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1.
PLoS One ; 15(7): e0236424, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32730292

RESUMO

Grapevines, although adapted to occasional drought or salt stress, are relatively sensitive to growth- and yield-limiting salinity stress. To understand the molecular mechanisms of salt tolerance and endoplasmic reticulum (ER) stress and identify genes commonly regulated by both stresses in grapevine, we investigated transcript profiles in leaves of the salt-tolerant grapevine rootstock 1616C under salt- and ER-stress. Among 1643 differentially expressed transcripts at 6 h post-treatment in leaves, 29 were unique to ER stress, 378 were unique to salt stress, and 16 were common to both stresses. At 24 h post-treatment, 243 transcripts were unique to ER stress, 1150 were unique to salt stress, and 168 were common to both stresses. GO term analysis identified genes in categories including 'oxidative stress', 'protein folding', 'transmembrane transport', 'protein phosphorylation', 'lipid transport', 'proteolysis', 'photosynthesis', and 'regulation of transcription'. The expression of genes encoding transporters, transcription factors, and proteins involved in hormone biosynthesis increased in response to both ER and salt stresses. KEGG pathway analysis of differentially expressed genes for both ER and salt stress were divided into four main categories including; carbohydrate metabolism, amino acid metabolism, signal transduction and lipid metabolism. Differential expression of several genes was confirmed by qRT-PCR analysis, which validated our microarray results. We identified transcripts for genes that might be involved in salt tolerance and also many genes differentially expressed under both ER and salt stresses. Our results could provide new insights into the mechanisms of salt tolerance and ER stress in plants and should be useful for genetic improvement of salt tolerance in grapevine.


Assuntos
Estresse do Retículo Endoplasmático/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Raízes de Plantas/genética , Estresse Salino/genética , Vitis/genética , Metabolismo dos Carboidratos/genética , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ontologia Genética , Análise de Sequência com Séries de Oligonucleotídeos , Osmose , Reguladores de Crescimento de Planta/farmacologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Caules de Planta/efeitos dos fármacos , Caules de Planta/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reprodutibilidade dos Testes , Estresse Salino/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Fatores de Transcrição/metabolismo , Tunicamicina/farmacologia
2.
PLoS One ; 15(6): e0234085, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32497140

RESUMO

Late embryogenesis abundant (LEA) proteins are widely involved in many adverse conditions among plants. In this study, we isolated a LEA4 gene from alfalfa (Medicago sativa L.) termed MsLEA4-4 via a homology cloning strategy. MsLEA4-4 encodes 166 amino acids, and the structural analysis showed that the gene contained five repeating TAQAAKEKTQQ amino acid motifs. There were a large number of α-helix in MsLEA4-4, and belongs to hydrophilic amino acid. Subcellular localization analysis showed that MsLEA4-4 was localized in the nucleus. The MsLEA4-4 promoter consisted of G-box and A-box elements, abscisic acid-responsive elements (ABREs), photo regulation and photoperiodic-controlling cis-acting elements, and endosperm expression motifs. The MsLEA4-4 overexpressing in Arabidopsis conferred late-germination phenotypes. Resistance of the overexpressed plants to abiotic stress significantly outperformed the wild-type (WT) plants. Under salt stress and abscisic acid treatment, with more lateral roots and higher chlorophyll content, the overexpressed plants has a higher survival rate measured against WT. Compared to those in the WT plants, the levels of soluble sugar and the activity of various antioxidant enzymes were elevated in the overexpressed plants, whereas the levels of proline and malondialdehyde were significantly reduced. The expression levels of several genes such as ABF3, ABI5, NCED5, and NCED9 increased markedly in the overexpressed plants compared to the WT under osmotic stress.


Assuntos
Arabidopsis/genética , Arabidopsis/fisiologia , Secas , Medicago sativa/genética , Estresse Oxidativo/genética , Proteínas de Plantas/genética , Estresse Salino/genética , Sequência de Aminoácidos , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Sequência de Bases , Clonagem Molecular , Expressão Gênica , Germinação , Pressão Osmótica , Proteínas de Plantas/química , Regiões Promotoras Genéticas/genética
3.
Gene ; 753: 144803, 2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32446917

