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1.
Sci Rep ; 13(1): 16863, 2023 10 06.
Artigo em Inglês | MEDLINE | ID: mdl-37803043

RESUMO

Pancreatic ductal adenocarcinoma (PDAC) cells have a great demand for nutrients in the form of sugars, amino acids, and lipids. Particularly, amino acids are critical for cancer growth and, as intermediates, connect glucose, lipid and nucleotide metabolism. PDAC cells meet these requirements by upregulating selective amino acid transporters. Here we show that SLC38A5 (SN2/SNAT5), a neutral amino acid transporter is highly upregulated and functional in PDAC cells. Using CRISPR/Cas9-mediated knockout of SLC38A5, we show its tumor promoting role in an in vitro cell line model as well as in a subcutaneous xenograft mouse model. Using metabolomics and RNA sequencing, we show significant reduction in many amino acid substrates of SLC38A5 as well as OXPHOS inactivation in response to SLC38A5 deletion. Experimental validation demonstrates inhibition of mTORC1, glycolysis and mitochondrial respiration in KO cells, suggesting a serious metabolic crisis associated with SLC38A5 deletion. Since many SLC38A5 substrates are activators of mTORC1 as well as TCA cycle intermediates/precursors, we speculate amino acid insufficiency as a possible link between SLC38A5 deletion and inactivation of mTORC1, glycolysis and mitochondrial respiration, and the underlying mechanism for PDAC attenuation. Overall, we show that SLC38A5 promotes PDAC, thereby identifying a novel, hitherto unknown, therapeutic target for PDAC.


Assuntos
Sistemas de Transporte de Aminoácidos Neutros , Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Humanos , Camundongos , Animais , Carcinógenos , Neoplasias Pancreáticas/patologia , Carcinoma Ductal Pancreático/patologia , Sistemas de Transporte de Aminoácidos , Alvo Mecanístico do Complexo 1 de Rapamicina/metabolismo , Aminoácidos , Linhagem Celular Tumoral , Proliferação de Células , Neoplasias Pancreáticas
2.
Front Microbiol ; 14: 1268051, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37886058

RESUMO

Date palm (Phoenixdactylifera) is the most significant crop across North Africa and the Middle East. However, the crop faces a severe threat from Bayoud disease caused by the fungal pathogen Fusarium oxysporum f. sp. albedinis (FOA). FOA is a soil-borne fungus that infects the roots and vascular system of date palms, leading to widespread destruction of date palm plantations in North Africa over the last century. This is considered the most devastating pathogen of oasis agriculture in North Africa and responsible for loss of 13 million trees in Algeria and Morocco alone. In this study, we present a chromosome-scale high-quality genome assembly of the virulent isolate Foa 44, which provides valuable insights into understanding the genetic basis of Bayoud disease. The genome assembly consists of 11 chromosomes and 40 unplaced contigs, totalling 65,971,825 base pairs in size. It exhibits a GC ratio of 47.77% and a TE (transposable element) content of 17.30%. Through prediction and annotation, we identified 20,416 protein-coding genes. By combining gene and repeat densities analysis with alignment to Fusarium oxysporum f. sp. lycopersici (FOL) 4287 isolate genome sequence, we determined the core and lineage-specific compartments in Foa 44, shedding light on the genome structure of this pathogen. Furthermore, a phylogenomic analysis based on the 3,292 BUSCOs core genome revealed a distinct clade of FOA isolates within the Fusarium oxysporum species complex (FOSC). Notably, the genealogies of the five identified Secreted In Xylem (SIX) genes (1, 6, 9, 11 and 14) in FOA displayed a polyphyletic pattern, suggesting a horizontal inheritance of these effectors. These findings provide a valuable genomics toolbox for further research aimed at combatting the serious biotic constraints posed by FOA to date palm. This will pave the way for a deeper understanding of Bayoud disease and facilitate the development of effective diagnostic tools and control measures.

