Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 6.473
Filtrar
1.
PLoS Pathog ; 16(8): e1008801, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32866183

RESUMO

Rice stripe virus (RSV) is one of the most destructive viral diseases affecting rice production. However, so far, only one RSV resistance gene has been cloned, the molecular mechanisms underlying host-RSV interaction are still poorly understood. Here, we show that increasing levels or signaling of brassinosteroids (BR) and jasmonic acid (JA) can significantly enhance the resistance against RSV. On the contrary, plants impaired in BR or JA signaling are more susceptible to RSV. Moreover, the enhancement of RSV resistance conferred by BR is impaired in OsMYC2 (a key positive regulator of JA response) knockout plants, suggesting that BR-mediated RSV resistance requires active JA pathway. In addition, we found that RSV infection suppresses the endogenous BR levels to increase the accumulation of OsGSK2, a key negative regulator of BR signaling. OsGSK2 physically interacts with OsMYC2, resulting in the degradation of OsMYC2 by phosphorylation and reduces JA-mediated defense to facilitate virus infection. These findings not only reveal a novel molecular mechanism mediating the crosstalk between BR and JA in response to virus infection and deepen our understanding about the interaction of virus and plants, but also suggest new effective means of breeding RSV resistant crops using genetic engineering.


Assuntos
Brassinosteroides/metabolismo , Ciclopentanos/metabolismo , Oryza , Oxilipinas/metabolismo , Plantas Geneticamente Modificadas , Transdução de Sinais , Tenuivirus , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Oryza/genética , Oryza/metabolismo , Oryza/virologia , Doenças das Plantas/genética , Doenças das Plantas/virologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/virologia , Tenuivirus/genética , Tenuivirus/metabolismo
2.
PLoS One ; 15(8): e0237448, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32790800

RESUMO

We established that Endosidin2 (ES2) affected the trafficking routes of both newly synthesized and endocytic pools of PIN-FORMED2 (PIN2) in Arabidopsis root epidermal cells. PIN2 populations accumulated in separated patches, which gradually merged into large and compact ES2 aggregates (ES2As). FM4-64 endocytic tracer labeled ES2As as well. Both PIN2 pools also appeared in vacuoles. Accelerated endocytosis of PIN2, its aggregation in the cytoplasm, and redirection of PIN2 flows to vacuoles led to a substantial reduction of the abundance of this protein in the plasma membrane. Whereas PIN-FORMED3 and PIN-FORMED4 also aggregated in the cytoplasm, SYT1 was not sensitive to ES2 treatment and did not appear either in the cytoplasmic aggregates or vacuoles. Ultrastructural analysis revealed that ES2 affects the Golgi apparatus so that stacks acquired cup-shape and even circular shape surrounded by several vesicles. Abnormally shaped Golgi stacks, stack remnants, multi-lamellar structures, separated Golgi cisterna rings, tubular structures, and vesicles formed discrete clusters.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Endocitose/efeitos dos fármacos , Limoninas/farmacologia , Proteínas de Membrana Transportadoras/metabolismo , Parede Celular/metabolismo , Citoplasma/metabolismo , Complexo de Golgi/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Transporte Proteico/efeitos dos fármacos , Sinaptotagmina I/metabolismo
3.
Ecotoxicol Environ Saf ; 202: 110918, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32800253

RESUMO

This work evaluates different generations of transgenic (cp4-EPSPS gene) and non-transgenic soybean plants through proteomics and metabolomics. For proteomics purpose, 24 differentially abundant protein spots were found through 2-D DIGE, being 4 belonging to transgenic plants. From this total, 19 were successfully identified, storage proteins as predominant class. Some identified proteins are involved in growing and cell division, and stress response, such as LEA and dehydrin. For metabolomics, 17 compounds were putatively annotated, mainly belonging to the secondary metabolism, such as flavonoids. From these analyzes, all generations and varieties of the soybean are prone to be differentiate by PLS-DA. According to our results, transgenic plants appear to be more stable than non-transgenic ones. In addition, the omics-based approaches allowed access some relations between those differential spot proteins and metabolites, mainly those storage proteins and flavonoid.


