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• 93 PavC2H2 proteins were identified from sweet cherry genome. • PavC2H2-4/16/28/54/69 were highly expressed in different sweet cherry cultivars. • PavC2H2-16/62/64/77 were induced by differnet abiotic stresses. Zinc finger protein is one of the important eukaryotic transcription factors involved in plant growth, development, biotic and abiotic stress response. Although the C2H2 gene family has been identified in many plants, little is known about it in sweet cherry (Prunus avium L.). In the present study, a total of 93 PavC2H2 proteins were identified and their distribution on chromosomes, gene structures, and conserved motifs were assessed. According to their protein structural and phylogenetic features, these 93 PavC2H2 proteins were classified into 8 distinct subclasses. Chromosome localization showed that the family members were located in 8 chromosomes of sweet cherry. According to the analysis of conserved motifs, 10 conserved motifs were identified, and motif 1 exists in 84 sequences. The expression of PavC2H2 genes in different sweet cherry cultivars and under abiotic stresses was detected. PavC2H2-4/16/28/54/69 were highly expressed in 'Hong Deng', 'Black Pearl', and Prunus tomentosa , while PavC2H2-16/62/64/77 was significantly responsive to low temperature, salt, and drought stresses. This study provides the systematic analysis of PavC2H2 genes in sweet cherry and found that some of them may play important roles in anthocyanin biosynthesis and abiotic stress response. [ABSTRACT FROM AUTHOR]

Drought is a major abiotic stress that limits maize production worldwide. Therefore, it is of great importance to improve drought tolerance in crop plants for sustainable agriculture. In this study, we examined the roles of Cys2/His2 zinc‐finger‐proteins (C2H2‐ZFPs) in maize's drought tolerance as C2H2‐ZFPs have been implicated for plant stress tolerance. By subjecting 150 Ac/Ds mutant lines to drought stress, we successfully identified a Ds‐insertion mutant, zmc2h2‐149, which shows increased tolerance to drought stress. Overexpression of ZmC2H2‐149 in maize led to a decrease in both drought tolerance and crop yield. DAP‐Seq, RNA‐Seq, Y1H and LUC assays additionally showed that ZmC2H2‐149 directly suppresses the expression of a positive drought tolerance regulator, ZmHSD1 (hydroxysteroid dehydrogenase 1). Consistently, the zmhsd1 mutants exhibited decreased drought tolerance and grain yield under water deficit conditions compared to their respective wild‐type plants. Our findings thus demonstrated that ZmC2H2‐149 can regulate ZmHSD1 for drought stress tolerance in maize, offering valuable theoretical and genetic resources for maize breeding programmes that aim for improving drought tolerance. [ABSTRACT FROM AUTHOR]

Maintaining global food security in the context of climate changes will be an important challenge in the next century. Improving abiotic stress tolerance of major crops such as wheat can contribute to this goal. This can be achieved by the identification of the genes involved and their use to develop tools for breeding programs aiming to generate better adapted cultivars. Recently, we identified the wheat TaZFP13D gene encoding Zinc Finger Protein 13D as a new gene improving water-stress tolerance. The current work analyzes the TaZFP13D-dependent transcriptome modifications that occur in well-watered and dehydration conditions to better understand its function during normal growth and during drought. Plants that overexpress TaZFP13D have a higher biomass under well-watered conditions, indicating a positive effect of the protein on growth. Survival rate and stress recovery after a severe drought stress are improved compared to wild-type plants. The latter is likely due the higher activity of key antioxidant enzymes and concomitant reduction of drought-induced oxidative damage. Conversely, down-regulation of TaZFP13D decreases drought tolerance and protection against drought-induced oxidative damage. RNA-Seq transcriptome analysis identified many genes regulated by TaZFP13D that are known to improve drought tolerance. The analysis also revealed several genes involved in the photosynthetic electron transfer chain known to improve photosynthetic efficiency and chloroplast protection against drought-induced ROS damage. This study highlights the important role of TaZFP13D in wheat drought tolerance, contributes to unravel the complex regulation governed by TaZFPs, and suggests that it could be a promising marker to select wheat cultivars with higher drought tolerance. [ABSTRACT FROM AUTHOR]

C2H2 zinc finger proteins are an important category of transcription factors in eukaryotes, which play important roles in plant growth and development and in response to abiotic stresses. OsZAT12, a C2H2 zinc finger protein in rice, which cloned in our previous study, was only expressed in rice roots and was localized in the nucleus. Overexpressing OsZAT12 in Arabidopsis thaliana exhibited dwarf phenotype. To further investigate the function of OsZAT12 in rice, qRT-PCR was used to analyze the response patterns of OsZAT12 under abiotic stresses and phytohormones treatment. The results were as follows:(1)OsZAT12 contained two typical C2H2 zinc finger domains and one EAR motif, and has transcriptional repressive activity. The promoter of the OsZAT12 contained elements related to abiotic stresses and phytohormones.(2)The results of abiotic stresses and phytohormones treatment in rice also revealed that low temperature stress(4 °C)and phytohormone abscisic acid(ABA)treatment significantly down-regulated OsZAT12 expression, while osmotic stress(20% PEG 6 000), phytohormone brassinosteroid(BR)or indole-3-acetic acid(IAA)treatment significantly up-regulated the expression of OsZAT12. These results showed that OsZAT12 involved in the changes in response to abiotic stresses and phytohormones in rice.(3)Homozygous OsZAT12 overexpression plants and OsZAT12 knockout plants were obtained using overexpression vector with 35S promoter and CRISPR/Cas9 gene editing technology, respectively.(4)Observation of the phenotype of OsZAT12 overexpression rice showed that compared with the wild type, the plant height of OsZAT12 overexpression plants was significantly shorter at tillering stage, heading stage and maturity stage. The plant height of OsZAT12 knockout plants did not change significantly compared with the wild type, while the panicle number and seed-setting rate of them were significantly lower than those of the wild type. These results indicated that OsZAT12 affected the establishment of agronomic traits such as rice plant type, panicle type and seed-setting rate.(5)The results in this study further showed that overexpression of OsZAT12 reduced the sensitivity of rice to exogenous ABA, while the opposite phenotype was observed in OsZAT12 knockout plants. Therefore, it is speculated that the effect of OsZAT12 on plant growth and development may be related to the regulation of this gene in response to abiotic stresses and hormonal signals, and this study provides an experimental basis of using OsZAT12 for molecular design breeding of stress-tolerant and stable yield in rice. [ABSTRACT FROM AUTHOR]

