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1.
Food Chem ; 308: 125707, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-31669943

RESUMO

The ripening of the apple (Malus × domestica Borkh.) fruit is regulated by the phytohormone ethylene, where degreening is an important physiological metabolism caused by chlorophyll (Chl) degradation. However, to date, research on how ethylene affects the Chl degradation pathway of apple peel during ripening remains scarce. In this study, the effects of ethylene on the expression of Chl catabolic genes (CCGs) of apple peel during ripening were studied by treating harvested commercial mature apples with 0.5 µL L-1 1-methylcyclopropene (1-MCP). The results showed that 1-MCP treatment led to a delayed climacteric peak of respiration and ethylene production, exhibiting higher Chl content and hue angle (H˚) compared to untreated fruit during ripening. Lower quantities of pheophorbide a oxygenase (PAO), pheophytinase (PPH) and red Chl catabolite reductase (RCCR) were also observed in peel tissues under 1-MCP treatment during ripening. Further study with quantitative real-time polymerase chain reaction (qPCR) revealed that the expression of CCGs, except for MdNYE1a, increased atdifferentdegrees upon ripening. Meanwhile, the apples treated with 1-MCP presented a downregulated expression of MdRCCR2, MdNYC1, MdNYC3 and MdNOL2 and a fluctuating expression of MdNYE1a, MdPPH1, MdPAO6, MdPAO8 and MdHCAR compared with the controls during ripening. Our results indicated the regulatory role of ethylene in the Chl degradation pathway of apple peel during ripening.


Assuntos
Clorofila/metabolismo , Ciclopropanos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Malus/metabolismo , Reguladores de Crescimento de Planta/farmacologia , Etilenos/metabolismo , Armazenamento de Alimentos , Frutas/efeitos dos fármacos , Frutas/metabolismo , Malus/efeitos dos fármacos , Proteínas de Plantas/metabolismo
2.
Food Chem ; 307: 125525, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31639577

RESUMO

This study investigated effects of different concentrations of methyl jasmonate (MeJA) on carotenoids accumulation, radical scavenging activity and proline content in germinated maize kernels. MeJA treatment promoted carotenoids accumulation, radical scavenging activity and proline accumulation, while salicyl hydroxamic acid (SHAM) reduced carotenoids accumulation. There was a significant increase of 42.5% in lutein content when treated with 0.5 µM MeJA. Furthermore, the transcriptional expression of seven carotenogenic genes were explored by MeJA and SHAM. The results showed that 0.5 µM MeJA significantly increased the gene expression levels of PSY, PDS, ZDS, LCYB, LCYE, BCH1, CYP97C, and their transcript levels, which were strongly associated with carotenoids content. Treatment of MeJA affected the carotenoids biosynthesis gene and led to the accumulation of carotenoids. These new findings would help to develop innovative approach for enrichment of lutein in germinated maize kernels for further development of functional food materials.


Assuntos
Acetatos/farmacologia , Carotenoides/metabolismo , Ciclopentanos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oxilipinas/farmacologia , Zea mays/crescimento & desenvolvimento , Carotenoides/análise , Cromatografia Líquida de Alta Pressão , Depuradores de Radicais Livres/química , Germinação , Luteína/análise , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Prolina/análise , Zea mays/metabolismo
3.
Food Chem ; 305: 125440, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31494496

RESUMO

Tea (Camellia sinensis) contains two active glutamate decarboxylases (CsGADs), whose unclear properties were examined here. CsGAD1 was 4-fold higher than CsGAD2 in activity. Their Km values for L-glutamate were around 5 mM. CsGAD1 and CsGAD2 performed best at 55 and 40 °C, respectively, and were both stimulated by calcium/calmodulin (Ca2+/CaM). Over 40 °C, their calmodulin-binding domains degraded. CsGADs were most active at pH 5.6, and were stimulated by Ca2+/CaM at pH 5.6-6.6, but inactivated at pH 3.6. Ca2+/CaM restored the CsGAD1 activity suppressed by inhibitors. CsGADs and CsCaM were localized to the cytosol. CsGAD1 was more highly expressed in most tissues, while CsGAD2 expression was more induced under stresses. The characteristics we first elucidated here revealed that CsGAD1 is the predominant isoform in tea plant, with CsGAD2 exhibiting a supplementary role under certain conditions. The information will contribute to regulation of GABA tea quality.


