Dof Genes Research Articles

Transcriptional profiling and in silico analysis of Dof transcription factor gene family for understanding their regulation during seed development of rice Oryza sativa L.

Seed development is a complex process controlled by temporal and spatial expression of many transcription factors (TF) inside the developing seed. In the present study, transcript profiles of all the 30 members of rice DofTFs from flowering to seed development stages were analyzed. It was found that 16 Dof genes besides a previously characterized Dof gene 'RPBF' are differentially expressed during the seed development and unlike RPBF are not seed specific. Based on the expression patterns, these rice DofTFs were categorized into four groups-6 genes were constitutive while 4 genes were up-regulated and 3 genes were down regulated and four genes were maximally expressed at specific stages of seed development viz. one gene at flowering, two genes at watery ripe and one gene at milky stage. The involvement of more than one gene at different stages of seed development is suggestive of combinatorial regulation of their downstream genes involved in seed development. In silico expression analysis of wheat and Arabidopsis Dof Tfs also revealed that more than 50% of the Dof genes are expressed during the seed development process. Further in silico study of regulatory elements present in the promoters of these genes revealed the presence of some unique and common motifs in the promoters of rice and wheat Dof genes which indicate that Dof genes are possibly involved in ethylene and jasmonate signaling pathways affecting grain filling and grain quality. These Dof genes containing ethylene responsive motifs in their promoter region could possibly be the targets of recently identified Sub1 gene which codes for a ethylene responsive factor.

Differential Expression of PBF Dof Transcription Factor in Different Tissues of Three Finger Millet Genotypes Differing in Seed Protein Content and Color

Finger millet is one of the most nutritious cereal crops, as it is rich in calcium, minerals, and phosphorus. It also contains high protein, with prolamins being the major seed storage proteins. Genes encoding prolamins are coordinately expressed in developing endosperm wherein they are under spatial and temporal transcription control, involving cis-acting and trans-acting motifs in their promoters and trans-acting transcription factors. The PBF Dof (prolamin-binding factor DNA binding with one finger only) transcription factor can be an important regulator for seed storage protein gene expression. In this study, the spatial distribution of the PBF Dof gene has been investigated in different tissues, including root, stem, and flag leaf during vegetative growth as well as S1, S2, S3, and S4 stages of developing spikes in three finger millet genotypes, including brown (PRM-1), golden (PRM-701), and white (PRM-801) differing in grain protein content and color using reverse transcriptase polymerase chain reaction (PCR) and real-time PCR. Semi- quantitative and quantitative PCR results revealed that PBF Dof is expressed in all tissues analyzed; however, expression of PBF Dof is relatively higher in developing stages of spikes than in other tissues in all three genotypes. Spatial expression of PBF Dof genes in all tissues analyzed is similar to that reported for wheat PBF genes but in contrast to that of barley and maize PBFs wherein expression is restricted to the endosperm. Moreover, a negative correlation is observed between prolamin content and each of grain yield and grain protein in all three finger millet genotypes. The grain protein content of the white (PRM-801) genotype is highest (9.56%), followed by golden (PRM-701; 9.0%) and brown (PRM-1; 7.5%) genotypes, and it is related to higher expression of PBF Dof in S2, S3, and S4 stages, respectively. This indicates that early induction of seed protein genes in the white genotype provides a longer period for accumulation of seed storage proteins in the endosperm when compared with those in other genotypes.

Expression of DOF genes identifies early stages of vascular development in Arabidopsis leaves

The sequence of events underlying the formation of vascular networks in the leaf has long fascinated developmental biologists. In Arabidopsis leaves, vascular-precursor procambial cells derive from the elongation of morphologically inconspicuous ground cells that selectively activate expression of the HD-ZIP III gene ATHB8. Inception of ATHB8 expression operationally defines acquisition of a typically irreversible preprocambial cell state that preludes to vein formation. A view of the constellation of genes whose expression is activated at preprocambial stages would therefore be particularly desirable; however, very few preprocambial gene expression profiles have been identified. Here, we show that expression of three genes encoding members of the DOF family of plant-specific transcription factors is activated at stages overlapping onset of ATHB8 expression. Expression of DOF genes is initiated in wide domains that become confined to sites of vein development. Congruence between DOF expression fields and zones of vein formation persists upon experimental manipulation of leaf vascular patterning, suggesting that DOF expression identifies consistently recurring steps in vein ontogeny. Our results contribute to defining preprocambial cell identity at the molecular level.

