blast2GO were > 92% in both ‘Suziblue’ and ‘Powderblue’ with overall equal distribution of gene ontology (GO) terms in the two cultivars. Analyses of alternative splicing events indicated that around 40% non-redundant sequences exhibited more than one isoform. Additionally, long non-coding RNAs were predicted to represent 5.6% and 7% of the transcriptomes in ‘Suziblue’ and ‘Powderblue’, respectively. Fruit ripening is regulated by several hormone-related genes and transcription factors. Among transcripts associated with phytohormone metabolism/signaling, the highest number of transcripts were related to abscisic acid (ABA) and auxin metabolism followed by those for brassinosteroid, jasmonic acid and ethylene metabolism. Among transcription factor-associated transcripts, those belonging to ripening-related APETALA2/ethylene-responsive element-binding factor (AP2/ERF), NAC (NAM, ATAF1/2 and CUC2), leucine zipper (HB-zip), basic helix-loop-helix (bHLH), MYB (v-MYB, discovered in avian myeloblastosis virus genome) and MADS-Box gene families, were abundant.Further we measured three fruit ripening quality traits and indicators [ABA, and anthocyanin concentration, and texture] during fruit development and ripening. ABA concentration increased during the initial stages of fruit ripening and then declined at the Ripe stage, whereas anthocyanin content increased during the final stages of fruit ripening in both cultivars. Fruit firmness declined during ripening in ‘Powderblue’. Genes associated with the above parameters were identified using the full-length transcriptome. Transcript abundance patterns of these genes were consistent with changes in the fruit ripening and quality-related characteristics.ConclusionsA full-length, well-annotated fruit transcriptome was generated for two blueberry species commonly cultivated in the southeastern United States. The robustness of the transcriptome was verified by the identification and expression analyses of multiple fruit ripening and quality–regulating genes. The full-length transcriptome is a valuable addition to the blueberry genomic resources and will aid in further improving the annotation. It will also provide a useful resource for the investigation of molecular aspects of ripening and postharvest processes.

Genome-wide identification of myeloblastosis gene family and its response to cadmium stress in Ipomoea aquatica.

The myeloblastosis (MYB) proteins perform key functions in mediating cadmium (Cd) tolerance of plants. Ipomoea aquatica has strong adaptability to Cd Stress, while the roles of the I. aquatica MYB gene family with respect to Cd stress are still unclear. Here, we identified a total of 183 MYB genes in the I. aquatica genome (laMYB), which were classified into 66 1R-type IaMYB, 112 2R-type IaMYB, four 3R-type IaMYB, and one 4R-type IaMYB based on the number of the MYB repeat in each gene. The analysis of phylogenetic tree indicated that most of IaMYB genes are associated with the diverse biological processes including defense, development and metabolism. Analysis of sequence features showed that the IaMYB genes within identical subfamily have the similar patterns of the motif distributions and gene structures. Analysis of gene duplication events revealed that the dispersed duplication (DSD) and whole-genome duplication (WGD) modes play vital roles in the expansion of the IaMYB gene family. Expression profiling manifests that approximately 20% of IaMYB genes had significant role in the roots of I. aquatica under Cd stress. Promoter profiling implied that the differentially expressed genes might be induced by environmental factors or inherent hormones and thereby execute their function in Cd response. Remarkably, the 2R-type IaMYB157 with abundant light-responsive element G-box and ABA-responsive element ABRE in its promoter region exhibited very strong response to Cd stress. Taken together, our findings provide an important candidate IaMYB gene for further deciphering the molecular regulatory mechanism in plant with respect to Cd stress.

