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Iron is an essential element for plants. Iron uptake by plants is highly regulated, but the underlying mechanism is poorly understood. Using a truncated fragment of the iron deficiency-responsive bHLH100 gene promoter, we screened the Arabidopsis transcription factor yeast one-hybrid (Y1H) library and identified the DOF family protein, OBP3, as a crucial component of the iron deficiency-signaling pathway. OBP3 is a transcriptional repressor with a C-terminal activation domain. Its expression is induced by iron deficiency. The transgenic lines that overexpress OBP3 exhibited iron overload and premature leaf necrosis, while the obp3 mutant was less tolerant of iron deficiency. It was discovered that OBP3 directly targets the Ib subgroup of bHLH gene promoters. OBP3 interacts with the bHLH transcription factor ILR3 (IAA-LEUCINE RESISTANT3), and their interaction enhances the DNA-binding ability and transcriptional promoting activity of OBP3, resulting in the positive regulation of iron deficiency-response genes. In addition, the E3 Ligase BRUTUS facilitates 26S proteasome-mediated degradation of OBP3 protein to prevent excessive iron uptake in plants. In conclusion, our research emphasizes the vital role of OBP3 in regulating plant iron homeostasis. Synopsis: Iron is an essential element for plant growth and development, with its homeostasis being tightly regulated at both transcriptional and post-transcriptional levels. This study identifies OBP3 as a transcription factor whose abundance changes with iron status, and that triggers iron-deficiency-responsive gene expression. Expression of OBP3 depends on iron availability. OBP3 overexpression results in iron overload in transgenic plants. OBP3 interacts with the bHLH transcription factor ILR3. The E3 ligase Brutus (BTS) regulates OBP3 abundance and maintains iron equilibrium. The DOF transcription factor OBP3 heterodimerizes with the bHLH family protein ILR3 and activates transcription under iron-deficient conditions. [ABSTRACT FROM AUTHOR]

SUMMARY: Cotton seed development and fiber elongation are the inseparable and overlapped development processes requiring the continuous supply of sucrose as the direct carbon source. However, little is known about the molecular mechanism of how sucrose is transported from the source tissues (leaves) into growing cotton seeds. Here, we identify the function of a sucrose transporter gene, Sugars Will Eventually be Exported Transporter 10, GhSWEET10 in cotton seed development. GhSWEET10 encodes a functional sucrose transporter, predominantly expressing in the funiculus, inner seedcoat, and endosperm during fiber elongation. GhSWEET10 RNAi plants (GhSWEET10i) accumulated less sucrose and glucose in growing seeds and that led to shorter fibers and smaller seeds, whereas GhSWEET10 overexpressed plants (GhSWEET10OE) had bigger seeds and longer fibers with more sugar accumulation during fiber elongation. GhSWEET10 gene is transcriptionally controlled by the transcription factor GhDOFD45. GhDOFD45 knockout plants (GhDOFD45‐KO) possessed the phenotypes of smaller seeds and shorter fibers like those of GhSWEET10i plants. Furthermore, GhSWEET10 mainly exports the sucrose from the funiculus into developing seeds according to the mimic‐analysis of sucrose transporting. Collectively, all these findings show that GhDOFD45 positively regulates GhSWEET10 expression to mainly transport sucrose from leaves into developing cotton seeds. Our findings also imply that the sucrose transport into enlarging seeds benefits fiber development, and thus GhSWEET10 can be selected as a target of breeding novel cotton varieties with larger and more vigorous seeds. Significance Statement: The results of this manuscript provide clues and gene candidates for cotton breeding of improving cotton yield (seed and fiber) by directed‐mutation strategy. The regulation of sucrose transportation and cotton seed development is essential for promoting fiber quality and cotton yield. [ABSTRACT FROM AUTHOR]

Salt stress is a common abiotic factor that restricts plant growth and development. As a halophyte, Tamarix hispida is a good model plant for exploring salt-tolerance genes and regulatory mechanisms. DNA-binding with one finger (DOF) is an important transcription factor (TF) that influences and controls various signaling substances involved in diverse biological processes related to plant growth and development, but the regulatory mechanisms of DOF TFs in response to salt stress are largely unknown in T. hispida. In the present study, a newly identified Dof gene, ThDOF8 , was cloned from T. hispida , and its expression was found to be induced by salt stress. Transient overexpression of ThDOF8 enhanced T. hispida salt tolerance by enhancing proline levels, and increasing the activities of the antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD). These results were also verified in stably transformed Arabidopsis. Results from TF-centered yeast one-hybrid (Y1H) assays and EMSAs showed that ThDOF8 binds to a newly identified cis -element (TGCG). Expression profiling by gene chip analysis identified four potential direct targets of ThDOF8, namely the cysteine-rich receptor-like kinases genes, CRK10 and CRK26, and two glutamate decarboxylase genes, GAD41 , and GAD42 , and these were further verified by ChIP–quantitative-PCR, EMSAs, Y1H assays, and β-glucuronidase enzyme activity assays. ThDOF8 can bind to the TGCG element in the promoter regions of its target genes, and transient overexpression of ThCRK10 also enhanced T. hispida salt tolerance. On the basis of our results, we propose a new regulatory mechanism model, in which ThDOF8 binds to the TGCG cis -element in the promoter of the target gene CRK10 to regulate its expression and improve salt tolerance in T. hispida. This study provides a basis for furthering our understanding the role of DOF TFs and identifying other downstream candidate genes that have the potential for improving plant salt tolerance via molecular breeding. [ABSTRACT FROM AUTHOR]

Abstract Background Chrysanthemum, one of the four major cut flowers all over the world, is very sensitive to salinity during cultivation. DNA binding with one finger (DOF) transcription factors play important roles in biological processes in plants. The response mechanism of CmDOF18 from chrysanthemum to salt stress remains unclear. Results In this study, CmDOF18 was cloned from Chrysanthemum morifolium, and its expression was induced by salinity stress. The gene encodes a 291-amino acid protein with a typical DOF domain. CmDOF18 was localized to the nucleus in onion epidermal cells and showed transcriptional activation in yeast. CmDOF18 transgenic plants were generated to identify the role of this gene in resistance to salinity treatment. Chrysanthemum plants overexpressing CmDOF18 were more resistant to salinity stress than wild-type plants. Under salinity stress, the malondialdehyde content and leaf electrolyte conductivity in CmDOF18-overexpressing transgenic plants were lower than those in wild-type plants, while the proline content, chlorophyll content, superoxide dismutase activity and peroxidase activity were higher than those in wild-type plants. The opposite findings were observed in gene-silenced plants compared with wild-type plants. The gene expression levels of oxidoreductase increased in CmDOF18-overexpressing transgenic plants but decreased in CmDOF18-SRDX gene-silenced transgenic plants. Conclusion In summary, we analyzed the function of CmDOF18 from chrysanthemum, which may regulate salinity stress in plants, possibly due to its role in the regulation of oxidoreductase.

