Trihelix Transcription Factor Research Articles

Genome wide identification of the trihelix transcription factors and overexpression of Gh_A05G2067 (GT-2), a novel gene contributing to increased drought and salt stresses tolerance in cotton.

We identified 102, 51 and 51 proteins encoded by the trihelix genes in Gossypium hirsutum, Gossypium arboreum and Gossypium raimondii, respectively. RNA sequence data and real‐time quantitative polymerase chain reaction analysis showed that Gh_A05G2067 (GT‐2) was highly upregulated under drought and salt stress conditions. Transient expression of GT‐2‐green fluorescent protein fusion protein in protoplast showed that GT‐2 was localized in the nucleus. The overexpression of GT‐2 conferred an enhanced drought tolerance to cotton, with lower malondialdehyde, hydrogen peroxide contents and higher reactive oxygen scavenging enzyme activities. Moreover, chlorophyll content, relative leaf water content (RLWC), excised leaf water loss (ELWL) and cell membrane stability (CMS) were relatively stable in the GT‐2‐overexpressed lines compared to wild‐type (WT). Similarly, stress‐responsive genes RD29A, SOS1, ABF4 and CBL1 were highly upregulated in the GT‐2‐overexpressed lines but were significantly downregulated in WT. In addition, the GT‐2‐silenced cotton plants exhibited a high level of oxidation injury, due to high levels of oxidant enzymes, in addition to negative effects on CMS, ELWL, RLWC and chlorophyll content. These results mark the foundation for future exploration of the trihelix genes in cotton, with an aim of developing more resilient, versatile and highly tolerant cotton genotypes.

Genome-Wide Characterization and Identification of Trihelix Transcription Factor and Expression Profiling in Response to Abiotic Stresses in Rice (Oryza sativa L.).

Trihelix transcription factors play a role in plant growth, development and various stress responses. Here, we identified 41 trihelix family genes in the rice genome. These OsMSLs (Myb/SANT-LIKE) were located on twelve chromosomes. Synteny analysis indicated only six duplicated gene pairs in the rice trihelix family. Phylogenetic analysis of these OsMSLs and the trihelix genes from other species divided them into five clusters. OsMSLs from different groups significantly diverged in terms of gene structure and conserved functional domains. However, all OsMSLs contained the same five cis-elements. Some of these were responsive to light and dehydration stress. All OsMSLs expressed in four tissues and six developmental stages of rice but with different expression patterns. Quantitative real-time PCR analysis revealed that the OsMSLs responded to abiotic stresses including drought and high salt stress and stress signal molecule including ABA (abscisic acid), hydrogen peroxide. OsMSL39 were simultaneously expressed under all treatments, while OsMSL28 showed high expression under hydrogen peroxide, drought, and high salt treatments. Moreover, OsMSL16/27/33 displayed significant expression under ABA and drought treatments. Nevertheless, their responses were regulated by light. The expression levels of the 12 chosen OsMSLs differed between light and dark conditions. In conclusion, our results helped elucidate the biological functions of rice trihelix genes and provided a theoretical basis for further characterizing their biological roles in responding to abiotic stresses.

The Trihelix transcription factor GT2-like 1 (GTL1) promotes salicylic acid metabolism, and regulates bacterial-triggered immunity.

The Trihelix Transcription factor GT2-like 1 (GTL1) was previously shown to be a key regulator of ploidy-dependent trichome growth and drought tolerance. Here, we report that GTL1 plays an important role in coordinating plant immunity. We show that gtl1 mutants are compromised in the regulation of basal immunity, microbial pattern-triggered immunity (PTI) and effector-triggered RIN4-mediated immunity. Transcriptome analysis revealed that GTL1 positively regulates defense genes and inhibits factors that mediate growth and development. By performing hormonal measurements and chromatin-immunoprecipitation studies, we found GTL1 to coordinate genes involved in salicylic acid metabolism, transport and response. Interaction studies and comparative transcriptomics to known data sets revealed that GTL1 is part of the MPK4 pathway and regulates oppositely the expression of differentially expressed genes in mpk4 plants. We introduced the gtl1 mutation in the mpk4 mutant and thereby partially suppressed its dwarfism and the high resistance against a bacterial invader. Our data show that GTL1 is part of the MPK4 pathway and acts as a positive regulator of bacterial-triggered immunity and SA homeostasis.