C2H2 zinc finger proteins (ZFPs) are a class of important transcriptional regulators in eukaryotes involved in multiple biological regulation processes. Here, MaNCP1, a C2H2 ZFP, was functionally characterized in the model entomopathogenic fungus Metarhizium acridum. Deletion of MaNCP1 delayed conidial germination and hyphal growth, decreased the conidial yield and reduced the tolerances to UV-B irradiation and heat-shock. The N-terminal zinc fingers (ZFs) of MaNCP1 made the main contributions to these traits. In addition, disruption of MaNCP1 altered the conidial surface structure and decreased the conidial hydrophobicity. Bioassays showed that the virulence of the MaNCP1 -disruption strain (Δ MaNCP1) was reduced in topical inoculation compared to the WT or the mutant complemented strain (CP), and the N-terminal C2H2 ZFs made a major contribution to virulence. Furthermore, the Δ MaNCP1 and C2H2 ZFs deletion mutants (MaNCP1∆ N and MaNCP1∆ N+C ) impaired cuticular penetration. RNA-seq showed that several cuticle-degrading genes were down-regulated in the Δ MaNCP1 background, suggesting that MaNCP1 plays vital roles in regulating insect cuticle penetration. In summary, MaNCP1 affected the growth, stress tolerances and virulence of M. acridum , and the N-terminal C2H2 ZFs played indispensable roles in these important biocontrol traits. These results provide further insights into the functions of C2H2 ZFPs in entomopathogenic fungi. • MaNCP1 is required for conidial germination, conidiation, and tolerances to UV-B and heat-shock. • MaNCP1 has crucial roles in conidial hydrophobicity. • MaNCP1 plays vital roles in the fungal virulence by regulating the insect cuticle penetration. • The N-terminal C2H2 zinc fingers play indispensable roles in these important biocontrol traits. [ABSTRACT FROM AUTHOR]

ABSTRACT Asexual sporulation is the most common reproduction mode of fungi. Most filamentous fungi have two conidiation patterns, normal conidiation and microcycle conidiation, which may be regulated by nutritional conditions. Nitrogen source can affect the fungal conidiation pattern, but the regulatory mechanism is not fully understood. In this study, we report a C2H2 zinc finger protein, MaNCP1, which has typical transcription factor characteristics and is screened from the subtractive library regulated by nitrate in the entomopathogenic fungus Metarhizium acridum. MaNCP1 and its N-terminal play critical roles in the conidiation pattern shift. Further study shows that MaNCP1 interacts with MaNmrA, which also contributes to the conidiation pattern shift and is involved in the reductive pathway of nitric oxide (NO) synthesis. Intriguingly, the conidiation pattern of the MaNCP1-disruption strain (ΔMaNCP1) can be restored to microcycle conidiation when grown on the microcycle conidiation medium, SYA, supplemented with NO donor or overexpressing MaNmrA in ΔMaNCP1. Here, we reveal that MaNCP1 governs the conidiation pattern shift through regulating the reductive synthesis of NO by physically targeting MaNmrA in M. acridum. This work provides new mechanistic insights into how changes in nitrogen utilization are linked to the regulation of fungal morphological changes. IMPORTANCE Fungal conidia play important roles in the response to environmental stimuli and evasion of the host immune system. The nitrogen source is one of the main factors affecting shifts in fungal conidiation patterns, but the regulatory mechanism involved is not fully understood. In this work, we report that the C2H2 zinc finger protein, MaNCP1, governs the conidiation pattern shift in M. acridum by targeting the MaNmrA gene, thereby altering the regulation of the reductive pathway for NO synthesis. This work provides further insights into how the nutritional environment can regulate the morphogenesis of filamentous fungi.

C2H2 zinc finger is one of the most common motifs found in the transcription factors (TFs) in eukaryotes organisms, which have a broad range of functions, such as regulation of growth and development, stress tolerance and pathogenicity. Here, PlCZF1 was identified to encode a C2H2 zinc finger in the litchi downy blight pathogen Peronophythora litchii. PlCZF1 is conserved in P. litchii and Phytophthora species. In P. litchii, PlCZF1 is highly expressed in sexual developmental and early infection stages. We generated Δplczf1 mutants using the CRISPR/Cas9 method. Compared with the wild type, the Δplczf1 mutants showed no significant difference in vegetative growth and asexual reproduction, but were defective in oospore development and virulence. Further experiments revealed that the transcription of PlM90, PlLLP and three laccase encoding genes were down-regulated in the Δplczf1 mutant. Our results demonstrated that PlCZF1 is a vital regulator for sexual development and pathogenesis in P. litchii.