Dof (DNA binding with one finger) proteins are plant-specific transcription factors involved in gene regulation during abiotic and biotic stresses. Our previous studies suggested that tobacco Dof proteins, including BBF3, can upregulate the expression of the virus resistance N gene as well as resistance-related genes. In this study, we generated and characterized transgenic tobacco lines carrying the N gene that overexpressed BBF3 cDNA constitutively. In the absence of virus challenge, the BBF3 overexpression caused no developmental defects and no changes in the N transcript level, although the transcript levels of some defense-related genes were affected compared with levels in non-transgenic plants. TMV infection induced a hypersensitive reaction (HR) with larger necrotic lesions in transgenic lines than in non-transgenic plants, but there was no change in total virus accumulation per lesion in a standardized area. Activation of the N gene expression in non-transgenic plants during synchronously-induced HR was followed by downregulation of BBF3 gene expression. Constitutive overexpression of the BBF3 transgene resulted in higher accumulation of the N gene as well as HR marker genes (Hin1 and Hsr203j) and lower accumulation of salicylic acid- and jasmonic acid-signaling marker genes (PR1-a and PR-1b). In particular, the expression of PR-1b was remarkably suppressed in the BBF3-overexpressing transgenic lines. Dof-binding motifs were confirmed in the upstream regions of all target genes examined. Our results suggest that BBF3 has a role in modulating the expression of genes associated with virus elicitor recognition and defense responses resulting in larger areas of HR-associated cell death. [ABSTRACT FROM AUTHOR]

Photosynthetic efficiency is the primary determinant of crop yield, including vegetative biomass and grain yield. Manipulation of key transcription factors known to directly control photosynthetic machinery can be an effective strategy to improve photosynthetic traits. In this study, we identified an Arabidopsis gain-of-function mutant, cogwheel1-3D , that shows a significantly enlarged rosette and increased biomass compared with wild-type plants. Overexpression of COG1 , a Dof transcription factor, recapitulated the phenotype of cogwheel1-3D , whereas knocking out COG1 and its six paralogs resulted in a reduced rosette size and decreased biomass. Transcriptomic and quantitative reverse transcription polymerase chain reaction analyses demonstrated that COG1 and its paralogs were required for light-induced expression of genes involved in photosynthesis. Further chromatin immunoprecipitation and electrophoretic mobility shift assays indicated that COG1 can directly bind to the promoter regions of multiple genes encoding light-harvesting antenna proteins. Physiological, biochemical, and microscopy analyses revealed that COG1 enhances photosynthetic capacity and starch accumulation in Arabidopsis rosette leaves. Furthermore, combined results of bioinformatic, genetic, and molecular experiments suggested that the functions of COG1 in increasing biomass are conserved in different plant species. These results collectively demonstrated that COG1 acts as a key regulator of plant biomass by promoting photosynthesis and starch accumulation. Manipulating COG1 to optimize photosynthetic capacity would create new strategies for future crop yield improvement. This study characterizes the functions of the Dof transcription factor COG1 in increasing plant biomass and reveals that COG1 can directly bind to the promoters of multiple genes encoding light-harvesting antenna proteins to promote their expression. Overexpression of COG1 significantly increased photosynthetic capacity, starch accumulation, and biomass production. [ABSTRACT FROM AUTHOR]

• Dof genes were identified in quinoa and its ancestral diploid relatives. • All the Dof genes were clustered into 6 sub-groups. • The duplication CqDof33 / CqDof34 was specific to quinoa. • Two CDF homologs CqDof31 / CqDof32 may be potential candidates for yield improvement. Dof (D NA binding with O ne F inger) proteins are plant-specific transcription factors characteristic of a highly conserved DNA binding domain. Dof family members have been shown to play important roles in growth, development, stress defense, and light and phytohormone responses. In this study, we identified 38 CqDof genes in quinoa (Chenopodium quinoa). Phylogenetic analysis of Dof genes from quinoa, Arabidopsis, rice, sugar beet, C. pallidicaule and C. suecicum indicated that those Dof s were clustered into 6 sub-groups. Gene structure and conserved motif analysis showed that motif composition varied between different sub-groups. Duplication analysis indicated CqDof family was expanded mainly by segmental duplication, and we found the duplication events CqDof31 / CqDof36 and CqDof31 / CqDof37 could be traced back to before the divergence of Chenopodium ancestor from sugar beet, and a recent duplication event CqDof33 / CqDof34 was detected specific to quinoa. Analysis of cis -acting elements showed that CqDof genes may be involved in light, phytohormone and stress response. Expression pattern analysis showed that CqDof27, CqDof31 / CqDof32, CqDof33 / CqDof34 and CqDof35 / CqDof36 in sub-group VI were affected by different photoperiods and they may act as important floral regulator. Based on the cis -element analysis and diurnal expression results, we speculated the recent duplication event CqDof33 / CqDof34 may render quinoa some growth habits to adapt to different latitudes, and CqDof31 / CqDof32 might be potential candidates for inflorescence branching and yield improvement. Together, the results obtained in this study will help understand the functions of Dof genes in quinoa. [ABSTRACT FROM AUTHOR]

Abiotic stress is the focus of passion fruit research since it harms the industry, in which high temperature is an important influencing factor. Dof transcription factors (TFs) act as essential regulators in stress conditions. TFs can protect against abiotic stress via a variety of biological processes. There is yet to be published a systematic study of the Dof (PeDof) family of passion fruit. This study discovered 13 PeDof family members by using high-quality genomes, and the members of this characterization were identified by bioinformatics. Transcriptome sequencing and qRT-PCR were used to analyze the induced expression of PeDofs under high-temperature stress during three periods, in which PeDof-11 was significantly induced with high expression. PeDof-11 was then chosen and converted into yeast, tobacco, and Arabidopsis, with the findings demonstrating that PeDof-11 could significantly respond to high-temperature stress. This research lays the groundwork for a better understanding of PeDof gene regulation under high-temperature stress. [ABSTRACT FROM AUTHOR]

Petal size is a key indicator of the ornamental value of plants, such as Petunia hybrida L., which is a popular ornamental species worldwide. Our previous study identified a flower-specific expression pattern of a DNA-binding one finger (Dof)-type transcription factor (TF) PhDof28, in the semi-flowering and full-flowering stages of petunia. In this study, subcellular localization and activation assays showed that PhDof28 was localized in the cell nucleus and could undergo in vitro self-activation. The expression levels of PhDof28 tended to be significantly up-regulated at the top parts of petals during petunia flower opening. Transgenic petunia 'W115' and tobacco plants overexpressing PhDof28 showed similar larger petal phenotypes. The cell sizes at the middle and top parts of transgenic petunia petals were significantly increased, along with higher levels of endogenous indole-3-acetic acid (IAA) hormone. Interestingly, the expression levels of two TFs, PhNAC100 and PhBPEp, which were reported as negative regulators for flower development, were dramatically increased, while the accumulation of jasmonic acid (JA), which induces PhBPEp expression, was also significantly enhanced in the transgenic petals. These results indicated that PhDof28 overexpression could increase petal size by enhancing the synthesis of endogenous IAA in petunias. Moreover, a JA-related feedback regulation mechanism was potentially activated to prevent overgrowth of petals in transgenic plants. This study will not only enhance our knowledge of the Dof TF family, but also provide crucial genetic resources for future improvements of plant ornamental traits. [ABSTRACT FROM AUTHOR]

The Dof transcription factor is a plant-specific transcription gene family that plays various biological functions in plant development and stress response. However, no relevant research has been conducted on Medicago polymorpha. Here, 36 MpDof genes were identified in the M. polymorpha genome and further divided into 10 groups based on the comparative phylogenetic analysis. The essential information of MpDof genes, such as chromosomal localization, gene structure, conserved motifs, and selective pressures were systematically analyzed. All 36 MpDof genes were predicted to contain more cis-acting elements related to hormone response. MpDof24 and MpDof25 were predicted to interact with MpDof11 and MpDof26 to involve in the photoperiod blooms process. The MpDof genes showed a diverse expression pattern in different tissues. Notably, MpDof29 and MpDof31 were specifically expressed in the large pod and root, respectively, suggesting their crucial role in the pod and root development. qRT-PCR analysis indicated that the expression levels of MpDof10, MpDof25, MpDof26, and MpDof29 were obviously up-regulated under drought, salt, and cold stress. Collectively, genome-wide identification, evolutionary, and expression analysis of the Dof transcription gene family in M. polymorpha will provide new information to further understand and utilize the function of these Dof genes in Medicago plants. [ABSTRACT FROM AUTHOR]

