RESUMO
Sporopollenin, as the main component of the pollen exine, is a highly resistant polymer that provides structural integrity under unfavourable environmental conditions. Tetraketone α-pyrone reductase 1 (TKPR1) is essential for sporopollenin formation, catalyzing the reduction of tetraketone carbonyl to hydroxylated α-pyrone. The functional role of TKPR1 in male sterility has been reported in flowering plants such as maize, rice, and Arabidopsis. However, the molecular cloning and functional characterization of TKPR1 in cotton remain unaddressed. In this study, we identified 68 TKPR1s from four cotton species, categorized into three clades. Transcriptomics and RT-qPCR demonstrated that GhTKPR1_8 exhibited typical expression patterns in the tetrad stage of the anther. GhTKPR1_8 was localized to the endoplasmic reticulum. Moreover, ABORTED MICROSPORES (GhAMS) transcriptionally activated GhTKPR1_8 as indicated by luciferase complementation tests. GhTKPR1_8-knockdown inhibited anther dehiscence and reduced pollen viability in cotton. Additionally, overexpression of GhTKPR1_8 in the attkpr1 mutant restored its male sterile phenotype. This study offers novel insights into the investigation of TKPR1 in cotton while providing genetic resources for studying male sterility.
Assuntos
Regulação da Expressão Gênica de Plantas , Gossypium , Proteínas de Plantas , Pólen , Pólen/genética , Pólen/fisiologia , Gossypium/genética , Gossypium/fisiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Flores/genética , Flores/fisiologia , Infertilidade das Plantas/genética , FilogeniaRESUMO
The sugar transporter protein (STP) family has been shown to play important roles in plant growth, development, and stress response. However, it has not been studied in cotton compared to other major crops. In this study, we identified 90 STP genes from four cotton species, performed bioinformatic analysis, and focused on the role of GhSTP18 in salt stress. According to our results, cotton STP proteins were divided into four subgroups according to the phylogenetic tree. A synteny analysis suggested that whole-genome duplication (WGD) and segmental duplication were key drivers in the expansion of the STP gene family. The transcriptomic data analysis showed that 29 GhSTP genes exhibited sink-specific expression. Quantitative real time-polymerase chain reaction (qRT-PCR) analyses revealed that expression of GhSTP18 was induced by salt treatment, heat treatment, cold treatment, and drought treatment, and continuously increased during a salt stress time course. Notably, GhSTP18 encodes a plasma membrane-localized galactose transporter. Suppression of GhSTP18 transcription by a virus-induced gene silencing (VIGS) assay reduced sensitivity to salt stress in cotton, indicating that GhSTP18 negatively regulates plant salt tolerance. These results provide an important reference and resource for further studying and deploying STP genes for cotton improvement.
Assuntos
Proteínas de Plantas , Estresse Fisiológico , Filogenia , Estresse Fisiológico/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Salino/genética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Açúcares , Gossypium/genética , Gossypium/metabolismo , Regulação da Expressão Gênica de PlantasRESUMO
Neutral/alkaline invertases (N/AINVs) are sucrose hydrolases with important roles in plants. In this study, 15, 15, 15, 29, and 30 N/AINVs were identified in the Gossypium species, G. raimondii, G. herbaceum, G. arboreum, G. hirsutum, and G. barbadense, respectively. Along with two previously discovered branches, α and ß, a new clade γ was first discovered in our study. Investigation of gene collinearity showed that whole-genome duplication (WGD) and polyploidization were responsible for the expansion of the N/AINV gene family in allopolyploid Gossypium. Moreover, expression patterns revealed that GhN/AINV3/13/17/23/24/28 from the ß clade is highly expressed during the period of fiber initiation. The invertase activity of GhN/AINV13 and GhN/AINV23 were confirmed by restoring defects of invertase-deficient yeast mutant SEY2102. Treatments of abiotic stress showed that most GhN/AINVs were induced in response to polyethylene glycol (PEG) or salt stress. A virus-induced gene-silencing (VIGS) experiment and yeast two-hybrid assay demonstrated that GhN/AINV13 may interact with their positive regulators Gh14-3-3 proteins and participate in the fiber initiation or stress tolerance of cotton. Our results provided fundamental information regarding N/AINVs and highlight their potential functions in cotton stress tolerance.