Assuntos
Camellia sinensis/enzimologia , Glutamato Descarboxilase/metabolismo , Proteínas de Plantas/metabolismo , Ácido gama-Aminobutírico/metabolismo , Secas , Estabilidade Enzimática , Ácido Glutâmico/metabolismo , Concentração de Íons de Hidrogênio , Isoenzimas/metabolismo , Cinética , Temperatura Ambiente
4.
Food Chem ; 305: 125450, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31499291

RESUMO

A round robin comparison was performed in order to test the performance of a recently developed LC-MS/MS method for quantification of 6 folate forms. Eighty-nine samples representing the food groups of fruits, vegetables, legumes, cereals, dairy products, meat, and offal were analyzed by two LC-MS/MS methods and a microbiological assay (MA). A plant-origin deconjugase enzyme (Arabidopsis thaliana) for deconjugation of folates (PE-LC-MS/MS), or animal-origin deconjugase (rat serum and chicken pancreas) (AE-LC-MS/MS) was used in the LC-MS/MS methods, each in a single enzymatic step. In contrast, the MA involved tri-enzyme extraction including human plasma as a deconjugase. A significant bias of 17% lower and 25% higher results was found when PE-LC-MS/MS was compared to MA and AE-LC-MS/MS, respectively. The PE-LC-MS/MS provides fast quantification of various folate vitamers and total folate content, which could be a proper substitute to the currently standardized but imprecise and time-consuming microbiological assay in the future.


Assuntos
Ácido Fólico/análise , Espectrometria de Massas em Tandem/métodos , Animais , Galinhas , Cromatografia Líquida de Alta Pressão , Análise de Alimentos , Frutas/metabolismo , Humanos , Proteínas de Plantas/metabolismo , Ratos , Verduras/metabolismo , gama-Glutamil Hidrolase/metabolismo
5.
Food Chem ; 305: 125485, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31522126

RESUMO

Mashua (Tropaeolum tuberosum) is an important food in certain areas of the Andean region, where it is popularly believed to possess medicinal properties. Several studies have previously shown the potential of this tuber as a source of bioactive compounds. Traditionally, the tuber is exposed to the sun before consumption, in order to reduce its bitterness. The present work aims to study, at the proteome level, the differential abundance of proteins in tubers subjected to different postharvest treatments: sun-exposure (SUN), shade (SHA), refrigeration (COLD) and shade combined with sun-exposure (SHA-SUN) compared to recently harvested tubers (INIT). Results showed that sun exposure for prolonged times (9 days) resulted in increased abundance of proteins classified as heat shock proteins, intracellular traffic, disease/defense and protein degradation. Our results reflect that the sun treatment activates defense systems and osmoprotection adjustment against water loss and reactive oxygen species.


Assuntos
Proteoma/análise , Proteômica/métodos , Tropaeolum/metabolismo , Cromatografia Líquida de Alta Pressão , Temperatura Baixa , Análise Discriminante , Proteínas de Choque Térmico/metabolismo , Análise dos Mínimos Quadrados , Espectrometria de Massas , Proteínas de Plantas/metabolismo , Tubérculos/metabolismo , Luz Solar
6.
J Agric Food Chem ; 67(46): 12709-12719, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31697495

RESUMO

The major components of wheat storage proteins are gliadins and glutenins, and as they contribute differently to baking quality, a balanced mixture of these components is essential. The application of foliar nitrogen (N) at anthesis is a common practice to improve protein concentration and composition. The aim of this study was to investigate the effects of a foliar N application at anthesis on storage protein gene expression during grain development and on the distribution of protein concentration and protein body size within the grain. In this experiment, an additional N application at anthesis stimulated the expression of genes of the majority of storage proteins when the N supply was low. Furthermore, it led to higher protein concentrations in the subaleurone layers, while in the center of the lobes, the protein concentrations were decreased. These changes will affect the protein recovery in white flours, as proportionally more protein might be lost during milling processes.