As a kind of plant-specific transcription factor (TF), DNA-Binding One Zinc Finger (Dof) is widely involved in the response to environmental change, and as an evolutionarily important perennial plant species, Akebia trifoliata is ideal for studying environmental adaptation. In this study, a total of 41 AktDofs were identified in the A. trifoliata genome. First, the characteristics, including the length, exon number, and chromosomal distribution of the AktDofs and the isoelectric point (PI), amino acid number, molecular weight (MW), and conserved motifs of their putative proteins, were reported. Second, we found that all AktDofs evolutionarily underwent strong purifying selection, and many (33, 80.5%) of them were generated by whole-genome duplication (WGD). Third, we outlined their expression profiles by the use of available transcriptomic data and RT-qPCR analysis. Finally, we identified four candidate genes (AktDof21, AktDof20, AktDof36, and AktDof17) and three other candidate genes (AktDof26, AktDof16, and AktDof12) that respond to long day (LD) and darkness, respectively, and that are closely associated with phytohormone-regulating pathways. Overall, this research is the first to identify and characterize the AktDofs family and is very helpful for further research on A. trifoliata adaptation to environmental factors, especially photoperiod changes. [ABSTRACT FROM AUTHOR]

DNA-binding with one finger (Dof) transcription factors play a crucial role in plant abiotic stress regulatory networks, although massive Dofs have been systematically characterized in plants, they have not been identified in the hexaploid crop sweetpotato. Herein, 43 IbDof genes were detected to be disproportionally dispersed across 14 of the 15 chromosomes of sweetpotato, and segmental duplications were discovered to be the major driving force for the expansion of IbDofs. The collinearity analysis of IbDofs with their related orthologs from eight plants revealed the potential evolutionary history of Dof gene family. Phylogenetic analysis displayed that IbDof proteins were assigned into nine subfamilies, and the regularity of gene structures and conserved motifs was consistent with the subgroup classification. Additionally, five chosen IbDof genes were shown to be substantially and variably induced under various abiotic conditions (salt, drought, heat, and cold), as well as hormone treatments (ABA and SA), according to their transcriptome data and qRT-PCR experiments. Consistently, the promoters of IbDofs contained a number of cis-acting elements associated with hormone and stress responses. Besides, it was noted that IbDof2 had transactivation activity in yeasts, while IbDof-11/-16/-36 did not, and protein interaction network analysis and yeast two-hybrid experiments revealed a complicated interaction connection amongst IbDofs. Collectively, these data lay a foundation for further functional explorations of IbDof genes, especially with regards to the possible application of multiple IbDof members in breeding the tolerant plants. [ABSTRACT FROM AUTHOR]

In plants, Dof transcription factors are involved in regulating the expression of a series of genes related to N uptake and utilization. Therefore, the present study investigated how DNA-binding with one finger (Dof) genes are expressed in response to nitrogen (N) form and concentration to clarify the role of Dof genes and their functions in promoting N assimilation and utilization in poplar. The basic characteristics and expression patterns of Dof genes in poplar were analyzed by the use of bioinformatics methods. Dof genes expressed in response to N were screened, after which the related genes were cloned and transformed into Arabidopsis thaliana; the physiological indexes and the expression of related genes were subsequently determined. The function of Dof genes was then verified in Arabidopsis thaliana plants grown in the presence of different N forms and concentrations. Forty-four Dof genes were identified, most of which were expressed in the roots and young leaves, and some of the Dof genes were expressed under ammonia- and nitrate-N treatments. Three genes related to N induction were cloned, their proteins were found to localize in the nucleus, and PnDof30 was successfully transformed into Arabidopsis thaliana for functional verification. On comparing Arabidopsis thaliana with WT Arabidopsis thaliana plants, Arabidopsis thaliana plants overexpressing the Dof gene grew better under low N levels; the contents of soluble proteins and chlorophyll significantly increased, while the soluble sugar content significantly decreased. The expressions of several AMT, NRT, and GS genes were upregulated, while the expressions of several others were downregulated, and the expression of PEPC and PK genes significantly increased. In addition, the activity of PEPC, PK, GS, and NR enzymes significantly increased. The results showed that overexpression of PnDof30 significantly increased the level of carbon and N metabolism and improved the growth of transgenic Arabidopsis thaliana plants under low-N conditions. The study revealed the biological significance of poplar Dof transcription factors in N response and regulation of related downstream gene expression and provided some meaningful clues to explain the huge difference between poplar and Arabidopsis thaliana transformed by exogenous Dof gene, which could promote the comprehensive understanding of the molecular mechanism of efficient N uptake and utilization in trees.

Aesculus chinensis is an important medicinal and horticultural plant. Its dried mature seeds, known as "Suoluozi", are a well-known traditional Chinese medicine. Aescins are its main active components, possessing multiple pharmacological activities such as anti-inflammatory and anti-exudative effects. They are widely used in the treatment of diseases such as lumbar disc herniation, postoperative edema, and sports injuries, leading to a continuous increase in market demand in recent years. The DNA binding with one finger(Dof) family is a unique transcription factor family found in the plant kingdom. It plays a crucial role in plant growth, development, stress responses, and regulation of secondary metabolism. However, research on the Dof gene family in A. chinensis is relatively scarce. In this study, we identified 36 AcDof genes from the genome of A. chinensis and analyzed their physicochemical properties, chromosomal localization, phylogenetic relationships, gene structures, cis-acting elements, and expression patterns in different tissues. The results showed that AcDof proteins ranged from 81 to 493 amino acids in length, with molecular weights ranging from 9 270.38 to 55 015.68 and isoelectric points ranging from 4.84 to 10.2. The subcellular localization analysis revealed that 34 AcDof proteins were located in the nucleus, while the remaining two AcDof proteins were located in the chloroplasts. Phylogenetic analysis divided AcDofs into nine subgroups, and gene structure analysis indicated that all AcDof genes possessed a C2-C2 type single zinc finger domain. Gene expression analysis using transcriptome data revealed tissue-specific expression patterns among AcDof family members. Specifically, AcDof04, AcDof02, and AcDof03 exhibited specific expression in seeds, suggesting their potential involvement in the regulation of aescin biosynthesis. This study not only enhances our understanding of the Dof gene family in A. chinensis but also provides important genetic resources for further investigation of the functions and regulatory mechanisms of Dof genes in this species.

Background. DNA binding with one finger (Dof) proteins are plant-specific tran- scription factor (TF) that plays a significant role in various biological processes such as plant growth and development, hormone regulation, and resistance to abiotic stress. The Dof genes have been identified and reported in multiple plants, but so far, the whole genome identification and analysis of Dof transcription factors in blueberry (Vaccinium corymbosum L.) have not been reported yet. Methods. Using the Vaccinium genome, we have identified 51 VcDof genes in blueberry. We have further analyzed their physicochemical properties, phylogenetic relationships, gene structure, collinear analysis, selective evolutionary pressure, cis- acting promoter elements, and tissue and abiotic stress expression patterns. Results. Fifty-one VcDof genes were divided into eight subfamilies, and the genes in each subfamily contained similar gene structure and motif ordering. A total of 24 pairs of colinear genes were screened; VcDof genes expanded mainly due to whole-genome duplication, which was subjected to strong purifying selection pressure during the evolution. The promoter of VcDof genes contains three types of cis-acting elements for plant growth and development, phytohormone and stress defense responsiveness. Expression profiles of VcDof genes in different tissues and fruit developmental stages of blueberry indicated that VcDof2 and VcDof45 might play a specific role in anthesis and fruit growth and development. Expression profiles of VcDof genes in different stress indicated that VcDof1, VcDof11, and VcDof15 were highly sensitive to abiotic stress. This study provides a theoretical basis for further clarifying the biological function of Dof genes in blueberry. [ABSTRACT FROM AUTHOR]