Assuntos
Gossypium/genética , Pressão Osmótica , Proteínas de Plantas/genética , Estresse Salino , beta-Frutofuranosidase/genética , Proteínas 14-3-3/metabolismo , Duplicação Gênica , Regulação da Expressão Gênica de Plantas , Gossypium/enzimologia , Gossypium/metabolismo , Proteínas de Plantas/metabolismo , beta-Frutofuranosidase/metabolismoRESUMO
Tubby-like protein genes (TULPs), present in the form of large multigene families, play important roles in environmental stress. However, little is known regarding the TULP family genes in cotton. In this study, we systematically identified and analyzed the membership, characterization, and evolutionary relationship of TULPs in four species of cotton. Transcriptome analysis indicated that GhTULPs participate in environmental stress and cotton tissue development. At the same time, we also predicted and analyzed the potential molecular regulatory mechanisms and functions of TULPs. GhTULP34, as a candidate gene, significantly reduced the germination rate of transgenic Arabidopsis plants under salt stress, and inhibited root development and stomatal closure under mannitol stress. The yeast two-hybrid and luciferase (LUC) systems showed that GhTULP34 can interact with GhSKP1A, a subunit of the SCF-type (Skp1-Cullin-1-F-box) complex. This study will provide a basis and reference for future research on their roles in stress tolerance.
Assuntos
Proteínas F-Box/metabolismo , Gossypium/genética , Pressão Osmótica , Proteínas de Plantas/metabolismo , Proteínas F-Box/genética , Gossypium/metabolismo , Proteínas de Plantas/genética , Ligação Proteica , Proteínas Ligases SKP Culina F-Box/genética , Proteínas Ligases SKP Culina F-Box/metabolismoRESUMO
Block of proliferation 1 (BOP1) is a key protein that helps in the maturation of ribosomes and promotes the progression of the cell cycle. However, its role in the leaf morphogenesis of cotton remains unknown. Herein, we report and study the function of GhBOP1 isolated from Gossypium hirsutum. The sequence alignment revealed that BOP1 protein was highly conserved among different species. The yeast two-hybrid experiments, bimolecular fluorescence complementation, and luciferase complementation techniques revealed that GhBOP1 interact with GhPES and GhWDR12. Subcellular localization experiments revealed that GhBOP1, GhPES and GhWDR12 were localized at the nucleolus. Suppression of GhBOP1 transcripts resulted in the uneven bending of leaf margins and the presence of young wrinkled leaves by virus-induced gene silencing assay. Abnormal palisade arrangements and the presence of large upper epidermal cells were observed in the paraffin sections of the wrinkled leaves. Meanwhile, a jasmonic acid-related gene, GhOPR3, expression was increased. In addition, a negative effect was exerted on the cell cycle and the downregulation of the auxin-related genes was also observed. These results suggest that GhBOP1 plays a critical role in the development of wrinkled cotton leaves, and the process is potentially modulated through phytohormone signaling.
Assuntos
Gossypium , Folhas de Planta , Regulação da Expressão Gênica de Plantas , Gossypium/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Acid invertase (AINV) is a kind of sucrose hydrolase with an important role in plants. Currently, the AINV genes have not been systematically studied in cotton. In this study, a total of 92 AINV genes were identified in five cotton species. The phylogenetic analysis revealed that the AINV proteins were divided into two subgroups in cotton: vacuolar invertase (VINV) and cell wall invertase (CWINV). The analysis of gene structures, conserved motifs, and three-dimensional protein structures suggested that GhAINVs were significantly conserved. The synteny analysis showed that whole-genome duplication was the main force promoting the expansion of the AINV gene family. The cis-element, transcriptome, and quantitative real time-polymerase chain reaction (qRT-PCR) showed that some GhAINVs were possibly associated with stress response. GhCWINV4, highly expressed in PEG treatment, was cloned, and subsequent virus-induced gene silencing assay confirmed that this gene was involved in the drought stress response. Overall, this study might be helpful for further analyzing the biological function of AINVs and provide clues for improving the resistance of cotton to stress.