RESUMO

R2R3-type MYBs are a key group of regulatory factors that control diverse developmental processes and stress tolerance in plants. Soybean is a major legume crop with the richness of seed protein and edible vegetable oil, and 244 R2R3-type MYBs have been identified in soybean. However, the knowledge regarding their functional roles has been greatly limited as yet. In this study, a novel R2R3-type MYB (GmMYB81) was functionally characterized in soybean, and it is closely related to two abiotic stress-associated regulators (AtMYB44 and AtMYB77). GmMYB81 transcripts not only differentially accumulated in soybean tissues and during embryo development, but also were significantly enhanced by drought, salt and cold stress. Histochemical GUS assay in Arabidopsis indicated that GmMYB81 promoter showed high activity in seedlings, rosette leaves, inflorescences, silique wall, mature anthers, roots, and germinating seeds. Further investigation indicated that over-expression of GmMYB81 in Arabidopsis caused auxin-associated phenotypes, including small flower and silique, more branch, and weakened apical dominance. Moreover, over-expression of GmMYB81 significantly elevated the rates of seed germination and green seedling under salt and drought stress, indicating that GmMYB81 might confer plant tolerance to salt and drought stress during seed germination. Additionally, protein interaction analysis showed that GmMYB81 interacts with the abiotic stress regulator GmSGF14l. Further observation indicated that they displayed similar expression patterns under drought and salt stress, suggesting GmMYB81 and GmSGF14l might cooperatively affect stress tolerance. These findings will facilitate future investigations of the regulatory mechanisms of GmMYB81 in response to plant stress tolerance, especially seed germination under abiotic stresses.


Assuntos
Proteínas de Arabidopsis/genética , Soja/genética , Estresse Fisiológico/genética , Fatores de Transcrição/genética , Ácido Abscísico/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Secas , Fabaceae/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Germinação/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Estresse Salino/genética , Tolerância ao Sal/genética , Sementes/metabolismo , Fatores de Transcrição/metabolismo
4.
Gene ; 753: 144802, 2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32454178

RESUMO

Synchronous and timely regulation of multiple genes results in an effective defense response that decides the fate of the host when challenged with pathogens or unexpected changes in environmental conditions. One such gene, which is downregulated in response to multiple bacterial pathogens, is a putative nonspecific lipid transfer protein (nsLTP) of unknown function that we have named DISEASE RELATED NONSPECIFIC LIPID TRANSFER PROTEIN 1 (DRN1). We show that upon pathogen challenge, DRN1 is strongly downregulated, while a putative DRN1-targeting novel microRNA (miRNA) named DRN1 Regulating miRNA (DmiR) is reciprocally upregulated. Furthermore, we provide evidence that DRN1 is required for defense against bacterial and fungal pathogens as well as for normal seedling growth under salinity stress. Although nsLTP family members from different plant species are known to be a significant source of food allergens and are often associated with antimicrobial properties, our knowledge on the biological functions and regulation of this gene family is limited. Our current work not only sheds light on the mechanism of regulation but also helps in the functional characterization of DRN1, a putative nsLTP family member of hitherto unknown function.


Assuntos
Arabidopsis/genética , Proteínas de Transferência de Fosfolipídeos/genética , Estresse Salino/genética , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Resistência à Doença/genética , Secas , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Proteínas de Transferência de Fosfolipídeos/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Patologia Vegetal , Plantas Geneticamente Modificadas , Salinidade , Tolerância ao Sal/genética , Plântula/genética , Estresse Fisiológico/genética
5.
Mol Genet Genomics ; 295(4): 877-890, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32239329

RESUMO

Basic helix-loop-helix (bHLH) gene family is a gene family of transcription factors that plays essential roles in plant growth and development, secondary metabolism and response to biotic and abiotic stresses. Therefore, a comprehensive knowledge of the bHLH gene family is paramount to understand the molecular mechanisms underlying these processes and develop advanced technologies to manipulate the processes efficiently. Ginseng, Panax ginseng C.A. Meyer, is a well-known medicinal herb; however, little is known  about the bHLH genes (PgbHLH) in the species. Here, we identified 137 PgbHLH genes from Jilin ginseng cultivar, Damaya, widely cultivated in Jilin, China, of which 50 are newly identified by pan-genome analysis. These 137 PgbHLH genes were phylogenetically classified into 26 subfamilies, suggesting their sequence diversification. They are alternatively spliced into 366 transcripts in a 4-year-old plant and involved in 11 functional subcategories of the gene ontology, indicating their functional differentiation in ginseng. The expressions of the PgbHLH genes dramatically vary spatio-temporally and across 42 genotypes, but they are still somehow functionally correlated. Moreover, the PgbHLH gene family, at least some of its genes, is shown to have roles in plant response to the abiotic stress of saline. These results provide a new insight into the evolution and functional differentiation of the bHLH gene family in plants, new bHLH genes to the PgbHLH gene family, and saline stress-responsive genes for genetic improvement in ginseng and other plant species.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Evolução Molecular , Panax/genética , Estresse Salino/genética , Processamento Alternativo/genética , China , Regulação da Expressão Gênica de Plantas/genética , Ontologia Genética , Família Multigênica/genética , Panax/efeitos dos fármacos , Panax/crescimento & desenvolvimento , Filogenia , Solução Salina/toxicidade , Fatores de Transcrição
6.
Artigo em Inglês | MEDLINE | ID: mdl-32251957