Lotus (Nelumbo nucifera Gaertn.) is a traditional Chinese aquatic flower with high ornamental and economic value, but water salinity seriously affects lotus cultivation and distribution. The Dof transcription factors (TFs) play a crucial function in the regulatory network of growth and defense in plants. However, no systematic investigations of the Dof TFs in lotus have been performed. In this study, comprehensive searches of the lotus genome yielded 29 potential NnDofs. We carried out a series of standardized analyses, which include physical properties, multiple sequence alignment, phylogenetic analysis, gene structure, motif composition, cis-acting element prediction, chromosome distribution, and synteny analysis. The results showed that segment duplication probably caused the NnDofs gene family expansion. The potential functions of NnDofs in lotus development and stress conditions are speculated by promoter analysis. Furthermore, a complete expression investigation of NnDofs utilizing an RNA-seq atlas and quantitative real-time polymerase chain reaction (qRT-PCR) was performed. The majority of the NnDofs exhibit tissue-specific expression patterns, and many genes have been identified as being extremely sensitive to salt stressors. Overall, this study is the first to report a genome-wide assessment of the Dof family in lotus, and the findings offer vital insights for prospective functional studies on lotus salinity stress. [ABSTRACT FROM AUTHOR]

This study examined the effects of overexpressing the DOF (DNA binding with one finger protein) transcription factor gene, TaDOF6 , on wheat grain native starch and Ⅲ-type resistant starch (RS 3). Overexpression of TaDOF6 in the endosperm resulted in a significant increase in amylopectin content, particularly in long-branched chains, as well as larger starch particle size. Additionally, there was an increase in both rapidly and slowly digestible starches, along with an elevated level of native resistant starch. The enhanced crystallinity and orderliness of native starch, along with improved pasting properties, were also observed. However, TaDOF6 overexpression negatively impacted RS 3 by reducing its crystal order, thermal stability, and pasting performance. Principal component analysis highlighted the substantial role of amylopectin in determining the crystal structure and physicochemical properties of starch. Moreover, a significant correlation was found between the particle size of RS 3 and its physicochemical characteristics. Overall, these findings demonstrate that TaDOF6 overexpression alters the composition of grain starch, leading to improvements in its molecular structure and physicochemical properties. • TaDOF6 is a transcription factor gene specifically expressed in wheat seeds and its overexpression altered the fine structure of starch. • Overexpression of TaDOF6 enhanced the relative crystallinity and orderliness of starch crystals, thereby improving their gelatinization properties. • TaDOF6 negatively affected the crystal order, thermal stability, and gelatinization performance of RS 3. [ABSTRACT FROM AUTHOR]

The Dof transcription factor is a plant-specific transcription gene family that plays various biological functions in plant development and stress response. However, no relevant research has been conducted on Medicago polymorpha. Here, 36 MpDof genes were identified in the M. polymorpha genome and further divided into 10 groups based on the comparative phylogenetic analysis. The essential information of MpDof genes, such as chromosomal localization, gene structure, conserved motifs, and selective pressures were systematically analyzed. All 36 MpDof genes were predicted to contain more cis-acting elements related to hormone response. MpDof24 and MpDof25 were predicted to interact with MpDof11 and MpDof26 to involve in the photoperiod blooms process. The MpDof genes showed a diverse expression pattern in different tissues. Notably, MpDof29 and MpDof31 were specifically expressed in the large pod and root, respectively, suggesting their crucial role in the pod and root development. qRT-PCR analysis indicated that the expression levels of MpDof10, MpDof25, MpDof26, and MpDof29 were obviously up-regulated under drought, salt, and cold stress. Collectively, genome-wide identification, evolutionary, and expression analysis of the Dof transcription gene family in M. polymorpha will provide new information to further understand and utilize the function of these Dof genes in Medicago plants.

In plants, Dof transcription factors are involved in regulating the expression of a series of genes related to N uptake and utilization. Therefore, the present study investigated how DNA-binding with one finger (Dof) genes are expressed in response to nitrogen (N) form and concentration to clarify the role of Dof genes and their functions in promoting N assimilation and utilization in poplar. The basic characteristics and expression patterns of Dof genes in poplar were analyzed by the use of bioinformatics methods. Dof genes expressed in response to N were screened, after which the related genes were cloned and transformed into Arabidopsis thaliana; the physiological indexes and the expression of related genes were subsequently determined. The function of Dof genes was then verified in Arabidopsis thaliana plants grown in the presence of different N forms and concentrations. Forty-four Dof genes were identified, most of which were expressed in the roots and young leaves, and some of the Dof genes were expressed under ammonia- and nitrate-N treatments. Three genes related to N induction were cloned, their proteins were found to localize in the nucleus, and PnDof30 was successfully transformed into Arabidopsis thaliana for functional verification. On comparing Arabidopsis thaliana with WT Arabidopsis thaliana plants, Arabidopsis thaliana plants overexpressing the Dof gene grew better under low N levels; the contents of soluble proteins and chlorophyll significantly increased, while the soluble sugar content significantly decreased. The expressions of several AMT, NRT, and GS genes were upregulated, while the expressions of several others were downregulated, and the expression of PEPC and PK genes significantly increased. In addition, the activity of PEPC, PK, GS, and NR enzymes significantly increased. The results showed that overexpression of PnDof30 significantly increased the level of carbon and N metabolism and improved the growth of transgenic Arabidopsis thaliana plants under low-N conditions. The study revealed the biological significance of poplar Dof transcription factors in N response and regulation of related downstream gene expression and provided some meaningful clues to explain the huge difference between poplar and Arabidopsis thaliana transformed by exogenous Dof gene, which could promote the comprehensive understanding of the molecular mechanism of efficient N uptake and utilization in trees. [ABSTRACT FROM AUTHOR]

Background DNA binding with one finger (Dof) proteins are plant-specific transcription factors playing vital roles in developmental processes and stress responses in plants. Nevertheless, the characterizations, expression patterns, and functions of the Dof family under drought stress (a key determinant of plant physiology and metabolic homeostasis) in woody plants remain unclear. Methods The birch (Betula platyphylla var. mandshuric) genome and plant TFDB database were used to identify Dof gene family members in birch plants. ClustalW2 of BioEdit v7.2.1, MEGA v7.0, ExPASy ProtParam tool, Subloc, TMHMM v2.0, GSDS v2.0, MEME, TBtools, KaKs Calculator v2.0, and PlantCARE were respectively used to align the BpDof sequences, build a phylogenetic tree, identify the physicochemical properties, analyze the chromosomal distribution and synteny, and identify the cis-elements in the promoter regions of the 26 BpDof genes. Additionally, the birch seedlings were exposed to PEG6000-simulated drought stress, and the expression patterns of the BpDof genes in different tissues were analyzed by qRT-PCR. The histochemical staining and the evaluation of physiological indexes were performed to assess the plant tolerance to drought with transient overexpression of BpDof4, BpDof11, and BpDof17 genes. SPSS software and ANOVA were used to conduct all statistical analyses and determine statistically significant differences between results. Results A total of 26 BpDof genes were identified in birch via whole-genome analysis. The conserved Dof domain with a C(x)2C(x)21C(x)2C zinc finger motif was present in all BpDof proteins. These birch BpDofs were classified into four groups (A to D) according to the phylogenetic analysis of Arabidopsis thaliana Dof genes. BpDof proteins within the same group mostly possessed similar motifs, as detected by conserved motif analysis. The exon–intron analysis revealed that the structures of BpDof genes differed, indicating probable gene gain and lose during the BpDof evolution. The chromosomal distribution and synteny analysis showed that the 26 BpDofs were unevenly distributed on 14 chromosomes, and seven duplication events among six chromosomes were found. Cis-acting elements were abundant in the promoter regions of the 26 BpDof genes. qRT-PCR revealed that the expression of the 26 BpDof genes was differentially regulated by drought stress among roots, stems, and leaves. Most BpDof genes responded to drought stress, and BpDof4, BpDof11, and BpDof17­ were significantly up-regulated. Therefore, plants overexpressing these three genes were generated to investigate drought stress tolerance. The BpDof4-, BpDof11-, and BpDof17­-overexpressing plants showed promoted reactive oxygen species (ROS) scavenging capabilities and less severe cell damage, suggesting that they conferred enhanced drought tolerance in birch. This study provided an in-depth insight into the structure, evolution, expression, and function of the Dof gene family in plants.