Characterizations of MYB Transcription Factors in Camellia oleifera Reveal the Key Regulators Involved in Oil Biosynthesis

MYB (myeloblastosis) transcription factors plays an important role in various physiological and biochemical processes in plants. However, little is known about the regulatory roles of MYB family genes underlying seed oil biosynthesis in Camellia oleifera. To identify potential regulators, we performed the genome-wide characterizations of the MYB family genes and their expression profiles in C. oleifera. A total of 186 CoMYB genes were identified, including 128 R2R3-type MYB genes that had conserved R2 and R3 domains. Phylogenetic analysis revealed the CoR2R3-MYBs formed 25 subgroups and possessed some highly conserved motifs outside the MYB DNA-binding domain. We investigated the promoter regions of CoR2R3-MYBs and revealed a series of cis-acting elements related to development, hormone response, and environmental stress response, suggesting a diversified regulatory mechanism of gene functions. In addition, we identified four tandem clusters containing eleven CoR2R3-MYBs, which indicated that tandem duplications played an important role in the expansion of the CoR2R3-MYB subfamily. Furthermore, we analyzed the global gene expression profiles at five stages during seed development and revealed seven CoR2R3-MYB genes that potentially regulated lipid metabolism and seed maturation in C. oleifera. These results provide new insights into understanding the function of the MYB genes and the genetic improvement of seed oil.

Genome-Wide Identification of R2R3-MYB Transcription Factor and Expression Analysis under Abiotic Stress in Rice.

The myeloblastosis (MYB) family comprises a large group of transcription factors (TFs) that has a variety of functions. Among them, the R2R3-MYB type of proteins are the largest group in plants, which are involved in controlling various biological processes such as plant growth and development, physiological metabolism, defense, and responses to abiotic and biotic stresses. In this study, bioinformatics was adopted to conduct genome-wide identification of the R2R3-MYB TFs in rice. We identified 190 MYB TFs (99 R2R3-MYBs), which are unevenly distributed on the 12 chromosomes of rice. Based on the phylogenetic clustering and protein sequence characteristics, OsMYBs were classified into five subgroups, and 59.6% of the Os2R_MYB genes contained two introns. Analysis of cis-acting elements in the 2000 bp upstream region of Os2R_MYB genes showed that all Os2R_MYB genes contained plant hormones-related or stress-responsive elements since 91.9%, 79.8%, 79.8%, and 58.6% of Os2R_MYB genes contain ABRE, TGACG, CGTCA, and MBS motifs, respectively. Protein–protein network analysis showed that the Os2R_MYBs were involved in metabolic process, biosynthetic process, and tissue development. In addition, some genes showed a tissue-specific or developmental-stage-specific expression pattern. Moreover, the transcription levels of 20 Os2R_MYB genes under polyethylene glycol (PEG) and cadmium chloride (CdCl2) stress inducers were dissected by qRT-PCR. The results indicated genes with an altered expression upon PEG or CdCl2 stress induction. These results potentially supply a basis for further research on the role that Os2R_MYB genes play in plant development and stress responses.

Genome-wide identification and expression analysis of MYB gene family under nitrogen stress in Panax notoginseng.

The myeloblastosis (MYB) gene family, involved in regulating many important physiological and biochemical processes, is one of the largest transcript factor superfamilies in plants. Since the identification of genome sequencing of Panax notoginseng has been completed, there was little known about the whole genome of its specific MYB gene family and the response to abiotic stresses, in consideration of the excessive application of nitrogen fertilizers in P. notoginseng. In this study, 123 PnMYB genes (MYB genes of P. notoginseng) have been identified and divided into 3 subfamilies by the phylogenetic analysis. These PnMYB genes were unevenly located on 12 chromosomes. Meanwhile, the gene structure and protein conserved domain were established by MEME Suite. The analysis of collinear relationships reflected that there were 121 homologous genes between P. notoginseng and Arabidopsis and 30 between P. notoginseng and rice. Moreover, cis-acting elements of PnMYB gene promoters were predicted which indicated that PnMYBs are involved in biotic, abiotic stress, and hormone induction. The expressions of PnMYB transcription factors in its roots, flowers, and leaves were detected by qRT-PCR and they had tissue-specific expressions and related to the growth of different tissues. Under nitrogen stress, MYB transcription factors had great feedback. Ten R2R3-MYB subfamily genes were significantly induced and indicated the possible function of protecting P. notoginseng from excess nitrogen. With further knowledge on identification of PnMYB gene related to tissue selectivity and abiotic stresses, this study laid the foundation for the functional development of PnMYB gene family and improved the cultivation of P. notoginseng.