C2H2 zinc finger proteins (ZFPs) play important roles in plant development and response to abiotic stresses, and have been studied extensively. However, there are few studies on ZFPs in mangroves and mangrove associates, which represent a unique plant community with robust stress tolerance. MpZFP1, which is highly induced by salt stress in the mangrove associate Millettia pinnata, was cloned and functionally characterized in this study. MpZFP1 protein contains two zinc finger domains with conserved QALGGH motifs and targets to the nucleus. The heterologous expression of MpZFP1 in Arabidopsis increased the seeds’ germination rate, seedling survival rate, and biomass accumulation under salt stress. The transgenic plants also increased the expression of stress-responsive genes, including RD22 and RD29A, and reduced the accumulation of reactive oxygen species (ROS). These results indicate that MpZFP1 is a positive regulator of plant responses to salt stress due to its activation of gene expression and efficient scavenging of ROS.

The gene family known as C2H2 zinc finger genes is widely recognized for its significant role in facilitating plant growth, development and reproduction. Many C2H2 zinc finger proteins (C2H2-ZFPs) have been identified and intensively studied in other plants, but little is known about their function in apples. In this study, the C2H2-ZFPs genes of apples were identified and analyzed by whole genome sequence analysis. In our study, 70 members of the C2H2-ZFPs gene family were identified in apples. Phylogenetic analyses divided the apple C2H2-ZFPs gene family into 6 subgroups, with the same subgroups having similar gene structures and motif patterns and speculating on their possible functions. After analysis based on numerous cis -elements on gene promoters, the apple C2H2-ZFPs family contained many light-responsive elements, phytohormone-responsive elements, plant growth and development-related elements, and stress-responsive elements, which indicated the diversity of their functions. The RNA-seq results of apples under P. expansum treatment revealed that fifteen C2H2-ZFPs genes were differentially expressed. The RT-qPCR further validated the same trend as transcriptome data. C2H2-ZFPs genes were significantly expressed in the process of P. expansum infestation. Our analysis evidenced that the apple C2H2-ZFPs gene family plays a vital role in the process of P. expansum infestation in apples. The identified genes can serve as potential candidates for further investigation. The present study provides new ideas for further research on the function of C2H2-ZFPs genes and the innovation of disease resistance technology in apples. • The C2H2-ZFPs genes of apples were identified and analyzed. • The transcriptome of apples under P. expansum treatment was studied. • The MdZFP genes expressed in the process of P. expansum infestation were screened. [ABSTRACT FROM AUTHOR]

Aluminum (Al) toxicity limits plant growth and has a major impact on the agricultural productivity in acidic soils. The zinc-finger protein (ZFP) family plays multiple roles in plant development and abiotic stresses. Although previous reports have confirmed the function of these genes, their transcriptional mechanisms in wild soybean (Glycine soja) are unclear. In this study, GsGIS3 was isolated from Al-tolerant wild soybean gene expression profiles to be functionally characterized in Arabidopsis. Laser confocal microscopic observations demonstrated that GsGIS3 is a nuclear protein, containing one C2H2 zinc-finger structure. Our results show that the expression of GsGIS3 was of a much higher level in the stem than in the leaf and root and was upregulated under AlCl3, NaCl or GA3 treatment. Compared to the control, overexpression of GsGIS3 in Arabidopsis improved Al tolerance in transgenic lines with more root growth, higher proline and lower Malondialdehyde (MDA) accumulation under concentrations of AlCl3. Analysis of hematoxylin staining indicated that GsGIS3 enhanced the resistance of transgenic plants to Al toxicity by reducing Al accumulation in Arabidopsis roots. Moreover, GsGIS3 expression in Arabidopsis enhanced the expression of Al-tolerance-related genes. Taken together, our findings indicate that GsGIS3, as a C2H2 ZFP, may enhance tolerance to Al toxicity through positive regulation of Al-tolerance-related genes.

Growing evidence indicates that some grass species are more tolerant to various abiotic and biotic stresses than many crops. Zinc finger proteins play important roles in plant abiotic and biotic stresses. Although genes coding for these proteins have been cloned and identified in various plants, their function and underlying transcriptional mechanisms in the halophyte Zoysia japonica are barely known. In the present study, ZjZFN1 was isolated from Z. japonica using RACE method. Quantitative real time PCR results revealed that the expression of ZjZFN1 was much higher in leaf than in root and stem tissues, and induced by salt, cold or ABA treatment. The subcellular localization assay demonstrated that ZjZFN1 was localized to the nucleus. Expression of the ZjZFN1 in Arabidopsis thaliana improved seed germination and enhanced plant adaption to salinity stress with improved percentage of green cotyledons and growth status under salinity stress. Physiological and transcriptional analyses suggested that ZjZFN1 might, at least in part, influence reactive oxygen species accumulation and regulate the transcription of salinity responsive genes. Furthermore, RNA-sequencing analysis of ZjZFN1-overexpressing plants revealed that ZjZFN1 may serve as a transcriptional activator in the regulation of stress responsive pathways, including phenylalanine metabolism, α-linolenic acid metabolism and phenylpropanoid biosynthesis pathways. Taken together, these results provide evidence that ZjZFN1 is a potential key player in plants’ tolerance to salt stress, and it could be a valuable gene in Z. japonica breeding projects.