Genomic, expressional, protein-protein interactional analysis of Trihelix transcription factor genes in Setaria italia and inference of their evolutionary trajectory

BackgroundTrihelix transcription factors (TTF) play important roles in plant growth and response to adversity stress. Until now, genome-wide identification and analysis of this gene family in foxtail millet has not been available. Here, we identified TTF genes in the foxtail millet and its grass relatives, and characterized their functional domains.ResultsAs to sequence divergence, TTF genes were previously divided into five subfamilies, I-V. We found that Trihelix family members in foxtail millet and other grasses mostly preserved their ancestral chromosomal locations during millions of years’ evolution. Six amino acid sites of the SIP1 subfamily possibly were likely subjected to significant positive selection. Highest expression level was observed in the spica, with the SIP1 subfamily having highest expression level. As to the origination and expansion of the gene family, notably we showed that a subgroup of subfamily IV was the oldest, and therefore was separated to define a new subfamily O. Overtime, starting from the subfamily O, certain genes evolved to form subfamilies III and I, and later from subfamily I to develop subfamilies II and V. The oldest gene, Si1g016284, has the most structural changes, and a high expression in different tissues. What’s more interesting is that it may have bridge the interaction with different proteins.ConclusionsBy performing phylogenetic analysis using non-plant species, notably we showed that a subgroup of subfamily IV was the oldest, and therefore was separated to define a new subfamily O. Starting from the subfamily O, certain genes evolved to form other subfamilies. Our work will contribute to understanding the structural and functional innovation of Trihelix transcription factor, and the evolutionary trajectory.

Erratum to: Transcriptome profiling reveals an IAA-regulated response to adventitious root formation in lotus seedling.

Adventitious roots (ARs) of lotus (Nelumbonucifera Gaertn.) play a critical role in water and nutrient uptake. We found that exogenously applied 10-μM indole-3-acetic acid (IAA) promoted the formation of ARs, while 150-μM IAA significantly inhibited the emergence of ARs. However, little is known about these different responses to various concentrations of IAA at the molecular level. This study, therefore, examined the gene expression profiling in four libraries treated with 10- and 150-μM IAA based on the high-throughout tag sequencing technique. Approximately 2.4×107 clean tags were obtained after the removal of low-quality tags from each library respectively, among which about 10% clean tags were unambiguous tag-mapped genes to the reference genes. We found that some genes involved in auxin metabolism showed a similar tendency for expression in the A/CK and C/CK libraries, while three genes were enhanced their expression only in the A/CK libraries. Two transcription factors including B3 domain-containing protein At2g36080-like and trihelix transcription factor were up-regulated for transcriptional level in the A/C libraries. The expressions of six important genes related to AR formation were significantly different in the A/CK and C/CK libraries. In summary, this study provides a comprehensive understanding of gene expression regulated by IAA involved in AR formation in lotus.

Genome-Wide Regulatory Network Mapping of miRNA and Transcription Factors in Banana Roots

MicroRNA (miRNA) are important regulators of gene expression. Plant miRNA have been reported both to target and to be regulated by transcription factors, however, genomic distribution of miRNA, transcription factor targets for miRNA, transcription factor binding sites (TFBS) of miRNA promoters and their regulatory networks have not been systematically mapped in banana. In this study, genome-wide annotation of miRNA in the most recently published banana genome sequence was used to predict miRNA promoter regions and to map TFBS of miRNA genes. A total of 183 mature miRNAs, comprising 144 orthologous miRNA and 39 Musa-specific miRNA were predicted. Following this, banana root degradome data was used to confirm miRNA targets and the transcription factor targets were placed into a predicted network together with their targeting miRNA using cytoscape. Gene ontology of the 20 transcription factors among the predicted miRNA targets, showed predominance for auxin-activated signalling and developmental processes. Profiling of TFBS motifs across miRNA promoter regions showed that binding site motifs for TCP, AP2/ERF, GATA, NF-YB, DOF, B3, bZIP, trihelix, ZF-HD, bHLH and Dehydrin transcription factor families are abundant in the Musa acuminata genome. Finally, we propose a regulatory network for the miRNA families miR156, miR164, miR166, miR171, miR319, miR396, miR528, mac-miR-new14 and mac-miR-new20 and their respective transcription factor targets.