Comprehensive analysis of 1R- and 2R-MYBs reveals novel genic and protein features, complex organisation, selective expansion and insights into evolutionary tendencies.

Myeloblastosis (MYB) family, the largest plant transcription factor family, has been subcategorised based on the number and type of repeats in the MYB domain. In spite of several reports, evolution of MYB genes and repeats remains enigmatic. Brassicaceae members are endowed with complex genomes, including dysploidy because of its unique history with multiple rounds of polyploidisation, genomic fractionations and rearrangements. The present study is an attempt to gain insights into the complexities of MYB family diversity, understand impacts of genome evolution on gene families and develop an evolutionary framework to understand the origin of various subcategories of MYB gene family. We identified and analysed 1129 MYBs that included 1R-, 2R-, 3R- and atypical-MYBs across sixteen species representing protists, fungi, animals and plants and exclude MYB identified from Brassicaceae except Arabidopsis thaliana; in addition, a total of 1137 2R-MYB genes from six Brassicaceae species were also analysed. Comparative analysis revealed predominance of 1R-MYBs in protists, fungi, animals and lower plants. Phylogenetic reconstruction and analysis of selection pressure suggested ancestral nature of R1-type repeat containing 1R-MYBs that might have undergone intragenic duplication to form multi-repeat MYBs. Distinct differences in gene structure between 1R-MYB and 2R-MYBs were observed regarding intron number, the ratio of gene length to coding DNA sequence (CDS) length and the length of exons encoding the MYB domain. Conserved as well as novel and lineage-specific intron phases were identified. Analyses of physicochemical properties revealed drastic differences indicating functional diversification in MYBs. Phylogenetic reconstruction of 1R- and 2R-MYB genes revealed a shared structure-function relationship in clades which was supported when transcriptome data was analysed in silico. Comparative genomics to study distribution pattern and mapping of 2R-MYBs revealed congruency and greater degree of synteny and collinearity among closely related species. Micro-synteny analysis of genomic segments revealed high conservation of genes that are immediately flanking the surrounding tandemly organised 2R-MYBs along with instances of local duplication, reorganisations and genome fractionation. In summary, polyploidy, dysploidy, reshuffling and genome fractionation were found to cause loss or gain of 2R-MYB genes. The findings need to be supported with functional validation to understand gene structure-function relationship along the evolutionary lineage and adaptive strategies based on comparative functional genomics in plants.

Independent Evolution of the MYB Family in Brown Algae.

Myeloblastosis (MYB) proteins represent one of the largest families of eukaryotic transcription factors and regulate important processes in growth and development. Studies on MYBs have mainly focused on animals and plants; however, comprehensive analysis across other supergroups such as SAR (stramenopiles, alveolates, and rhizarians) is lacking. This study characterized the structure, evolution, and expression of MYBs in four brown algae, which comprise the biggest multicellular lineage of SAR. Subfamily 1R-MYB comprised heterogeneous proteins, with fewer conserved motifs found outside the MYB domain. Unlike the SHAQKY subgroup of plant 1R-MYB, THAQKY comprised the largest subgroup of brown algal 1R-MYBs. Unlike the expansion of 2R-MYBs in plants, brown algae harbored more 3R-MYBs than 2R-MYBs. At least ten 2R-MYBs, fifteen 3R-MYBs, and one 6R-MYB orthologs existed in the common ancestor of brown algae. Phylogenetic analysis showed that brown algal MYBs had ancient origins and a diverged evolution. They showed strong affinity with stramenopile species, while not with red algae, green algae, or animals, suggesting that brown algal MYBs did not come from the secondary endosymbiosis of red and green plastids. Sequence comparison among all repeats of the three types of MYB subfamilies revealed that the repeat of 1R-MYBs showed higher sequence identity with the R3 of 2R-MYBs and 3R-MYBs, which supports the idea that 1R-MYB was derived from loss of the first and second repeats of the ancestor MYB. Compared with other species of SAR, brown algal MYB proteins exhibited a higher proportion of intrinsic disordered regions, which might contribute to multicellular evolution. Expression analysis showed that many MYB genes are responsive to different stress conditions and developmental stages. The evolution and expression analyses provided a comprehensive analysis of the phylogeny and functions of MYBs in brown algae.