Growing evidence indicates that some grass species are more tolerant to various abiotic and biotic stresses than many crops. Zinc finger proteins play important roles in plant abiotic and biotic stresses. Although genes coding for these proteins have been cloned and identified in various plants, their function and underlying transcriptional mechanisms in the halophyte Zoysia japonica are barely known. In the present study, ZjZFN1 was isolated from Z. japonica using RACE method. Quantitative real time PCR results revealed that the expression of ZjZFN1 was much higher in leaf than in root and stem tissues, and induced by salt, cold or ABA treatment. The subcellular localization assay demonstrated that ZjZFN1 was localized to the nucleus. Expression of the ZjZFN1 in Arabidopsis thaliana improved seed germination and enhanced plant adaption to salinity stress with improved percentage of green cotyledons and growth status under salinity stress. Physiological and transcriptional analyses suggested that ZjZFN1 might, at least in part, influence reactive oxygen species accumulation and regulate the transcription of salinity responsive genes. Furthermore, RNA-sequencing analysis of ZjZFN1-overexpressing plants revealed that ZjZFN1 may serve as a transcriptional activator in the regulation of stress responsive pathways, including phenylalanine metabolism, a-linolenic acid metabolism and phenylpropanoid biosynthesis pathways. Taken together, these results provide evidence that ZjZFN1 is a potential key player in plants' tolerance to salt stress, and it could be a valuable gene in Z. japonica breeding projects. [ABSTRACT FROM AUTHOR]

Plant C2H2-type zinc finger proteins (ZFPs) play essential roles in developmental control and stress responses. The whole complement of ZFP genes has been identified in Arabidopsis and rice, while the genome-scale identification and functional analysis of maize ZFPs is not yet reported. Hence, we performed a comprehensive analysis, including gene structure, chromosome location, duplicated event, selective pressure, phylogeny, gene ontology annotation, and expression profiling under developmental stages and abiotic stresses. Phylogenetic analyses suggested that the ZmZFP gene family can be grouped into three classes (A, B, and C). The analysis of differential gene expression in different developmental stages and stress treatments (drought, salt, and cold) was conducted based on microarray and RNA-seq data. A total of 99.05 % (209 genes) of the total ZmZFP genes (211 genes) were detected in 60 different tissues in microarray data. Under drought stress, 13 differentially expressed genes were found in leaf, of which 7 and 6 genes were up-regulated and down-regulated, respectively. For salt stress, crown root (CR), primary root (PR) and seed root (SR) each had one significantly elevated gene, while 2, 1, and 7 genes were obviously down-regulated in CR, PR and SR, respectively. Additionally, 8 and 3 genes were significantly up-regulated and down-regulated, respectively, in the cold-tolerant line ETH-DH7. This study will lay the foundation for understanding the roles of ZFPs in maize growth and stress resistance, contributing to the molecular breeding of maize for food. [ABSTRACT FROM AUTHOR]

C2H2 zinc finger proteins (C2H2-ZFPs), as one of the largest TF families in eukaryotes, play an essential role in plant development and stress responses, but their involvement in the ripening of economically important fruits, as well as their transcriptional regulatory mechanisms, remain largely unclear. In this study, two C2H2-ZFPs, named as MaC2H2-1/2 , were identified and characterized from banana fruit. MaC2H2-1/2 contain a typical ERF-associated amphiphilic repression (EAR) motif at their C-terminus. Subcellular localization demonstrated that MaC2H2-1/2 proteins were both localized to the nucleus. Dual-luciferase reporter assay (DLR) showed that MaC2H2-1/2 were transcriptional repressors in vivo . Gene expression analysis revealed a significant induction of MaC2H2-1/2 transcripts during the ripening of banana fruit with three different ripening characteristics caused by natural, ethylene-induced, and 1-methylcyclopropene (1-MCP)-delayed treatments, which correlated well with ethylene production. More importantly, electrophoretic mobility shift assay (EMSA) and transient expression showed that MaC2H2-1/2 bound to the promoters of the key ethylene biosynthetic genes MaACS1 and MaACO1 , and repressed their activities. Collectively, these results suggest that MaC2H2-1/2 are transcriptional repressors and may mediate a finely tuned regulation of ethylene production during banana fruit ripening, possibly via transcriptional repression of ethylene biosynthetic genes, expanding the knowledgement of the involvement of C2H2 zinc finger proteins in fruit ripening. [ABSTRACT FROM AUTHOR]

The C1-2i wheat Q-type C2H2 zinc finger protein (ZFP) transcription factor subclass has been reported to play important roles in plant stress responses. This subclass of ZFPs has not been studied in hexaploid wheat ( Triticum aestivum) and we aimed to identify all members of this subclass and evaluate their responses to different abiotic stresses causing oxidative stress. Exploiting the recently published wheat draft genome sequence, we identified 53 members (including homoeologs from A, B and D genomes) of the C1-2i wheat Q-type C2H2 ZFPs ( TaZFPs) representing 21 genes. Evolution analysis revealed that 9 TaZFPs members are directly inherited from the parents Triticum urartu and Aegilops tauschii, while 15 diverged through neoploidization events. This TaZFP subclass is responsive to the oxidative stress generator HO and to high light, drought stress and flooding. Most TaZFPs are responsive to HO (37/53), high light (44/53), flooding (31/53) or drought (37/53); 32 TaZFPs were up-regulated by at least 3 stresses and 16 were responsive to all stresses tested. A large number of these TaZFPs were physically mapped on different wheat draft genome sequences with known markers useful for QTL mapping. Our results show that the C1-2i subclass of TaZFPs is associated with responses to different abiotic stresses and that most TaZFPs (30/53 or 57 %) are located on group 5 chromosomes known to be involved in environment adaptation. Detailed characterization of these novel wheat TaZFPs and their association to QTL or eQTL may help to design wheat cultivars with improved tolerance to abiotic stress. [ABSTRACT FROM AUTHOR]