Assuntos
Plantas Geneticamente Modificadas/fisiologia , Soja/fisiologia , Metabolômica , Plantas Geneticamente Modificadas/metabolismo , Proteômica , Sementes/metabolismo , Soja/genética , Soja/metabolismo
4.
PLoS One ; 15(8): e0236246, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32804956

RESUMO

K+ is an essential nutrient for plant growth and is responsible for many important physiological processes. K+ deficiency leads to crop yield losses, and overexpression of K+ transporter genes has been proven to be an effective way to resolve this problem. However, current research on the overexpression of K+ transporter genes is limited to plant sources. TrkH is a bacterial K+ transporter whose function generally depends on the regulation of TrkA. To date, whether TrkH can improve K+ uptake in eukaryotic organisms is still unknown. In this study, a novel MbtrkH gene was cloned from marine microbial metagenomic DNA. Functional complementation and K+-depletion analyses revealed that MbTrkH functions in K+ uptake in the K+-deficient yeast strain CY162. Moreover, K+-depletion assays revealed that MbtrkH overexpression improves plant K+ uptake. K+ hydroponic culture experiments showed that, compared with WT tobacco lines, MbtrkH transgenic tobacco lines had significantly greater fresh weights, dry weights and K+ contents. These results indicate that MbTrkH promotes K+ uptake independently of TrkA in eukaryotes and provide a new strategy for improving K+-use efficiency in plants.


Assuntos
Organismos Aquáticos/genética , Potássio/metabolismo , Saccharomyces cerevisiae/metabolismo , Água do Mar/microbiologia , Tabaco/metabolismo , Clonagem Molecular , Metagenoma , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Canais de Potássio/genética , Canais de Potássio/metabolismo , Saccharomyces cerevisiae/genética , Tabaco/genética
5.
PLoS One ; 15(7): e0236943, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32735612

RESUMO

Halophyte Lobularia maritima LmSAP encodes an A20AN1 zinc-finger stress-associated protein which expression is up-regulated by abiotic stresses and heavy metals in transgenic tobacco. To deepen our understanding of LmSAP function, we isolated a 1,147 bp genomic fragment upstream of LmSAP coding sequence designated as PrLmSAP. In silico analyses of PrLmSAP revealed the presence of consensus CAAT and TATA boxes and cis-regulatory elements required for abiotic stress, phytohormones, pathogen, and wound responses, and also for tissue-specific expression. The PrLmSAP sequence was fused to the ß-glucuronidase (gusA) reporter gene and transferred to rice. Histochemical GUS staining showed a pattern of tissue-specific expression in transgenic rice, with staining observed in roots, coleoptiles, leaves, stems and floral organs but not in seeds or in the root elongation zone. Wounding strongly stimulated GUS accumulation in leaves and stems. Interestingly, we observed a high stimulation of the promoter activity when rice seedlings were exposed to NaCl, PEG, ABA, MeJA, GA, cold, and heavy metals (Al3+, Cd2+, Cu2+ and Zn2+). These results suggest that the LmSAP promoter can be a convenient tool for stress-inducible gene expression and is a potential candidate for crop genetic engineering.


Assuntos
Regulação da Expressão Gênica de Plantas/genética , Regiões Promotoras Genéticas , Plantas Tolerantes a Sal/genética , Estresse Fisiológico/genética , Dedos de Zinco/genética , Produtos Agrícolas/genética , Engenharia Genética , Glucuronidase/metabolismo , Metais Pesados/metabolismo , Especificidade de Órgãos , Oryza/genética , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Tabaco/genética
6.
PLoS One ; 15(7): e0236571, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32730284