3.
Genet Mol Biol ; 46(2): e20220021, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37276609

RESUMO

The genetic diversity between 23 Moroccan date palm cultivars collected from the National Palm Collection at the INRA (National Agricultural Research Institute) experimental field in Zagora was assessed using SSR markers that are specifically designed for date palm. Among the 16 tested SSR, 13 were successfully amplified, and were selected to carry out this study. 208 bands were amplified, ranging from 10 to 25 bands per cultivar with an average of 16 alleles per cultivar. The value of heterozygosity of the studied markers ranged from 0.11 to 0.30. The pairwise genetic distances between those cultivars ranged from 0.06 to 0.46. The hierarchical cluster analysis distributed the 23 genotypes into four different groups of one to ten cultivars.

4.
J Genet Eng Biotechnol ; 21(1): 66, 2023 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-37217693

RESUMO

BACKGROUND: Date palm, oasis pivot, plays a vital socio-economic part in the southern area of Morocco. However, with climate change and drought intensity and frequency increasing, the Moroccan palm grove is threatened with significant genetic degradation. Genetic characterization of this resource is key element for the development of effective conservation and management strategies in the current circumstances of climate change and various biotic and abiotic stresses. To evaluate the genetic diversity of date palm populations collected from different Moroccan oases, we used simple sequence repeats (SSR) and directed amplification of mini-satellite DNA (DAMD) markers. Our results showed that used markers could efficiently assess genetic diversity in Phoenix dactylifera L. RESULTS: A total of 249 and 471 bands were respectively scored for SSR and DAMD, of which 100% and 92.9% were polymorphic. The polymorphic information content (PIC = 0.95), generated by the SSR primer was nearly identical to that generated by the DAMD primer (PIC = 0.98). The resolving power (Rp) was higher in DAMD than SSR (29.46 and 19.51, respectively). Analysis of the molecular variance (AMOVA) based on the combined data sets for both markers revealed a higher variance within populations (75%) than among populations (25%). Principal coordinate analysis (PCoA) and the ascendant hierarchical classification showed that the population of Zagora and Goulmima regions were the closest populations. The STRUCTURE analysis clustering of the 283 tested samples into seven clusters based on their genetic composition. CONCLUSION: The results drawn from this study will orient genotypes selection strategies for a successful future breeding and conservation program, particularly under climate change context.

5.
J Pers Med ; 13(3)2023 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-36983633

RESUMO

Breast cancer is one of the main global priorities in terms of public health. It remains the most frequent cancer in women and is the leading cause of their death. The human microbiome plays various roles in maintaining health by ensuring a dynamic balance with the host or in the appearance of various pathologies including breast cancer. In this study, we performed an analysis of bacterial signature differences between tumor and adjacent tissues of breast cancer patients in Morocco. Using 16S rRNA gene sequencing, we observed that adjacent tissue contained a much higher percentage of the Gammaproteobacteria class (35.7%) while tumor tissue was characterized by a higher percentage of Bacilli and Actinobacteria classes, with about 18.8% and 17.2% average abundance, respectively. Analysis of tumor subtype revealed enrichment of genus Sphingomonodas in TNBC while Sphingomonodas was predominant in HER2. The LEfSe and the genus level heatmap analysis revealed a higher abundance of the Rothia genus in tumor tissues. The identified microbial communities can therefore serve as potential biomarkers for prognosis and diagnosis, while also helping to develop new strategies for the treatment of breast cancer patients.

6.
Biomolecules ; 12(11)2022 10 27.
Artigo em Inglês | MEDLINE | ID: mdl-36358924

RESUMO

Aiming to reduce the gap between in vitro and in vivo environment, a complex culture medium, Plasmax, was introduced recently, which includes nutrients and metabolites with concentrations normally found in human plasma. Herein, to study the influence of this medium on cellular behaviors, we utilized Plasmax to cultivate two cancer cell lines, including one breast cancer cell line, MDA-MB-231BR, and one brain cancer cell line, CRL-1620. Cancer cells were harvested and prepared for transcriptomics and proteomics analyses to assess the discrepancies caused by the different nutritional environments of Plasmax and two commercial media: DMEM, and EMEM. Total RNAs of cells were extracted using mammalian total RNA extract kits and analyzed by next-generation RNA sequencing; proteomics analyses were performed using LC-MS/MS. Gene oncology and pathway analysis were employed to study the affected functions. The cellular invasion and cell death were inhibited in MDA-MB-231BR cell line when cultured in Plasmax compared to DMEM and EMEM, whereas the invasion, migration and protein synthesis of CRL-1620 cell line were activated in Plasmax in relative to both commercial media. The expression changes of some proteins were more significant compared to their corresponding transcripts, indicating that Plasmax has more influence upon regulatory processes of proteins after translation. This work provides complementary information to the original study of Plasmax, aiming to facilitate the selection of appropriate media for in vitro cancer cell studies.