Cucumber fruits that have tubercules and spines (trichomes) are known to possess a warty (Wty) phenotype. In this study, the tuberculate fruit gene Tu was identified by map-based cloning, and was found to encode a transcription factor (TF) with a single C2H2 zinc finger domain. Tu was identified in all 38 Wty lines examined, and was completely absent from all 56 non-warty ( nWty) lines. Cucumber plants transgenic for Tu (TCP) revealed that Tu was required for the Wty fruit phenotype. Subcellular localization showed that the fusion protein GFP-Tu was localized mainly to the nucleus. Based on analyses of semi-quantitative and quantitative reverse transcription polymerase chain reaction (RT-PCR), and mRNA in situ hybridization, we found that Tu was expressed specifically in fruit spine cells during development of fruit tubercules. Moreover, cytokinin ( CTK) content measurements and cytological observations in Wty and nWty fruits revealed that the Wty fruit phenotype correlated with high endogenous CTK concentrations. As a result of further analyses on the transcriptomic profile of the nWty fruit epidermis and TCP fruit warts, expression of CTK-associated genes, and hormone content in nWty fruit epidermis, Wty fruit warts and epidermis, and TCP fruit warts and epidermis, we found that Tu probably promoted CTK biosynthesis in fruit warts. Here we show that Tu could not be expressed in the glabrous and tubercule-free mutant line gl that contained Tu, this result that futher confirmed the epistatic effect of the trichome (spine) gene Gl over Tu. Taken together, these data led us to propose a genetic pathway for the Wty fruit trait that could guide future mechanistic studies. [ABSTRACT FROM AUTHOR]

C2H2 zinc finger is one of the most common motifs found in the transcription factors (TFs) in eukaryotes organisms, which have a broad range of functions, such as regulation of growth and development, stress tolerance and pathogenicity. Here, PlCZF1 was identified to encode a C2H2 zinc finger in the litchi downy blight pathogen Peronophythora litchii. PlCZF1 is conserved in P. litchii and Phytophthora species. In P. litchii, PlCZF1 is highly expressed in sexual developmental and early infection stages. We generated Δplczf1 mutants using the CRISPR/Cas9 method. Compared with the wild type, the Δplczf1 mutants showed no significant difference in vegetative growth and asexual reproduction, but were defective in oospore development and virulence. Further experiments revealed that the transcription of PlM90, PlLLP and three laccase encoding genes were down-regulated in the Δplczf1 mutant. Our results demonstrated that PlCZF1 is a vital regulator for sexual development and pathogenesis in P. litchii. [ABSTRACT FROM AUTHOR]

Background and aims: Al tolerance is one of the most important trait for worldwide crop production. Using microarrays, we previously identified a transcription factor belonging to the C2H2 zinc finger protein (ZFP) family associated with Al tolerance in wheat (Houde and Diallo, BMC Genomics 9:400, ). The current work aimed to identify specific members of the C2H2 ZFP family that are associated with Al tolerance. Methods: Wheat ESTs were used to assemble C2H2 ZFP family members that do not contain a classical EAR repressor domain. Specific primers were designed for qRT-PCR expression profiling of wheat root tips exposed to Al and HO Two Al-tolerant and sensitive wheat cultivars including a pair of near-isogenic lines differing in Al tolerance were used. Results: We reconstituted 16 wheat Q-type C2H2 ZFP. Expression profiling identified two transcripts ( TaZFP2 and TaZFP3) that accumulate rapidly upon exposure to Al or in response to HO in two tolerant wheat cultivars, including the tolerant near-isogenic line. Conclusion: The responsiveness of these transcripts to HO suggests that they may represent the wheat orthologs of ZFP transcription factors ZAT7 and ZAT12 that were shown to improve ROS tolerance in Arabidopsis. Thus, they may play a crucial role in the improvement of oxidative stress tolerance in wheat. [ABSTRACT FROM AUTHOR]

Asexual sporulation is the most common reproduction mode of fungi. Most filamentous fungi have two conidiation patterns, normal conidiation and microcycle conidiation, which may be regulated by nutritional conditions. Nitrogen source can affect the fungal conidiation pattern, but the regulatory mechanism is not fully understood. In this study, we report a C2H2 zinc finger protein, MaNCP1, which has typical transcription factor characteristics and is screened from the subtractive library regulated by nitrate in the entomopathogenic fungus Metarhizium acridum. MaNCP1 and its N-terminal play critical roles in the conidiation pattern shift. Further study shows that MaNCP1 interacts with MaNmrA , which also contributes to the conidiation pattern shift and is involved in the reductive pathway of nitric oxide (NO) synthesis. Intriguingly, the conidiation pattern of the MaNCP1 -disruption strain (Δ MaNCP1 ) can be restored to microcycle conidiation when grown on the microcycle conidiation medium, SYA, supplemented with NO donor or overexpressing MaNmrA in Δ MaNCP1 . Here, we reveal that MaNCP1 governs the conidiation pattern shift through regulating the reductive synthesis of NO by physically targeting MaNmrA in M. acridum. This work provides new mechanistic insights into how changes in nitrogen utilization are linked to the regulation of fungal morphological changes. IMPORTANCE Fungal conidia play important roles in the response to environmental stimuli and evasion of the host immune system. The nitrogen source is one of the main factors affecting shifts in fungal conidiation patterns, but the regulatory mechanism involved is not fully understood. In this work, we report that the C2H2 zinc finger protein, MaNCP1, governs the conidiation pattern shift in M. acridum by targeting the MaNmrA gene, thereby altering the regulation of the reductive pathway for NO synthesis. This work provides further insights into how the nutritional environment can regulate the morphogenesis of filamentous fungi.

