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1.
Gene ; 758: 144954, 2020 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-32683079

RESUMO

Teosinte branched1/cycloidea/proliferating cell factor1 (TCP) is a plant-specific protein family member involved in plant growth and development. However, the functions of most members of the cotton TCP family are unknown. In this study, the GbTCP5 gene encodes a sea-island cotton class II TCP CIN subclass transcription factor. The GbTCP5 transcription factor is located in the nucleus, has transcriptional activation activity, and can bind to TCP II cis-acting elements. GbTCP5 was widely expressed in tissues with the highest transcript level in the calyx. GbTCP5 is expressed at different developmental stages of the fiber and has significantly high transcriptional level expression in the fibers at 20, 30 and 35 days post anthesis (DPA). Heterologous overexpression of the GbTCP5 gene increased root hair length, root hair and stem trichome density, and stem lignin content in transgenic Arabidopsis compared to the wild type (WT). GbTCP5 binds the promoters of the GL3, EGL3, CPC, MYB46, LBD30, CesA4, VND7, CCOMT1, and CAD5 genes to upregulate their expression. Moreover, the homologous genes of these genes are expressed in the fibers of different developmental stages of the sea-island cotton fiber. These results indicate that GbTCP5 regulates root hair development and secondary wall formation in Arabidopsis and may be a candidate gene for improving cotton fiber quality.


Assuntos
Arabidopsis/genética , Gossypium/genética , Lignina/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Fatores de Transcrição/genética , Tricomas/crescimento & desenvolvimento , Arabidopsis/crescimento & desenvolvimento , Fibra de Algodão/análise , Proteínas de Ligação a DNA/genética , Raízes de Plantas/fisiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Regiões Promotoras Genéticas/genética , Ativação Transcricional/genética
2.
PLoS One ; 15(7): e0235344, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32628728

RESUMO

A Glycine max (soybean) hemicellulose modifying gene, xyloglucan endotransglycoslase/hydrolase (XTH43), has been identified as being expressed within a nurse cell known as a syncytium developing within the soybean root undergoing the process of defense to infection by the parasitic nematode, Heterodera glycines. The highly effective nature of XTH43 overexpression in suppressing H. glycines parasitism in soybean has led to experiments examining whether the heterologous expression of XTH43 in Gossypium hirsutum (upland cotton) could impair the parasitism of Meloidogyne incognita, that form a different type of nurse cell called a giant cell that is enclosed within a swollen root structure called a gall. The heterologous transgenic expression of XTH43 in cotton resulted in an 18% decrease in the number of galls, 70% decrease in egg masses, 64% decrease in egg production and a 97% decrease in second stage juvenile (J2) production as compared to transgenic controls. The heterologous XTH43 expression does not significantly affect root mass. The results demonstrate XTH43 expression functions effectively in impairing the development of M. incognita at numerous life cycle stages occurring within the cotton root. The experiments reveal that there are highly conserved aspects of the defense response of G. max that can function effectively in G. hirsutum to impair M. incognita having a different method of parasitism.


Assuntos
Glicosiltransferases/genética , Gossypium/parasitologia , Doenças das Plantas/prevenção & controle , Proteínas de Soja/genética , Soja/enzimologia , Tylenchoidea , Animais , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Glicosiltransferases/metabolismo , Gossypium/genética , Doenças das Plantas/parasitologia , Raízes de Plantas/parasitologia , Plantas Geneticamente Modificadas , Proteínas de Soja/metabolismo , Soja/genética
3.
Plant Mol Biol ; 104(1-2): 67-79, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32621165

RESUMO

Acetylation and deacetylation of histones are important for regulating a series of biological processes in plants. Histone deacetylases (HDACs) control the histone deacetylation that plays an important role in plant response to abiotic stress. In our study, we show the evidence that GhHDT4D (a member of the HD2 subfamily of HDACs) is involved in cotton (Gossypium hirsutum) response to drought stress. Overexpression of GhHDT4D in Arabidopsis increased plant tolerance to drought, whereas silencing GhHDT4D in cotton resulted in plant sensitivity to drought. Simultaneously, the H3K9 acetylation level was altered in the GhHDT4D silenced cotton, compared with the controls. Further study revealed that GhHDT4D suppressed the transcription of GhWRKY33, which plays a negative role in cotton defense to drought, by reducing its H3K9 acetylation level. The expressions of the stress-related genes, such as GhDREB2A, GhDREB2C, GhSOS2, GhRD20-1, GhRD20-2 and GhRD29A, were significantly decreased in the GhHDT4D silenced cotton, but increased in the GhWRKY33 silenced cotton. Given these data together, our findings suggested that GhHDT4D may enhance drought tolerance by suppressing the expression of GhWRKY33, thereby activating the downstream drought response genes in cotton.