Assuntos
Fertilizantes/análise , Nitrogênio/farmacologia , Proteínas de Plantas/genética , Triticum/efeitos dos fármacos , Farinha/análise , Proteínas de Grãos/metabolismo , Nitrogênio/metabolismo , Proteínas de Plantas/metabolismo , Triticum/genética , Triticum/crescimento & desenvolvimento , Triticum/metabolismo
7.
Biol Res ; 52(1): 56, 2019 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-31699158

RESUMO

BACKGROUND: ADP-glucose pyrophosphorylase (AGPase), the key enzyme in plant starch biosynthesis, is a heterotetramer composed of two identical large subunits and two identical small subunits. AGPase has plastidial and cytosolic isoforms in higher plants, whereas it is mainly detected in the cytosol of grain endosperms in cereal crops. Our previous results have shown that the expression of the TaAGPL1 gene, encoding the cytosolic large subunit of wheat AGPase, temporally coincides with the rate of starch accumulation and that its overexpression dramatically increases wheat AGPase activity and the rate of starch accumulation, suggesting an important role. METHODS: In this study, we performed yeast one-hybrid screening using the promoter of the TaAGPL1 gene as bait and a wheat grain cDNA library as prey to screen out the upstream regulators of TaAGPL1 gene. And the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method was used to verify the functional characterization of the identified regulators in starch biosynthesis. RESULTS: Disulfide isomerase 1-2 protein (TaPDIL1-2) was screened out, and its binding to the TaAGPL1-1D promoter was further verified using another yeast one-hybrid screen. Transiently silenced wheat plants of the TaPDIL1-2 gene were obtained by using BSMV-VIGS method under field conditions. In grains of BSMV-VIGS-TaPDIL1-2-silenced wheat plants, the TaAGPL1 gene transcription levels, grain starch contents, and 1000-kernel weight also significantly increased. CONCLUSIONS: As important chaperones involved in oxidative protein folding, PDIL proteins have been reported to form hetero-dimers with some transcription factors, and thus, our results suggested that TaPDIL1-2 protein could indirectly and negatively regulate the expression of the TaAGPL1 gene and function in starch biosynthesis.


Assuntos
Pão , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Glucose-1-Fosfato Adenililtransferase/metabolismo , Proteínas de Plantas/metabolismo , Triticum/metabolismo , Glucose-1-Fosfato Adenililtransferase/genética , Proteínas de Plantas/genética , Fatores de Transcrição , Triticum/genética
8.
Plant Mol Biol ; 101(4-5): 499-506, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31621004

RESUMO

A simple, accessible, and inexpensive assay to quantify the strength of DNA-protein interactions was developed. The assay relies on capturing DNA-protein complexes using an affinity resin that binds tagged, recombinant proteins. Sequential washes with filtration spin cups and centrifugation remove non-specific interactions in a gentle, uniform manner and a final elution isolates specific DNA-protein complexes. SYBR Gold nucleic acid stain is added to the eluted product and the fluorescence intensity accurately quantifies the amount of captured DNA, ultimately illustrating the relative strength of the DNA-protein interaction. The major utility of the assay resides in the versatility and quantitative nature of the SYBR Gold:nucleic acid interaction, eliminating the need for customized or labeled oligos and permitting relatively inexpensive quantification of binding capacity. The assay also employs DNA-protein complex capture by the very common purification tag, 6xHis, but other tags could likely be utilized. Further, SYBR Gold fluorescence is compatible with a wide variety of instruments, including UV transilluminators, a staple to any molecular biology laboratory. This assay was used to compare the binding capacities of different auxin response factor (ARF) transcription factors to various dsDNA targets, including the classical AuxRE motif and several divergent sequences. Results from dose-response assays suggest that different ARF proteins might show distinct comparative affinities for AuxRE variants, emphasizing that specific ARF-AuxRE binding strengths likely contribute to the complex and fine-tuned cellular auxin response.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , DNA de Plantas/metabolismo , Proteínas de Ligação a DNA/metabolismo , Compostos Orgânicos , Proteínas de Plantas/metabolismo , Soja/metabolismo , Fatores de Transcrição/metabolismo , Fluorescência , Ligação Proteica , Sensibilidade e Especificidade
9.
Plant Mol Biol ; 101(4-5): 343-354, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31621005

RESUMO

KEY MESSAGE: Short review focussing on the role and targeting of vacuolar substructure in plant immunity and pathogenesis. Plants lack specialized immune cells, therefore each plant cell must defend itself against invading pathogens. A typical plant defense strategy is the hypersensitive response that results in host cell death at the site of infection, a process largely regulated by the vacuole. In plant cells, the vacuole is a vital organelle that plays a central role in numerous fundamental processes, such as development, reproduction, and cellular responses to biotic and abiotic stimuli. It shows divergent membranous structures that are continuously transforming. Recent technical advances in visualization and live-cell imaging have significantly altered our view of the vacuolar structures and their dynamics. Understanding the active nature of the vacuolar structures and the mechanisms of vacuole-mediated defense responses is of great importance in understanding plant-pathogen interactions. In this review, we present an overview of the current knowledge about the vacuole and its internal structures, as well as their role in plant-microbe interactions. There is so far limited information on the modulation of the vacuolar structures by pathogens, but recent research has identified the vacuole as a possible target of microbial interference.