Although root hair development in Arabidopsis thaliana has been extensively studied, it remains unknown whether the zinc finger proteins, the largest family of transcription factors in plants, are involved in this process. Here we report that the C2H2 zinc finger protein ZINC FINGER PROTEIN 5 (ZFP5) is a key regulator of root hair initiation and morphogenesis in Arabidopsis. ZFP5 is mainly expressed in root and preferentially in root hair cells. Using both zfp5 mutants and ZFP5 RNAi lines, we show that reduction in ZFP5 function leads to fewer and much shorter root hairs compared to wild-type. Genetic and molecular experiments demonstrate that ZFP5 exerts its effect on root hair development by directly promoting expression of the CAPRICE ( CPC) gene. Furthermore, we show that ZFP5 expression is induced by cytokinin, and that ZFP5 mediates cytokinin and ethylene effects on the formation and growth of root hairs. These results suggest that ZFP5 integrates various plant hormone cues to control root epidermal cell development in Arabidopsis. [ABSTRACT FROM AUTHOR]

The goal of this work was to characterize an Arabidopsis C2H2 single zinc finger gene, named AtZFP11, that is similar to SUPERMAN ( SUP) and RABBIT EARS ( RBE). No altered phenotype was observed in mutants analyzed that were derived through TILLING nor a T-DNA insertion into the exon of At ZFP11. Plants derived by intercrossing AtZFP11 to SUP and RBE alleles did not display any phenotype that could be attributed to At ZFP11. Transgenic plants constitutively overexpressing the AtZFP11 protein displayed severe abnormalities. Phenotypically normal transgenic plants that contained an ecdysone-inducible promoter system fused to the AtZFP11 gene were recovered. These transgenic plants displayed similar abnormalities once induced with methoxyfenozide. Microarray and RT-PCR analysis of gene expression in plants overexpressing At ZFP11 after induction revealed changes in gene expression of a number of genes involved in auxin, jasmonic acid, and stress responses. These results suggest that AtZFP11 plays a role in regulation of gene expression and may be functionally redundant or that conditions needed to observe phenotypic differences in the mutant plants were not provided in the course of these experiments. [ABSTRACT FROM AUTHOR]

During their development, plants are subjected to repeated and fluctuating wind loads, an environmental factor predicted to increase in importance by scenarios of global climatic change. Notwithstanding the importance of wind stress on plant growth and development, little is known about plant acclimation to the bending stresses imposed by repeated winds. The time-course of acclimation of young poplars (Populus tremula L.×P. alba L.) to multiple stem bendings is studied here by following diameter growth and the expression of four genes PtaZFP2, PtaTCH2, PtaTCH4, and PtaACS6, previously described to be involved in the mechanical signalling transduction pathway. Young trees were submitted either to one transient bending per day for several days or to two bendings, 1–14 days apart. A diminution of molecular responses to subsequent bending was observed as soon as a second bending was applied. The minimum rest periods between two successive loadings necessary to recover a response similar to that observed after a single bending, were 7 days and 5 days for growth and molecular responses, respectively. Taken together, our results show a desensitization period of a few days after a single transitory bending, indicating a day-scale acclimation of sensitivity to the type of wind conditions plants experience in their specific environment. This work establishes the basic kinetics of acclimation to low bending frequency and these kinetic analyses will serve as the basis of ongoing work to investigate the molecular mechanisms involved. Future research will also concern plant acclimation to higher wind frequencies. [ABSTRACT FROM PUBLISHER]

Abstract: The C2H2 zinc finger proteins form one of the largest families of transcriptional regulators in eukaryotes. We identified a Phytophthora sojae C2H2 zinc finger (PsCZF1), that is highly conserved in sequenced oomycete pathogens. In transformants of P. sojae containing the PsCZF1 promoter fused to the β-glucuronidase (GUS) reporter gene, GUS activity was highly induced in the P. sojae oospore stage and upregulated after infection. To elucidate the function of PsCZF1, its expression was silenced by introducing anti-sense constructs into P sojae. PsCZF1-silenced transformants did not exhibit altered cell size or morphology of sporangia and hyphae; however, hyphal growth rate was reduced by around 50% in the mutants. PsCZF1-deficient mutants were also impaired in production of oospores, swimming zoospores and germinating cysts, indicating that the gene is involved in various stages of the life cycle. Furthermore, we found that PsCZF1-deficient mutants lost virulence on host soybean cultivars. Our results suggest that this oomycete-specific C2H2-type zinc finger protein plays an important role in growth, development, and pathogenesis; therefore, PsCZF1 might be an attractive oomycete-specific target for chemical fungicide screening. [Copyright &y& Elsevier]

Abstract: The cis-regulatory region of target genes is key elements in the transcriptional regulation of gene expression. Many of these cis-regulatory regions have not been identified by either biological experiments or computational methods. Recently, a few additional C2H2 zinc finger transcription factor binding sites have been discovered. The majority of the zinc finger binding sites, however, are still unknown. In this study, we used publically available data to evaluate possible interaction patterns between nucleotides and the amino acids of zinc finger domains. We calculated the most probable state path of three nucleotides sequences using a Hidden Markov Model (HMM). We used these computations to predict C2H2 zinc finger transcription factor binding sites in cis-regulatory regions of their target genes (http://bioinfo.hanyang.ac.kr/ZIFIBI/frameset.php). [Copyright &y& Elsevier]

Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1. [ABSTRACT FROM AUTHOR]

Zinc finger proteins play important roles in a variety of cellular functions, including cell growth, proliferation, apoptosis, and intracellular signal transduction, and the zinc finger-containing transcription factor has been implicated as a critical regulator of multiple cardiac-expressed genes as well as a regulator of inducible gene expression in response to hypertrophic stimulation. With the aim of identifying the genes involved in human heart development and diseases, we have isolated a novel LER-related zinc finger gene named ZNF394 from human heart cDNA library. ZNF394 gene has a predicted 561-amino acid open reading frame, encoding a 64 kDa zinc finger protein. The N-terminus of ZNF394 protein has a leucine-rich region (LER or SCAN domain), followed by a well-conserved krűppel-associated box domain. The C-terminus of the protein contains 7 C2H2 zinc finger motifs in tandem arrays with the highly conserved space region of the H/C-link. ZNF394 gene is mapped to chromosome 7q11.21. Northern blot analysis indicates that a 2.18 kb transcript specific for ZNF394 is specifically expressed in the heart, skeletal muscle, and brain in human adult tissues. ZNF394 protein is expressed in cell nucleus. Overexpression of ZNF394 in the cell inhibits the transcriptional activities of c-Jun and AP-1 reporters, suggesting that ZNF394 is a new transcriptional repressor in mitogen-activated protein kinase signaling pathways and may play an important role in cardiac development and/or cardiac function. [Copyright &y& Elsevier]