Members of the Dof transcription factor family in Triticum aestivum are associated with light-mediated gene regulation

DNA binding with One Finger (Dof) protein is a plant-specific transcription factor implicated in the regulation of many important plant-specific processes, including photosynthesis and carbohydrate metabolism. This study has identified 31 Dof genes (TaDof) in bread wheat through extensive analysis of current nucleotide databases. Phylogenetic analysis suggests that the TaDof family can be divided into four clades. Expression analysis of the TaDof family across all major organs using quantitative RT-PCR and searches of the wheat genome array database revealed that the majority of TaDof members were predominately expressed in vegetative organs. A large number of TaDof members were down-regulated by drought and/or were responsive to the light and dark cycle. Further expression analysis revealed that light up-regulated TaDof members were highly correlated in expression with a number of genes that are involved in photosynthesis or sucrose transport. These data suggest that the TaDof family may have an important role in light-mediated gene regulation, including involvement in the photosynthetic process.

Functional characterization of rice OsDof12

DNA-binding with one finger (Dof) proteins are a large family of transcription factors involved in a variety of biological processes in plants. In rice, 30 different Dof genes have been identified through genome analysis. Here we report the functional characteristics of a rice Dof gene, OsDof12, which encodes a predicted Dof protein. The nuclear localization of OsDof12 was investigated by the transient expression assays of the OsDof12-GFP fusion protein in onion epidermal cells. Trans-activation assays in a yeast one-hybrid system indicated that OsDof12 had transcriptional activity. RNA expression analyses showed that the expression of OsDof12 was not tissue-specific in general and fluctuated at different development stages in rice. In addition, OsDof12 was strongly inhibited by dark treatments. The transgenic lines overexpressing OsDof12 showed early flowering under long-day (LD) conditions, whereas OsDof12 overexpression had no effect on flowering time under short-day (SD) conditions. In transgenic lines overexpressing OsDof12, the transcription levels of Hd3a and OsMADS14 were up-regulated under LD conditions but not SD conditions, whereas the expression of Hd1, OsMADS51, Ehd1 and OsGI did not change under LD and SD conditions. These results suggested that OsDof12 might regulate flowering by controlling the expression of Hd3a and OsMADS14.

Functional genomics of the Dof transcription factor family genes in suspension-cultured cells of Arabidopsis thaliana

The Dof (DNA-binding with one finger) genes are members of a family of plant-specific transcription factors that have a highly conserved DNA-binding domain, namely, Dof domain. The Dof domain is a particular class of zinc finger domain that has been demonstrated to bind specifically to DNA sequences with a T/AAAAG core. In the Arabidopsis genomic database, 36 Dof genes have been identified, whereas the functions of most of the members still remain to be studied. Therefore, we attempted to comprehensively and systematically investigate functions of Arabidopsis Dof genes. As the first step, we isolated cDNAs of all the Arabidopsis Dof genes based on the coding sequences identified on the genomic database. Then, we selected genes, which are subjected to further functional analysis, through a phylogenetic analysis, and transformed Arabidopsis cultured cells (line T87) using cDNAs corresponding to the selected genes. After that, we examined transcriptional profiles in the Dof gene-overexpressed calli using an Arabidopsis DNA microarray. From the results, possible involvements of the Dof genes in regulation of metabolic pathways are discussed.