Assuntos
Secas , Gossypium/metabolismo , Histona Desacetilases/metabolismo , Estresse Fisiológico/fisiologia , Acetilação , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Gossypium/genética , Histona Desacetilases/genética , Histonas/genética , Histonas/metabolismo , Filogenia , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Estresse Fisiológico/genética , Transcriptoma
4.
Mol Genet Genomics ; 295(6): 1393-1400, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32651630

RESUMO

Anthocyanins are a group of important secondary metabolites, functioning as colorant in plant organs as well as protective agents against several stresses. Sub-red plant (Rs) cottons, accumulating moderate level of anthocyanins in shoots, had increased photosynthesis efficiency compared to green- (GL) and red-plant (R1) cottons. The present work aimed to clarify the molecular base of anthocyanin regulation in Rs cotton. It was found that GhPAP1A was significantly up-regulated in Rs plants compared to GL cottons, but its expression level is lower than that of GhPAP1D in R1 plants. Virus induced gene silencing of GhPAP1s inhibited the red pigmentation in Rs plants. Comparative cloning revealed a 50-bp tandem repeat in the promoter of GhPAP1A in Rs cotton, which showed stronger activity to drive the expression of downstream genes in petals. Considered that the coding sequence of GhPAP1As from Rs and GL cottons had similar functions to promote anthocyanin biosynthesis in transgenic tobaccos, we attributed moderate anthocyanin accumulation in Rs cotton to increased transcription of GhPAP1A, resulted from varied promoter structure. Our works suggested GhPAP1s as useful tool to manipulate anthocyanin level and several breeding targets, including herbivore- and pathogen- resistance, high photosynthesis efficiency and colored fibers.


Assuntos
Antocianinas/biossíntese , Regulação da Expressão Gênica de Plantas , Gossypium/metabolismo , Pigmentação/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Gossypium/genética , Gossypium/crescimento & desenvolvimento , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/genética
5.
PLoS One ; 15(6): e0235317, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32598401

RESUMO

The Dof (DNA-binding one zinc finger) transcription factor family is a representative of plant-specific classes of transcription factors. In this study, we performed a genome-wide screening and characterization of the Dof gene family within two tetraploid species Gossypium barbadense, Gossypium hirsutum, and two diploid species Gossypium arboreum, Gossypium raimondii. 115, 116, 55 and 56 Dof genes were identified respectively and all of the genes contain a sequence encoding the Dof DNA-binding domain. Those genes were unevenly distributed across 13/26 chromosomes of the cotton. Genome comparison revealed that segmental duplication may have played crucial roles in the expansion of the cotton Dof gene family, and tandem duplication also played a minor role. Analysis of RNA-Seq data indicated that cotton Dof gene expression levels varied across different tissues and in response to different abiotic stress. Overall, our results could provide valuable information for better understanding the evolution of cotton Dof genes, and lays a foundation for future investigation in cotton.


Assuntos
Proteínas de Ligação a DNA/genética , Diploide , Genoma de Planta , Gossypium/genética , Proteínas de Plantas/genética , Tetraploidia , Dedos de Zinco/genética , Cromossomos de Plantas/genética , Proteínas de Ligação a DNA/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Gossypium/crescimento & desenvolvimento , Gossypium/metabolismo , Família Multigênica , Filogenia , Proteínas de Plantas/metabolismo
6.
PLoS One ; 15(4): e0231733, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32339186