RESUMO

The occurrence of genotype by environment interaction (G x E), which is defined as the differential response of genotypes to environmental variation, is frequently reported in maize cultures, making it challenging to recommend cultivars. Methods allowing to study the potential nonlinear pattern of genotype responses to environmental variation allied to prior beliefs on unknown parameters are interesting to evaluate the phenotypic adaptability and stability of genotypes. In this context, the present study aimed to assess the adaptability and stability of maize hybrids, by using the Bayesian segmented regression model, and evaluate the efficacy of using informative and minimally informative prior distributions for the selection of cultivars. Randomized complete-block design experiments were carried out to study the yield (kg/ha) of 25 maize hybrids, in 22 different environments, in Northeastern Brazil. The Bayesian segmented regression model fitted using informative prior distributions presented lower credibility intervals and Deviance Criterium of Information values, compared to those obtained by fitting using minimally informative distributions. Therefore, the model using informative prior distributions was considered for the adaptability and stability evaluation of maize genotypes. Once most northeastern farmers in Brazil have limited capital, the genotype P4285HX should be considered for planting, due to its high yield performance and adaptability to unfavorable environments.


Assuntos
Modelos Genéticos , Zea mays/genética , Adaptação Fisiológica/genética , Teorema de Bayes , Interação Gene-Ambiente , Genótipo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismo
7.
Nat Commun ; 11(1): 3439, 2020 07 10.
Artigo em Inglês | MEDLINE | ID: mdl-32651385

RESUMO

Various stress conditions induce the nuclear translocation of cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC), but its nuclear function in plant stress responses remains elusive. Here we show that GAPC interacts with a transcription factor to promote the expression of heat-inducible genes and heat tolerance in Arabidopsis. GAPC accumulates in the nucleus under heat stress. Overexpression of GAPC enhances heat tolerance of seedlings and the expression of heat-inducible genes whereas knockout of GAPCs has opposite effects. Screening of Arabidopsis transcription factors identifies nuclear factor Y subunit C10 (NF-YC10) as a GAPC-binding protein. The effects of GAPC overexpression are abolished when NF-YC10 is deficient, the heat-induced nuclear accumulation of GAPC is suppressed, or the GAPC-NF-YC10 interaction is disrupted. GAPC overexpression also enhances the binding ability of NF-YC10 to its target promoter. The results reveal a cellular and molecular mechanism for the nuclear moonlighting of a glycolytic enzyme in plant response to environmental changes.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Fator de Ligação a CCAAT/genética , Fator de Ligação a CCAAT/metabolismo , Núcleo Celular/genética , Núcleo Celular/metabolismo , Núcleo Celular/fisiologia , Citosol/metabolismo , Citosol/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Gliceraldeído-3-Fosfato Desidrogenases/genética , Resposta ao Choque Térmico/genética , Resposta ao Choque Térmico/fisiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Plântula/genética , Plântula/metabolismo , Plântula/fisiologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
8.
Nat Commun ; 11(1): 2965, 2020 06 11.
Artigo em Inglês | MEDLINE | ID: mdl-32528082

RESUMO

Trajectories of cellular ontogeny are tightly controlled and often involve feedback-regulated molecular antagonism. For example, sieve element differentiation along developing protophloem cell files of Arabidopsis roots requires two antagonistic regulators of auxin efflux. Paradoxically, loss-of-function in either regulator triggers similar, seemingly stochastic differentiation failures of individual sieve element precursors. Here we show that these patterning defects are distinct and non-random. They can be explained by auxin-dependent bistability that emerges from competition for auxin between neighboring cells. This bistability depends on the presence of an auxin influx facilitator, and can be triggered by either flux enhancement or repression. Our results uncover a hitherto overlooked aspect of auxin uptake, and highlight the contributions of local auxin influx, efflux and biosynthesis to protophloem formation. Moreover, the combined experimental-modeling approach suggests that without auxin efflux homeostasis, auxin influx interferes with coordinated differentiation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Arabidopsis/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Ácidos Indolacéticos/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Transformação Genética/genética
9.
Plant Mol Biol ; 104(1-2): 55-65, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32572798

RESUMO

Increase in atmospheric carbon dioxide (CO2) has a significant effect on plant growth and development. To explore the elevated-CO2 response, we generated transcriptional profiles over a time course (2 h-14 days) of exposure to elevated CO2 in Arabidopsis thaliana. Genes related to photosynthesis were down-regulated and circadian rhythm-related genes were abnormally regulated in the early to middle phase of elevated CO2 exposure. To understand the novel mechanism of elevated CO2 signaling, we focused on 42 unknown small coding genes that showed differential expression patterns under elevated CO2 conditions. Four transgenic plants overexpressing the small coding gene exhibited a growth-defective phenotype under elevated CO2 but not under current CO2. Transcriptome analysis showed that circadian rhythm-related genes were commonly regulated in four transgenic plants. These circadian rhythm-related genes were transcribed in the dark when CO2 concentrations in the leaf was high. Taken together, our identified four small coding genes are likely to participate in elevated CO2 signaling to the circadian rhythm.