ShCIGT, a Trihelix family gene, mediates cold and drought tolerance by interacting with SnRK1 in tomato

Abiotic stress, such as drought and cold stress, have a major impact on plant growth and development. The trihelix transcription factor family plays important roles in plant morphological development and adaptation to abiotic stresses. In this study, we isolated a cold-induced gene named ShCIGT from the wild tomato species Solanum habrochaites and found that it contributes to abiotic stress tolerance. ShCIGT belongs to the GT-1 subfamily of the trihelix transcription factors. It was constitutively expressed in various tissues. Its expression was induced by multiple abiotic stresses and abscisic acid (ABA). Overexpression of ShCIGT in cultivated tomato enhanced cold and drought stress tolerance. In addition, the transgenic plants displayed a reduced sensitivity to ABA during post-germination growth. We found that ShCIGT interacts with SnRK1, an energy sensor in the metabolic signaling network, which controls plant metabolism, growth and development, and stress tolerance. Based on these data, we conclude ShCIGT may improve abiotic-stress tolerance in tomato by interacting with SnRK1.

Transcriptome profiling reveals an IAA-regulated response to adventitious root formation in lotus seedling.

Adventitious roots (ARs) of lotus (Nelumbonucifera Gaertn.) play a critical role in water and nutrient uptake. We found that exogenously applied 10-μM indole-3-acetic acid (IAA) promoted the formation of ARs, while 150-μM IAA significantly inhibited the emergence of ARs. However, little is known about these different responses to various concentrations of IAA at the molecular level. This study, therefore, examined the gene expression profiling in four libraries treated with 10- and 150-μM IAA based on the high-throughout tag sequencing technique. Approximately 2.4×107 clean tags were obtained after the removal of low-quality tags from each library respectively, among which about 10% clean tags were unambiguous tag-mapped genes to the reference genes. We found that some genes involved in auxin metabolism showed a similar tendency for expression in the A/CK and C/CK libraries, while three genes were enhanced their expression only in the A/CK libraries. Two transcription factors including B3 domain-containing protein At2g36080-like and trihelix transcription factor were up-regulated for transcriptional level in the A/C libraries. The expressions of six important genes related to AR formation were significantly different in the A/CK and C/CK libraries. In summary, this study provides a comprehensive understanding of gene expression regulated by IAA involved in AR formation in lotus.

Arabidopsis thaliana Trihelix Transcription Factor AST1 Mediates Salt and Osmotic Stress Tolerance by Binding to a Novel AGAG-Box and Some GT Motifs.

Trihelix transcription factors are characterized by containing a conserved trihelix (helix-loop-helix-loop-helix) domain that binds to GT elements required for light response, and they play roles in light stress and in abiotic stress responses. However, only a few of them have been functionally characterized. In the present study, we characterized the function of AST1 (Arabidopsis SIP1 clade Trihelix1) in response to salt and osmotic stress. AST1 shows transcriptional activation activity, and its expression is induced by osmotic and salt stress. A conserved sequence highly present in the promoters of genes regulated by AST1 was identified, which was bound by AST1, and termed the AGAG-box with the sequence [A/G][G/A][A/T]GAGAG. Additionally, AST1 also binds to some GT motifs including the sequence of GGTAATT, TACAGT, GGTAAAT and GGTAAA, but failed in binding to the sequence of GTTAC and GGTTAA. Chromatin immunoprecipitation combined with quantitative real-time reverse transcription-PCR analysis suggested that AST1 binds to the AGAG-box and/or some GT motifs to regulate the expression of stress tolerance genes, resulting in reduced reactive oxygen species, Na+ accumulation, stomatal apertures, lipid peroxidation, cell death and water loss rate, and increased proline content and reactive oxygen species scavenging capability. These physiological changes affected by AST1 finally improve salt and osmotic tolerance.