The DNA- binding with one finger (Dof) transcription factors plays an important role in various biological functions in plants such as photosynthetic carbon assimilation, stress response, and flowering time. The information regarding the Dof protein is limited in wheat due to lack of wheat genomic data. In this regard, we conducted TaDof gene identification in chromosome 2 and identified eighteen TaDof proteins. These putative genes were mapped on the 2A, 2B and 2D chromosome. The motif analysis confirms the presence of the DNA-binding domain containing C2C2-type zinc figure motif at N-terminal and a transcriptional regulation domain at the C-terminal in the identified sequence. We compared TaDof in rice, wheat, maize, sorghum and performed phylogenetic analysis. These genes were classified into five groups based on phylogenetic analysis supported by high bootstrap values. Tissue- and developmental stage -specific expression analysis of RNA-seq datasets revealed the constitutive as well as the inductive response of TaDof genes in different tissues of wheat. RT-PCR analysis of five selected TaDof TFs were confirmed their responsiveness to drought, heat and salt stress. This study provides valuable information for understanding not only the potential function of Dof genes in regulating abiotic stresses, but also will be basis for exploring the function and evolution studies of more candidate TaDof genes. [ABSTRACT FROM AUTHOR]

DNA-binding One Zinc Finger (Dof) transcription factor (TF) family plays a pivotal role in various plant-specific biological processes. Sugar beet, Beta vulgaris L., is an important crop widely grown for sugar production, animal feed, and has many industrial and biotechnology applications. Although genome-wide analyses of the Dof family have been conducted in several plant species, information regarding Dof genes in sugar beet is extremely restricted. The present study analyzed 22 B. vulgaris Dof proteins (BvDofs) on the basis of conserved motifs, chromosomal locations, phylogenetic relationships, gene duplication, and expression patterns. A total of 15 conserved motifs were identified from all BvDof proteins and their distribution was supported by phylogeny. All genes were mapped on seven of the nine sugar beet chromosomes, except chromosomes 3 and 4. We showed that the BvDof family expanded mainly via ancient, segmental duplications; although more recent, tandem duplications were also involved. Comparative phylogeny using the Dof family members of Spinacia oleracea and Arabidopsis thaliana, divided Dof genes in Caryophyllales into seven major clades. Synteny analysis provided evidence for the substantial chromosome rearrangements during the evolution of the Caryophyllales genomes. Gene expression profiling of the BvDofs by RNA-seq analysis revealed differential expression of BvDofs during response to viral stress and between cultivars with distinct taproot traits, during different developmental stages. Taken together, these results offer an extensive resource for understanding the structure, evolution and function of Dof genes in sugar beet, which will help to improve crop productivity and biotechnological potential of this legume under stress conditions. [ABSTRACT FROM AUTHOR]

Abstract Background The regulation of genes related to lipid metabolism by genetic engineering is an important way to increase the accumulation of lipids in microalgae. DNA binding with one finger (DOF) is a plant-specific transcription factor in higher plants, where it regulates carbon and nitrogen metabolic pathways by regulating key genes involved in these pathways. Overexpression of DOF can increase lipid production in plants; however, it is not clear whether overexpression of DOF can increase lipids in microalgae. Results In this study, we cloned a DOF transcription factor, crDOF, from Chlamydomonas reinhardtii. The sequence of this transcription factor is 1875 bp and encodes a peptide of 624 amino acids with a conserved DOF domain. Overexpression of crDOF in C. reinhardtii significantly increased the intracellular lipid content. The content of total fatty acids in the transgenic algae line Tranc-crDOF-12 was 126.01 μg/mg (dry weight), which was 23.24% higher than that of the wild type. Additionally, the content of unsaturated fatty acids in the transgenic Tranc-crDOF-12 line increased significantly. Fluorescence quantitative PCR analysis showed that in the transgenic line Tranc-crDOF-12, the expression levels of BCC1, FAT1, SQD1, MGD1, DGD1 and PGP1 genes were significantly upregulated, while the expression levels of ACP1, ACS1, CIS1 and SQD2 were downregulated. Conclusions Our results confirm that crDOF increases intracellular lipids in C. reinhardtii by regulating key genes involved in lipid metabolism. According to these findings, we propose that enhancing the lipid content in microalgae by overexpressing DOF may be achieved in other industrial strains of microalgae and be employed for the industrial production of biodiesel.

Lotus (Nelumbo nucifera Gaertn.) is a traditional Chinese aquatic flower with high ornamental and economic value, but water salinity seriously affects lotus cultivation and distribution. The Dof transcription factors (TFs) play a crucial function in the regulatory network of growth and defense in plants. However, no systematic investigations of the Dof TFs in lotus have been performed. In this study, comprehensive searches of the lotus genome yielded 29 potential NnDofs. We carried out a series of standardized analyses, which include physical properties, multiple sequence alignment, phylogenetic analysis, gene structure, motif composition, cis-acting element prediction, chromosome distribution, and synteny analysis. The results showed that segment duplication probably caused the NnDofs gene family expansion. The potential functions of NnDofs in lotus development and stress conditions are speculated by promoter analysis. Furthermore, a complete expression investigation of NnDofs utilizing an RNA-seq atlas and quantitative real-time polymerase chain reaction (qRT-PCR) was performed. The majority of the NnDofs exhibit tissue-specific expression patterns, and many genes have been identified as being extremely sensitive to salt stressors. Overall, this study is the first to report a genome-wide assessment of the Dof family in lotus, and the findings offer vital insights for prospective functional studies on lotus salinity stress.

Dof (DNA binding with one finger) proteins play important roles in plant development and defense regulatory networks. In the present study, we report a genome-wide analysis of rose Dof genes (RchDof), including phylogenetic inferences, gene structures, chromosomal locations, gene duplications, and expression diversity. A total of 24 full-length RchDof genes were identified in Rosa chinensis, which were assigned to nine distinct subgroups. These RchDof genes were unevenly distributed on rose chromosomes. The genome-scale analysis of synteny indicated that segmental duplication events may have played a major role in the evolution of the RchDof gene family. Analysis of cis-acting elements revealed putative functions of Dofs in rose during development as well as under numerous biotic and abiotic stress conditions. Moreover, the expression profiles derived from qRT-PCR experiments demonstrated distinct expression patterns in various tissues, and gene expression divergence existed among the duplicated RchDof genes, suggesting a fundamentally functional divergence of the duplicated Dof paralogs in rose. The gene expression analysis of RchDofs under drought and salt stress conditions was also performed. The present study offered novel insights into the evolution of RchDofs and can aid in the further functional characterization of its candidate genes.