Assuntos
Arabidopsis/genética , Arabidopsis/metabolismo , Dióxido de Carbono/metabolismo , Ritmo Circadiano/genética , Ritmo Circadiano/fisiologia , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação para Baixo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Fenótipo , Fotossíntese/genética , Desenvolvimento Vegetal , Folhas de Planta/genética , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , RNA de Plantas/genética , RNA de Plantas/isolamento & purificação , Transcriptoma
10.
Food Chem ; 327: 127048, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32454285

RESUMO

In our previous study, a novel LMW-GS designated as LMW-N13 with a unique molecular structure was identified from Aegilops uniaristata. LMW-N13 has been characterized as the largest LMW-GS, so far, and possesses an extra cysteine residue compared with typical LMW-GS. In order to analyze the contribution of LMW-N13 to dough quality, in this work, three transgenic wheat lines overexpressing LMW-N13 were generated. Compared with non-transformation (NT) lines, transgenic (TG) lines demonstrated superior dough properties. These superior dough properties were accompanied by the higher contents of glutenin macropolymer (GMP) and total protein. The microstructure of the dough was further investigated by scanning electron microscopy; starch granules in NT lines were smaller than those in transgenic lines. The protein matrix in NT lines was relatively loose and discontinuous. Conversely, the protein matrix in transgenic lines was more continuous and tight. The application of LMW-N13 in wheat breeding is also discussed.


Assuntos
Farinha/análise , Glutens/química , Plantas Geneticamente Modificadas/química , Triticum/química , Aegilops/genética , Dissulfetos/química , Glutens/genética , Glutens/metabolismo , Microscopia Eletrônica de Varredura , Peso Molecular , Plantas Geneticamente Modificadas/metabolismo , Amido/química , Triticum/metabolismo , Água/química
11.
Nat Commun ; 11(1): 2170, 2020 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-32358503

RESUMO

Plants as non-mobile organisms constantly integrate varying environmental signals to flexibly adapt their growth and development. Local fluctuations in water and nutrient availability, sudden changes in temperature or other abiotic and biotic stresses can trigger changes in the growth of plant organs. Multiple mutually interconnected hormonal signaling cascades act as essential endogenous translators of these exogenous signals in the adaptive responses of plants. Although the molecular backbones of hormone transduction pathways have been identified, the mechanisms underlying their interactions are largely unknown. Here, using genome wide transcriptome profiling we identify an auxin and cytokinin cross-talk component; SYNERGISTIC ON AUXIN AND CYTOKININ 1 (SYAC1), whose expression in roots is strictly dependent on both of these hormonal pathways. We show that SYAC1 is a regulator of secretory pathway, whose enhanced activity interferes with deposition of cell wall components and can fine-tune organ growth and sensitivity to soil pathogens.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Citocininas/metabolismo , Resistência à Doença/genética , Ácidos Indolacéticos/metabolismo , Proteínas de Membrana/metabolismo , Reguladores de Crescimento de Planta/metabolismo , Raízes de Plantas/metabolismo , Transcriptoma/genética , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Parede Celular/química , Parede Celular/metabolismo , Endossomos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Complexo de Golgi/metabolismo , Proteínas de Membrana/genética , Raízes de Plantas/microbiologia , Plantas Geneticamente Modificadas/metabolismo , Plasmodioforídeos/patogenicidade , Via Secretória/genética , Solo , Proteínas de Transporte Vesicular/metabolismo
12.
Plant Mol Biol ; 103(4-5): 561-580, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32405802