Assuntos
Interações Hospedeiro-Patógeno , Imunidade Vegetal , Plantas/ultraestrutura , Vacúolos/ultraestrutura , Biomarcadores/metabolismo , Morte Celular , Membranas Intracelulares/imunologia , Membranas Intracelulares/microbiologia , Membranas Intracelulares/ultraestrutura , Proteínas de Plantas/metabolismo , Plantas/imunologia , Plantas/microbiologia , Vacúolos/imunologia , Vacúolos/microbiologia
10.
J Agric Food Chem ; 67(45): 12382-12392, 2019 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-31635461

RESUMO

Protoporphyrinogen oxidase (PPO, EC 1.3.3.4) is a promising target for herbicide discovery. Search for new compounds with novel chemotypes is a key objective for agrochemists. Here, we describe the discovery and systematic SAR-based structure optimization of novel N-isoxazolinylphenyltriazinones 5-9 as PPO inhibitors. The in vivo herbicidal activity and in vitro Nicotiana tabacum PPO (NtPPO) inhibitory activity were explored in detail. A number of the new synthetic compounds displayed strong PPO inhibitory activity with Ki values in the nanomolar range. Some compounds exhibited excellent and broad-spectrum weed control at the rate of 9.375-37.5 g ai/ha by postemergence application and showed improved monocotyledonous weed control compared to saflufenacil. Most promisingly, ethyl 3-(2-chloro-5-(3,5-dimethyl-2,6-dioxo-4-thioxo-1,3,5-triazinan-1-yl)-4-fluorophenyl)-5-methyl-4,5-dihydroisoxazole-5-carboxylate, 5a, with a Ki value of 4.9 nM, displayed over 2- and 6-fold higher potency than saflufenacil (Ki = 10 nM) and trifludimoxazin (Ki = 31 nM), respectively. Moreover, 5a showed excellent and broad-spectrum weed control against 32 kinds of weeds at 37.5-75 g ai/ha. Rice exhibited relative tolerance to 5a at 150 g ai/ha by postemergence application, indicating that 5a could be a potential herbicide candidate for weed control in paddy fields.


Assuntos
Inibidores Enzimáticos/farmacologia , Herbicidas/química , Herbicidas/farmacologia , Proteínas de Plantas/antagonistas & inibidores , Protoporfirinogênio Oxidase/antagonistas & inibidores , Descoberta de Drogas , Inibidores Enzimáticos/química , Cinética , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Daninhas/química , Plantas Daninhas/efeitos dos fármacos , Plantas Daninhas/enzimologia , Protoporfirinogênio Oxidase/química , Protoporfirinogênio Oxidase/metabolismo , Relação Quantitativa Estrutura-Atividade , Tabaco/química , Tabaco/efeitos dos fármacos , Tabaco/enzimologia , Controle de Plantas Daninhas
11.
J Agric Food Chem ; 67(44): 12219-12227, 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31613626

RESUMO

Quantification, using an accurate analytical approach, of capsinoids and capsaicinoids was performed on three chili pepper (Capsicum spp.) genotypes: "Chiltepín", "Tampiqueño 74", and "Bhut Jolokia" at various stages of fruit development. The accumulation of capsinoids, in all these peppers started between 10 to 20 days post-anthesis (dpa), increased and reached the highest capsinoid amount at 40 dpa, and then decreased until 60 dpa. Conversely, capsaicinoids could already be determined at 10 dpa in "Bhut Jolokia" and their accumulation pattern was different from that of the capsinoids in this genotype. The capsiate/dihydrocapsiate ratio presented a higher variation between genotypes and developmental stages than the capsaicin/dihydrocapsaicin ratio. Capsinoid ratios (4-24%) and Pun1/pAMT genotyping were determined. These results provide information on the progress of the accumulation of capsinoids in the aforementioned pungent and superhot cultivars and could support future breeding studies toward the understanding of the factors affecting their accumulation.