GTL1 and DF1 regulate root hair growth through transcriptional repression of ROOT HAIR DEFECTIVE 6-LIKE 4 in Arabidopsis

ABSTRACTHow plants determine the final size of growing cells is an important, yet unresolved, issue. Root hairs provide an excellent model system with which to study this as their final cell size is remarkably constant under constant environmental conditions. Previous studies have demonstrated that a basic helix-loop helix transcription factor ROOT HAIR DEFECTIVE 6-LIKE 4 (RSL4) promotes root hair growth, but how hair growth is terminated is not known. In this study, we demonstrate that a trihelix transcription factor GT-2-LIKE1 (GTL1) and its homolog DF1 repress root hair growth in Arabidopsis. Our transcriptional data, combined with genome-wide chromatin-binding data, show that GTL1 and DF1 directly bind the RSL4 promoter and regulate its expression to repress root hair growth. Our data further show that GTL1 and RSL4 regulate each other, as well as a set of common downstream genes, many of which have previously been implicated in root hair growth. This study therefore uncovers a core regulatory module that fine-tunes the extent of root hair growth by the orchestrated actions of opposing transcription factors.

Ubiquitin-related genes are differentially expressed in isogenic lines contrasting for pericarp cell size and grain weight in hexaploid wheat

BackgroundThere is an urgent need to increase global crop production. Identifying and combining specific genes controlling distinct biological processes holds the potential to enhance crop yields. Transcriptomics is a powerful tool to gain insights into the complex gene regulatory networks that underlie such traits, but relies on the availability of a high-quality reference sequence and accurate gene models. Previously, we identified a grain weight QTL on wheat chromosome 5A (5A QTL) which acts during early grain development to increase grain length through cell expansion in the pericarp. In this study, we performed RNA-sequencing on near isogenic lines (NILs) segregating for the 5A QTL and used the latest gene models to identify differentially regulated genes and pathways that potentially influence pericarp cell size and grain weight in wheat.ResultsWe sampled grains at 4 and 8 days post anthesis and found that genes associated with metabolism, biosynthesis, proteolysis and the defence response are upregulated during this stage of grain development in both NILs. We identified a specific set of 112 transcripts differentially expressed (DE) between 5A NILs at either time point, including eight potential candidates for the causal 5A gene and its downstream targets. The 112 DE transcripts had functional annotations including non-coding RNA, transposon-associated, cell-cycle control, ubiquitin-related, heat-shock, transcription and histone-related. Many of the genes identified belong to families that have been previously associated with seed/grain development in other species. Notably, we identified DE transcripts at almost all steps of the pathway associated with ubiquitin-mediated protein degradation. In the promoters of a subset of DE transcripts we identified enrichment of binding sites associated with C2H2, MYB/SANT, YABBY, AT-HOOK and Trihelix transcription factor families.ConclusionsIn this study, we identified DE transcripts with a diverse range of predicted biological functions, reflecting the complex nature of the pathways that control early grain development. Few of these are the direct orthologues of grain size genes in other species and none have been previously characterised in wheat. Further functional characterisation of these candidates and how they interact will provide novel insights into the control of grain size in cereals.

Ectopic expression of Limonium bicolor (Bag.) Kuntze DREB (LbDREB) results in enhanced salt stress tolerance of transgenic Populus ussuriensis Kom

High salinity is one of the most common abiotic stresses that plants have to confront, and selecting salt-tolerant plants is becoming a very important task in breeding. In this study, non-transgenic (NT) and three LbDREB-transgenic Populus ussuriensis Kom. (Dr22, Dr8, and Dr2) lines were treated with 150 mM NaCl. The salt tolerance of NT, Dr2, Dr8, and Dr22 were evaluated and compared. Phenotypic observation, chlorophyll content detection and leaf salt injury index calculation suggested NT exhibited more severe salt injury symptoms and weaker photosynthetic capacity than Dr2, Dr8, and Dr22. LbDREB gene exhibited opposite temporal expression patterns with no apical meristem (NAM), trihelix transcription factor (GT-1) and WRKY transcription factor 70 (WRKY70), and consistent expression patterns with Cu–Zn superoxide dismutase and peroxidase-PO1 genes, in Dr2, Dr8, and Dr22 under salt stress. The superoxide dismutase and peroxidase activities of transgenic lines were higher than NT plants during the salt stress treatment. Malonaldehyde accumulation in NT leaves was more significant than in transgenic leaves, while proline accumulation in NT leaves was much less than that in transgenic leaves under salt stress. The root to shoot ratios were significantly increased in Dr2, Dr8, and Dr22 plants, but were slightly decreased in NT plants, 12 days after salt stress treatment. The relative water content of NT leaves decreased more than transgenic leaves, meanwhile, the relative electrolytic leakage of NT leaves increased more significantly than transgenic leaves after salt stress. In summary, Dr2, Dr8, and Dr22 exhibited stronger salt stress tolerance than NT. The transgenic lines Dr2, Dr8, and Dr22 can serve as alternative salt-tolerant germplasm resources in P. ussuriensis breeding.