Assuntos
Neoplasias da Mama , Proteômica , Animais , Humanos , Feminino , Cromatografia Líquida , Transcriptoma , Linhagem Celular Tumoral , Espectrometria de Massas em Tandem , Neoplasias da Mama/genética , Mamíferos
7.
Front Cell Dev Biol ; 10: 1025240, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36313560

RESUMO

Adipose tissue (AT) is an endocrine organ with a central role on whole-body energy metabolism and development of metabolic diseases. Single-cell and single-nuclei RNA sequencing (scRNA-seq and snRNA-seq, respectively) analyses in mice and human AT have revealed vast cell heterogeneity and functionally distinct subtypes that are potential therapeutic targets to metabolic disease. In periparturient dairy cows, AT goes through intensive remodeling and its dysfunction is associated with metabolic disease pathogenesis and decreased productive performance. The contributions of depot-specific cells and subtypes to the development of diseases in dairy cows remain to be studied. Our objective was to elucidate differences in cellular diversity of visceral (VAT) and subcutaneous (SAT) AT in dairy cows at the single-nuclei level. We collected matched SAT and VAT samples from three dairy cows and performed snRNA-seq analysis. We identified distinct cell types including four major mature adipocytes (AD) and three stem and progenitor cells (ASPC) subtypes, along with endothelial cells (EC), mesothelial cells (ME), immune cells, and pericytes and smooth muscle cells. All major cell types were present in both SAT and VAT, although a strong VAT-specificity was observed for ME, which were basically absent in SAT. One ASPC subtype was defined as adipogenic (PPARG+) while the other two had a fibro-adipogenic profile (PDGFRA+). We identified vascular and lymphatic EC subtypes, and different immune cell types and subtypes in both SAT and VAT, i.e., macrophages, monocytes, T cells, and natural killer cells. Not only did VAT show a greater proportion of immune cells, but these visceral immune cells had greater activation of pathways related to immune and inflammatory response, and complement cascade in comparison with SAT. There was a substantial contrast between depots for gene expression of complement cascade, which were greatly expressed by VAT cell subtypes compared to SAT, indicating a pro-inflammatory profile in VAT. Unprecedently, our study demonstrated cell-type and depot-specific heterogeneity in VAT and SAT of dairy cows. A better understanding of depot-specific molecular and cellular features of SAT and VAT will aid in the development of AT-targeted strategies to prevent and treat metabolic disease in dairy cows, especially during the periparturient period.

8.
Biomolecules ; 12(6)2022 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-35740868

RESUMO

A complex physiological culture medium (Plasmax) was introduced recently, composed of nutrients and metabolites at concentrations normally found in human plasma to mimic the in vivo environment for cell line cultivation. As glycosylation has been proved to be involved in cancer development, it is necessary to investigate the glycan expression changes in media with different nutrients. In this study, a breast cancer cell line, MDA-MB-231BR, and a brain cancer cell line, CRL-1620, were cultivated in Plasmax and commercial media to reveal cell line glycosylation discrepancies prompted by nutritional environments. Glycomics analyses of cell lines were performed using LC-MS/MS. The expressions of multiple fucosylated N-glycans, such as HexNAc4Hex3DeoxyHex1 and HexNAc5Hex3DeoxyHex1, derived from both cell lines exhibited a significant increase in Plasmax. Among the O-glycans, significant differences were also observed. Both cell lines cultivated in EMEM had the lowest amounts of O-glycans expressed. The original work described the development of Plasmax, which improves colony formation, and resulted in transcriptomic and metabolomic alterations of cancer cell lines, while our results indicate that Plasmax can significantly impact protein glycosylation. This study also provides information to guide the selection of media for in vitro cancer cell glycomics studies.