RESUMO

Soil salinity of fields is often non-uniform. To obtain a better understanding of molecular response to non-uniform salt stress, we conducted transcriptomic analysis on the leaves and roots of alfalfa grown under 0/0, 200/200, and 0/200 mM NaCl treatments. A total of 233,742 unigenes were obtained from the assembled cDNA libraries. There were 98 and 710 unigenes identified as significantly differentially expressed genes (DEGs) in the leaves of non-uniform and uniform salt treatment, respectively. Furthermore, there were 5178 DEGs in the roots under uniform salt stress, 273 DEGs in the non-saline side and 4616 in the high-saline side roots under non-uniform salt stress. Alfalfa treated with non-uniform salinity had greater dry weight and less salt damage compared to treatment with uniform salinity. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the DEGs in roots revealed that both sides of the non-uniform salinity were enriched in pathways related to "phenylpropanoid biosynthesis" and "linoleic acid metabolism"; and "MAPK signaling pathway-plant" was also indicated as a key pathway in the high-saline roots. We also combined a set of important salt-response genes and found that roots from the non-saline side developed more roots with increased water uptake by altering the expression of aquaporins and genes related to growth regulation. Moreover, the hormone signal transduction and the antioxidant pathway probably play important roles in inducing more salt-related genes and increasing resistance to non-uniform salt stress on both sides of the roots.


Assuntos
Medicago sativa , Tolerância ao Sal , Transcriptoma , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Medicago sativa/genética , Medicago sativa/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Salinidade , Estresse Salino/genética , Tolerância ao Sal/genética
7.
Artigo em Inglês | MEDLINE | ID: mdl-32315911

RESUMO

Arundo donax L. is an invasive grass species with high tolerance to a wide range of environmental stresses. The response of potted A. donax plants to soil stress characterized by prolonged exposure (43 days) to salinity (+Na), to high concentration of phosphorus (+P), and to the combination of high Na and P (+NaP) followed by 14 days of recovery under optimal nutrient solution, was investigated along the entire time-course of the experiment. After an exposure of 43 days, salinity induced a progressive decline in stomatal conductance that hampered A. donax growth through diffusional limitations to photosynthesis and, when combined with high P, reduced the electron transport rate. Isoprene emission from A. donax leaves was stimulated as Na+ concentration raised in leaves. Prolonged growth in P-enriched substrate did not significantly affect A. donax performance, but decreased isoprene emission from leaves. Prolonged exposure of A. donax to + NaP increased the leaf level of H2O2, stimulated the production of carbohydrates, phenylpropanoids, zeaxanthin and increased the de-epoxidation state of the xanthophylls. This might have resulted in a higher stress tolerance that allowed a fast and full recovery following stress relief. Moreover, the high amount of ABA-glucose ester accumulated in leaves of A. donax exposed to + NaP might have favored stomata re-opening further sustaining the observed prompt recovery of photosynthesis. Therefore, prolonged exposure to high P exacerbated the negative effects of salt stress in A. donax plants photosynthetic performances, but enhanced activation of physiological mechanisms that allowed a prompt and full recovery after stress.


Assuntos
Fósforo , Poaceae , Estresse Salino , Solo , Peróxido de Hidrogênio , Fósforo/farmacologia , Fotossíntese , Folhas de Planta/efeitos dos fármacos , Poaceae/efeitos dos fármacos , Estresse Salino/genética , Solo/química
8.
PLoS One ; 15(2): e0228400, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32027667

RESUMO

Obesity is a multifactorial disease caused by complex interactions between genes and dietary factors. Salt-rich diet is related to the development and progression of several chronic diseases including obesity. However, the molecular basis of how salt sensitivity genes (SSG) contribute to adiposity in obesity patients remains unexplored. In this study, we used the microarray expression data of visceral adipose tissue samples and constructed a complex protein-interaction network of salt sensitivity genes and their co-expressed genes to trace the molecular pathways connected to obesity. The Salt Sensitivity Protein Interaction Network (SSPIN) of 2691 differentially expressed genes and their 15474 interactions has shown that adipose tissues are enriched with the expression of 23 SSGs, 16 hubs and 84 bottlenecks (p = 2.52 x 10-16) involved in diverse molecular pathways connected to adiposity. Fifteen of these 23 SSGs along with 8 other SSGs showed a co-expression with enriched obesity-related genes (r ≥ 0.8). These SSGs and their co-expression partners are involved in diverse metabolic pathways including adipogenesis, adipocytokine signaling pathway, renin-angiotensin system, etc. This study concludes that SSGs could act as molecular signatures for tracing the basis of adipogenesis among obese patients. Integrated network centered methods may accelerate the identification of new molecular targets from the complex obesity genomics data.


Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Redes Reguladoras de Genes/efeitos dos fármacos , Obesidade Pediátrica/genética , Estresse Salino/genética , Cloreto de Sódio na Dieta/farmacologia , Adiposidade/efeitos dos fármacos , Adiposidade/genética , Adolescente , Estudos de Casos e Controles , Epistasia Genética/efeitos dos fármacos , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Análise em Microsséries/métodos , Obesidade Pediátrica/metabolismo , Obesidade Pediátrica/patologia , Estresse Salino/efeitos dos fármacos , Integração de Sistemas , Adulto Jovem
9.
Gene ; 738: 144460, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32045659

RESUMO

MicroRNA390 (miR390), an ancient and highly conserved miRNA family in land plants, plays multiple roles in plant growth, development and stress responses. In this study, we isolated and identified MIR390, miR390, TAS3a/b/c, tasiARF-1/2/3 (trans-acting small interfering RNAs influencing Auxin Response Factors) and ARF2/3/4 in Jerusalem artichoke (Helianthus tuberosus L.). Treatment with 100 mM NaCl induced expression of miR390, increased cleavage of TAS3, produced high levels of tasiARFs, and subsequently enhanced cleavage of ARF3/4, which was most likely associated with salt tolerance of the plants. In contrast, treatment with 300 mM NaCl inhibited expression of miR390, attenuated cleavage of TAS3, produced a small amount of tasiARFs, and reduced cleavage of ARF3/4. We proposed that ARF2, one of the targets of tasiARFs, induced under salinity was likely to play an active role in salt tolerance of Jerusalem artichoke. The study of the miR390-TAS3-ARF model in Jerusalem artichoke may broaden our understanding of salt tolerance mechanisms, and provides a theoretical support for further genetic identification and breeding crops with increased tolerance to salt stress.


Assuntos
Helianthus/metabolismo , MicroRNAs/metabolismo , Estresse Salino/fisiologia , Regulação da Expressão Gênica de Plantas , Helianthus/genética , MicroRNAs/genética , Raízes de Plantas/genética , RNA de Plantas/genética , RNA Interferente Pequeno/genética , Salinidade , Estresse Salino/genética , Tolerância ao Sal/genética
10.
Artigo em Inglês | MEDLINE | ID: mdl-32005379

RESUMO

SnRK2 (sucrose non-fermenting 1-related protein kinases 2) protein kinase family involves in several abiotic stress response in plants. Although the regulatory mechanism of SnRK2 have been well demonstrated in Arabidopsis thaliana, their functions in rice are still largely unknown. Here, we report a SnRK2 family gene, OsSAPK8, can be strongly induced by abiotic stresses, including low-temperature, drought and high salt stress. The ossapk8 mutants showed lower tolerance to low-temperature, high salinity and drought stresses at the vegetative stages. Moreover, the expressions of marker genes for those abiotic stresses, e.g. OsDREB1, OsDREB2, OsNCED and OsRAB21, were downregulated in the ossapk8 mutants. We further confirmed that the yield was reduced in ossapk8 mutant lines compared with the wild type. Our results provide evidence for OsSAPK8 acting as a positive regulator in cold, drought, and salt stress responses.


Assuntos
Regulação da Expressão Gênica de Plantas/fisiologia , Oryza/fisiologia , Proteínas de Plantas/genética , Proteínas Quinases/genética , Estresse Fisiológico/genética , Temperatura Baixa , Secas , Oryza/genética , Proteínas de Plantas/metabolismo , Proteínas Quinases/metabolismo , Estresse Salino/genética
11.
Artigo em Inglês | MEDLINE | ID: mdl-32058898

RESUMO

NACs are one of the largest transcription factor families in plants and are involved in the response to abiotic stress. A new stress-responsive NAC transcription factor gene, LpNAC13, was isolated from Lilium pumilum bulbs. The expression of LpNAC13 was induced by drought, salt, cold and ABA treatments. LpNAC13 overexpressing plants were generated to explore the function of LpNAC13 in response to drought and salt stress. Overexpression of LpNAC13 in tobacco displayed a reduced drought tolerance but exhibited an enhanced salt tolerance. The LpNAC13 overexpression plants had decreased antioxidant enzyme activities, content of proline and chlorophyll, increased MDA content under drought condition, the results in the LpNAC13 plants under salt condition were opposite to those under drought condition. The seed germination and root length assays of overexpression of LpNAC13 showed decreased sensitivity to ABA. Functional analyses demonstrate that LpNAC13 plays opposite roles in drought and salt stress tolerance, acting as a negative regulator of drought response but as a positive regulator of salt response in tobacco.