RESUMO

Genetically modified (GM) crops are one of the most valuable tools of modern biotechnology that secure yield potential needed to sustain the global agricultural demands for food, feed, fiber, and energy. Crossing single GM events through conventional breeding has proven to be an effective way to pyramid GM traits from individual events and increase yield protection in the resulting combined products. Even though years of research and commercialization of GM crops show that these organisms are safe and raise no additional biosafety concerns, some regulatory agencies still require risk assessments for these products. We sought out to investigate whether stacking single GM events would have a significant impact on agronomic and phenotypic plant characteristics in soybean, maize, and cotton. Several replicated field trials designed as randomized complete blocks were conducted by Monsanto Regulatory Department from 2008 to 2017 in field sites representative of cultivation regions in Brazil. In total, twenty-one single and stacked GM materials currently approved for in-country commercial use were grown with the corresponding conventional counterparts and commercially available GM/non-GM references. The generated data were presented to the Brazilian regulatory agency CTNBio (National Biosafety Technical Committee) over the years to request regulatory approvals for the single and stacked products, in compliance with the existing normatives. Data was submitted to analysis of variance and differences between GM and control materials were assessed using t-test with a 5% significance level. Data indicated the predominance of similarities and neglectable differences between single and stacked GM crops when compared to conventional counterpart. Our results support the conclusion that combining GM events through conventional breeding does not alter agronomic or phenotypic plant characteristics in these stacked crops. This is compatible with a growing weight of evidence that indicates this long-adopted strategy does not increase the risks associated with GM materials. It also provides evidence to support the review and modernization of the existing regulatory normatives to no longer require additional risk assessments of GM stacks comprised of previously approved single events for biotechnology-derived crops. The data analyzed confirms that the risk assessment of the individual events is sufficient to demonstrate the safety of the stacked products, which deliver significant benefits to growers and to the environment.


Assuntos
Gossypium/genética , Plantas Geneticamente Modificadas/genética , Soja/genética , Zea mays/genética , Biofortificação/legislação & jurisprudência , Biotecnologia/legislação & jurisprudência , Brasil , Fenótipo , Melhoramento Vegetal/métodos , Plantas Geneticamente Modificadas/efeitos adversos , Locos de Características Quantitativas , Distribuição Aleatória , Medição de Risco , Desenvolvimento Sustentável
7.
Nat Genet ; 52(5): 516-524, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32284579

RESUMO

Upon assembling the first Gossypium herbaceum (A1) genome and substantially improving the existing Gossypium arboreum (A2) and Gossypium hirsutum ((AD)1) genomes, we showed that all existing A-genomes may have originated from a common ancestor, referred to here as A0, which was more phylogenetically related to A1 than A2. Further, allotetraploid formation was shown to have preceded the speciation of A1 and A2. Both A-genomes evolved independently, with no ancestor-progeny relationship. Gaussian probability density function analysis indicates that several long-terminal-repeat bursts that occurred from 5.7 million years ago to less than 0.61 million years ago contributed compellingly to A-genome size expansion, speciation and evolution. Abundant species-specific structural variations in genic regions changed the expression of many important genes, which may have led to fiber cell improvement in (AD)1. Our findings resolve existing controversial concepts surrounding A-genome origins and provide valuable genomic resources for cotton genetic improvement.


Assuntos
Genoma de Planta/genética , Gossypium/genética , Mapeamento Cromossômico/métodos , Fibra de Algodão , Diploide , Evolução Molecular , Genômica/métodos , Filogenia , Análise de Sequência de DNA/métodos
8.
Nat Genet ; 52(5): 525-533, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32313247

RESUMO

Polyploidy is an evolutionary innovation for many animals and all flowering plants, but its impact on selection and domestication remains elusive. Here we analyze genome evolution and diversification for all five allopolyploid cotton species, including economically important Upland and Pima cottons. Although these polyploid genomes are conserved in gene content and synteny, they have diversified by subgenomic transposon exchanges that equilibrate genome size, evolutionary rate heterogeneities and positive selection between homoeologs within and among lineages. These differential evolutionary trajectories are accompanied by gene-family diversification and homoeolog expression divergence among polyploid lineages. Selection and domestication drive parallel gene expression similarities in fibers of two cultivated cottons, involving coexpression networks and N6-methyladenosine RNA modifications. Furthermore, polyploidy induces recombination suppression, which correlates with altered epigenetic landscapes and can be overcome by wild introgression. These genomic insights will empower efforts to manipulate genetic recombination and modify epigenetic landscapes and target genes for crop improvement.