Assuntos
Capsaicina/análogos & derivados , Capsaicina/metabolismo , Capsicum/genética , Capsicum/metabolismo , Aromatizantes/metabolismo , Frutas/crescimento & desenvolvimento , Proteínas de Plantas/genética , Alelos , Sequência de Aminoácidos , Capsaicina/análise , Capsicum/química , Capsicum/crescimento & desenvolvimento , Aromatizantes/análise , Frutas/química , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Genótipo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Alinhamento de Sequência
12.
J Agric Food Chem ; 67(44): 12164-12171, 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31600067

RESUMO

Plant biostimulants (PBS) increase crop productivity and induce beneficial processes in plants. Although PBS can stimulate plant tolerance to some abiotic stresses, their effect in improving crop resistance to herbicide injuries has barely been investigated. Therefore, a study on the effect of a biostimulant (Megafol) on maize (Zea mays L.) tolerance to a chloro-acetanilide herbicide (metolachlor) was carried out. We found that Megafol reduced the negative effects of metolachlor on maize. Indeed, biostimulated samples showed increases in germination, biomass production, Vigor index, and EC50 (effective concentration causing 50% reductions to roots and aerial biomass) with respect to the samples treated with metolachlor alone. Furthermore, plants treated with the herbicide in combination with Megafol showed lower levels of malondialdehyde (MDA). Antioxidant enzymes, namely, ascorbate peroxidase (APX), guaiacol peroxidase (GPX), and catalase (CAT), were assayed in samples treated with metolachlor alone or in combination with Megafol, and higher enzymes activities were found in biostimulated plants. The results of this study open the perspective of using Megafol, as well as other suitable plant biostimulants, in improving the crop's capacity to cope with injuries and unwanted effects that herbicide could cause to these species.


Assuntos
Acetamidas/farmacologia , Herbicidas/farmacologia , Reguladores de Crescimento de Planta/farmacologia , Zea mays/efeitos dos fármacos , Ascorbato Peroxidases/metabolismo , Catalase/metabolismo , Germinação/efeitos dos fármacos , Malondialdeído/metabolismo , Peroxidase/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismo
13.
J Agric Food Chem ; 67(42): 11805-11814, 2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31566383

RESUMO

The impact of cross-breeding two low phytic acid (lpa) rice mutants on the content of phytic acid and the metabolite profile of the resulting double mutant was investigated. Progenies resulting from the cross of Os-lpa-XS110-1, a rice mutant carrying the myo-inositol kinase (OsMIK) mutated gene, and Os-lpa-XS110-2, with the multidrug resistance-associated protein ABC transporter gene 5 (OsMRP5) as the mutation target, were subjected to high-pressure ion chromatography. The reduction of the phytic acid content in the double mutant (-63%) was much more pronounced than in the single mutants (-26 and -47%). Gas chromatography-based metabolite profiling revealed a superimposition of the metabolite profiles inherited from the lpa progenitors in the double mutant progenies; the resulting metabolite signature was predominated by the OsMIK mutation effect. The study demonstrated that cross-breeding of two single lpa mutants can be employed to generate double lpa rice mutants showing both a significant reduction in the content of phytic acid and the imprinting of a specific mutation-induced metabolite signature.


Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Hibridização Genética , Oryza/química , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Ácido Fítico/análise , Proteínas de Plantas/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Mutação , Oryza/genética , Oryza/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Ácido Fítico/metabolismo , Proteínas de Plantas/metabolismo , Sementes/química , Sementes/genética , Sementes/metabolismo
14.
J Agric Food Chem ; 67(45): 12408-12418, 2019 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-31644287

RESUMO

Vegetables are an ideal source of human Se intake; it is important to understand selenium (Se) speciation in plants due to the distinct biological functions of selenocompounds. In this hydroponic study, the accumulation and assimilation of selenite and selenate in pak choi (Brassica rapa), a vastly consumed vegetable, were investigated at 1-168 h with HPLC speciation and RNA-sequencing. The results showed that the Se content in shoots and Se translocation factors with selenate addition were at least 10.81 and 11.62 times, respectively, higher than those with selenite addition. Selenite and selenate up-regulated the expression of SULT1;1 and PHT1;2 in roots by over 240% and 400%, respectively. Selenite addition always led to higher proportions of seleno-amino acids, while SeO42- was dominant under selenate addition (>49% of all Se species in shoots). However, in roots, SeO42- proportions declined substantially by 51% with a significant increase of selenomethionine proportions (63%) from 1 to 168 h. Moreover, with enhanced transcript of methionine gamma-lyase (60% of up-regulation compared to the control) plus high levels of methylselenium in shoots (approximately 70% of all Se species), almost 40% of Se was lost during the exposure under the selenite treatment. This work provides evidence that pak choi can rapidly transform selenite to methylselenium, and it is promising to use the plant for Se biofortification.