Molecular cloning and characterisation of a novel putative MYB-related transcription factor (ClMYB1R1) from Curcuma longa L.

Turmeric (Curcuma longa L.) is gaining immense global importance due to the presence of curcumin that contributes to its immunomodulatory properties. Curcumin, a phenylpropanoid derivative, is the most important secondary metabolite present in turmeric rhizomes and understanding the regulatory mechanism of curcumin biosynthesis is hence important. In plants, activator and repressor type transcription factors (TFs) of the myeloblastosis (MYB) family have been found to regulate the biosynthesis of secondary metabolites. MYB TF genes of the R2R3 class were identified by RNA-Seq based approach from turmeric. The present study involves RNA seq based identification, cloning and characterisation of a novel MYB TF of the 1R class. Gene-specific primers were used to amplify 1R MYB from the rhizome, and it was sequenced to give a full-length gene of 1056 bp. The identified MYB gene was designated as ClMYB1R1 and consisted of a 693 bp open reading frame (ORF) which encodes a 230 amino acid length protein with a molecular mass of 25 kDa and pI 9.51. The protein was predicted to consist of a single helix-turn-helix MYB-like motif and other highly conserved residues. ClMYB1R1 expression varied in different genotypes and tissues. UV-C light was found to upregulate, while NaCl and nutrient stresses down-regulated the expression. The expression showed a positive correlation with a candidate pathway gene ClPKS11 under all of the above experimental conditions, indicating the putative role of ClMYB1R1 in regulating curcumin biosynthesis.

Characterization of the MYB Genes Reveals Insights Into Their Evolutionary Conservation, Structural Diversity, and Functional Roles in Magnaporthe oryzae.

The myeloblastosis (MYB) transcription factor family is evolutionarily conserved among plants, animals, and fungi, and contributes to their growth and development. We identified and analyzed 10 putative MYB genes in Magnaporthe oryzae (MoMYB) and determined their phylogenetic relationships, revealing high divergence and variability. Although MYB domains are generally defined by three tandem repeats, MoMYBs contain one or two weakly conserved repeats embedded in extensive disordered regions. We characterized the secondary domain organization, disordered segments, and functional contributions of each MoMYB. During infection, MoMYBs are distinctively expressed and can be subdivided into two clades of being either up- or down-regulated. Among these, MoMYB1 and MoMYB8 are up-regulated during infection and vegetative growth, respectively. We found MoMYB1 localized predominantly to the cytosol during the formation of infection structures. ΔMomyb1 exhibited reduced virulence on intact rice leaves corresponding to the diminished ability to form hypha-driven appressorium (HDA). We discovered that MoMYB1 regulates HDA formation on hard, hydrophobic surfaces, whereas host surfaces partially restored HDA formation in ΔMomyb1. Lipid droplet accumulation in hyphal tips and expression of HDA-associated genes were strongly perturbed in ΔMomyb1 indicating genetic interaction of MoMYB1 with downstream components critical to HDA formation. We also found that MoMYB8 is necessary for fungal growth, dark-induced melanization of hyphae, and involved in higher abiotic stress tolerance. Taken together, we revealed a multifaceted picture of the MoMYB family, wherein a low degree of conservation has led to the development of distinct structures and functions, ranging from fungal growth to virulence.

Capsicum chinense MYB Transcription Factor Genes: Identification, Expression Analysis, and Their Conservation and Diversification With Other Solanaceae Genomes.