RESUMO

KEY MESSAGE: CmHKT1;1 selectively exports Na+ from plant cells. Upon NaCl stress, its expression increased in a salt-tolerant melon cultivar. Overexpression of CmHKT1;1 increased transgenic Arabidopsis salt tolerance through improved K+/Na+ balance. High-affinity K+ transporters (HKTs) are thought to be involved in reducing Na+ in plant shoots under salt stress and modulating salt tolerance, but their function in a moderately salt-tolerant species of melon (Cucumis melo L.) remains unclear. In this study, a Na+ transporter gene, CmHKT1;1 (GenBank accession number: MK986658), was isolated from melons based on genome data. The transcript of CmHKT1;1 was relatively more abundant in roots than in stems or leaves from melon seedlings. The tobacco transient expression system showed that CmHKT1;1 was plasma-membrane localized. Upon salt stress, CmHKT1;1 expression was more strongly upregulated in a salt-tolerant melon cultivar, 'Bingxuecui' (BXC) compared with a salt-sensitive cultivar, 'Yulu' (YL). Electrophysiological evidence demonstrated that CmHKT1;1 only transported Na+, rather than K+, when expressed in Xenopus laevis oocytes. Overexpression of CmHKT1;1 increased salt sensitivity in Saccharomyces cerevisiae and salt tolerance in Arabidopsis thaliana. Under NaCl treatments, transgenic Arabidopsis plants accumulated significantly lower concentrations of Na+ in shoots than wild type plants and showed a better K+/Na+ balance, leading to better Fv/Fm, root length, biomass, and enhanced plant growth. The CmHKT1;1 gene may serve as a useful candidate for improving crop salt tolerance.


Assuntos
Arabidopsis/metabolismo , Cucumis melo/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Potássio/metabolismo , Sódio/metabolismo , Arabidopsis/genética , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Clorofila/análise , Clonagem Molecular , Cucumis melo/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Membrana Transportadoras/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Raízes de Plantas/metabolismo , Brotos de Planta/genética , Saccharomyces cerevisiae/genética , Tolerância ao Sal , Plântula/genética , Plântula/metabolismo , Alinhamento de Sequência , Análise de Sequência de Proteína , Cloreto de Sódio/metabolismo , Estresse Fisiológico/genética , Estresse Fisiológico/fisiologia , Simportadores/genética , Simportadores/metabolismo , Tabaco/genética , Tabaco/metabolismo
13.
Plant Mol Biol ; 103(6): 689-704, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32472480

RESUMO

KEY MESSAGE: CmCAD2 and CmCAD3 function more positively than CmCAD1 in oriental melon for lignin synthesis which is important to ensure internal water status and thus for drought tolerance. Well-lignification may be the guarantee of efficient axial water transport and barrier of lateral water flow in oriental melon tolerating drought stress, however remains to be verified. As an important enzyme in monolignol synthesis pathway, five cinnamyl alcohol dehydrogenase (CAD) genes were generally induced in melon seedlings by drought. Here we further revealed the roles of CmCAD1, 2, and 3 in lignin synthesis and for drought tolerance. Results found that overexpressing CmCAD2 or 3 strongly recovered CAD activities, lignin synthesis and composition in Arabidopsis cadc cadd, whose lignin synthesis is disrupted, while CmCAD1 functioned modestly. In melon seedlings, silenced CmCAD2 and 3 individually or collectively decreased CAD activities and lignin depositions drastically, resulting in dwarfed phenotypes. Reduced lignin, mainly composed by guaiacyl units catalyzed by CmCAD3, is mainly due to the limited lignification in tracheary elements and development of Casparion strip. While CmCAD1 and 2 exhibited catalysis to p-coumaraldehyde and sinapaldehyde, respectively. Compared with CmCAD1, drought treatments revealed higher sensitivity of CmCAD2 and/or 3 silenced melon seedlings, accompanying with lower relative water contents, water potentials and relatively higher total soluble sugar contents. Slightly up-regulated expressions of aquaporin genes together with limited lignification might imply higher lateral water loss in stems of silenced lines. In Arabidopsis, CmCAD2 and 3 transgenic lines enhanced cadc cadd drought tolerance through recovering lignin synthesis and root development, accompanying with decreased electrolyte leakage ratios and increased RWCs, thus improved survival rates. Briefly, lignin synthesized by CmCAD2 and 3 functions importantly for drought tolerance in melon.