Dof (DNA binding with one finger) proteins play important roles in plant development and defense regulatory networks. In the present study, we report a genome-wide analysis of rose Dof genes (RchDof), including phylogenetic inferences, gene structures, chromosomal locations, gene duplications, and expression diversity. A total of 24 full-length RchDof genes were identified in Rosa chinensis , which were assigned to nine distinct subgroups. These RchDof genes were unevenly distributed on rose chromosomes. The genome-scale analysis of synteny indicated that segmental duplication events may have played a major role in the evolution of the RchDof gene family. Analysis of cis -acting elements revealed putative functions of Dofs in rose during development as well as under numerous biotic and abiotic stress conditions. Moreover, the expression profiles derived from qRT-PCR experiments demonstrated distinct expression patterns in various tissues, and gene expression divergence existed among the duplicated RchDof genes, suggesting a fundamentally functional divergence of the duplicated Dof paralogs in rose. The gene expression analysis of RchDof s under drought and salt stress conditions was also performed. The present study offered novel insights into the evolution of RchDof s and can aid in the further functional characterization of its candidate genes. [ABSTRACT FROM AUTHOR]

Abstract Background GRAS transcription factor (TF) family is unique and numerous in higher plants with diverse functions that involving in plant growth and development processes, such as gibberellin (GA) signal transduction, root development, root nodule formation, and mycorrhiza formation. Walnut tree is exposed to various environmental stimulus that causing concern about its resistance mechanism. In order to understand the molecular mechanism of walnut to adversity response, a GRAS TF (JrGRAS2) was cloned and characterized from Juglans regia in this study. Results A 1500 bp promoter fragment of JrGRAS2 was identified from the genome of J. regia, in which the cis-elements were screened. This JrGRAS2 promoter displayed expression activity that was enhanced significantly by high temperature (HT) stress. Yeast one-hybrid assay, transient expression and chromatin immunoprecipitation (Chip)-PCR analysis revealed that JrDof3 could specifically bind to the DOFCOREZM motif and share similar expression patterns with JrGRAS2 under HT stress. The transcription of JrGRAS2 was induced by HT stress and up-regulated to 6.73-~11.96-fold in the leaf and 2.53-~4.50-fold in the root to control, respectively. JrGRAS2 was overexpressed in Arabidopsis, three lines with much high expression level of JrGRAS2 (S3, S7, and S8) were selected for HT stress tolerance analysis. Compared to the wild type (WT) Arabidopsis, S3, S7, and S8 exhibited enhanced seed germination rate, fresh weight accumulation, and activities of catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and glutathione-S-transferase (GST) under HT stress. In contrast, the Evans blue staining, electrolyte leakage (EL) rates, hydrogen dioxide (H2O2) and malondialdehyde (MDA) content of transgenic seedlings were all lower than those of WT exposed to HT stress. Furthermore, the expression of heat shock proteins (HSPs) in S3, S7, and S8 was significant higher than those in WT plants. The similar results were obtained in JrGRAS2 transient overexpression walnut lines under normal and HT stress conditions. Conclusions Our results suggested that JrDof3 TF contributes to improve the HT stress response of JrGRAS2, which could effectively control the expression of HSPs to enhance HT stress tolerance. JrGRAS2 is an useful candidate gene for heat response in plant molecular breeding.

Copyright of Acta Agriculturae Zhejiangensis is the property of Acta Agriculturae Zhejiangensis Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Stem solidness is an important agronomic trait of durum (Triticum turgidum L. var. durum) and bread (Triticum aestivum L.) wheat that provides resistance to the wheat stem sawfly. This dominant trait is conferred by the SSt1 locus on chromosome 3B. However, the molecular identity and mechanisms underpinning stem solidness have not been identified. Here, we demonstrate that copy number variation of TdDof, a gene encoding a putative DNA binding with one finger protein, controls the stem solidness trait in wheat. Using map-based cloning, we localized TdDof to within a physical interval of 2.1 Mb inside the SSt1 locus. Molecular analysis revealed that hollow-stemmed wheat cultivars such as Kronos carry a single copy of TdDof, whereas solid-stemmed cultivars such as CDC Fortitude carry multiple identical copies of the gene. Deletion of all TdDof copies from CDC Fortitude resulted in the loss of stem solidness, whereas the transgenic overexpression of TdDof restored stem solidness in the TdDof deletion mutant pithless1 and conferred stem solidness in Kronos. In solidstemmed cultivars, increased TdDof expression was correlated with the down-regulation of genes whose orthologs have been implicated in programmed cell death (PCD) in other species. Anatomical and histochemical analyses revealed that hollow-stemmed lines had stronger PCD-associated signals in the pith cells compared to solid-stemmed lines, which suggests copy numberdependent expression of TdDof could be directly or indirectly involved in the negative regulation of PCD. These findings provide opportunities to manipulate stem development in wheat and other monocots for agricultural or industrial purposes. [ABSTRACT FROM AUTHOR]

Key message: Plant-specific Dof transcription factors VDOF1 and VDOF2 are novel regulators of vascular cell differentiation through the course of a lifetime in Arabidopsis, with shifting their transcriptional target genes. Vascular system is one of critical tissues for vascular plants to transport low-molecular compounds, such as water, minerals, and the photosynthetic product, sucrose. Here, we report the involvement of two Dof transcription factors, named VASCULAR-RELATED DOF1 (VDOF1)/VDOF4.6 and VDOF2/VDOF1.8, in vascular cell differentiation and lignin biosynthesis in Arabidopsis. VDOF genes were expressed in vascular tissues, but the detailed expression sites were partly different between VDOF1 and VDOF2. Vein patterning and lignin analysis of VDOF overexpressors and double mutant vdof1 vdof2 suggested that VDOF1 and VDOF2 would function as negative regulators of vein formation in seedlings, and lignin deposition in inflorescence stems. Interestingly, effects of VDOF overexpression in lignin deposition were different by developmental stages of inflorescence stems, and total lignin contents were increased and decreased in VDOF1 and VDOF2 overexpressors, respectively. RNA-seq analysis of inducible VDOF overexpressors demonstrated that the genes for cell wall biosynthesis, including lignin biosynthetic genes, and the transcription factor genes related to stress response and brassinosteroid signaling were commonly affected by VDOF1 and VDOF2 overexpression. Taken together, we concluded that VDOF1 and VDOF2 are novel regulators of vascular cell differentiation through the course of a lifetime, with shifting their transcriptional target genes: in seedlings, the VDOF genes negatively regulate vein formation, while at reproductive stages, the VDOF proteins target lignin biosynthesis. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Henan Agricultural Sciences is the property of Editorial Board of Journal of Henan Agricultural Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Leaf senescence is a highly complex developmental process that is tightly controlled by multiple layers of regulation. Abscisic acid (ABA) and reactive oxygen species (ROS) are two well‐known factors that promote leaf senescence. We show here that the transcription factor CDF4 positively regulates leaf senescence. Constitutive and inducible overexpression of CDF4 accelerates leaf senescence, while knockdown of CDF4 delays it. CDF4 increases endogenous ABA levels by upregulating the transcription of the ABA biosynthesis genes 9‐cis‐epoxycarotenoid dioxygenase 2, 3 (NCED2, 3) and suppresses H2O2 scavenging by repressing expression of the catalase2 (CAT2) gene. NCED2, 3 knockout and CAT2 overexpression partially rescue premature leaf senescence caused by CDF4 overexpression. We also show that CDF4 promotes floral organ abscission by activating the polygalacturonase PGAZAT gene. Based on these results, we propose that the levels of CDF4, ABA, and ROS undergo a gradual increase driven by their interlinking positive feedback loops during the leaf senescence and floral organ abscission processes. Synopsis: This study discovers an amplification loop, involving CDF4, ABA, and ROS, and provides insights into the molecular regulatory network of leaf senescence and floral organ abscission in Arabidopsis. CDF4 is a senescence‐associated gene whose expression is induced by ABA and ROS, two well‐known leaf senescence inducers.CDF4 promotes ABA biosynthesis by activating the transcription of NCED2 and NCED3 and repressing H2O2 scavenging by inhibiting CAT2 transcription. At the same time, CDF4 activates the PGAZAT promoter, thus promoting the floral organ abscission process.The levels of CDF4, ABA and ROS undergo a gradual increase driven by their interlinking positive feedback loops during the leaf senescence and floral organ abscission processes. [ABSTRACT FROM AUTHOR]

Copyright of Bulletin of Botanical Research is the property of Bulletin of Botanical Research Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In a changing climate, extreme weather events such as heatwaves will be more frequent and could affect grain weight and the quality of crops such as wheat, one of the most significant crops in terms of global food security. In this work, we characterized the response of Triticum turgidum L. spp. durum wheat to short-term heat stress (HS) treatment at transcriptomic and physiological levels during early grain filling in glasshouse experiments. We found a significant reduction in grain weight (23.9%) and grain dimensions from HS treatment. Grain quality was also affected, showing a decrease in starch content (20.8%), in addition to increments in grain protein levels (14.6%), with respect to the control condition. Moreover, RNA-seq analysis of durum wheat grains allowed us to identify 1590 differentially expressed genes related to photosynthesis, response to heat, and carbohydrate metabolic process. A gene regulatory network analysis of HS-responsive genes uncovered novel transcription factors (TFs) controlling the expression of genes involved in abiotic stress response and grain quality, such as a member of the DOF family predicted to regulate glycogen and starch biosynthetic processes in response to HS in grains. In summary, our results provide new insights into the extensive transcriptome reprogramming that occurs during short-term HS in durum wheat grains.