Assuntos
Neoplasias Encefálicas , Espectrometria de Massas em Tandem , Linhagem Celular , Cromatografia Líquida , Humanos , Polissacarídeos/metabolismo , Espectrometria de Massas em Tandem/métodos
9.
Phytopathology ; 112(4): 862-871, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34622696

RESUMO

With the progressive loss of fungicide efficacy against Phakopsora pachyrhizi, the causal agent of Asian soybean rust (ASR), alternative methods to protect soybean crops are needed. Resistance induction is a low impact alternative and/or supplement to fungicide applications that fortifies innate plant defenses against pathogens. Here, we show that a microbial fermentation product (MFP) induces plant defenses in soybean, and transcriptional induction is enhanced with the introduction of ASR. MFP-treated plants exhibited 1,011 and 1,877 differentially expressed genes (DEGs) 12 and 60 h after treatment, respectively, compared with water controls. MFP plants exposed to the pathogen 48 h after application and sampled 12 h later (for a total of 60 h) had 2,401 DEGs compared with control. The plant defense genes PR1, PR2, IPER, PAL, and CHS were induced with MFP application, and induction was enhanced with ASR. Enriched pathways associated with pathogen defense included plant-pathogen interactions, MAPK signaling pathways, phenylpropanoid biosynthesis, glutathione metabolism, flavonoid metabolism, and isoflavonoid metabolism. In field conditions, elevated antioxidant peroxidase activities and phenolic accumulation were measured with MFP treatment; however, improved ASR control or enhanced crop yield were not observed. MFP elicitation differences between field and laboratory grown plants necessitates further testing to identify best practices for effective disease management with MFP-treated soybean.


Assuntos
Glycine max , Phakopsora pachyrhizi , Fermentação , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/genética , Glycine max/genética
10.
BMC Ecol Evol ; 21(1): 218, 2021 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-34872490

RESUMO

BACKGROUND: Airborne environmental DNA (eDNA) research is an emerging field that focuses on the detection of species from their genetic remnants in the air. The majority of studies into airborne eDNA of plants has until now either focused on single species detection, specifically only pollen, or human health impacts, with no previous studies surveying an entire plant community through metabarcoding. We therefore conducted an airborne eDNA metabarcoding survey and compared the results to a traditional plant community survey. RESULTS: Over the course of a year, we conducted two traditional transect-based visual plant surveys alongside an airborne eDNA sampling campaign on a short-grass rangeland. We found that airborne eDNA detected more species than the traditional surveying method, although the types of species detected varied based on the method used. Airborne eDNA detected more grasses and forbs with less showy flowers, while the traditional method detected fewer grasses but also detected rarer forbs with large showy flowers. Additionally, we found the airborne eDNA metabarcoding survey required less sampling effort in terms of the time needed to conduct a survey and was able to detect more invasive species than the traditional method. CONCLUSIONS: Overall, we have demonstrated that airborne eDNA can act as a sensitive and efficient plant community surveying method. Airborne eDNA surveillance has the potential to revolutionize the way plant communities are monitored in general, track changes in plant communities due to climate change and disturbances, and assist with the monitoring of invasive and endangered species.


Assuntos
DNA Ambiental , Biodiversidade , Código de Barras de DNA Taxonômico , Monitoramento Ambiental , Humanos , Inquéritos e Questionários
11.
Mitochondrial DNA B Resour ; 6(10): 3059-3061, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34589591

RESUMO

The complete mitogenome of Fusarium oxysporum f. sp. albedinis (FOA), the causal agent of the destructive fusarium wilt in date palm, is sequenced and assembled. The circular mitogenome of isolate Foa44 is 51,601 bp in length and contains 26 transfer RNA (tRNA) genes, one ribosomal RNA (rRNA), and 28 protein-coding genes. A mitogenome-based phylogenetic analysis of Fusarium revealed that FOA is congruent with previous nuclear-gene phylogenetic results.