Assuntos
Genoma de Planta/genética , Gossypium/genética , Fibra de Algodão , Domesticação , Epigenômica/métodos , Evolução Molecular , Regulação da Expressão Gênica de Plantas/genética , Genômica/métodos , Filogenia , Poliploidia
9.
Plant Mol Biol ; 103(1-2): 211-223, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32172495

RESUMO

The mitogen-activated protein kinase (MAPK) cascade pathway, which has three components, MAP3Ks, MKKs and MPKs, is involved in diverse biological processes in plants. In the current study, MAPK cascade genes were identified in three cotton species, based on gene homology with Arabidopsis. Selection pressure analysis of MAPK cascade genes revealed that purifying selection occurred among the cotton species. Expression pattern analysis showed that some MAPK cascade genes differentially expressed under abiotic stresses and phytohormones treatments, and especially under drought stress. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) experiments showed extensive interactions between different MAPK cascade proteins. Virus-induced gene silencing (VIGS) assays showed that some MAPK cascade modules play important roles in the drought stress response, and the GhMAP3K14-GhMKK11-GhMPK31 signal pathway was demonstrated to regulate drought stress tolerance in cotton. This study provides new information on the function of MAPK cascade genes in the drought response, and will help direct molecular breeding for improved drought stress tolerance in cotton.


Assuntos
Gossypium/genética , Sistema de Sinalização das MAP Quinases/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Aclimatação/genética , Secas , Perfilação da Expressão Gênica , Genes de Plantas , Genoma de Planta , Gossypium/enzimologia , Gossypium/fisiologia , Filogenia
10.
Mol Genet Genomics ; 295(3): 645-660, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32172356

RESUMO

Genetic male sterility (GMS) facilitates hybrid seed production in crops including cotton (Gossypium hirsutum). However, the genetic and molecular mechanisms specifically involved in this developmental process are poorly understood. In this study, small RNA sequencing, degradome sequencing, and transcriptome sequencing were performed to analyze miRNAs and their target genes during anther development in a GMS mutant ('Dong A') and its fertile wildtype (WT). A total of 80 known and 220 novel miRNAs were identified, 71 of which showed differential expressions during anther development. A further degradome sequencing revealed a total of 117 candidate target genes cleaved by 16 known and 36 novel miRNAs. Based on RNA-seq, 24, 11, and 21 predicted target genes showed expression correlations with the corresponding miRNAs at the meiosis, tetrad and uninucleate stages, respectively. In addition, a large number of differentially expressed genes were identified, most of which were involved in sucrose and starch metabolism, carbohydrate metabolism, and plant hormone signal transduction based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The results of our study provide valuable information for further functional investigations of the important miRNAs and target genes involved in genetic male sterility and advance our understanding of miRNA regulatory functions during cotton anther development.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Gossypium/crescimento & desenvolvimento , Gossypium/genética , MicroRNAs/genética , Mutação , Infertilidade das Plantas/genética , Proteínas de Plantas/genética , Flores/genética , Flores/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Transcriptoma
11.
PLoS One ; 15(3): e0230519, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32187234

RESUMO

Promoters are specified segments of DNA that lead to the initiation of transcription of a specific gene. The designing of a gene cassette for plant transformation is significantly dependent upon the specificity of a promoter. Constitutive Cauliflower mosaic virus promoter, CaMV35S, due to its developmental role, is the most commonly used promoter in plant transformation. While Gossypium hirsutum (Gh) being fiber-specific promoter (GhSCFP) specifically activates transcription in seed coat and fiber associated genes. The Expansin genes are renowned for their versatile roles in plant growth. The overexpression of Expansin genes has been reported to enhance fiber length and fineness. Thus, in this study, a local Cotton variety was transformed with Expansin (CpEXPA1) gene, in the form of two separate cassettes, each with a different promoter, named as 35SEXPA1 and FSEXPA1 expressed under CaMV35S and GhSCFP promoters respectively. Integration and Spatiotemporal relative expression of the transgene were studied in an advanced generation. GhSCFP bearing transgene expression was significantly higher in Cotton fiber than other plant parts. While transgene with CaMV35S promoter was found to be continually expressing in all tissues but the expression was lower in fiber than that expressed under GhSCFP. The temporal expression profile was quite interesting with a gradual increasing pattern of both constructs from 1DPA (days post anthesis) to 18DPA and decreased expression from 24 to 30 DPA. Besides the relative expression of promoters, fiber cellulose quantification and fluorescence intensity were also observed. The study significantly compared the two most commonly used promoters and it is deduced from the results that the GhSCFP promoter could be used more efficiently in fiber when compared with CaMV35S which being constitutive in nature preferred for expression in all parts of the plant.