The heterotrophic marine microalgae Schizochytrium sp. is an important industrial producer of docosahexaenoic acid (DHA). Increased production of DHA and lipids in Schizochytrium sp. has been achieved by standard fermentation optimization and metabolic engineering methods; however, regulatory mechanisms for DHA and lipid biosynthesis remain unknown. In this study, the C 2 H 2 zinc finger protein LipR was identified in Schizochytrium sp. ATCC 20888 by transcriptional analysis. Deletion of the lipR gene significantly ( P <  0.001) increased production of total lipids and DHA by 33% and 48%, respectively. LipR repressed DHA and lipid production by directly inhibiting transcription of polyunsaturated fatty acid (PUFA) and fatty acid synthase (FAS) genes ( pfa1 , pfa2 , pfa3 , and fas ). Specific binding of LipR to 9-bp recognition sequence 5'-(C/A)(A/G)CCATCTT-3' in upstream regions of target genes was demonstrated by electrophoretic mobility shift assays (EMSAs) and DNase I footprinting assays. Expression of several key genes ( acc , acl , ampD , fabD , mae , zwf , and dga1 ) related to levels of precursors and NADPH, and to triacylglycerol storage rate, were also directly repressed by LipR. Our findings, taken together, indicate that the evolutionarily unique regulator LipR is an essential repressor of DHA and saturated fatty acid biosynthesis in Schizochytrium sp. IMPORTANCE Regulatory mechanisms for DHA and saturated fatty acid biosynthesis in the heterotrophic marine microalgae Schizochytrium sp. are unclear. We demonstrate here that deletion of the gene ( lipR ) encoding the C 2 H 2 zinc finger protein LipR promotes DHA and saturated fatty acid production in this genus. LipR acts as a key repressor of such production by binding to 9-bp consensus sequence 5'-(C/A)(A/G)CCATCTT-3' in the upstream regions of polyunsaturated fatty acid and fatty acid synthase genes ( pfa1 , pfa2 , pfa3 , and fas ), and genes related to levels of precursors and NADPH ( acc , acl , ampD , fabD , mae , and zwf ), and to triacylglycerol storage rate ( dga1 ). This is the first demonstration that a regulator inhibits synthesis of DHA and lipids in Schizochytrium sp. by directly controlling transcription of PUFA synthase and fas genes. Manipulation of the lipR gene provides a potential strategy for enhancing accumulation of polyunsaturated fatty acids and lipids in thraustochytrids.

• Hydrophobins differ between two haplotype C. trogii genomes. • Clustering pattern and six-cysteine hydrophobins are found in C. trogii. • Hydrophobins participate in growth and development of C. trogii. Hydrophobins are small, secreted proteins with important physiological functions in mycelial growth and fungal development. Here, 1 nucleus-specific and 35 allelic hydrophobin genes were identified in the genome of a white rot fungus, Coriolopsis trogii. Among these, 22 were eight-cysteine class I hydrophobin genes and the other 14 were uncommon six-cysteine hydrophobin genes. The six-cysteine hydrophobins were speculated to have originated from a common ancestor. The hydrophobin genes favored a clustering distribution and two recent duplication pairs were identified. The genes had conserved gene structures with three exons and two introns. Cthyd18 , Cthyd19 , and Cthyd32 were constitutively highly expressed in all developmental stages. Cthyd20 , Cthyd21 , Cthyd22 , Cthyd28 , Cthyd30 , Cthyd31 , and Cthyd33 were highly expressed in mycelia, and Cthyd12 and Cthyd35 in the reproductive stages. Sixteen hydrophobin genes were regulated differently in the transition from mycelia to primordia; Cthyd35 showed maximal upregulation of 1922-fold, and Cthyd23 showed maximal downregulation of 552-fold. Most (32) hydrophobin genes showed significant differential expression between mycelia cultured in different media (potato dextrose agar or broth). Weighted gene co-expression network analysis and promoter analysis revealed that C2H2 zinc finger proteins may regulate hydrophobin genes. These results may support further research into the function and evolution of hydrophobins. [ABSTRACT FROM AUTHOR]

C2H2 zinc finger proteins (ZFPs) play important roles in plant development and response to abiotic stresses, and have been studied extensively. However, there are few studies on ZFPs in mangroves and mangrove associates, which represent a unique plant community with robust stress tolerance. MpZFP1, which is highly induced by salt stress in the mangrove associate Millettia pinnata, was cloned and functionally characterized in this study. MpZFP1 protein contains two zinc finger domains with conserved QALGGH motifs and targets to the nucleus. The heterologous expression of MpZFP1 in Arabidopsis increased the seeds' germination rate, seedling survival rate, and biomass accumulation under salt stress. The transgenic plants also increased the expression of stress-responsive genes, including RD22 and RD29A, and reduced the accumulation of reactive oxygen species (ROS). These results indicate that MpZFP1 is a positive regulator of plant responses to salt stress due to its activation of gene expression and efficient scavenging of ROS. [ABSTRACT FROM AUTHOR]

In animal systems the C2H2 zinc finger protein (ZFP) genefamily is the largest group of regulatory proteins and its members havea wide and important role in growth and development. It is likely thatthis family of ZFP transcription factors will also be important inplants. We have used a PCR approach employing highly degenerateoligonucleotide primers to isolate several Arabidopsis genomic andcDNA clones encoding potential ZFPs. In addition we have used thesequence information from these clones to identify two ESTs as membersof this family. Five two-fingered and one three-fingered ZFPs have beenidentified. Outside of the zinc finger regions, there is considerablesequence diversity, including the sequence and length of the interfingerregion; the expression pattern of each gene is different. This is thefirst report of the isolation of a three-fingered plant C2H2ZFP gene and we discuss its possible evolutionary origin fromtwo-fingered ZFP genes. [ABSTRACT FROM AUTHOR]