Myeloblastosis (MYB) genes are important transcriptional regulators of plant growth, development, and secondary metabolic biosynthesis pathways, such as capsaicinoid biosynthesis in Capsicum. Although MYB genes have been identified in Capsicum annuum, no comprehensive study has been conducted on other Capsicum species. We identified a total of 251 and 240 MYB encoding genes in Capsicum chinense MYBs (CcMYBs) and Capsicum baccatum MYBs (CbMYBs). The observation of twenty tandem and 41 segmental duplication events indicated expansion of the MYB gene family in the C. chinense genome. Five CcMYB genes, i.e., CcMYB101, CcMYB46, CcMYB6, CcPHR8, and CcRVE5, and two CaMYBs, i.e., CaMYB3 and CaHHO1, were found within the previously reported capsaicinoid biosynthesis quantitative trait loci. Based on phylogenetic analysis with tomato MYB proteins, the Capsicum MYBs were classified into 24 subgroups supported by conserved amino acid motifs and gene structures. Also, a total of 241 CcMYBs were homologous with 225 C. annuum, 213 C. baccatum, 125 potato, 79 tomato, and 23 Arabidopsis MYBs. Synteny analysis showed that all 251 CcMYBs were collinear with C. annuum, C. baccatum, tomato, potato, and Arabidopsis MYBs spanning over 717 conserved syntenic segments. Using transcriptome data from three fruit developmental stages, a total of 54 CcMYBs and 81 CaMYBs showed significant differential expression patterns. Furthermore, the expression of 24 CcMYBs from the transcriptome data was validated by quantitative real-time (qRT) PCR analysis. Eight out of the 24 CcMYBs validated by the qRT-PCR were highly expressed in fiery hot C. chinense than in the lowly pungent C. annuum. Furthermore, the co-expression analysis revealed several MYB genes clustered with genes from the capsaicinoid, anthocyanin, phenylpropanoid, carotenoid, and flavonoids biosynthesis pathways, and related to determining fruit shape and size. The homology modeling of 126 R2R3 CcMYBs showed high similarity with that of the Arabidopsis R2R3 MYB domain template, suggesting their potential functional similarity at the proteome level. Furthermore, we have identified simple sequence repeat (SSR) motifs in the CcMYB genes, which could be used in Capsicum breeding programs. The functional roles of the identified CcMYBs could be studied further so that they can be manipulated for Capsicum trait improvement.

Integrated metabolome and transcriptome analysis reveals candidate genes involved in metabolism of terpenoids and phthalides in celery seeds

Celery (Apium graveolens L.) seeds have a unique aroma and can be used in the pharmaceutical industry. Terpenoids are the main volatile organic compounds (VOCs) of celery seed essential oil, and phthalides are the source of its aroma. However, thus far, no studies have been conducted on the metabolism of terpenoids and phthalides in celery. In this study, 195 VOCs were detected in celery seeds by using headspace injection combined with gas chromatography–mass spectrometry (GC–MS). The content of 85 VOCs, including 20 terpenoids and 6 phthalides, was found to be significantly different during the five stages of celery seed development. Based on the pattern of changes in the concentrations of terpenoids and phthalides, an integrated transcriptome and metabolome analysis of three developmental celery seed stages was performed. The results showed that 11 differentially expressed structural genes involved in the metabolism of terpenoids have a positive correlation coefficient with 16 terpenoids, while 103 differentially expressed transcription factor genes are correlated with these 11 differentially expressed genes (DEGs). Among these differentially expressed transcription factor genes, two basic helix-loop-helix (bHLH) genes are similar with Arabidopsis thaliana bHLH gene (MYC2), which has already known to be involved in the regulation of terpenoid metabolism. The results of the integrated analysis also revealed that 88 differentially expressed transcription factor genes are related to 6 phthalides, and one myeloblastosis (MYB) gene is similar with petunia MYB gene (ODORANT1), which is involved in the regulation of the metabolism of volatile benzene compounds in petunia (Pharbitis nil) flowers. Therefore, this study has laid a foundation for further research related to the identification of key genes that regulate the metabolism of terpenoids and phthalides in celery. Moreover, these results can contribute significantly to the improvement in the quality of celery seed essential oil.

Comparative genomic analysis of MYB transcription factors for cuticular wax biosynthesis and drought stress tolerance in Helianthus annuus L.

Sunflower is an important oil-seed crop in Pakistan, it is mainly cultivated in the spring season. It is severely affected by drought stress resulting in lower yield. Cuticular wax acts as the first defense line to protect plants from drought stress condition. It seals the aerial parts of plants and reduce the water loss from leaf surfaces. Various myeloblastosis (MYB) transcription factors (TFs) are involved in biosynthesis of epicuticular waxes under drought-stress. However, less information is available for MYB, TFs in drought stress and wax biosynthesis in sunflower. We used different computational tools to compare the Arabidopsis MYB, TFs involved in cuticular wax biosynthesis and drought stress tolerance with sunflower genome. We identified three putative MYB genes (MYB16, MYB94 and MYB96) in sunflower along with their seven homologs in Arabidopsis. Phylogenetic association of MYB TFs in Arabidopsis and sunflower indicated strong conservation of TFs in plant species. From gene structure analysis, it was observed that intron and exon organization was family-specific. MYB TFs were unevenly distributed on sunflower chromosomes. Evolutionary analysis indicated the segmental duplication of the MYB gene family in sunflower. Quantitative Real-Time PCR revealed the up-regulation of three MYB genes under drought stress. The gene expression of MYB16, MYB94 and MYB96 were found many folds higher in experimental plants than control. The present study provided the first insight into MYB TFs family's characterization in sunflower under drought stress conditions and wax biosynthesis TFs.

Functional identification of PsMYB57 involved in anthocyanin regulation of tree peony

BackgroundR2R3 myeloblastosis (MYB) genes are widely distributed in plants and comprise one of the largest transcription factor gene families. They play important roles in the regulatory networks controlling development, metabolism, and stress responses. Researches on functional genes in tree peony are still in its infancy. To date, few MYB genes have thus far been reported.ResultsIn this study, we constructed a comprehensive reference gene set by transcriptome sequencing to obtain R2R3 MYB genes. The transcriptomes of eight different tissues were sequenced, and 92,837 unigenes were obtained with an N50 of 1662 nt. A total of 48,435 unigenes (77.98%) were functionally annotated in public databases. Based on the assembly, we identified 57 R2R3 MYB genes containing full-length open reading frames, which clustered into 35 clades by phylogenetic analysis. PsMYB57 clustered with anthocyanin regulation genes in Arabidopsis and was mainly transcribed in the buds and young leaves. The overexpression of PsMYB57 induced anthocyanin accumulation in tobacco, and four detected anthocyanin structural genes, including NtCHS, NtF3’H, NtDFR, and NtANS, were upregulated. The two endogenous bHLH genes NtAn1a and NtAn1b were also upregulated and may work in combination with PsMYB57 in regulating anthocyanin structural genes.ConclusionsOur study offers a useful reference to the selection of candidate MYB genes for further functional studies in tree peony. Function analysis of PsMYB57 is helpful to understand the color accumulation in vegetative organs of tree peony. PsMYB57 is also a promising resource to improve plant color in molecular breeding.

Molecular Characterization and Expression Analysis of MYB Transcription Factors Involved in the Glucosinolate Pathway in Chinese Cabbage (Brassica rapa ssp. pekinensis)

Chinese cabbage (Brassica rapa) is a perennial crucifer vegetable that has long been used for forage. Crucifers are rich sources of glucosinolates (GSLs), which are anti-carcinogenic in humans and involved in plant defense responses. Myeloblastosis (MYB) proteins are a large family of transcription factors (TFs) in plants and play major regulatory roles in many biological processes. We identified 14 functional R2R3-MYB genes involved in glucosinolate biosynthesis in B. rapa ssp. pekinensis. Bioinformatic analysis of their phylogeny, protein motifs, gene interaction network, and molecular characteristics showed that Chinese cabbage MYB genes are comparable to those of Arabidopsis thaliana. The expression levels of the 14 BrMYB genes under fluorescent lamp, blue, and red light were quantitated using qRT-PCR analysis. Almost all of the R2R3-BrMYBs were upregulated and expressed more under red light than under fluorescent lamp or blue light, except BrMYB34s. We also calculated the total GSLs under each light condition. The total GSL content was higher under red light than under fluorescent lamp or blue light. Furthermore, the individual glucosinolates, comprised of four aliphatic GSLs (progoitrin, sinigrin, gluconapin, and glucobrassicanapin) and one indolic GSL (glucobrassicin), were higher under red light than the other light conditions. The relationships between light quality and glucosinolate biosynthesis require further investigation.