Assuntos
Plântula/metabolismo , Cucumis melo/metabolismo , Cucumis melo/fisiologia , Secas , Regulação da Expressão Gênica de Plantas/fisiologia , Lignina/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Plântula/fisiologia
14.
PLoS One ; 15(5): e0231355, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32437389

RESUMO

The overexploitation of medicinal plants is depleting gene pool at an alarming rate. In this scenario inducing the genetic variability through targeted mutations could be beneficial in generating varieties with increased content of active compounds. The present study aimed to develop a reproducible protocol for in vitro multiplication and mutagenesis of Hyoscyamus niger targeting putrescine N-methyltransferase (PMT) and 6ß-hydroxy hyoscyamine (H6H) genes of alkaloid biosynthetic pathway. In vitro raised callus were treated with different concentrations (0.01% - 0.1%) of Ethyl Methane Sulfonate (EMS). Emerging multiple shoots and roots were obtained on the MS media supplemented with cytokinins and auxins. Significant effects on morphological characteristics were observed following exposure to different concentrations of EMS. EMS at a concentration of 0.03% was seen to be effective in enhancing the average shoot and root number from 14.5±0.30 to 22.2 ±0.77 and 7.2±0.12 to 8.8±0.72, respectively. The lethal dose (LD50) dose was calculated at 0.08% EMS. The results depicted that EMS has an intense effect on PMT and H6H gene expression and metabolite accumulation. The transcripts of PMT and H6H were significantly upregulated at 0.03-0.05% EMS compared to control. EMS treated explants showed increased accumulation of scopolamine (0.639 µg/g) and hyoscyamine (0.0344µg/g) compared to untreated.


Assuntos
Metanossulfonato de Etila/toxicidade , Hiosciamina/metabolismo , Hyoscyamus/crescimento & desenvolvimento , Metiltransferases/genética , Oxigenases de Função Mista/genética , Mutagênese , Mutação , Escopolamina/metabolismo , Vias Biossintéticas , Regulação da Expressão Gênica de Plantas , Hyoscyamus/efeitos dos fármacos , Hyoscyamus/genética , Hyoscyamus/metabolismo , Mutagênicos/toxicidade , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo
15.
Ecotoxicol Environ Saf ; 200: 110721, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32464438

RESUMO

Glutaredoxins (Grxs) are small (10-15 kDa) glutathione (GSH) - dependent redox proteins. The role of Grxs are well documented in tolerance to heavy metal stress in prokaryotic and mammalian systems and a few plant genera, but is poorly understood in plants against drought. In the present study, two rice glutaredoxin (Osgrx) genes (LOC_Os02g40500 and LOC_Os01g27140) responsible for tolerance against heavy metal stress have been studied for investigating their role against drought. Each glutaredoxin gene was over-expressed in Arabidopsis thaliana to reveal their role in drought stress. The relative expression of both Osgrx genes was higher in the transgenic lines. Transgenic lines of both Osgrxs showed longer roots, higher seed germination, and survival efficiency during drought stress. The physiological parameters (PN, gs, E, WUE, qP, NPQ and ETR), antioxidant enzymes (GRX, GR, GPX, GST, APX, POD, SOD, CAT, DHAR, and MDHAR), antioxidant molecules (ascorbate and GSH) and stress-responsive amino acids (cysteine and proline) levels were additionally increased in transgenic lines of both Osgrxs to provide drought tolerance. The outcomes from this study strongly determined that each Osgrx gene participated in the moderation of drought and might be utilized in biological engineering strategies to overcome drought conditions in different crops.