Assuntos
Gossypium , beta-Frutofuranosidase , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Gossypium/genética , Gossypium/metabolismo , Família Multigênica , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Fisiológico/genéticaRESUMO
Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents, and metastatic OS is the major cause of OS-related death. Carboxypeptidase E (CPE) is known to be highly expressed in some cancer types, and its N-terminal truncated form, CPE-ΔN, is implicated in tumor metastasis and poor prognosis. In this study, we investigated the effect of CPE-ΔN on cell migration, invasiveness, and the epithelial-mesenchymal transition (EMT) of OS cells, and illustrated the molecular mechanisms. We first constructed CPE-ΔN overexpressing human OS cell lines (143B and U2OS cells), and found that ectopic CPE-ΔN expression in OS cells enhanced cell migration and invasiveness, and promoted the EMT process. Further, overexpression of CPE-ΔN increased the levels of c-myc and nuclear ß-catenin in OS cells, which suggested the CPE-ΔN promotes activation of the Wnt-ß-catenin pathway in OS cells. Treatment with ß-catenin small interfering RNA (siRNA) inhibited the migration and invasiveness of CPE-ΔN-overexpressing cells, and reduced the expression of E-cadherin. Together, these results suggest that CPE-ΔN promotes migration, invasiveness, and the EMT of OS cells via the Wnt-ß-catenin signaling pathway.
Assuntos
Carboxipeptidase H/metabolismo , Movimento Celular , Transição Epitelial-Mesenquimal , Invasividade Neoplásica , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Via de Sinalização Wnt , Carboxipeptidase H/biossíntese , Humanos , Osteossarcoma/enzimologia , Células Tumorais CultivadasRESUMO
The young leaves of virescent mutants are yellowish and gradually turn green as the plants reach maturity. Understanding the genetic basis of virescent mutants can aid research of the regulatory mechanisms underlying chloroplast development and chlorophyll biosynthesis, as well as contribute to the application of virescent traits in crop breeding. In this study, fine mapping was employed, and a recessive gene (v 1) from a virescent mutant of Upland cotton was narrowed to an 84.1-Kb region containing ten candidate genes. The GhChlI gene encodes the cotton Mg-chelatase I subunit (CHLI) and was identified as the candidate gene for the virescent mutation using gene annotation. BLAST analysis showed that the GhChlI gene has two copies, Gh_A10G0282 and Gh_D10G0283. Sequence analysis indicated that the coding region (CDS) of GhChlI is 1269 bp in length, with three predicted exons and one non-synonymous nucleotide mutation (G1082A) in the third exon of Gh_D10G0283, with an amino acid (AA) substitution of arginine (R) to lysine (K). GhChlI-silenced TM-1 plants exhibited a lower GhChlI expression level, a lower chlorophyll content, and the virescent phenotype. Analysis of upstream regulatory elements and expression levels of GhChlI showed that the expression quantity of GhChlI may be normal, and with the development of the true leaf, the increase in the Gh_A10G0282 dosage may partially make up for the deficiency of Gh_D10G0283 in the v 1 mutant. Phylogenetic analysis and sequence alignment revealed that the protein sequence encoded by the third exon of GhChlI is highly conserved across diverse plant species, in which AA substitutions among the completely conserved residues frequently result in changes in leaf color in various species. These results suggest that the mutation (G1082A) within the GhChlI gene may cause a functional defect of the GhCHLI subunit and thus the virescent phenotype in the v1 mutant. The GhChlI mutation not only provides a tool for understanding the associations of CHLI protein function and the chlorophyll biosynthesis pathway but also has implications for cotton breeding.