Assuntos
Fibra de Algodão , Gossypium/genética , Gossypium/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Regiões Promotoras Genéticas/genética
12.
Artigo em Inglês | MEDLINE | ID: mdl-32114400

RESUMO

Protein acetylation (KAC) is a significant post-translational modification, which plays an essential role in the regulation of growth and development. Unfortunately, related studies are inadequately available in angiosperms, and to date, there is no report providing insight on the role of protein acetylation in cotton fiber development. Therefore, we first compared the lysine-acetylation proteome (acetylome) of upland cotton ovules in the early fiber development stages by using wild-type as well as its fuzzless-lintless mutant to identify the role of KAC in the fiber development. A total of 1696 proteins with 2754 acetylation sites identified with the different levels of acetylation belonging to separate subcellular compartments suggesting a large number of proteins differentially acetylated in two cotton cultivars. About 80% of the sites were predicted to localize in the cytoplasm, chloroplast, and mitochondria. Seventeen significantly enriched acetylation motifs were identified. Serine and threonine and cysteine located downstream and upstream to KAC sites. KEGG pathway enrichment analysis indicated oxidative phosphorylation, fatty acid, ribosome and protein, and folate biosynthesis pathways enriched significantly. To our knowledge, this is the first report of comparative acetylome analysis to compare the wild-type as well as its fuzzless-lintless mutant acetylome data to identify the differentially acetylated proteins, which may play a significant role in cotton fiber development.


Assuntos
Fibra de Algodão , Gossypium , Óvulo Vegetal , Acetilação , Fibra de Algodão/análise , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Gossypium/metabolismo , Mutação , Óvulo Vegetal/genética , Óvulo Vegetal/metabolismo , Proteínas de Plantas/metabolismo , Processamento de Proteína Pós-Traducional
13.
Plant Mol Biol ; 103(4-5): 409-423, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32189187

RESUMO

Cotton fibers are initiated from the epidermal cells of the ovule before or on the day of anthesis. Gossypium arboreum SMA-4 mutant contains recessive mutation (sma-4(ha)) and has the phenotypes of fibreless seeds and glabrous stems. In this study, fine mapping and alternative splicing analysis indicated a nucleotide substitution (AG → AC) at splicing site in a homeodomain-leucine zipper IV family gene (GaHD1) might cause gene A3S (Alternative 3' splicing) mistake, suggested that GaHD1 was the candidate gene of sma-4(ha). Many genes related to the fiber initiation are identified to be differentially expressed in the mutant which could result in the blocked fiber initiation signals such as H2O2, or Ca in the mutant. Further comparative physiological analysis of H2O2 production and Ca2+ flux in the SMA-4 and wide type cotton confirmed that H2O2 and Ca were important fiber initiation signals and regulated by GaHD1. The in vitro ovule culture of the mutant with hormones recovered the fibered phenotype coupled with the restoration of these signals. Overexpressing of GaHD1 in Arabidopsis increased trichome densities on the sepal, leaf, and stem tissues while transient silencing of the GaHD1 gene in G. arboreum reduced the trichome densities. These phenotypes indicated that GaHD1 is the candidate gene of SMA-4 with a crucial role in acting upstream molecular switch of signal transductions for cotton trichome and fiber initiations.