Abstract Background Dof (DNA binding with one finger) proteins, a class of plant-specific transcription factors which contain a conserved C2-C2-type zinc finger domain, are involved in many fundamental processes. In the Arabidopsis photoperiod response pathway, CDF (CYCLING DOF FACTOR) proteins have a primary role as acting via transcriptional repression of the direct FLOWERING LOCUS T (FT) activator CONSTANS (CO). Our previous study indicated that one of CDF homologs, OsDOf12, was involved in photoperiodic flowering. However, the functional characterization of other rice CDF like genes is still in progress. Here, we characterized the function of OsDof4 in rice. Results Phylogenic analysis indicated that OsDof4 is closely clustered into the same subgroup with CDFs and OsDof12. The subcellular localization experiment and transcriptional activity assay suggested that OsDof4 may function as a transcription factor. The diurnal expression pattern indicated that OsDof4 was regulated by endogenous circadian clock. Overexpression of OsDof4 led to earlier flowering under natural long-day field conditions (NLDs) and late flowering under natural short-day field conditions (NSDs), respectively. We compared the expression level of key floral genes in vector line and OsDof4-ox lines grown under long-day conditions (LDs) and short-day conditions (SDs). Real-time q-PCR results demonstrated that under LDs, Hd3a, RFT1 and Ehd1 were up-regulated whereas under SDs they were down-regulated. Hd1 was down-regulated at dusk period independent of photoperiods. Conclusions Taken these results together, we may speculate that the abnormal flowering responses in OsDof4-ox plants under LDs and SDs might be mediated by Ehd1 and Hd1.

• The first genome-wide identification of Dof family genes was performed in Hevea brasiliensis, a rubber-producing tree in Euphorbiaceae. • The family number of 46 in rubber is comparable to 45 in cassava and almost twice as 24 in castor, another two Euphorbiaceous plants. Dof transcription factors comprise a plant-specific family that plays diverse roles in gene regulation of many plant processes. Despite the importance of this family, little information is available in rubber (Hevea brasiliensis), a natural rubber-producing tree of the Euphorbiaceae family. This study presents the first genome-wide analysis of Dof family genes in rubber. The family number of 46 in rubber is comparable to 45 in cassava (Manihot esculenta) and almost twice as 24 in castor (Ricinus communis), another two Euphorbiaceous plants. In contrast to no recent duplicate present in castor, 22 duplicated HbDofs were identified and they were all shown to result from the so-called ρ whole-genome duplication (WGD). Despite sharing this recent WGD, comparison revealed species-specific gene loss between rubber and cassava. According to phylogenetic analysis, these genes were divided into nine groups, and 24 orthologous groups (OGs) were also proposed based on comparison of rubber, cassava, castor, arabidopsis, and rice Dofs. Tissue-specific expression profiles of HbDofs were further investigated, supporting expression divergence of HbDof duplicates. Moreover, HbDofs associated with leaf development, ethylene response, and tapping panel dryness were also identified. Results obtained from this study provide valuable information for further studies of Dof genes in rubber and other species. [ABSTRACT FROM AUTHOR]

Summary: In early seedlings, the primary root adapts rapidly to environmental changes through the modulation of endogenous hormone levels. The phytohormone ethylene inhibits primary root elongation, but the underlying molecular mechanism of how ethylene‐reduced root growth is modulated in environmental changes remains poorly understood.Here, we show that a novel rice (Oryza sativa) DOF transcription factor OsDOF15 positively regulates primary root elongation by regulating cell proliferation in the root meristem, via restricting ethylene biosynthesis. Loss‐of‐function of OsDOF15 impaired primary root elongation and cell proliferation in the root meristem, whereas OsDOF15 overexpression enhanced these processes, indicating that OsDOF15 is a key regulator of primary root elongation.This regulation involves the direct interaction of OsDOF15 with the promoter of OsACS1, resulting in the repression of ethylene biosynthesis. The control of ethylene biosynthesis by OsDOF15 in turn regulates cell proliferation in the root meristem. OsDOF15 transcription is repressed by salt stress, and OsDOF15‐mediated ethylene biosynthesis plays a role in inhibition of primary root elongation by salt stress.Thus, our data reveal how the ethylene‐inhibited primary root elongation is finely controlled by OsDOF15 in response to environmental signal, a novel mechanism of plants responding to salt stress and transmitting the information to ethylene biosynthesis to restrict root elongation. [ABSTRACT FROM AUTHOR]

Abstract Dof proteins comprise a plant-specific transcription factor family involved in regulation of many plant processes. Despite their diverse roles, little information is available in cassava (Manihot esculenta , Euphorbiaceae), the sixth major staple crop in the world. This study presents the first genome-wide identification of Dof family genes in cassava, resulting in 45 family members that are unevenly distributed across 17 out of the 18 chromosomes. Phylogenetic analysis assigned these genes into eight groups representing four subfamilies. Synteny analysis and BRH (Best Reciprocal Hit) method were also adopted to study evolutionary relationships. Among 21 MeDof duplicates identified, the majority of them were derived from the recent whole-genome duplication (WGD), whereas only one from local duplication. Transcriptional profiling revealed diverse expression patterns among various tissues, and further comparison of conserved motifs and expression profiles revealed divergence of MeDof duplicates. These findings provide valuable information for functional analysis of Dof genes in cassava. Highlights • The first genome-wide identification of Dof family genes was performed in cassava, resulting in a relatively high number of 45 members. • These MeDof genes are unevenly distributed across 17 out of the 18 chromosomes. • A high number of MeDof duplicates (21) were identified. • The majority of MeDof paralogs were derived from the recent whole-genome duplication and only one from local duplication. • Comparison of conserved motifs and expression profiles revealed divergence of MeDof duplicates. [ABSTRACT FROM AUTHOR]

Gossypium hirsutum L. is a worldwide economical crop; however, premature leaf senescence reduces its production and quality which is regulated by stresses, hormones, and genes. DNA binding with the one zinc finger (Dof) transcription factors (TFs) participate widely in plant development and responses to biotic and abiotic stresses, but there have been few reports of these TFs in cotton. Here, we perform a genome-wide study of G. Hirsutum L. Dof (GhDof) genes and analyze their phylogeny, duplication, and expression. In total, 114 GhDof genes have been identified and classified into nine subgroups (A, B1, B2.2, B2.1, C1, C2.1, C2.2, D1, and D2) based on phylogenetic analysis. An MCScanX analysis showed that the GhDof genes expanded due to segmental duplications. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that GhDofD9.6 was not only differentially expressed between CCRI10 (with premature senescence) and Liao4086 (without premature senescence) but also responded to salinity stress; GhDofA5.7, GhDofA7.4, GhDofA8.2, GhDof11.1, GhDofD7.2, and GhDofD11.3 significantly responded to cold (4 °C) stress. This work lays the foundation for further analysis of the function of GhDof genes in G. hirsutum, which will be helpful for improving the production and quality of cotton. [ABSTRACT FROM AUTHOR]