Assuntos
Clorofila/genética , Gossypium/genética , Filogenia , Proteínas de Plantas/genética , Clorofila/biossíntese , Cloroplastos/genética , Mapeamento Cromossômico , Regulação da Expressão Gênica de Plantas , Estudos de Associação Genética , Genoma de Planta , Fenótipo , Folhas de Planta/genética , Alinhamento de SequênciaRESUMO
Gossypium hirsutum L. is the most important fiber crop worldwide and contributes to more than 95% of global cotton production. Marker-assisted selection (MAS) is an effective approach for improving fiber quality, and quantitative trait loci (QTL) mapping of fiber quality traits is important for cotton breeding. In this study, a permanent intra-specific recombinant inbred line (RIL) population containing 137 families was used for fiber quality testing. Based on a previously reported high-density genetic map with an average marker distance of 0.63 cM, 186 additive QTLs were obtained for five fiber quality traits over five consecutive years, including 39 for fiber length (FL), 36 for fiber strength (FS), 50 for fiber uniformity (FU), 33 for micronaire (MC) and 28 for fiber elongation (FE). Three stable QTLs, qMC-A4-1, qMC-D2-3 and qFS-D9-1, were detected in four datasets, and another eight stable QTLs, qMC-A4-2, qMC-D11-2, qFU-A9-1, qFU-A10-4, qFS-D11-1, qFL-D9-2, qFL-D11-1 and qFE-A3-2, were detected in three datasets. The annotated genes in these 11 stable QTLs were collected, and these genes included many transcription factors with functions during fiber development. 33 QTL coincidence regions were found, and these involved nearly half of the total QTLs. Four chromosome regions containing at least 6 QTLs were promising for fine mapping. In addition, 41 pairs of epistatic QTLs (e-QTLs) were screened, including 6 for FL, 30 for FS, 2 for FU and 3 for MC. The identification of stable QTLs adds valuable information for further QTL fine mapping and gene positional cloning for fiber quality genetic detection and provides useful markers for further molecular breeding in enhancing fiber quality.
Assuntos
Mapeamento Cromossômico , Fibra de Algodão , Gossypium/genética , Endogamia , Locos de Características Quantitativas , Gossypium/metabolismoRESUMO
One of the major causes of damage to column-supported concrete structures, such as bridges and highways, are collisions from moving vehicles, such as cars and ships. It is essential to quantify the collision damage of the column so that appropriate actions can be taken to prevent catastrophic events. A widely used method to assess structural damage is through the root-mean-square deviation (RMSD) damage index established by the collected data; however, the RMSD index does not truly provide quantitative information about the structure. Conversely, the damage volume ratio that can only be obtained via simulation provides better detail about the level of damage in a structure. Furthermore, as simulation can also provide the RMSD index relating to that particular damage volume ratio, the empirically obtained RMSD index can thus be related to the structural damage degree through comparison of the empirically obtained RMSD index to numerically-obtained RMSD. Thus, this paper presents a novel method in which the impact-induced damage to a structure is simulated in order to obtain the relationship between the damage volume ratio to the RMSD index, and the relationship can be used to predict the true damage degree by comparison to the empirical RMSD index. In this paper, the collision damage of a bridge column by moving vehicles was simulated by using a concrete beam model subjected to continuous impact loadings by a freefalling steel ball. The variation in admittance signals measured by the surface attached lead zirconate titanate (PZT) patches was used to establish the RMSD index. The results demonstrate that the RMSD index and the damage ratio of concrete have a linear relationship for the particular simulation model.
RESUMO
Bolted spherical joints are widely used to form space steel structures. The stiffness and load capacity of the structures are affected by the looseness of bolted spherical joint connections in the structures. The looseness of the connections, which can be caused by fabrication error, low modeling accuracy, and "false twist" in the installation process, may negatively impact the load capacity of the structure and even lead to severe accidents. Furthermore, it is difficult to detect bolted spherical joint connection looseness from the outside since the bolts connect spheres with rods together from the inside. Active sensing methods are proposed in this paper to monitor the tightness status of the bolted spherical connection using piezoceramic transducers. A triangle-on-triangle offset grid composed of bolted spherical joints and steel tube bars was fabricated as the specimen and was used to validate the active sensing methods. Lead Zirconate Titanate (PZT) patches were used as sensors and actuators to monitor the bolted spherical joint tightness status. One PZT patch mounted on the central bolted sphere at the upper chord was used as an actuator to generate a stress wave. Another PZT patch mounted on the bar was used as a sensor to detect the propagated waves through the bolted spherical connection. The looseness of the connection can impact the energy of the stress wave propagated through the connection. The wavelet packet analysis and time reversal (TR) method were used to quantify the energy of the transmitted signal between the PZT patches by which the tightness status of the connection can be detected. In order to verify the effectiveness, repeatability, and consistency of the proposed methods, the experiments were repeated six times in different bolted spherical connection positions. The experimental results showed that the wavelet packet analysis and TR method are effective in detecting the tightness status of the connections. The proposed active monitoring method using PZT transducers can monitor the tightness levels of bolted spherical joint connections efficiently and shows its potential to guarantee the safety of space steel structures in construction and service.