Assuntos
Regulação da Expressão Gênica de Plantas/fisiologia , Gossypium/fisiologia , Peróxido de Hidrogênio/metabolismo , Proteínas de Plantas/metabolismo , Tricomas/crescimento & desenvolvimento , Processamento Alternativo , Sinalização do Cálcio , Mapeamento Cromossômico , Cromossomos de Plantas , Fibra de Algodão , Ligação Genética , Gossypium/genética , Mutação , Proteínas de Plantas/genética
14.
Plant Mol Biol ; 103(4-5): 391-407, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32193788

RESUMO

Mitogen-activated protein kinases (MAPKs) are important in regulating plant development as well as stress response. In this study, we genome-widely identified 56 MAPK genes in upland cotton. These MAPK genes unequally distribute on 22 chromosomes of cotton genome, but no MAPK gene is located on At_Chr6, Dt_Chr6, At_Chr13 and Dt_Chr13. The exons and introns in GhMAPK gene family vary widely at the position, number and length. Furthermore, GhMAPK family can be divided into 4 groups (A, B, C and D), and the TEY type of T-loop exists in three groups (A, B and C), but the TDY type of T-loop is only in group D. Further study revealed that some GhMAPK genes (including GhMPK6) are preferentially expressed in elongating fibers. GhMPK6 maintains a high phosphorylation level in elongating fibers, and its phosphorylation was enhanced in fibers by phytohormones brassinosteroid (BR), ethylene and indole-3-acetic acid (IAA). Additionally, GhMPK6 could interact with GhMKK2-2 and GhMKK4, suggesting that GhMKK2-2/4-GhMPK6 module may be involved in phosphorylation of its downstream proteins for regulating fiber elongation of cotton.


Assuntos
Regulação da Expressão Gênica de Plantas/fisiologia , Genoma de Planta , Gossypium/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Proteínas de Plantas/metabolismo , Fibra de Algodão , Regulação Enzimológica da Expressão Gênica , Estudo de Associação Genômica Ampla , Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Proteínas de Plantas/genética
15.
Planta ; 251(2): 56, 2020 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-32006110

RESUMO

MAIN CONCLUSION: The structure of the cotton uceA1.7 promoter and its modules was analyzed; the potential of their key sequences has been confirmed in different tissues, proving to be a good candidate for the development of new biotechnological tools. Transcriptional promoters are among the primary genetic engineering elements used to control genes of interest (GOIs) associated with agronomic traits. Cotton uceA1.7 was previously characterized as a constitutive promoter with activity higher than that of the constitutive promoter from the Cauliflower mosaic virus (CaMV) 35S gene in various plant tissues. In this study, we generated Arabidopsis thaliana homozygous events stably overexpressing the gfp reporter gene driven by different modules of the uceA1.7 promoter. The expression level of the reporter gene in different plant tissues and the transcriptional stability of these modules was determined compared to its full-length promoter and the 35S promoter. The full-length uceA1.7 promoter exhibited higher activity in different plant tissues compared to the 35S promoter. Two modules of the promoter produced a low and unstable transcription level compared to the other promoters. The other two modules rich in cis-regulatory elements showed similar activity levels to full-length uceA1.7 and 35S promoters but were less stable. This result suggests the location of a minimal portion of the promoter that is required to initiate transcription properly (the core promoter). Additionally, the full-length uceA1.7 promoter containing the 5'-untranslated region (UTR) is essential for higher transcriptional stability in various plant tissues. These findings confirm the potential use of the full-length uceA1.7 promoter for the development of new biotechnological tools (NBTs) to achieve higher expression levels of GOIs in, for example, the root or flower bud for the efficient control of phytonematodes and pest-insects, respectively, in important crops.


Assuntos
Gossypium/genética , Regiões 5' não Traduzidas , Arabidopsis/genética , Caulimovirus/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Genes Reporter , Engenharia Genética , Gossypium/anatomia & histologia , Gossypium/crescimento & desenvolvimento , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas
16.
J Biosci ; 452020.
Artigo em Inglês | MEDLINE | ID: mdl-32020913