Assuntos
Secas , Lilium , Estresse Fisiológico , Tabaco , Regulação da Expressão Gênica de Plantas , Lilium/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Estresse Salino/genética , Estresse Fisiológico/genética , Tabaco/genética , Tabaco/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
12.
Sci Rep ; 10(1): 1383, 2020 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-31992813

RESUMO

Small heat shock proteins (sHSPs) have been thought to function as chaperones, protecting their targets from denaturation and aggregation when organisms are subjected to various biotic and abiotic stresses. We previously reported an sHSP from Oryza sativa (OsHSP20) that homodimerizes and forms granules within the cytoplasm but its function was unclear. We now show that OsHSP20 transcripts were significantly up-regulated by heat shock and high salinity but not by drought. A recombinant protein was purified and shown to inhibit the thermal aggregation of the mitochondrial malate dehydrogenase (MDH) enzyme in vitro, and this molecular chaperone activity suggested that OsHSP20 might be involved in stress resistance. Heterologous expression of OsHSP20 in Escherichia coli or Pichia pastoris cells enhanced heat and salt stress tolerance when compared with the control cultures. Transgenic rice plants constitutively overexpressing OsHSP20 and exposed to heat and salt treatments had longer roots and higher germination rates than those of control plants. A series of assays using its truncated mutants showed that its N-terminal arm plus the ACD domain was crucial for its homodimerization, molecular chaperone activity in vitro, and stress tolerance in vivo. The results supported the viewpoint that OsHSP20 could confer heat and salt tolerance by its molecular chaperone activity in different organisms and also provided a more thorough characterization of HSP20-mediated stress tolerance in O. sativa.


Assuntos
Escherichia coli/metabolismo , Proteínas de Choque Térmico HSP20 , Microrganismos Geneticamente Modificados/metabolismo , Oryza/genética , Pichia/metabolismo , Proteínas de Plantas , Multimerização Proteica , Tolerância ao Sal , Escherichia coli/genética , Proteínas de Choque Térmico HSP20/biossíntese , Proteínas de Choque Térmico HSP20/genética , Microrganismos Geneticamente Modificados/genética , Pichia/genética , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Estresse Salino/genética
13.
BMC Plant Biol ; 20(1): 20, 2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31931714

RESUMO

BACKGROUND: Plant genomes contain a large number of HAK/KUP/KT transporters, which play important roles in potassium uptake and translocation, osmotic potential regulation, salt tolerance, root morphogenesis and plant development. Potassium deficiency in the soil of a sugarcane planting area is serious. However, the HAK/KUP/KT gene family remains to be characterized in sugarcane (Saccharum). RESULTS: In this study, 30 HAK/KUP/KT genes were identified in Saccharum spontaneum. Phylogenetics, duplication events, gene structures and expression patterns were analyzed. Phylogenetic analysis of the HAK/KUP/KT genes from 15 representative plants showed that this gene family is divided into four groups (clades I-IV). Both ancient whole-genome duplication (WGD) and recent gene duplication contributed to the expansion of the HAK/KUP/KT gene family. Nonsynonymous to synonymous substitution ratio (Ka/Ks) analysis showed that purifying selection was the main force driving the evolution of HAK/KUP/KT genes. The divergence time of the HAK/KUP/KT gene family was estimated to range from 134.8 to 233.7 Mya based on Ks analysis, suggesting that it is an ancient gene family in plants. Gene structure analysis showed that the HAK/KUP/KT genes were accompanied by intron gain/loss in the process of evolution. RNA-seq data analysis demonstrated that the HAK/KUP/KT genes from clades II and III were mainly constitutively expressed in various tissues, while most genes from clades I and IV had no or very low expression in the tested tissues at different developmental stages. The expression of SsHAK1 and SsHAK21 was upregulated in response to low-K+ stress. Yeast functional complementation analysis revealed that SsHAK1 and SsHAK21 could rescue K+ uptake in a yeast mutant. CONCLUSIONS: This study provided insights into the evolutionary history of HAK/KUP/KT genes. HAK7/9/18 were mainly expressed in the upper photosynthetic zone and mature zone of the stem. HAK7/9/18/25 were regulated by sunlight. SsHAK1 and SsHAK21 played important roles in mediating potassium acquisition under limited K+ supply. Our results provide valuable information and key candidate genes for further studies on the function of HAK/KUP/KT genes in Saccharum.