RESUMO
Leaf senescence is defined as a deterioration process that continues to the final developmental stage of leaf. This process is usually regulated by both external and internal factors. There are about 5356 senescence associated genes belonging to 44 plant species. A great number of these genes were identified in Arabidopsis. Leaf senescence can be regulated by many transcription factors. In this study, nine gene families were selected according to their expression levels during leaf senescence from our laboratory database. Phylogenetic tree was constructed by MEGA6. Cultivated cotton CCRI-10 seeds were sown in the experimental field of Institute of Cotton Research of CAAS for profiling and leaf development stages analysis. For abiotic (drought and salt) stress and phytohormone (ABA, SA, ET and JA) treatments, CCRI-10 seeds were sown in potting soil at 25 °C in a chamber room. Total RNA was isolated from various samples and the cDNA prepared for qRT-PCR. The comparative CT method was applied to calculate the relative expression levels of genes. For phylogenetic tree, nine cotton genes were divided into two groups, most of homologous genes in previous studies showed roles in phytohormones and abiotic stress. Expression profiling of the nine genes showed different patterns of tissue specific expression. In leaf development stages, majority of cotton genes showed high expression in early and complete senescence stage. Furthermore, most of cotton genes have positive or negative response to phytohormones and abiotic stress. Based on the results of this study, we found four cotton genes CotAD_07559, CotAD_37422, CotAD_21204 and CotAD_54353 as candidate genes for leaves senescence and abiotic stress.
RESUMO
Premature leaf senescence occurs in the ultimate phase of the plant, and it occurs through a complex series of actions regulated by stress, hormones and genes. In this study, a proteomic analysis was performed to analyze the factors that could induce premature leaf senescence in two cotton cultivars. We successfully identified 443 differential abundant proteins (DAPs) from 7388 high-confidence proteins at four stages between non-premature senescence (NS) and premature senescence (PS), among which 158 proteins were over-accumulated, 238 proteins were down-accumulated at four stages, and 47 proteins displayed overlapped accumulation. All the DAPs were mapped onto 21 different categories on the basis of a Clusters of Orthologous Groups (COG) analysis, and 9 clusters were based on accumulation. Gene Ontology (GO) enrichment results show that processes related to stress responses, including responses to cold temperatures and responses to hormones, are significantly differentially accumulated. More importantly, the enriched proteins were mapped in The Arabidopsis Information Resource (TAIR), showing that 58 proteins play an active role in abiotic stress, hormone signaling and leaf senescence. Among these proteins, 26 cold-responsive proteins (CRPs) are significantly differentially accumulated. The meteorological data showed that the median temperatures declined at approximately 15 days before the onset of aging, suggesting that a decrease in temperature is tightly linked to an onset of cotton leaf senescence. Because accumulations of H2O2 and increased jasmonic acid (JA) were detected during PS, we speculate that two pathways associated with JA and H2O2 are closely related to premature leaf senescence in cotton.