12.
EMBO Rep ; 22(6): e50600, 2021 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-33860601

RESUMO

Dishevelled (DVL) critically regulates Wnt signaling and contributes to a wide spectrum of diseases and is important in normal and pathophysiological settings. However, how it mediates diverse cellular functions remains poorly understood. Recent discoveries have revealed that constitutive Wnt pathway activation contributes to breast cancer malignancy, but the mechanisms by which this occurs are unknown and very few studies have examined the nuclear role of DVL. Here, we have performed DVL3 ChIP-seq analyses and identify novel target genes bound by DVL3. We show that DVL3 depletion alters KMT2D binding to novel targets and changes their epigenetic marks and mRNA levels. We further demonstrate that DVL3 inhibition leads to decreased tumor growth in two different breast cancer models in vivo. Our data uncover new DVL3 functions through its regulation of multiple genes involved in developmental biology, antigen presentation, metabolism, chromatin remodeling, and tumorigenesis. Overall, our study provides unique insight into the function of nuclear DVL, which helps to define its role in mediating aberrant Wnt signaling.


Assuntos
Neoplasias , Via de Sinalização Wnt , Proteínas Desgrenhadas/genética , Proteínas Desgrenhadas/metabolismo , Humanos , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Sequências Reguladoras de Ácido Nucleico , Via de Sinalização Wnt/genética
13.
Front Oncol ; 10: 576362, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33363010

RESUMO

Abnormal regulation of DNA methylation and its readers has been associated with a wide range of cellular dysfunction. Disruption of the normal function of DNA methylation readers contributes to cancer progression, neurodevelopmental disorders, autoimmune disease and other pathologies. One reader of DNA methylation known to be especially important is MeCP2. It acts a bridge and connects DNA methylation with histone modifications and regulates many gene targets contributing to various diseases; however, much remains unknown about how it contributes to cancer malignancy. We and others previously described novel MeCP2 post-translational regulation. We set out to test the hypothesis that MeCP2 would regulate novel genes linked with tumorigenesis and that MeCP2 is subject to additional post-translational regulation not previously identified. Herein we report novel genes bound and regulated by MeCP2 through MeCP2 ChIP-seq and RNA-seq analyses in two breast cancer cell lines representing different breast cancer subtypes. Through genomics analyses, we localize MeCP2 to novel gene targets and further define the full range of gene targets within breast cancer cell lines. We also further examine the scope of clinical and pre-clinical lysine deacetylase inhibitors (KDACi) that regulate MeCP2 post-translationally. Through proteomics analyses, we identify many additional novel acetylation sites, nine of which are mutated in Rett Syndrome. Our study provides important new insight into downstream targets of MeCP2 and provide the first comprehensive map of novel sites of acetylation associated with both pre-clinical and FDA-approved KDACi used in the clinic. This report examines a critical reader of DNA methylation and has important implications for understanding MeCP2 regulation in cancer models and identifying novel molecular targets associated with epigenetic therapies.

14.
Microbiol Resour Announc ; 9(29)2020 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-32675183

RESUMO

Fusarium oxysporum f. sp. albedinis is the causal agent of vascular wilt of date palm. Here, we report the genome assembly of the Foa 133 strain, which consists of 3,325 contigs with a total length of 56,228,901 bp, a GC content of 47.42%, an N 50 value of 131,587 bp, and 3,684 predicted genes.

15.
BMC Plant Biol ; 18(1): 147, 2018 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-30012094

RESUMO

BACKGROUND: SEVEN IN ABSENTIA (SINA) is a RING domain-containing ubiquitin ligase involved in Drosophila eye formation. SINA-like proteins in plants are involved in several signaling pathways. Of the 18 SINA-like proteins identified in Arabidopsis, SEVEN IN ABSENTIA 2 (SINA2) lacks a canonical RING domain and is thought to lack ubiquitin ligase activity. RESULTS: Our results show that SINA2 has E3 ligase activity in vitro, raising the possibility that a modified B-box domain may compensate for its lack of a RING domain. SINA2 physically interacts with the nuclear protein CYCLIN-DEPENDENT KINASE G1 (CDKG1), which acts as a positive regulator of plant responses to abiotic stress. CDKG1 is expressed in multiple tissues and its expression increased in response to abscisic acid (ABA) and osmotic stress. Transgenic Arabidopsis plants that ectopically express CDKG1 exhibit increased tolerance to ABA and osmotic stress treatments during seed germination and cotyledon development, while the loss-of-function cdkg1 mutant plants show reduced tolerance to ABA and osmotic stress treatments. Moreover, CDKG1-dependent phosphorylation of SINA2 positively affects its E3 ubiquitin ligase activity. CONCLUSIONS: Based on these results, we propose that CDKG1 modulates SINA2 ubiquitin ligase activity to regulate its effect on plant responses to ABA and osmotic stress.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Quinases Ciclina-Dependentes/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Ácido Abscísico/metabolismo , Arabidopsis/enzimologia , Proteínas de Arabidopsis/fisiologia , Quinases Ciclina-Dependentes/fisiologia , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Pressão Osmótica , Fosforilação , Reguladores de Crescimento de Plantas/metabolismo , Plantas Geneticamente Modificadas
16.
Plant Cell Environ ; 40(5): 702-716, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28039858