Assuntos
Proteínas de Transporte de Cátions/genética , Proteínas de Plantas/genética , Potássio/metabolismo , Saccharum , Proteínas de Transporte de Cátions/metabolismo , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Família Multigênica , Filogenia , Desenvolvimento Vegetal/fisiologia , Proteínas de Plantas/metabolismo , RNA-Seq , Saccharum/genética , Saccharum/metabolismo , Estresse Salino/genética , Estresse Salino/fisiologia
14.
Biochim Biophys Acta Gen Subj ; 1864(3): 129514, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31911239

RESUMO

BACKGROUND: Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker is maintained as an epiphyte on citrus leaves until entering the plant tissue. During epiphytic survival, bacteria may encounter low water availability that challenges the infection process. Proteomics analyses of Xcc under saline stress, mimicking the conditions found during epiphytic survival, showed increased abundance of a putative NAD(P)H dehydrogenase encoded by XAC2229. METHODS: Expression levels of XAC2229 and a Xcc mutant in XAC2229 were analyzed in salt and oxidative stress and during plant-pathogen interaction. An Escherichia coli expressing XAC2229 was obtained, and the role of this protein in oxidative stress resistance and in reactive oxygen species production was studied. Finally, Xac2229 protein was purified, spectrophotometric and cofactor analyses were done and enzymatic activities determined. RESULTS: XAC2229 was expressed under salt stress and during plant-pathogen interaction. ΔXAC2229 mutant showed less number of cankers and impaired epiphytic survival than the wild type strain. ΔXAC2229 survived less in the presence of H2O2 and produced more reactive oxygen species and thiobarbituric acid-reactive substances than the wild type strain. Similar results were observed for E. coli expressing XAC2229. Xac2229 is a FAD containing flavoprotein, displays diaphorase activity with an optimum at pH 6.0 and has quinone reductase activity using NADPH as an electron donor. CONCLUSIONS: A FAD containing flavoprotein from Xcc is a new NADPH quinone reductase required for bacterial virulence, particularly in Xcc epiphytic survival on citrus leaves. GENERAL SIGNIFICANCE: A novel protein involved in the worldwide disease citrus canker was characterized.


Assuntos
NAD(P)H Desidrogenase (Quinona)/metabolismo , Xanthomonas/enzimologia , Benzoquinonas/metabolismo , Citrus/metabolismo , Citrus/microbiologia , Peróxido de Hidrogênio/metabolismo , NAD(P)H Desidrogenase (Quinona)/genética , NADP/metabolismo , Estresse Oxidativo , Folhas de Planta/metabolismo , Estresse Salino/genética , Estresse Salino/fisiologia , Virulência , Xanthomonas/metabolismo , Xanthomonas/patogenicidade , Xanthomonas/fisiologia
15.
Appl Environ Microbiol ; 86(6)2020 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-31900307

RESUMO

Listeria monocytogenes causes the severe foodborne illness listeriosis and survives in food-associated environments due to its high stress tolerance. A data assembly and analysis protocol for microbial growth experiments was compiled to elucidate the strain variability of L. monocytogenes stress tolerance. The protocol includes measurement of growth ability under stress (step 1), selection of a suitable method for growth parameter calculation (step 2), comparison of growth patterns between strains (step 3), and biological interpretation of the discovered differences (step 4). In step 1, L. monocytogenes strains (n = 388) of various serovars and origins grown on media with 9.0% NaCl were measured using a Bioscreen C microbiology reader. Technical variability of the growth measurements was assessed and eliminated. In step 2, the growth parameters determined by Gompertz, modified-Gompertz, logistic, and Richards models and model-free splines were compared, illustrating differences in the suitability of these methods to describe the experimental data. In step 3, hierarchical clustering was used to describe the NaCl tolerance of L. monocytogenes measured by strain-specific variation in growth ability; tolerant strains had higher growth rates and maximum optical densities and shorter lag phases than susceptible strains. The spline parameter area under the curve best classified "poor," "average," and "good" growers. In step 4, the tested L. monocytogenes lineage I strains (serovars 4b and 1/2b) proved to be significantly more tolerant toward 9.0% NaCl than lineage II strains (serovars 1/2a, 1/2c, and 3a). Our protocol provides systematic tools to gain comparable data for investigating strain-specific variation of bacterial growth under stress.IMPORTANCE The pathogen Listeria monocytogenes causes the foodborne disease listeriosis, which can be fatal in immunocompromised individuals. L. monocytogenes tolerates several environmental stressors and can persist in food-processing environments and grow in foodstuffs despite traditional control measures such as high salt content. Nonetheless, L. monocytogenes strains differ in their ability to withstand stressors. Elucidating the intraspecies strain variability of L. monocytogenes stress tolerance is crucial for the identification of particularly tolerant strains. To enhance reliable identification of variability in bacterial stress tolerance phenotypes, we compiled a large-scale protocol for the entire data assembly and analysis of microbial growth experiments, providing a systematic approach and checklist for experiments on strain-specific growth ability. Our study illustrated the diversity and strain-specific variation of L. monocytogenes stress tolerance with an unprecedented scope and discovered biologically relevant serovar- and lineage-dependent phenotypes of NaCl tolerance.