Trihelix family transcription factors in tea plant (Camellia sinensis): identification, classification, and expression profiles response to abiotic stress

Tea plant is an economic crop in China. Trihelix family transcription factor, also known as GT factor, is one of plant-specific TF families in plant. Its functions have been recognized in the development and stress tolerance in various plants. However, the trihelix TFs and its roles in abiotic stress tolerance in tea plant remains unclear. In this study, ten putative trihelix family TFs were identified from the transcriptome of tea plant, and divided into five clades. The leaves of two tea plant cultivars (‘Yingshuang’ and ‘Huangjinya’) under four abiotic stress treatments (38, 4 °C, salt and drought) were collected. Then, the expression profiles of two genes under abiotic stress treatments were detected and analyzed by qRT-PCR. The protein–protein interactions of trihelix family between C. sinensis and Arabidopsis were also performed. The expression levels of CsGT1-3 and CsGT2-1 were varied in the two tea plant cultivars. Meanwhile, CsGT1-3 and CsGT2-1 were differentially induced by the abiotic stresses. Up-regulation of CsGT1-3 and CsGT2-1 expressions by the induction of salt stress was observed. Our results might suggest that the transcriptome-based analysis was considered to be an available method to identify the identification and characterization of trihelix family TFs in C. sinensis.

PETAL LOSS and ROXY1 Interact to Limit Growth Within and between Sepals But to Promote Petal Initiation in Arabidopsis thaliana.

The activity of genes controlling organ development may be associated with the redox state of subregions within the meristem. Glutaredoxins react to the level of oxidative potential and can reduce cysteine dithiols, in some cases to activate specific transcription factors. In Arabidopsis, loss of function of the glutaredoxin ROXY1 or the trihelix transcription factor PETAL LOSS (PTL) each results in reduced numbers of petals. Here, genetic studies have revealed that loss of petals in ptl mutant plants depends on ROXY1 function. The two genes also act together to restrain stamen-identifying C function from entering the outer whorls. On the other hand, they suppress growth between sepals and in sepal margins, with ROXY1 action partially redundant to that of PTL. Genetic interactions with aux1 mutations indicate that auxin activity is reduced in the petal whorl of roxy1 mutants as in ptl mutants. However, it is apparently increased in the sepal whorl of triple mutants associated with the ectopic outgrowth of sepal margins, and of finger-like extensions of inter-sepal zones that in 20% of cases are topped with bunches of ectopic sepals. These interactions may be indirect, although PTL and ROXY1 proteins can interact directly when co-expressed in a transient assay. Changes of conserved cysteines within PTL to similar amino acids that cannot be oxidized did not block its function. It may be in some cases that under reducing conditions ROXY1 binds PTL and activates it by reducing specific conserved cysteines, thus resulting in growth suppression.

A single base change explains the independent origin of and selection for the nonshattering gene in African rice domestication.