RESUMO

High-yielding Indian cotton varieties are not amenable for regeneration and transformation because they are recalcitrant in nature. In this work, we have developed Narasimha (NA1325) cotton variety by introducing three Cry genes driven by three different promoters conferring insect resistance. The meristematic region of embryo axis explants were infected and co-cultivated with Agrobacterium tumefacience (LBA4404) harbouring pMDC100 vector with three Cry gene cassettes (alpha-globulin : Cry2Ab, DECaMV35s : Cry1F and nodulin : Cry1Ac) with Npt II as a selectable marker gene. Out of 1010 embryo axes explants infected, 121 (T0) regenerated under two rounds of kanamycin selectionmedium.About 2551T1 seedswere collected from111T0 plants and these seeds screened again with kanamycin at seedling stage. The transgenic plants were characterized by PCR, real time quantitative PCR, lateral flow strip protein assay and insect bioassay. Out of 145 kanamycin resistant plants (T1), twelve showed amplification of all four transgenes: Npt II, Cry2Ab, Cry1F and Cry1Ac through PCR with expected amplicons as 395, 870, 840 and 618 bp, respectively. Further, lateral flow strip test revealed Cry1F and Cry1Ac proteins accumulated in 12 plants, whereas Cry2Ab protein was detected in eight only. The transcripts of all three Cry genes were accumulated significantly higher in transgenic plants at T2 generation. The transgenic lines showed effective resistance againstHelicoverpa armigera and Spodoptera litura larvae. The T2 line L-3 exhibited highest percentage of insect mortality, in which transcripts of all cry genes were accumulated higher than other plants. The transgenic cotton plants carrying triple Cry genes could be an excellent germplasmresource for the breeders for introgressions.


Assuntos
Agrobacterium tumefaciens/metabolismo , Proteínas de Bactérias/genética , Gossypium/genética , Gossypium/metabolismo , Larva/virologia , Nucleopoliedrovírus/patogenicidade , Plantas Geneticamente Modificadas/genética , Animais , Proteínas de Insetos , Receptores de Superfície Celular , Transformação Genética , Transgenes
17.
Artigo em Inglês | MEDLINE | ID: mdl-32005380

RESUMO

The soil-born vascular disease Verticillium wilt, which is caused by fungal pathogen Verticillium dahliae, is a devastating disease of cotton worldwide. In the last decade, a large number of genes have been found to participate in cotton-V. dahliae interactions, but the detailed mechanisms of cotton resistance to V. dahliae remain unclear. Here, we functionally characterized MPK3, a MAPK gene from cotton. MPK3 was induced in the roots of both resistant and susceptible cotton cultivars by V. dahliae inoculation. Transgenic cotton and tobacco with constitutively higher GbMPK3 expression conferred higher V. dahliae susceptibility, while MPK3 knockdown in cotton has limited effect on cotton resistance to V. dahliae. Expression profiling revealed that SA-mediated defense pathway genes (WRKY70, PR1, and PR5) accumulated after V. dahliae inoculation in roots of both wild-type and transgenic cotton, and the expression levels of these genes were higher in GbMPK3-overexpressing plants than in wild-type plants, indicating that GbMPK3 upregulation may reduce plant resistance to V. dahliae through regulating salicylic acid signaling transduction.


Assuntos
Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Ácido Salicílico/metabolismo , Transdução de Sinais , Verticillium/fisiologia , Gossypium/metabolismo , Gossypium/microbiologia , Lycopersicon esculentum/genética , Lycopersicon esculentum/metabolismo , Lycopersicon esculentum/microbiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/microbiologia
18.
Artigo em Inglês | MEDLINE | ID: mdl-32050119

RESUMO

Crops, including cotton, are sensitive to nitrogen (N) and excessive use can lead to an increase in production costs and environmental problems. We hypothesized that the use of cotton genotypes with substantial root systems and high genetic potentials for nitrogen-use efficiency (NUE) would best address these problems. Therefore, the interspecific variations and traits contributing to NUE in six cotton genotypes having contrasting NUEs were studied in response to various nitrate concentrations. Large genotypic variations were observed in morphophysiological and biochemical traits, especially shoot dry weight, root traits, and N-assimilating enzyme levels. The roots of all the cotton genotypes were more sensitive to low-than high-nitrate concentrations, and the genotype CCRI-69 had the largest root system irrespective of the nitrate concentration. The root morphological traits were positively correlated with N-utilization efficiency and were more affected by genotype than nitrate concentration. Conversely, growth and N-assimilating enzyme levels were more affected by nitrate concentration and were positively correlated with N-uptake efficiency. Based on shoot dry weight, CCRI-69 and XLZ-30 were identified as N-efficient and N-inefficient genotypes, respectively, and these results were confirmed by their contrasting root systems, N metabolism, and NUEs. In the future, multi-omics techniques will be performed to identify key genes/pathways involved in N metabolism, which may have the potential to improve root architecture and increase NUE.