Assuntos
Listeria monocytogenes/fisiologia , Estresse Salino/genética , Cloreto de Sódio/efeitos adversos , Ensaios de Triagem em Larga Escala , Listeria monocytogenes/genética , Fenótipo , Sorotipagem
16.
Artigo em Inglês | MEDLINE | ID: mdl-31928685

RESUMO

Salt stress is one of the major environmental factors limiting crop productivity. Although physiological and molecular characterization of salt stress response in plants has been the focus for many years, research on transporters for sodium ion (Na+) uptake, translocation and accumulation in plants, particularly in food crops like rice is limited. In this study, we functionally identified an uncharacterized sodium ion transporter named OsNHAD which encodes a putative Na+ / H+ antiporter in rice. Homology search shows its close relation to the Arabidopsis Na+/H+ antiporter AtNHD1 with 72.74% identity of amino acids. OsNHAD transcripts mainly express in leaves and are induced by Na+ stress. Confocal laser scanning microscopy analysis of OsNHAD::GFP fusion in tobacco leaves shows that OsNHAD resides in the chloroplast envelop. Knock-down of OsNHAD by RNA interference led to increased rice sensitivity to Na+, manifested by stunted plant growth, enhanced cellular damage, reduced PSII activity and changed chloroplast morphology. Mutation of OsNHAD also resulted in accumulation of more Na+ in chloroplasts and in shoots as well, suggesting that OsNHAD is involved in mediating efflux and detoxification of Na+ but does not affect K+ accumulation in plant cells. Complementation test reveals that OsNHAD was able to functionally restore the Arabidopsis mutant atnhd1-1 growth phenotype. These results suggest that OsNHAD possibly mediates homeostasis of sodium ions in the subcellular compartments and tissues of the plants when challenged to salt stress.


Assuntos
Cloroplastos/metabolismo , Oryza/fisiologia , Proteínas de Plantas/genética , Estresse Salino/genética , Trocadores de Sódio-Hidrogênio/genética , Oryza/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Sódio/metabolismo , Trocadores de Sódio-Hidrogênio/metabolismo
17.
Genes Genomics ; 42(3): 325-335, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31894476

RESUMO

BACKGROUND: DEAD-box protein family is the largest subfamily of RNA helicases and plays an important role in RNA metabolism and plant growth, development, and stress responses. Although DEAD-box genes have been characterized in various major crop plants, their identification and characterization in Convolvulaceae is still in infancy. Sweet potato (Ipomoea batatas, in Convolvulaceae) is the seventh most important crop in the world. Ipomoea trifida is one of the ancestors of sweet potato and is an effective resource for sweet potato cross-breeding. OBJECTIVE: Identification and characterisation of DEAD-box transcription factors in sweet potato wild ancestor I. trifida genome. METHOD: A systematic genome-wide analysis was conducted to identify the DEAD-box transcription factors in the I. trifida genome. RESULTS: We identified 17 ItfDEAD-box genes which distributed unevenly on the nine chromosomes of I. trifida and encoded 29 DEAD transcripts. The phylogenetic analysis classified the DEAD-box proteins into nine groups named I-IX. Homology model prediction of ItfDEAD-box proteins obtained 14 models which lay a preliminary foundation for the further functional exploration of the ItfDEAD-box proteins. The tissue-specific and abiotic stress-responsive expression profiles of ItfDEAD-box genes were analyzed in different tissues and under abiotic stress responses by RNA-seq data and confirmed by quantitative PCR analysis. Some genes were significantly up- or down-regulated by different abiotic stress, suggesting that ItfDEAD-box plays a crucial role in stress responses in I. trifida. CONCLUSION: The identification and gene expression of the ItfDEAD-box gene family might shed light on the function exploration of DEAD-box gene in I. trifida and promote the molecular breeding of sweet potato.


Assuntos
RNA Helicases DEAD-box/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Ipomoea/metabolismo , Estresse Fisiológico/genética , Mapeamento Cromossômico , Resposta ao Choque Frio/genética , RNA Helicases DEAD-box/química , RNA Helicases DEAD-box/genética , Bases de Dados Genéticas , Secas , Genes de Plantas , Genoma de Planta , Estudo de Associação Genômica Ampla , Resposta ao Choque Térmico/genética , Ipomoea/enzimologia , Ipomoea/genética , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Modelos Moleculares , Especificidade de Órgãos/genética , Filogenia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , RNA-Seq , Estresse Salino/genética
18.
Artigo em Inglês | MEDLINE | ID: mdl-31841963