Assuntos
Resposta ao Choque Frio , Gossypium/genética , Proteínas de Plantas/genética , Proteoma/genética , Ciclopentanos/metabolismo , Regulação da Expressão Gênica de Plantas , Gossypium/crescimento & desenvolvimento , Gossypium/metabolismo , Oxilipinas/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Proteoma/metabolismoRESUMO
BACKGROUND: Gossypium hirsutum L., or upland cotton, is an important renewable resource for textile fiber. To enhance understanding of the genetic basis of cotton earliness, we constructed an intra-specific recombinant inbred line population (RIL) containing 137 lines, and performed linkage map construction and quantitative trait locus (QTL) mapping. RESULTS: Using restriction-site associated DNA sequencing, a genetic map composed of 6,434 loci, including 6,295 single nucleotide polymorphisms and 139 simple sequence repeat loci, was developed from RIL population. This map spanned 4,071.98 cM, with an average distance of 0.63 cM between adjacent markers. A total of 247 QTLs for six earliness-related traits were detected in 6 consecutive years. In addition, 55 QTL coincidence regions representing more than 60 % of total QTLs were found on 22 chromosomes, which indicated that several earliness-related traits might be simultaneously improved. Fine-mapping of a 2-Mb region on chromosome D3 associated with five stable QTLs between Marker25958 and Marker25963 revealed that lines containing alleles derived from CCRI36 in this region exhibited smaller phenotypes and earlier maturity. One candidate gene (EMF2) was predicted and validated by quantitative real-time PCR in early-, medium- and late-maturing cultivars from 3- to 6-leaf stages, with highest expression level in early-maturing cultivar, CCRI74, lowest expression level in late-maturing cultivar, Bomian1. CONCLUSIONS: We developed an SNP-based genetic map, and this map is the first high-density genetic map for short-season cotton and has the potential to provide deeper insights into earliness. Cotton earliness-related QTLs and QTL coincidence regions will provide useful materials for QTL fine mapping, gene positional cloning and MAS. And the gene, EMF2, is promising for further study.
Assuntos
Mapeamento Cromossômico , Ligação Genética , Gossypium/genética , Locos de Características Quantitativas , DNA de Plantas/genética , DNA de Plantas/metabolismo , Marcadores Genéticos , Repetições de Microssatélites/genética , Fenótipo , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo RealRESUMO
BACKGROUND: Early maturity is one of the most important and complex agronomic traits in upland cotton (Gossypium hirsutum L). To dissect the genetic architecture of this agronomically important trait, a population consisting of 355 upland cotton germplasm accessions was genotyped using the specific-locus amplified fragment sequencing (SLAF-seq) approach, of which a subset of 185 lines representative of the diversity among the accessions was phenotypically characterized for six early maturity traits in four environments. A genome-wide association study (GWAS) was conducted using the generalized linear model (GLM) and mixed linear model (MLM). RESULTS: A total of 81,675 SNPs in 355 upland cotton accessions were discovered using SLAF-seq and were subsequently used in GWAS. Thirteen significant associations between eight SNP loci and five early maturity traits were successfully identified using the GLM and MLM; two of the 13 associations were common between the models. By computing phenotypic effect values for the associations detected at each locus, 11 highly favorable SNP alleles were identified for five early maturity traits. Moreover, dosage pyramiding effects of the highly favorable SNP alleles and significant linear correlations between the numbers of highly favorable alleles and the phenotypic values of the target traits were identified. Most importantly, a major locus (rs13562854) on chromosome Dt3 and a potential candidate gene (CotAD_01947) for early maturity were detected. CONCLUSIONS: This study identified highly favorable SNP alleles and candidate genes associated with early maturity traits in upland cotton. The results demonstrate that GWAS is a powerful tool for dissecting complex traits and identifying candidate genes. The highly favorable SNP alleles and candidate genes for early maturity traits identified in this study should be show high potential for improvement of early maturity in future cotton breeding programs.