RESUMO

Arabidopsis thaliana Stress Associated Protein 9 (AtSAP9) is a member of the A20/AN1 zinc finger protein family known to play important roles in plant stress responses and in the mammalian immune response. Although SAPs of several plant species were shown to be involved in abiotic stress responses, the underlying molecular mechanisms are largely unknown, and little is known about the involvement of SAPs in plant disease responses. Expression of SAP9 in Arabidopsis is up-regulated in response to dehydration, cold, salinity and abscisic acid (ABA), as well as pathogen infection. Constitutive expression of AtSAP9 in Arabidopsis leads to increased sensitivity to ABA and osmotic stress during germination and post-germinative development. Plants that overexpress AtSAP9 also showed increased susceptibility to infection by non-host pathogen Pseudomonas syringae pv. phaseolicola, indicating a potential role of AtSAP9 in disease resistance. AtSAP9 was found to interact with RADIATION SENSITIVE23d (Rad23d), a shuttle factor for the transport of ubiquitinated substrates to the proteasome, and it is co-localized with Rad23d in the nucleus. Thus, AtSAP9 may promote the protein degradation process by mediating the interaction of ubiquitinated targets with Rad23d. Taken together, these results indicate that AtSAP9 regulates abiotic and biotic stress responses, possibly via the ubiquitination/proteasome pathway.


Assuntos
Ácido Abscísico/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Transdução de Sinais , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Núcleo Celular/metabolismo , Flores/genética , Flores/fisiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Germinação/genética , Interações Hospedeiro-Patógeno/genética , Pressão Osmótica , Imunidade Vegetal/genética , Plantas Geneticamente Modificadas , Poliubiquitina/metabolismo , Ligação Proteica , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Pseudomonas syringae/fisiologia , Estresse Fisiológico/genética , Fatores de Tempo , Ubiquitina-Proteína Ligases/metabolismo
17.
J Plant Physiol ; 169(13): 1261-70, 2012 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-22633820

RESUMO

The regulation of gene expression is a key factor in plant acclimation to stress, and it is thought that manipulation of the expression of critical stress-responsive genes should ultimately provide increased protection against abiotic stress. The aim of this study was to test the hypothesis that the ectopic expression of the AtSAP5 (AT3G12630) gene in transgenic cotton (Gossypium hirsutum, cv. Coker 312) will improve tolerance to drought and heat stress by up-regulating the expression of endogenous stress-responsive genes. The SAP5 gene is a member of the stress-associated family of genes that encode proteins containing A20/AN1 zinc finger domains. Under non-stressful conditions, cotton plants that expressed the AtSAP5 gene showed elevated expression of at least four genes normally induced during water deficit or heat stress. The rate of net CO(2) assimilation A for three of four transgenic lines tested was less sensitive to rapidly developing water deficit over 4d than untransformed wild-type plants, but the recovery of A following drought was not significantly affected. The enhanced protection of photosynthesis during drought was determined to be primarily at the biochemical level, since the extent of stomatal closure was not significantly different for all genotypes. Expression of AtSAP5 resulted in the complete protection of photosystem (PS) II complexes from photodamage at mid-day after 4 d of drought, whereas wild-type plants experienced a 20% decline in active photosystem II (PSII) complexes. In addition, enhanced protection of seedling growth and leaf viability was associated with the expression of AtSAP5. Since A for the transgenic plants was significantly more heat tolerant than A for wild-type plants, we conclude that ectopic expression of SAP genes is a potentially viable approach to improving carbon gain and productivity for cotton grown in semi-arid regions with severe drought and heat stress.