RESUMO

Aquaporins (AQPs) transport water and other small molecules; however, their precise role in abiotic stress responses is not fully understood. In this study, we cloned and characterized the PIP2 group AQP gene, MaPIP2-7, in banana. MaPIP2-7 expression was upregulated after osmotic (mannitol), cold, and salt treatments. Overexpression of MaPIP2-7 in banana improved tolerance to multiple stresses such as drought, cold, and salt. MaPIP2-7 transgenic plants showed lower levels of malondialdehyde (MDA) and ion leakage (IL), but higher contents of chlorophyll, proline, soluble sugar, and abscisic acid (ABA) compared with wild type (WT) plants under stress and recovery conditions. Additionally, MaPIP2-7 overexpression decreased cellular contents of Na+ and K+ under salt and recovery conditions, and produced an elevated K+/Na+ ratio under recovery conditions. Finally, ABA biosynthetic and responsive genes exhibited higher expression levels in transgenic lines relative to WT under stress conditions. Taken together, our results demonstrate that MaPIP2-7 confers tolerance to drought, cold, and salt stresses by maintaining osmotic balance, reducing membrane injury, and improving ABA levels.


Assuntos
Aquaporinas , Secas , Musa , Proteínas de Plantas , Plantas Geneticamente Modificadas , Estresse Fisiológico , Regulação da Expressão Gênica de Plantas , Musa/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estresse Salino/genética , Estresse Fisiológico/genética
19.
Artigo em Inglês | MEDLINE | ID: mdl-31863135

RESUMO

KEY MESSAGE: PpCKX1 localizes to vacuoles and is dominantly expressed in the stem cells. PpCKX1 regulates developmental changes with increased growth of the rhizoid and enhances dehydration and salt tolerance. Cytokinins (CKs) are plant hormones that regulate plant development as well as many physiological processes, such as cell division, leaf senescence, control of shoot/root ratio, and reproductive competence. Cytokinin oxidases/dehydrogenases (CKXs) control CK concentrations by degradation, and thereby influence plant growth and development. In the moss Physcomitrella patens, an evolutionarily early divergent plant, we identified six putative CKXs that, by phylogenetic analysis, form a monophyletic clade. We also observed that ProPpCKX1:GUS is expressed specifically in the stem cells and surrounding cells and that CKX1 localizes to vacuoles, as indicated by Pro35S:PpCKX1-smGFP. Under normal growth conditions, overexpression of PpCKX1 caused many phenotypic changes at different developmental stages, and we suspected that increased growth of the rhizoid could affect those changes. In addition, we present evidence that the PpCKX1-overexpressor plants show enhanced dehydration and salt stress tolerance. Taken together, we suggest that PpCKX1 plays regulatory roles in development and adaptation to abiotic stresses in this evolutionarily early land plant species.


Assuntos
Bryopsida/enzimologia , Bryopsida/crescimento & desenvolvimento , Oxirredutases/metabolismo , Tolerância ao Sal , Bryopsida/genética , Citocininas/metabolismo , Desidratação , Regulação da Expressão Gênica de Plantas , Fenótipo , Filogenia , Plantas Geneticamente Modificadas , Estresse Salino/genética , Tolerância ao Sal/genética , Células-Tronco/metabolismo , Vacúolos/metabolismo
20.
PLoS One ; 14(12): e0225926, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31794584

RESUMO

Selecting appropriate reference genes is vital to normalize gene expression analysis in birch (Betula platyphylla) under different abiotic stress conditions using quantitative real-time reverse transcription PCR (qRT-PCR). In this study, 11 candidate birch reference genes (ACT, TUA, TUB, TEF, 18S rRNA, EF1α, GAPDH, UBC, YLS8, SAND, and CDPK) were selected to evaluate the stability of their expression in different tissues and under different abiotic stress conditions. Three statistical algorithms (GeNorm, NormFinder, and BestKeeper) were used to analyze the stability of the 11 candidate reference genes to identify the most appropriate one. The results indicated that EF-1α was the most stable reference gene in different birch tissues, ACT was the most stable reference gene for normal conditions, ACT and TEF were the most stable reference genes for salt stress treatment, TUB was the most stable reference gene for osmotic stress treatment, and ACT was the most appropriate choice in all samples of birch. In conclusion, the most appropriate reference genes varied among different experimental conditions. However, in this study, ACT was the optimum reference gene in all experimental groups, except in the different tissues group. GAPDH was the least stable candidate reference gene in all experimental conditions. In addition, three stress-induced genes (BpGRAS1, BpGRAS16, and BpGRAS19) were chosen to verify the stability of the selected reference genes in different tissues and under salt stress. This study laid the foundation for the selection of appropriate reference gene(s) for future gene expression pattern studies in birch.


Assuntos
Adaptação Biológica/genética , Betula/fisiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Pressão Osmótica , Estresse Salino/genética , Biologia Computacional/métodos , Reprodutibilidade dos Testes , Transcriptoma
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