Assuntos
Glutarredoxinas/genética , Oryza/enzimologia , Estresse Fisiológico , Antioxidantes/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Ácido Ascórbico/metabolismo , Cisteína/metabolismo , Secas , Genes de Plantas , Glutarredoxinas/metabolismo , Glutationa/metabolismo , Oryza/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Prolina/metabolismo , Estresse Fisiológico/genética
16.
PLoS One ; 15(5): e0232747, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32374765

RESUMO

The potential effects of Bt (Bacillus thuringiensis) maize on non-target organisms must be conducted before the Bt maize is commercially planted. Folsomia candida is one of the non-target organisms of Bt maize, also as an important indicator of soil quality and environmental pollution. In this study, a 90-day F. candida feeding test were conducted to evaluate the potential effects of two Bt maize lines IE09S034 and BT799 and their non-Bt conventional isolines Zong 31 and Zheng 58. The results show that Bt maize lines had no significant effects on the survival rate, reproduction, adult body length, larval body length, and the activities of acetyl cholinesterase, catalase and superoxide dismutase on the F. candida. Namely, Bt maize had no toxic effects on the F. candida.


Assuntos
Bacillus thuringiensis/genética , Candida/metabolismo , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Zea mays/genética , Acetilcolinesterase/metabolismo , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bioensaio/métodos , Catalase/metabolismo , Endotoxinas/genética , Endotoxinas/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Larva , Reprodução/genética , Microbiologia do Solo , Superóxido Dismutase/metabolismo , Zea mays/microbiologia
17.
Plant Mol Biol ; 103(6): 597-608, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32346812

RESUMO

KEY MESSAGE: Nanobody-heavy chain (VHH-Fc) antibody formats have the potential to immunomodulate even highly accumulating proteins and provide a valuable tool to experimentally modulate the subcellular distribution of seed storage proteins. Recombinant antibodies often obtain high accumulation levels in plants, and thus, besides being the actual end-product, antibodies targeting endogenous host proteins can be used to interfere with the localization and functioning of their corresponding antigens. Here, we compared the effect of a seed-expressed nanobody-heavy chain (VHH-Fc) antibody against the highly abundant Arabidopsis thaliana globulin seed storage protein cruciferin with that of a VHH-Fc antibody without endogenous target. Both antibodies reached high accumulation levels of around 10% of total soluble protein, but strikingly, another significant part was present in the insoluble protein fraction and was recovered only after extraction under denaturing conditions. In seeds containing the anti-cruciferin antibodies but not the antibody without endogenous target, the amount of soluble, processed globulin subunits was severely reduced and a major part of the cruciferin molecules was found as precursor in the insoluble fraction. Moreover, in these seeds, aberrant vacuolar phenotypes were observed that were different from the effects caused by the depletion of globulins in knock-out seeds. Remarkably, the seeds with strongly reduced globulin amounts are fully viable and germinate with frequencies similar to wild type, illustrating how flexible seeds can retrieve amino acids from the stored proteins to start germination.


Assuntos
Anticorpos/imunologia , Anticorpos/metabolismo , Globulinas/imunologia , Proteínas de Armazenamento de Sementes/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Proteínas de Armazenamento de Sementes/genética , Vacúolos/metabolismo
18.
Nat Commun ; 11(1): 1846, 2020 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-32296056

RESUMO

Seed size is a key agronomic trait that greatly determines plant yield. Elucidating the molecular mechanism underlying seed size regulation is also an important question in developmental biology. Here, we show that the KIX-PPD-MYC-GIF1 pathway plays a crucial role in seed size control in Arabidopsis thaliana. Disruption of KIX8/9 and PPD1/2 causes large seeds due to increased cell proliferation and cell elongation in the integuments. KIX8/9 and PPD1/2 interact with transcription factors MYC3/4 to form the KIX-PPD-MYC complex in Arabidopsis. The KIX-PPD-MYC complex associates with the typical G-box sequence in the promoter of GRF-INTERACTING FACTOR 1 (GIF1), which promotes seed growth, and represses its expression. Genetic analyses support that KIX8/9, PPD1/2, MYC3/4, and GIF1 function in a common pathway to control seed size. Thus, our results reveal a genetic and molecular mechanism by which the transcription factors MYC3/4 recruit KIX8/9 and PPD1/2 to the promoter of GIF1 and repress its expression, thereby determining seed size in Arabidopsis.


Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Transativadores/genética , Transativadores/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Diester Fosfórico Hidrolases/genética , Diester Fosfórico Hidrolases/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Regiões Promotoras Genéticas , Pirofosfatases/genética , Pirofosfatases/metabolismo
19.
PLoS Pathog ; 16(4): e1008518, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32324832

RESUMO

Competition is one of the fundamental driving forces of natural selection. Beauveria bassiana is a soil and plant phylloplane/root fungus capable of parasitizing insect hosts. Soil and plant environments are often enriched with other fungi against which B. bassiana competes for survival. Here, we report an antifungal peptide (BbAFP1), specifically expressed and localized to the conidial cell wall and is released into the surrounding microenvironment inhibiting growth of competing fungi. B. bassiana strains expressing BbAFP1, including overexpression strains, inhibited growth of Alternaria brassicae in co-cultured experiments, whereas targeted gene deletion of BbAFP1 significantly decreased (25%) this inhibitory effect. Recombinant BbAFP1 showed chitin and glucan binding abilities, and growth inhibition of a wide range of phytopathogenic fungi by disrupting membrane integrity and eliciting reactive oxygen species (ROS) production. A phenylalanine residue (F50) contributes to chitin binding and antifungal activity, but was not required for the latter. Expression of BbAFP1 in tomato resulted in transgenic plants with enhanced resistance to plant fungal pathogens. These results highlight the importance of fungal competition in shaping primitive competition strategies, with antimicrobial compounds that can be embedded in the spore cell wall to be released into the environment during the critical initial phases of germination for successful growth in its environmental niche. Furthermore, these peptides can be exploited to increase plant resistance to fungal pathogens.


Assuntos
Antifúngicos/metabolismo , Beauveria/metabolismo , Esporos Fúngicos/metabolismo , Animais , Antifúngicos/farmacologia , Beauveria/genética , Parede Celular/metabolismo , Quitina/metabolismo , Proteínas Fúngicas/metabolismo , Glucanos/metabolismo , Insetos/microbiologia , Peptídeos , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Espécies Reativas de Oxigênio , Estresse Fisiológico/efeitos dos fármacos , Virulência
20.
Artigo em Inglês | MEDLINE | ID: mdl-32114415

RESUMO

Rice tillering, which determines the panicle number per plant, is an important agronomic trait for grain production. In higher plants, ascorbic acid (Asc) plays a major role in ROS-scavenging activity. l-Galactono-1, 4-lactone dehydrogenase (GalLDH, EC1.3.2.3) is an enzyme that catalyzes the last step of Asc biosynthesis in plants. Previously, we have reported that homozygous L-GalLDH-suppressed transgenic rice plants (GI) display a reduced tiller number and a lower level of foliar carotenoids (Car) compared with wild type. Strigolactones (SL), which play an important role in the suppression of shoot branching, are synthesized in the roots of rice plant using Car as substrates. In this paper, the relationship between Asc, SL, the accumulation of H2O2, changes in antioxidant capacity, enzyme activities, and gene transcriptions related to the synthesis of SL were analyzed in transgenic rice plants for L-GalLDH suppressed (GI-1 and GI-2) and overexpressing (GO-2). The results showed that the altered level of Asc in the L-GalLDH transgenic rice plants leads to a change in redox homeostasis, resulting in a marked accumulation of H2O2 and decreased antioxidant capacity in GI-1 and GI-2, but lower H2O2 content and increased antioxidant capacity in GO-2. Meanwhile, the altered level of Asc also leads to altered enzyme activities and gene transcript abundances related to SL synthesis in L-GalLDH transgenics. These observations support the conclusion that Asc influences tiller number in the L-GalLDH transgenics by affecting H2O2 accumulation and antioxidant capacity, and altering those enzyme activities and gene transcript abundances related to SL synthesis.


Assuntos
Ácido Ascórbico/metabolismo , Regulação da Expressão Gênica de Plantas , Compostos Heterocíclicos com 3 Anéis/metabolismo , Lactonas/metabolismo , Oryza/metabolismo , Antioxidantes/metabolismo , Peróxido de Hidrogênio/metabolismo , Oryza/enzimologia , Oryza/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA