The Evolutionary History of R2R3-MYB Proteins Across 50 Eukaryotes: New Insights Into Subfamily Classification and Expansion.

R2R3-MYB proteins (2R-MYBs) are one of the main transcription factor families in higher plants. Since the evolutionary history of this gene family across the eukaryotic kingdom remains unknown, we performed a comparative analysis of 2R-MYBs from 50 major eukaryotic lineages, with particular emphasis on land plants. A total of 1548 candidates were identified among diverse taxonomic groups, which allowed for an updated classification of 73 highly conserved subfamilies, including many newly identified subfamilies. Our results revealed that the protein architectures, intron patterns, and sequence characteristics were remarkably conserved in each subfamily. At least four subfamilies were derived from early land plants, 10 evolved from spermatophytes, and 19 from angiosperms, demonstrating the diversity and preferential expansion of this gene family in land plants. Moreover, we determined that their remarkable expansion was mainly attributed to whole genome and segmental duplication, where duplicates were preferentially retained within certain subfamilies that shared three homologous intron patterns (a, b, and c) even though up to 12 types of patterns existed. Through our integrated distributions, sequence characteristics, and phylogenetic tree analyses, we confirm that 2R-MYBs are old and postulate that 3R-MYBs may be evolutionarily derived from 2R-MYBs via intragenic domain duplication.

[Systematically induced effects of Tetranychus cinnabarinus infestation on chemical defense in Zea mays inbred lines].

In the present study, we investigated the systematically induced production of defense-related compounds, including DIMBOA, total phenol, trypsin inhibitors (TI) and chymotrypsin inhibitor (CI), by Tetranychus cinnabarinus infestation in Zea mays. The first leaves of two corn in-bred line seedlings, the mite-tolerant line ' H1014168' and the mite-sensitive line 'H1014591', were sucked by T. cinnabarinus adult female for seven days, and then the contents of DIMBOA, total phenol, TI and CI were measured in the second leaf and in the roots, respectively. Results showed that as compared to the unsucked control, all contents of DIMBOA, total phenol, TI and CI induced by T. cinnabarinus sucking were significantly higher in the second leaf of both inbred lines as well as in the roots of the mite-tolerant 'H1014168'. However, in the roots of 'H1014591', these defense compounds had different trends, where there was a higher induction of TI and a lower level of total phenol than that of the healthy control, while had almost no difference in DIMBOA and CI. These findings suggested that the infestation of T. cinnabarinus could systematically induce accumulation of defense-related compounds, and this effect was stronger in the mite-tolerant inbred line than in the mite-sensitive inbred line.

Genome-wide identification and evolutionary and expression analyses of MYB-related genes in land plants.

MYB proteins constitute one of the largest transcription factor families in plants. Recent evidence revealed that MYB-related genes play crucial roles in plants. However, compared with the R2R3-MYB type, little is known about the complex evolutionary history of MYB-related proteins in plants. Here, we present a genome-wide analysis of MYB-related proteins from 16 species of flowering plants, moss, Selaginella, and algae. We identified many MYB-related proteins in angiosperms, but few in algae. Phylogenetic analysis classified MYB-related proteins into five distinct subgroups, a result supported by highly conserved intron patterns, consensus motifs, and protein domain architecture. Phylogenetic and functional analyses revealed that the Circadian Clock Associated 1-like/R-R and Telomeric DNA-binding protein-like subgroups are >1 billion yrs old, whereas the I-box-binding factor-like and CAPRICE-like subgroups appear to be newly derived in angiosperms. We further demonstrated that the MYB-like domain has evolved under strong purifying selection, indicating the conservation of MYB-related proteins. Expression analysis revealed that the MYB-related gene family has a wide expression profile in maize and soybean development and plays important roles in development and stress responses. We hypothesize that MYB-related proteins initially diversified through three major expansions and domain shuffling, but remained relatively conserved throughout the subsequent plant evolution.

Genome-wide analysis of the MYB transcription factor superfamily in soybean.

BACKGROUND: The MYB superfamily constitutes one of the most abundant groups of transcription factors described in plants. Nevertheless, their functions appear to be highly diverse and remain rather unclear. To date, no genome-wide characterization of this gene family has been conducted in a legume species. Here we report the first genome-wide analysis of the whole MYB superfamily in a legume species, soybean (Glycine max), including the gene structures, phylogeny, chromosome locations, conserved motifs, and expression patterns, as well as a comparative genomic analysis with Arabidopsis. RESULTS: A total of 244 R2R3-MYB genes were identified and further classified into 48 subfamilies based on a phylogenetic comparative analysis with their putative orthologs, showed both gene loss and duplication events. The phylogenetic analysis showed that most characterized MYB genes with similar functions are clustered in the same subfamily, together with the identification of orthologs by synteny analysis, functional conservation among subgroups of MYB genes was strongly indicated. The phylogenetic relationships of each subgroup of MYB genes were well supported by the highly conserved intron/exon structures and motifs outside the MYB domain. Synonymous nucleotide substitution (dN/dS) analysis showed that the soybean MYB DNA-binding domain is under strong negative selection. The chromosome distribution pattern strongly indicated that genome-wide segmental and tandem duplication contribute to the expansion of soybean MYB genes. In addition, we found that ~ 4% of soybean R2R3-MYB genes had undergone alternative splicing events, producing a variety of transcripts from a single gene, which illustrated the extremely high complexity of transcriptome regulation. Comparative expression profile analysis of R2R3-MYB genes in soybean and Arabidopsis revealed that MYB genes play conserved and various roles in plants, which is indicative of a divergence in function. CONCLUSIONS: In this study we identified the largest MYB gene family in plants known to date. Our findings indicate that members of this large gene family may be involved in different plant biological processes, some of which may be potentially involved in legume-specific nodulation. Our comparative genomics analysis provides a solid foundation for future functional dissection of this family gene.

The R2R3-MYB transcription factor gene family in maize.

MYB proteins comprise a large family of plant transcription factors, members of which perform a variety of functions in plant biological processes. To date, no genome-wide characterization of this gene family has been conducted in maize (Zea mays). In the present study, we performed a comprehensive computational analysis, to yield a complete overview of the R2R3-MYB gene family in maize, including the phylogeny, expression patterns, and also its structural and functional characteristics. The MYB gene structure in maize and Arabidopsis were highly conserved, indicating that they were originally compact in size. Subgroup-specific conserved motifs outside the MYB domain may reflect functional conservation. The genome distribution strongly supports the hypothesis that segmental and tandem duplication contribute to the expansion of maize MYB genes. We also performed an updated and comprehensive classification of the R2R3-MYB gene families in maize and other plant species. The result revealed that the functions were conserved between maize MYB genes and their putative orthologs, demonstrating the origin and evolutionary diversification of plant MYB genes. Species-specific groups/subgroups may evolve or be lost during evolution, resulting in functional divergence. Expression profile study indicated that maize R2R3-MYB genes exhibit a variety of expression patterns, suggesting diverse functions. Furthermore, computational prediction potential targets of maize microRNAs (miRNAs) revealed that miR159, miR319, and miR160 may be implicated in regulating maize R2R3-MYB genes, suggesting roles of these miRNAs in post-transcriptional regulation and transcription networks. Our comparative analysis of R2R3-MYB genes in maize confirm and extend the sequence and functional characteristics of this gene family, and will facilitate future functional analysis of the MYB gene family in maize.

Biochemical and molecular characterization of plant MYB transcription factor family.

MYB genes are widely distributed in higher plants and comprise one of the largest transcription factors, which are characterized by the presence of a highly conserved MYB domain at their N-termini. Over recent decades, biochemical and molecular characterizations of MYB have been extensively studied and reported to be involved in many physiological and biochemical processes. This review describes current knowledge of their structure characteristic, classification, multi-functionality, mechanism of combinational control, evolution, and function redundancy. It shows that the MYB transcription factors play a key role in plant development, such as secondary metabolism, hormone signal transduction, disease resistance, cell shape, organ development, etc. Furthermore, the expression of some members of the MYB family shows tissue-specificity.

[The roles of MYB transcription factors on plant defense responses and its molecular mechanism.].

Transcriptional regulation of defense gene expression is a crucial part of plant defense responses in plant defense environment stresses. As one of the largest plant transcription factor families, MYB (v-myb avian myeloblastosis viral on-cogene homolog) transcription factors play an important role in plant stress tolerance. In this paper, we review the structural features, functional characterization and molecular mechanism of MYB transcription factor family, and discuss the regula-tory roles of transcription factors in plant defense responses.

[Study of hrpN(CSDS001) and the gene expression profile of Arabidopsis thaliana induced by Harpin(CSDS001)].

Erwinia carotovora subsp. carotovora CSDS001 elicits hypersensitive reaction (HR) in tobacco. From the genomic libraries of Erwinia carotovora subsp. carotovora CSDS001, the hrpNCSDS001 gene (GenBank number AY939927), was isolated. The hrpNCSDS001 fusion protein was produced in Escherichia coli, and was used to induce HR by injecting into tobacco. We further examined the global regulation of Arabidopsis thaliana genes in response to HarpinCSDS001 at a concentration of 30 miccrog/mL. We indicated that 912, 1787, 2393, 1833 and 1,755 genes that were regulated significantly (log ratio or=1) at 3 h, 12 h, 24 h, 36 h and 48 h respectively after the treatment. Analysis of some transcription factors (TF) showed that 13 TF families responded to HarpinCSDS001 including ZIM, BES1, TCP, C2C2, AP2/EREBP, WRKY, bHLH, bZIP, GARP, MYB, NAC, HB, C2H2. These families mainly function in biological processes of plant defense, pho-tosynthesisdevelopment and flowering.

Starch accumulation and activities of key enzymes involved in starch synthesis in the grains of maize inbred lines with different starch contents.

Amylose, amylopectin and starch dynamic accumulation and key enzymes activities in the grains of 4 maize inbred lines (two high-starch ones and two low-starch ones) were studied. The amounts of amylose, amylopectin and starch in the grains of 4 maize inbred lines increased as sigmoid curves during grain filling period. The changes in amylose, amylopectin and starch accumulation rates followed single-peaked curves, and reached their peaks in the 25-30 days after pollination (DAP). Changes in activities of adenosine diphosphoglucose pyrophosphorylase (ADPG-PPase, EC 2.7.7.27), soluble starch synthase (SSS, EC 2.4.1.21) and starch granule-bound synthase (GBSS, EC 2.4.1.21) in the grains of 4 inbred lines appeared single-peaked curves with the peaks appearing 20-30 DAP. Changes in activities of starch-branching enzyme (Q-enzyme, EC 2.4.1.18) in the grains of high-starch inbred lines appeared single-peaked curves with the peak values at 20 DAP, while the two low-starch inbred lines showed double-peaked curves with the peak values in the 15-20 DAP and 30-35 DAP. There was significant correlation between ADPG-PPase, SSS and GBSS activities. The results indicated that the Q-enzyme had different expression in high- and low-starch maize inbred lines, and verified that ADPG-PPase, SSS and GBSS activities were significantly and positively correlated with amylose, amylopectin and starch accumulation rates.

[Analyses of Streptomyces coelicolor inner membrane proteome by multidimentional protein identification technology].

Streptomyces coelicolor is the model species among streptomycetes. Until now, proteomic analyses of S. coelicolor have been conducted using two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry method, few integral membrane proteins were identified due to the hydrophobic and low-abundance nature of these proteins. In this work, 154 possible inner membrane proteins from S. coelicolor were identified using high pH-proteinase K sample preparation method and multidimensional protein identification technology, among them 44 are integral membrane proteins containing at least one transmembrane domain, most peptides and their corresponding proteins were identified experimentally for the first time.

[The production and multi-color genomic in situ hybridization identification of maize-Z. perennis substituted material].

Genus Zea. L was composed of two sections: sect. Luxuriantes Doebley & Iltis including Z. dipperennis, Z. luxurians and Z. perennis, and sect. Zea. mays consisting only one species, annual Z. mays. To improve the biodiversity of germ-plasm in maize breeding, the study of transferring maize relatives gene into common maize were performed. Firstly, interspecific hybrids between maize (Zea. L) (2n = 20) and Z. perennis (2n = 40) were produced with the aim of transferring desirable horticultural traits from Z. perennis to maize. The F1 of maize x Z. perennis (2n = 30) plant had the most frequent configuration of 5 III + 5 II + 5 I, which were sterile and difficult to produce progeny because of genomic affinities. However,few F2 individuals of maize x Z. perennis could be obtained by some specially treatments, and one maize-like F2 plant were obtained, which were used as a female parent in backcrossing with maize parent. Twelve F2 x P1 ( BC1 F2 ) plants were obtained and then self-crossed to produce self-crossing generation of F2 x P1 (2n = 20) ( BC1 F3). The phenotypic characters of parents, F1 (2n = 30) hybrids, F2 and F2 x P1 were investigated, such as plant height, flowering, leaf shape and tillers. To further verify the genomic organization of maize-Z. perennis material, maize (inbred line 48-2) and BC1 F3 chromosomes, the root tip cells were analyzed by Multi-color GISH. We probed maize and BC1 F3 chromosomes with the probe mixture containing biotin-labeled Z. perennis genomic DNA and digoxigenin-labeled maize genomic DNA, the maize spread exhibited 10 chromosomes with yellow signals and the other 10 chromosomes carried green fluorescing bands. However, although the BC1 F3 was 2n = 20 in all cases. Multi-color GISH images revealed that 17 chromosomes had uniform signals similar to maize chromosomes, but dispersed red signals over the remaining three chromosomes were observed, which indicated that the 3 chromosomes originated from Z. perennis, and they were smaller than maize chromosomes. Data obtained from multi-color GISH images indicated that BC1 F3 was probably a substituted material from maize-Z. perennis.