Assuntos
Gossypium , Nitrogênio , Raízes de Plantas , Genótipo , Gossypium/genética , Gossypium/crescimento & desenvolvimento , Gossypium/metabolismo , Nitratos/metabolismo , Nitrogênio/metabolismo , Fenótipo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo
19.
Plant Mol Biol ; 102(4-5): 553-567, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31989373

RESUMO

KEY MESSAGE: Overexpression of K2-NhaD in transgenic cotton resulted in phenotypes with strong salinity and drought tolerance in greenhouse and field experiments, increased expression of stress-related genes, and improved regulation of metabolic pathways, such as the SOS pathway. Drought and salinity are major abiotic stressors which negatively impact cotton yield under field conditions. Here, a plasma membrane Na+/H+ antiporter gene, K2-NhaD, was introduced into upland cotton R15 using an Agrobacterium tumefaciens-mediated transformation system. Homozygous transgenic lines K9, K17, and K22 were identified by PCR and glyphosate-resistance. TAIL-PCR confirmed that T-DNA carrying the K2-NhaD gene in transgenic lines K9, K17 and K22 was inserted into chromosome 3, 19 and 12 of the cotton genome, respectively. Overexpression of K2-NhaD in transgenic cotton plants grown in greenhouse conditions and subjected to drought and salinity stress resulted in significantly higher relative water content, chlorophyll, soluble sugar, proline levels, and SOD, CAT, and POD activity, relative to non-transgenic plants. The expression of stress-related genes was significantly upregulated, and this resulted in improved regulation of metabolic pathways, such as the salt overly sensitive pathway. K2-NhaD transgenic plants growing under field conditions displayed strong salinity and drought tolerance, especially at high levels of soil salinity and drought. Seed cotton yields in transgenic line were significantly higher than in wild-type plants. In conclusion, the data indicate that K2-NhaD transgenic lines have great potential for the production of stress-tolerant cotton under field conditions.


Assuntos
Gossypium/metabolismo , Proteínas de Plantas/fisiologia , Tolerância ao Sal/genética , Trocadores de Sódio-Hidrogênio/fisiologia , Secas , Gossypium/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Trocadores de Sódio-Hidrogênio/genética , Trocadores de Sódio-Hidrogênio/metabolismo , Estresse Fisiológico , Água/metabolismo
20.
Int J Mol Sci ; 21(1)2020 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-31947720

RESUMO

Histone deacetylases (HDACs) play a significant role in a plant's development and response to various environmental stimuli by regulating the gene transcription. However, HDACs remain unidentified in cotton. In this study, a total of 29 HDACs were identified in allotetraploid Gossypium hirsutum, while 15 and 13 HDACs were identified in Gossypium arboretum and Gossypium raimondii, respectively. Gossypium HDACs were classified into three groups (reduced potassium dependency 3 (RPD3)/HDA1, HD2-like, and Sir2-like (SRT) based on their sequences, and Gossypium HDACs within each subgroup shared a similar gene structure, conserved catalytic domains and motifs. Further analysis revealed that Gossypium HDACs were under a strong purifying selection and were unevenly distributed on their chromosomes. Gene expression data revealed that G. hirsutum HDACs were differentially expressed in various vegetative and reproductive tissues, as well as at different developmental stages of cotton fiber. Furthermore, some G. hirsutum HDACs were co-localized with quantitative trait loci (QTLs) and single-nucleotide polymorphism (SNPs) of fiber-related traits, indicating their function in fiber-related traits. We also showed that G. hirsutum HDACs were differentially regulated in response to plant hormones (abscisic acid (ABA) and auxin), DNA damage agent (methyl methanesulfonate (MMS)), and abiotic stresses (cold, salt, heavy metals and drought), indicating the functional diversity and specification of HDACs in response to developmental and environmental cues. In brief, our results provide fundamental information regarding G. hirsutum HDACs and highlight their potential functions in cotton growth, fiber development and stress adaptations, which will be helpful for devising innovative strategies for the improvement of cotton fiber and stress tolerance.


Assuntos
Dano ao DNA , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Histona Desacetilases/genética , Proteínas de Plantas/genética , Diploide , Genes de Plantas , Genoma de Planta , Gossypium/fisiologia , Família Multigênica , Filogenia , Reguladores de Crescimento de Planta/metabolismo , Poliploidia , Estresse Fisiológico
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