Assuntos
Proteínas de Arabidopsis/metabolismo , Gossypium/genética , Gossypium/fisiologia , Fotossíntese/genética , Fotossíntese/fisiologia , Estresse Fisiológico/fisiologia , Ubiquitina-Proteína Ligases/metabolismo , Adaptação Fisiológica , Secas , Regulação da Expressão Gênica de Plantas , Variação Genética , Genótipo , Resposta ao Choque Térmico/genética , Resposta ao Choque Térmico/fisiologia , Temperatura Alta , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia , Regulação para Cima
18.
Plant Mol Biol ; 75(4-5): 451-66, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21293909

RESUMO

AtSAP5, one of approximately 14 members of the Stress Associated Protein gene family in Arabidopsis, was identified by its expression in response to salinity, osmotic, drought and cold stress. AtSAP5 shows strong homology to OSISAP1, an A20/AN1-type zinc finger protein implicated in stress tolerance in rice. To evaluate the function of AtSAP5 in the regulation of abiotic stress responses, transgenic Arabidopsis plants that over-express AtSAP5 (35S::AtSAP5) were characterized, along with wild-type and T-DNA knock-down plants. Plants that over-express AtSAP5 showed increased tolerance to environmental challenges including salt stress, osmotic stress and water deficit. Comparison of gene expression patterns between 35S::AtSAP5 transgenic plants and wild-type plants under normal conditions and water deficit stress indicated that over-expression of AtSAP5 correlates with up-regulation of drought stress responsive gene expression. Analysis of transgenic plants that express GFP-AtSAP5 showed that it is localized primarily in nuclei of root cells and recombinant AtSAP5 has E3 ubiquitin ligase activity in vitro. These results indicate that AtSAP5 has E3 ligase activity and acts as a positive regulator of stress responses in Arabidopsis.


Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiologia , Arabidopsis/genética , Arabidopsis/fisiologia , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/fisiologia , Aclimatação/genética , Aclimatação/fisiologia , Sequência de Aminoácidos , Proteínas de Arabidopsis/química , Sequência de Bases , Clima Frio , DNA de Plantas/genética , Secas , Expressão Gênica , Técnicas de Silenciamento de Genes , Genes de Plantas , Dados de Sequência Molecular , Pressão Osmótica , Plantas Geneticamente Modificadas , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Salinidade , Homologia de Sequência de Aminoácidos , Estresse Fisiológico , Ubiquitina-Proteína Ligases/química , Dedos de Zinco
19.
Planta ; 232(5): 1191-205, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20711605

RESUMO

Plant cell wall extensibility is mediated, in part, by xyloglucan endotransglycosylases/hydrolases (XTH) that are able to cleave and reattach xyloglucan polymers that make up the hemicelluloses matrix of type I cell walls. In Arabidopsis and other plants, XTHs are encoded by relatively large gene families that are regulated in specific spatial and temporal patterns. In silico screening of a cotton expressed sequence tag (EST) database identified 23 sequences with close sequence similarity to Arabidopsis XTH coding sequences. Analysis of full-length cotton cDNAs derived from these ESTs allow for the identification of three distinct GhXTH cDNAs (denoted GhXTH1, GhXTH2 and GhXTH3) based primarily on their 3' untranslated sequences. The three GhXTH genes were expressed differently with GhXTH1 predominantly expressed in elongating cotton fibers. The function of GhXTH1 in mediating cotton fiber elongation was analyzed in transgenic cotton plants that express a transgene consisting of the GhXTH1 coding sequence under transcriptional control of the CaMV 35S promoter. Plants that over-expressed GhXTH1 had increased XTH activity and produced mature cotton fibers that were between 15 and 20% longer than wild-type cotton plants under both greenhouse and field growth conditions. Segregation analysis showed that the 35S::GhXTH1 transgene acts as a dominant fiber length allele in transgenic cotton. These results confirm that GhXTH1 is the predominant XTH in elongating fibers and its expression limits cotton fiber elongation.


Assuntos
Fibra de Algodão , Glicosiltransferases/metabolismo , Gossypium/enzimologia , Gossypium/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Sequência de Aminoácidos , Glicosiltransferases/química , Glicosiltransferases/genética , Gossypium/genética , Dados de Sequência Molecular , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Homologia de Sequência de Aminoácidos
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