Background: Dof (DNA binding with one finger) proteins, a class of plant-specific transcription factors which contain a conserved C2-C2-type zinc finger domain, are involved in many fundamental processes. In the Arabidopsis photoperiod response pathway, CDF (CYCLING DOF FACTOR) proteins have a primary role as acting via transcriptional repression of the direct FLOWERING LOCUS T (FT) activator CONSTANS (CO). Our previous study indicated that one of CDF homologs, OsDOf12, was involved in photoperiodic flowering. However, the functional characterization of other rice CDF like genes is still in progress. Here, we characterized the function of OsDof4 in rice. Results: Phylogenic analysis indicated that OsDof4 is closely clustered into the same subgroup with CDFs and OsDof12. The subcellular localization experiment and transcriptional activity assay suggested that OsDof4 may function as a transcription factor. The diurnal expression pattern indicated that OsDof4 was regulated by endogenous circadian clock. Overexpression of OsDof4 led to earlier flowering under natural long-day field conditions (NLDs) and late flowering under natural short-day field conditions (NSDs), respectively. We compared the expression level of key floral genes in vector line and OsDof4-ox lines grown under long-day conditions (LDs) and short-day conditions (SDs). Real-time q-PCR results demonstrated that under LDs, Hd3a, RFT1 and Ehd1 were up-regulated whereas under SDs they were down-regulated. Hd1 was down-regulated at dusk period independent of photoperiods. Conclusions: Taken these results together, we may speculate that the abnormal flowering responses in OsDof4-ox plants under LDs and SDs might be mediated by Ehd1 and Hd1. [ABSTRACT FROM AUTHOR]

Only a few transcription factors have been described in the regulation of the strawberry (Fragaria x ananassa) fruit ripening process. Using a transcriptomic approach, we identified and functionally characterized FaDOF2, a DOF-type ripening-related transcription factor, which is hormonally regulated and specific to the receptacle, though high expression levels were also found in petals. The expression pattern of FaDOF2 correlated with eugenol content, a phenylpropanoid volatile, in both fruit receptacles and petals. When FaDOF2 expression was silenced in ripe strawberry receptacles, the expression of FaEOBII and FaEGS2, two key genes involved in eugenol production, were down-regulated. These fruits showed a concomitant decrease in eugenol content, which confirmed that FaDOF2 is a transcription factor that is involved in eugenol production in ripe fruit receptacles. By using the yeast two-hybrid system and bimolecular fluorescence complementation, we demonstrated that FaDOF2 interacts with FaEOBII, a previously reported regulator of eugenol production, which determines fine-tuning of the expression of key genes that are involved in eugenol production. These results provide evidence that FaDOF2 plays a subsidiary regulatory role with FaEOBII in the expression of genes encoding enzymes that control eugenol production. Taken together, our results provide new insights into the regulation of the volatile phenylpropanoid pathway in ripe strawberry receptacles. [ABSTRACT FROM AUTHOR]

Gossypium hirsutum L. is a worldwide economical crop; however, premature leaf senescence reduces its production and quality which is regulated by stresses, hormones, and genes. DNA binding with the one zinc finger (Dof) transcription factors (TFs) participate widely in plant development and responses to biotic and abiotic stresses, but there have been few reports of these TFs in cotton. Here, we perform a genome-wide study of G. Hirsutum L. Dof (GhDof) genes and analyze their phylogeny, duplication, and expression. In total, 114 GhDof genes have been identified and classified into nine subgroups (A, B1, B2.2, B2.1, C1, C2.1, C2.2, D1, and D2) based on phylogenetic analysis. An MCScanX analysis showed that the GhDof genes expanded due to segmental duplications. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that GhDofD9.6 was not only differentially expressed between CCRI10 (with premature senescence) and Liao4086 (without premature senescence) but also responded to salinity stress; GhDofA5.7, GhDofA7.4, GhDofA8.2, GhDof11.1, GhDofD7.2, and GhDofD11.3 signfificantly responded to cold (4 °C) stress. This work lays the foundation for further analysis of the function of GhDof genes in G. hirsutum, which will be helpful for improving the production and quality of cotton.

Despite their essential and multiple roles in biological processes, the molecular mechanism of Dof transcription factors (TFs) for responding to abiotic stresses is rarely reported in plants. We identified a soybean Dof gene GmDof41 which was involved in the responses to drought, salt, and exogenous ABA stresses. Overexpression of GmDof4 1 in soybean transgenic hairy roots attenuated H 2 O 2 accumulation and regulated proline homeostasis, resulting in the drought and salt tolerance. Yeast one-hybrid and electrophoretic mobility shift assay (EMSA) illustrated that GmDof41 was regulated by the DREB1-type protein GmDREB1B;1 that could improve drought and salt tolerance in plants. Further studies illustrated GmDof41 can directly bind to the promoter of GmDREB2A which encodes a DREB2-type protein and affects abiotic stress tolerance in plants. Collectively, our results suggested that GmDof41 positively regulated drought and salt tolerance by correlating with GmDREB1B;1 and GmDREB2A. This study provides an important basis for further exploring the abiotic stress-tolerance mechanism of Dof TFs in soybean. [ABSTRACT FROM AUTHOR]

Dof (DNA binding with one finger) proteins, a class of plant-specific transcription factors, are involved in plant growth and developmental processes and stress responses. However, their biological functions remain to be elucidated, especially in rice (Oryza sativa L.). Previously, we have reported that OsDof12 can promote rice flowering under long-day conditions. Here, we further investigated the other important agronomical traits of the transgenic plants overexpressing OsDof12 and found that overexpressing OsDof12 could lead to reduced plant height, erected leaf, shortened leaf blade, and smaller panicle resulted from decreased primary and secondary branches number. These results implied that OsDof12 is involved in rice plant architecture formation. Furthermore, we performed a series of Brassinosteroid (BR)-responsive tests and found that overexpression of OsDof12 could also result in BR hyposensitivity. Of note, in WT plants the expression of OsDof12 was found upregulated by BR treatment while in OsDof12 overexpression plants two positive BR signaling regulators, OsBRI1 and OsBZR1, were significantly down-regulated, indicating that OsDof12 may act as a negative BR regulator in rice. Taken together, our results suggested that overexpression of OsDof12 could lead to altered plant architecture by suppressing BR signaling. Thus, OsDof12 might be used as a new potential genetic regulator for future rice molecular breeding.

Key message: To explore the function of Dof transcription factors during kernel development in maize, we first identified Dof genes in the maize genome. We found that ZmDof3 was exclusively expressed in the endosperm of maize kernel and had the features of a Dof transcription factor. Suppression of ZmDof3 resulted in a defective kernel phenotype with reduced starch content and a partially patchy aleurone layer. The expression levels of starch synthesis-related genes and aleurone differentiation-associated genes were down-regulated in ZmDof3 knockdown kernels, indicating that ZmDof3 plays an important role in maize endosperm development. Abstract: The maize endosperm, occupying a large proportion of the kernel, plays an important role in seed development and germination. Current knowledge regarding the regulation of endosperm development is limited. Dof proteins, a family of plant-specific transcription factors, play critical roles in diverse biological processes. In this study, an endosperm-specific Dof protein gene, ZmDof3, was identified in maize through genome-wide screening. Suppression of ZmDof3 resulted in a defective kernel phenotype. The endosperm of ZmDof3 knockdown kernels was loosely packed with irregular starch granules observed by electronic microscope. Through genome-wide expression profiling, we found that down-regulated genes were enriched in GO terms related to carbohydrate metabolism. Moreover, ZmDof3 could bind to the Dof core element in the promoter of starch biosynthesis genes Du1 and Su2 in vitro and in vivo. In addition, the aleurone at local position in mature ZmDof3 knockdown kernels varied from one to three layers, which consisted of smaller and irregular cells. Further analyses showed that knockdown of ZmDof3 reduced the expression of Nkd1, which is involved in aleurone cell differentiation, and that ZmDof3 could bind to the Dof core element in the Nkd1 promoter. Our study reveals that ZmDof3 functions in maize endosperm development as a positive regulator in the signaling system controlling starch accumulation and aleurone development. [ABSTRACT FROM AUTHOR]