Reduced seed shattering was a critical evolutionary step in crop domestication. Two cultivated rice species, Oryza sativa and Oryza glaberrima, were independently domesticated from the wild species Oryza rufipogon in Asia and Oryza barthii in Africa, respectively. A single nucleotide polymorphism (SNP) in the c gene, which encodes a trihelix transcription factor, causes nonshattering in O.sativa. However, the genetic mechanism of nonshattering in O.glaberrima is poorly understood. We conducted an association analysis for the coding sequences of SH3/SH4 in AA- genome rice species and the mutation suggested to cause nonshattering was demonstrated to do so using a positional-cloning approach in the O.sativa genetic background. We found that the loss of seed shattering in O.glaberrima was caused by an SNP resulting in a truncated SH3/SH4 protein. This mutation appears to be endemic and to have spread in the African gene pool by hybridization with some O.barthii accessions. We showed that interaction between the O.sativa and O.glaberrima domestication alleles of SH3 in heterozygotes induces a 'throwback' seed-shattering phenotype similar to that in the wild species. Identification of the causative SNP provides new insights into the molecular basis of seed shattering in crops and may facilitate investigation of the history of African rice domestication.

Identification of GT Factors in Response to Stresses and Leaf Senescence in Gossypium hirsutum L.

GT factors, also called trihelix transcription factors (TFs), belong to a small family of TFs and were first discovered because of their unique binding to GT elements. Recent studies have demonstrated the functions of GT factors in plant development and stress. Cotton is a major source of fiber and oilseed. However, studies about the GT factors in cotton are limited, especially with regard to their function and evolution. Therefore, it is important to study the GT factors in cotton. This study was based on three genome databases of cotton (A, D, and AD) and the GT factors in Arabidopsis. We retrieved 39 putative GhGT genes in Gossypium hirsutum, 14 GaGT in G. arboreum, and 14 GrGT in G. raimondii. For 39 GhGT genes, we characterized their motif features, domain sequence alignments, and phylogenetic relationship with other species. To understand the functions of GT factors in the senescent cotton leaves, we isolated eight GhGT genes from yellow leaves. In addition, we detected the expression patterns of these eight genes in that particular tissue as well as their effects on fiber development and responses to stresses and plant hormones, especially in leaf development, using real-time quantitative PCR. In addition, we also studied the function of GhGT31 in Arabidopsis. The results not only laid the foundation for an understanding of the functions of the GT factors in cotton but also provided significant information about the effects of GT factors on cotton development and on the responses to various stresses.

The Wheat GT Factor TaGT2L1D Negatively Regulates Drought Tolerance and Plant Development.

GT factors are trihelix transcription factors that specifically regulate plant development and stress responses. Recently, several GT factors have been characterized in different plant species; however, little is known about the role of GT factors in wheat. Here, we show that TaGT2L1A, TaGT2L1B, and TaGT2L1D are highly homologous in hexaploid wheat, and are localized to wheat chromosomes 2A, 2B, and 2D, respectively. These TaGT2L1 genes encode proteins containing two SANT domains and one central helix. All three homologs were ubiquitously expressed during wheat development and were responsive to osmotic stress. Functional analyses demonstrated that TaGT2L1D acts as a transcriptional repressor; it was able to suppress the expression of AtSDD1 in Arabidopsis by binding directly to the GT3 box in its promoter that negatively regulates drought tolerance. TaGT2L1D overexpression markedly increased the number of stomata and reduced drought tolerance in gtl1-3 plants. Notably, ectopic expression of TaGT2L1D also affected floral organ development and overall plant growth. These results demonstrate that TaGT2L1 is an ortholog of AtGTL1, and that it plays an evolutionarily conserved role in drought resistance by fine tuning stomatal density in wheat. Our data also highlight the role of TaGT2L1 in plant growth and development.

Transcriptome-Wide Identification and Expression Profiling Analysis of Chrysanthemum Trihelix Transcription Factors.

Trihelix transcription factors are thought to feature a typical DNA-binding trihelix (helix-loop-helix-loop-helix) domain that binds specifically to the GT motif, a light-responsive DNA element. Members of the trihelix family are known to function in a number of processes in plants. Here, we characterize 20 trihelix family genes in the important ornamental plant chrysanthemum (Chrysanthemum morifolium). Based on transcriptomic data, 20 distinct sequences distributed across four of five groups revealed by a phylogenetic tree were isolated and amplified. The phylogenetic analysis also identified four pairs of orthologous proteins shared by Arabidopsis and chrysanthemum and five pairs of paralogous proteins in chrysanthemum. Conserved motifs in the trihelix proteins shared by Arabidopsis and chrysanthemum were analyzed using MEME, and further bioinformatic analysis revealed that 16 CmTHs can be targeted by 20 miRNA families and that miR414 can target 9 CmTHs. qPCR results displayed that most chrysanthemum trihelix genes were highly expressed in inflorescences, while 20 CmTH genes were in response to phytohormone treatments and abiotic stresses. This work improves our understanding of the various functions of trihelix gene family members in response to hormonal stimuli and stress.