The family of DOF transcription factors: from green unicellular algae to vascular plants

This article deals with the origin and evolution of the DOF transcription factor family through a phylogenetic analysis of those DOF sequences identified from a variety of representative organisms from different taxonomic groups: the green unicellular alga Chlamydomonas reinhardtii, the moss Physcomitrella patens, the fern Selaginella moellendorffii, the gymnosperm Pinus taeda, the dicotyledoneous Arabidopsis thaliana and the monocotyledoneous angiosperms Oryza sativa and Hordeum vulgare. In barley, we have identified 26 different DOF genes by sequence analyses of clones isolated from the screening of genomic libraries and of ESTs, whereas a single DOF gene was identified by bioinformatics searches in the Chlamydomonas genome. The phylogenetic analysis groups all these genes into six major clusters of orthologs originated from a primary basal grade. Our results suggest that duplications of an ancestral DOF, probably formed in the photosynthetic eukaryotic ancestor, followed by subsequent neo-, sub-functionalization and pseudogenization processes would have triggered the expansion of the DOF family. Loss, acquisition and shuffling of conserved motifs among the new DOFs likely underlie the mechanism of formation of the distinct subfamilies.

Divergence of the Dof Gene Families in Poplar, Arabidopsis, and Rice Suggests Multiple Modes of Gene Evolution after Duplication

It is widely accepted that gene duplication is a primary source of genetic novelty. However, the evolutionary fate of duplicated genes remains largely unresolved. The classical Ohno's Duplication-Retention-Non/Neofunctionalization theory, and the recently proposed alternatives such as subfunctionalization or duplication-degeneration-complementation, and subneofunctionalization, each can explain one or more aspects of gene fate after duplication. Duplicated genes are also affected by epigenetic changes. We constructed a phylogenetic tree using Dof (DNA binding with one finger) protein sequences from poplar (Populus trichocarpa) Torr. & Gray ex Brayshaw, Arabidopsis (Arabidopsis thaliana), and rice (Oryza sativa). From the phylogenetic tree, we identified 27 pairs of paralogous Dof genes in the terminal nodes. Analysis of protein motif structure of the Dof paralogs and their ancestors revealed six different gene fates after gene duplication. Differential protein methylation was revealed between a pair of duplicated poplar Dof genes, which have identical motif structure and similar expression pattern, indicating that epigenetics is involved in evolution. Analysis of reverse transcription-PCR, massively parallel signature sequencing, and microarray data revealed that the paralogs differ in expression pattern. Furthermore, analysis of nonsynonymous and synonymous substitution rates indicated that divergence of the duplicated genes was driven by positive selection. About one-half of the motifs in Dof proteins were shared by non-Dof proteins in the three plants species, indicating that motif co-option may be one of the forces driving gene diversification. We provided evidence that the Ohno's Duplication-Retention-Non/Neofunctionalization, subfunctionalization/duplication-degeneration-complementation, and subneofunctionalization hypotheses are complementary with, not alternative to, each other.

Identification of genes of the plant-specific transcription-factor families cooperatively regulated by ethylene and jasmonate in Arabidopsis thaliana

The analysis of expression patterns of transcription-factor genes will be the basis for a better understanding of their biological functions in plants. In this study, we designed and developed an oligo-DNA macroarray consisting of gene-specific probes of 60-65 nucleotides for 288 transcription-factor genes, which cover COL, DOF, ERF, and NAC family genes. To investigate transcription-factor genes that are cooperatively regulated by jasmonate and ethylene in arabidopsis (Arabidopsis thaliana (L.) Heynh.) plants, we analyzed the expression profile of transcription-factor genes using the oligo-DNA macroarray technique in arabidopsis plants treated with methyl jasmonate and 1-aminocyclopropane-1-carboxylic acid. Then, transcript levels of candidate genes-which were selected based on the result of macroarray analysis-were evaluated by the quantitative real-time RT-PCR method. Finally, we identified an ERF family gene that is cooperatively regulated by both hormones, and designated as cooperatively regulated by ethylene and jasmonate 1 (CEJ1).