Aluminum (Al) toxicity limits plant growth and has a major impact on the agricultural productivity in acidic soils. The zinc-finger protein (ZFP) family plays multiple roles in plant development and abiotic stresses. Although previous reports have confirmed the function of these genes, their transcriptional mechanisms in wild soybean (Glycine soja) are unclear. In this study, GsGIS3 was isolated from Al-tolerant wild soybean gene expression profiles to be functionally characterized in Arabidopsis. Laser confocal microscopic observations demonstrated that GsGIS3 is a nuclear protein, containing one C2H2 zinc-finger structure. Our results show that the expression of GsGIS3 was of a much higher level in the stem than in the leaf and root and was upregulated under AlCl3, NaCl or GA3 treatment. Compared to the control, overexpression of GsGIS3 in Arabidopsis improved Al tolerance in transgenic lines with more root growth, higher proline and lower Malondialdehyde (MDA) accumulation under concentrations of AlCl3. Analysis of hematoxylin staining indicated that GsGIS3 enhanced the resistance of transgenic plants to Al toxicity by reducing Al accumulation in Arabidopsis roots. Moreover, GsGIS3 expression in Arabidopsis enhanced the expression of Al-tolerance-related genes. Taken together, our findings indicate that GsGIS3, as a C2H2 ZFP, may enhance tolerance to Al toxicity through positive regulation of Al-tolerance-related genes. [ABSTRACT FROM AUTHOR]

Cucumber is one of the most important vegetables in the world. The C2H2 zinc finger protein (C2H2-ZFP) family plays an important role in the growth development and abiotic stress responses of plants. However, there have been no systematic studies on cucumber. In this study, we performed a genome-wide study of C2H2-ZFP genes and analyzed their chromosomal location, gene structure, conservation motif, and transcriptional expression. In total, 101 putative cucumber C2H2-ZFP genes were identified and divided into six groups (I–VI). RNA-seq transcriptome data on different organs revealed temporal and spatial expression specificity of the C2H2-ZFP genes. Expression analysis of sixteen selected C2H2-ZFP genes in response to cold, drought, salt, and abscisic acid (ABA) treatments by real-time quantitative polymerase chain reaction showed that C2H2-ZFP genes may be involved in different signaling pathways. These results provide valuable information for studying the function of cucumber C2H2-ZFP genes in the future. [ABSTRACT FROM AUTHOR]

Growing evidence indicates that some grass species are more tolerant to various abiotic and biotic stresses than many crops. Zinc finger proteins play important roles in plant abiotic and biotic stresses. Although genes coding for these proteins have been cloned and identified in various plants, their function and underlying transcriptional mechanisms in the halophyte Zoysia japonica are barely known. In the present study, ZjZFN1 was isolated from Z. japonica using RACE method. Quantitative real time PCR results revealed that the expression of ZjZFN1 was much higher in leaf than in root and stem tissues, and induced by salt, cold or ABA treatment. The subcellular localization assay demonstrated that ZjZFN1 was localized to the nucleus. Expression of the ZjZFN1 in Arabidopsis thaliana improved seed germination and enhanced plant adaption to salinity stress with improved percentage of green cotyledons and growth status under salinity stress. Physiological and transcriptional analyses suggested that ZjZFN1 might, at least in part, influence reactive oxygen species accumulation and regulate the transcription of salinity responsive genes. Furthermore, RNA-sequencing analysis of ZjZFN1 -overexpressing plants revealed that ZjZFN1 may serve as a transcriptional activator in the regulation of stress responsive pathways, including phenylalanine metabolism, α-linolenic acid metabolism and phenylpropanoid biosynthesis pathways. Taken together, these results provide evidence that ZjZFN1 is a potential key player in plants' tolerance to salt stress, and it could be a valuable gene in Z. japonica breeding projects.

Cucumber fruits that have tubercules and spines (trichomes) are known to possess a warty (Wty) phenotype. In this study, the tuberculate fruit gene Tu was identified by map-based cloning, and was found to encode a transcription factor (TF) with a single C2 H2 zinc finger domain. Tu was identified in all 38 Wty lines examined, and was completely absent from all 56 non-warty (nWty) lines. Cucumber plants transgenic for Tu (TCP) revealed that Tu was required for the Wty fruit phenotype. Subcellular localization showed that the fusion protein GFP-Tu was localized mainly to the nucleus. Based on analyses of semi-quantitative and quantitative reverse transcription polymerase chain reaction (RT-PCR), and mRNA in situ hybridization, we found that Tu was expressed specifically in fruit spine cells during development of fruit tubercules. Moreover, cytokinin (CTK) content measurements and cytological observations in Wty and nWty fruits revealed that the Wty fruit phenotype correlated with high endogenous CTK concentrations. As a result of further analyses on the transcriptomic profile of the nWty fruit epidermis and TCP fruit warts, expression of CTK-associated genes, and hormone content in nWty fruit epidermis, Wty fruit warts and epidermis, and TCP fruit warts and epidermis, we found that Tu probably promoted CTK biosynthesis in fruit warts. Here we show that Tu could not be expressed in the glabrous and tubercule-free mutant line gl that contained Tu, this result that futher confirmed the epistatic effect of the trichome (spine) gene Gl over Tu. Taken together, these data led us to propose a genetic pathway for the Wty fruit trait that could guide future mechanistic studies., (© 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.)