Assuntos
Genoma de Planta , Gossypium/genética , Polimorfismo de Nucleotídeo Único/genética , Locos de Características Quantitativas/genética , Cruzamento , Mapeamento Cromossômico , Fibra de Algodão , Estudo de Associação Genômica Ampla , Genótipo , Desequilíbrio de Ligação , FenótipoRESUMO
BACKGROUND: Leaf senescence is an important developmental programmed degeneration process that dramatically affects crop quality and yield. The regulation of senescence is highly complex. Although senescence regulatory genes have been well characterized in model species such as Arabidopsis and rice, there is little information on the control of this process in cotton. Here, the senescence process in cotton (Gossypium hirsutum L.) leaves was investigated over a time course including young leaf, mature leaf and leaf samples from different senescence stages using RNA-Seq. RESULTS: Of 24,846 genes detected by mapping the tags to Gossypium genomes, 3,624 genes were identified as differentially expressed during leaf senescence. There was some overlap between the genes identified here and senescence-associated genes previously identified in other species. Most of the genes related to photosynthesis, chlorophyll metabolism and carbon fixation were downregulated; whereas those for plant hormone signal transduction were upregulated. Quantitative real-time PCR was used to evaluate the results of RNA-Seq for gene expression profiles. Furthermore, 519 differentially expressed transcription factors were identified, notably WRKY, bHLH and C3H. In addition, 960 genes involved in the metabolism and regulation of eight hormones were identified, of which many genes involved in the abscisic acid, brassinosteroid, jasmonic acid, salicylic acid and ethylene pathways were upregulated, indicating that these hormone-related genes might play crucial roles in cotton leaf development and senescence. However, most auxin, cytokinin and gibberellin pathway-related genes were downregulated, suggesting that these three hormones may act as negative regulators of senescence. CONCLUSIONS: This is the first high-resolution, multiple time-course, genome-wide comprehensive analysis of gene expression in cotton. These data are the most comprehensive dataset currently available for cotton leaf senescence, and will serve as a useful resource for unraveling the functions of many specific genes involved in cotton leaf development and senescence.
Assuntos
Regulação da Expressão Gênica de Plantas , Genoma de Planta , Gossypium/genética , Transcriptoma , Regulação da Expressão Gênica no Desenvolvimento , Gossypium/crescimento & desenvolvimento , Gossypium/metabolismo , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNARESUMO
SQUAMOSA promoter binding protein-like (SPL) genes encode plant-specific transcription factors that are involved in many fundamental developmental processes. Certain SPL genes contain sequences complementary to miR156, a microRNA (miRNA) that plays a role in modulating plant gene expression. In this study, 30 SPL genes were identified in the reference genome of Gossypium raimondii and 24 GhSPLs were cloned from Gossypium hirsutum. G. raimondii is regarded as the putative contributor of the D-subgenome of G. hirsutum. Comparative analysis demonstrated sequence conservation between GhSPLs and other plant species. GhSPL genes could be classified into seven subclades based on phylogenetic analysis, diverse intron-exon structure, and motif prediction. Within each subclade, genes shared a similar structure. Sequence and experimental analysis predicted that 18 GhSPL genes are putative targets of GhmiR156. Additionally, tissue-specific expression analysis of GhSPL genes showed that their spatiotemporal expression patterns during development progressed differently, with most genes having high transcript levels in leaves, stems, and flowers. Finally, overexpression of GhSPL3 and GhSPL18 in Arabidopsis plants demonstrated that these two genes are involved in the development of leaves and second shoots and play an integral role in promoting flowering. The flowering integrator GhSOC1 may bind to the promoter of GhSPL3 but not GhSPL18 to regulate flowering. In conclusion, our analysis of GhSPL genes will provide some gene resources and a further understanding of GhSPL3 and GhSPL18 function in flowering promotion. Furthermore, the comparative genomics and functional analysis deepened our understanding of GhSPL genes during upland cotton vegetative and reproductive growth.
Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Genômica , Gossypium/genética , Família Multigênica , Arabidopsis/genética , Sequência de Bases , Cromossomos de Plantas/genética , Éxons/genética , Flores/fisiologia , Íntrons/genética , Dados de Sequência Molecular , Motivos de Nucleotídeos/genética , Especificidade de Órgãos/genética , Fenótipo , Filogenia , Folhas de Planta/anatomia & histologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas/genética , Ligação Proteica , Alinhamento de SequênciaRESUMO
Family 1 GT, designated as UGT, is the largest and most functionally important multigene family in the plant kingdom. In this study, we carried out a genome-wide identification, analysis, and comparison of 142, 146, and 196 putative UGTs from Gossypium raimondii, Gossypium arboreum, and Gossypium hirsutum, respectively. All members present the 44 amino-acid conserved consensus sequence termed the plant secondary product glycosyltransferase motif. According to the phylogenetic relationship among the cotton UGT proteins and those from other species, GrUGTs and GaUGTs could be classified into 16 major phylogenetic groups (A-P), whereas GhUGTs are classified into 15 major phylogenetic groups with a lack of group C. All cotton UGTs are dispersed throughout the chromosomes and are displayed in clusters with the same open reading frame orientation. The expansion of them appears to result from genome duplication and rearrangement. Two conserved introns, A and B, are detected in most of the intron-containing-UGTs in G. raimondii and G. arboreum, whereas only intron A is detected in the intron-containing-UGTs in G. hirsutum. Furthermore, expression patterns of the UGT genes in G. hirsutum wild type and its near isogenic fuzzless-lintless mutant at the stage of fiber initiation were analyzed using the RNA-seq data. Overall, this study not only deepens our understanding of the structure, phylogeny, evolution, and expression of cotton UGT genes, but also provides a solid foundation for further cloning and functional studies of the UGT family genes.