Extensive Transcriptome Changes During Natural Onset and Release of Vegetative Bud Dormancy in Populus.

To survive winter, many perennial plants become endodormant, a state of suspended growth maintained even in favorable growing environments. To understand vegetative bud endodormancy, we collected paradormant, endodormant, and ecodormant axillary buds from Populus trees growing under natural conditions. Of 44,441 Populus gene models analyzed using NimbleGen microarrays, we found that 1,362 (3.1%) were differentially expressed among the three dormancy states, and 429 (1.0%) were differentially expressed during only one of the two dormancy transitions (FDR p-value < 0.05). Of all differentially expressed genes, 69% were down-regulated from paradormancy to endodormancy, which was expected given the lower metabolic activity associated with endodormancy. Dormancy transitions were accompanied by changes in genes associated with DNA methylation (via RNA-directed DNA methylation) and histone modifications (via Polycomb Repressive Complex 2), confirming and extending knowledge of chromatin modifications as major features of dormancy transitions. Among the chromatin-associated genes, two genes similar to SPT (SUPPRESSOR OF TY) were strongly up-regulated during endodormancy. Transcription factor genes and gene sets that were atypically up-regulated during endodormancy include a gene that seems to encode a trihelix transcription factor and genes associated with proteins involved in responses to ethylene, cold, and other abiotic stresses. These latter transcription factors include ETHYLENE INSENSITIVE 3 (EIN3), ETHYLENE-RESPONSIVE ELEMENT BINDING PROTEIN (EBP), ETHYLENE RESPONSE FACTOR (ERF), ZINC FINGER PROTEIN 10 (ZAT10), ZAT12, and WRKY DNA-binding domain proteins. Analyses of phytohormone-associated genes suggest important changes in responses to ethylene, auxin, and brassinosteroids occur during endodormancy. We found weaker evidence for changes in genes associated with salicylic acid and jasmonic acid, and little evidence for important changes in genes associated with gibberellins, abscisic acid, and cytokinin. We identified 315 upstream sequence motifs associated with eight patterns of gene expression, including novel motifs and motifs associated with the circadian clock and responses to photoperiod, cold, dehydration, and ABA. Analogies between flowering and endodormancy suggest important roles for genes similar to SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL), DORMANCY ASSOCIATED MADS-BOX (DAM), and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1).

Cloning and functional analysis of the cotton Trihelix transcription factor GhGT29

Trihelix transcription factors are important proteins involved in response to abiotic stresses in plants. Understanding the molecular mechanisms of Trihelix in cottons will lay the foundation to improve stress tolerance by gene engineering. In this study, a gene encoding Trihelix transcription factor was isolated in upland cottons using reverse transcription PCR according to bioinformatic analysis. The gene was named as GhGT29 (GenBank accession No. JQ013097), which was 1 092 bp, contained a 1 089 bp open reading frame and encoded a protein of 363 amino acids with a predicted molecular weight of 40.9 kDa and a isoelectric point of 5.45. SMART analysis showed GhGT29 contained one typical SANT motif. Phylogenetic analysis showed that GhGT29 belonged to the SH4 subfamily of the Trihelix family and was most closely related to AtSH4-like1 and AtSH4-like2. Quantitative real-time PCR (qRT-PCR) analysis revealed that GhGT29 was induced by high salt, drought, cold and abscisic acid. The expression profile also revealed that GhGT29 was constitutively expressed in all tested tissues, such as roots, stems, leaves, flowers, ovules (0 DPA) and fibers (12 DPA). The expression level of GhGT29 was the highest in flowers and the lowest in stems. Using the Arabidopsis protoplasts assay system, we found that the GhGT29 protein was located in cell nuclei and had trans-activation activity. These results revealed that GhGT29 might be involved in the regulation of stress resistance-related genes in stress signaling pathways in upland cottons.