Assuntos
Genoma de Planta , Estudo de Associação Genômica Ampla , Glicosiltransferases/genética , Gossypium/genética , Sequência de Aminoácidos , Cromossomos de Plantas , Éxons , Glicosiltransferases/química , Gossypium/classificação , Íntrons , Dados de Sequência Molecular , FilogeniaRESUMO
Many existing studies have demonstrated that common polymorphisms in the ABCA1 gene may play important roles in the development and progression of coronary heart disease (CHD), but individually published results are inconclusive. This meta-analysis aimed to derive a more precise estimation of the relationship between the ABCA1 rs4149313 polymorphism and CHD risk. We searched the CISCOM, CINAHL, Web of Science, PubMed, Google Scholar, EBSCO, Cochrane Library, and CBM databases from inception through 1 September 2013. Meta-analysis was performed using the STATA 12.0 software. Odds ratios (OR) and their 95% confidence intervals (CI) were estimated. Eleven case-control studies were included with a total of 5416 CHD patients and 20,897 healthy controls. Our meta-analysis results revealed that the ABCA1 rs4149313 polymorphism may be associated with an increased risk of CHD. Subgroup analysis by ethnicity suggested that there were significant associations between the ABCA1 rs4149313 polymorphism and an increased risk of CHD in Asian populations, but not in Caucasian populations (all P > 0.05). Meta-regression analyses showed that ethnicity may be a main source of heterogeneity. The present meta-analysis suggests that the ABCA1 rs4149313 polymorphism may contribute to the risk of CHD, especially in Asian populations.
Assuntos
Transportador 1 de Cassete de Ligação de ATP/genética , Alelos , Doença das Coronárias/genética , Predisposição Genética para Doença , Polimorfismo Genético , Estudos de Casos e Controles , Etnicidade/genética , Frequência do Gene , Genótipo , Humanos , Razão de Chances , Viés de PublicaçãoRESUMO
BACKGROUND: Male sterility is a common phenomenon in flowering plant species, and it has been successfully developed in several crops by taking advantage of heterosis. Using space mutation breeding of upland cotton, a novel photosensitive genetic male sterile (PGMS) mutant was isolated. To take advantage of the PGMS lines in cotton hybrid breeding, it is of great importance to study the molecular mechanisms of its male sterility. RESULTS: Delayed degradation of the PGMS anther tapetum occurred at different developmental stages as shown by analysis of anther cross-sections. To gain detailed insights into the cellular defects that occurred during PGMS pollen development, we used a differential proteomic approach to investigate the protein profiles of mutant and wild-type anthers at the tetrad, uninucleate and binucleate pollen stages. This approach identified 62 differentially expressed protein spots, including 19 associated with energy and metabolic pathways, 7 involved with pollen tube growth, 5 involved with protein metabolism, and 4 involved with pollen wall development. The remaining 27 protein spots were classified into other functional processes, such as protein folding and assembly (5 spots), and stress defense (4 spots). These differentially expressed proteins strikingly affected pollen development in the PGMS mutant anther and resulted in abnormal pollen grain formation, which may be the key reason for its male sterility. CONCLUSIONS: This work represents the first study using comparative proteomics between fertile and PGMS cotton plants to identify PGMS-related proteins. The results demonstrate the presence of a complicated metabolic network in anther development and advance our understanding of the molecular mechanisms of microgamete formation, providing insights into the molecular mechanisms of male sterility.