Assuntos
Brassica rapa/genética , Brassica rapa/metabolismo , Ácido Selênico/metabolismo , Ácido Selenioso/metabolismo , Selênio/metabolismo , Biotransformação , Brassica rapa/química , Brassica rapa/crescimento & desenvolvimento , Cromatografia Líquida de Alta Pressão , Hidroponia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/química , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Ácido Selênico/análise , Ácido Selenioso/análise , Selênio/análise , Análise de Sequência de RNA
15.
J Agric Food Chem ; 67(42): 11568-11576, 2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31584809

RESUMO

Tribenuron-methyl (TM), as one of the sulfonylurea (SU) herbicides, has been widely and effectively applied for many kinds of plants. SUs inhibit plant growth by restraining the biosynthetic pathway of branched-chain amino acids (BCAAs) catalyzed by acetolactate synthase (ALS). Safeners are agrochemicals that protect crops from herbicide injuries. To improve the crop tolerance under TM toxicity stress, this paper evaluated the protective effect of N-tosyloxazolidine-3-carboxamide. It turned out that most of the tested compounds showed significant protection against TM via enhancing the glutathione (GSH) content and glutathione S-transferase (GST) activity. Among all of the tested compounds, compound 16 exhibited more excellent protection than the contrast safener R-28725 and other target compounds. A positive correlation between the growth level, endogenous GSH content, and GST activity was observed in this research. The GST kinetic parameter Vmax of the maize was increased by 29.6% after treatment with compound 16, while Km was decreased by 51.9% compared to the untreated control. The molecular docking model indicated that compound 16 could compete with TM in the active site of ALS, which could interpret the protective effects of safeners. The present work demonstrated that N-tosyloxazolidine-3-carboxamide derivatives could be considered as potential candidates for developing new safeners in the future.


Assuntos
Herbicidas/toxicidade , Proteínas de Plantas/metabolismo , Substâncias Protetoras/farmacologia , Zea mays/efeitos dos fármacos , Zea mays/enzimologia , Acetolactato Sintase/química , Acetolactato Sintase/metabolismo , Glutationa Transferase/química , Glutationa Transferase/metabolismo , Cinética , Simulação de Acoplamento Molecular , Proteínas de Plantas/química , Compostos de Sulfonilureia/toxicidade , Zea mays/química
16.
J Agric Food Chem ; 67(40): 11077-11088, 2019 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-31525039

RESUMO

Cuticular wax accumulation in plants contributes to drought tolerance. Here, we compared the drought levels on two varieties with different genotypes in turnip (Brassica rapa var. rapa) and found that the drought tolerance was higher in the waxy KTRG-B48a than in the wax-free KTRG-B48b. A combination of PacBio and Illumina sequencing analyses revealed that differential transcripts were mainly enriched in the wax synthesis pathway, and a splice variant (BrrWSD1-X2) was identified in the waxy KTRG-B48a. BrrWSD1-X2 had a stronger ability to synthesize wax esters than BrrWSD1-X1 using heterologous expression in yeast (Saccharomyces cerevisiae) mutant H1246a. Then, we speculated that the T to C transversion of the third intron and the higher number of TA repeats in the third intron of BrrWSD1 DNA in the waxy KTRG-B48a may result in a lower efficiency of splicing recognition of the third intron, resulting in the emergence of BrrWSD1-X2 in waxy varieties.


Assuntos
Brassica rapa/fisiologia , Ésteres/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ceras/metabolismo , Brassica rapa/genética , Secas , Processamento de RNA , Água/análise , Água/metabolismo
17.
J Agric Food Chem ; 67(39): 10844-10852, 2019 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-31525997

RESUMO

The discovery of 4-hydroxyphenylpyruvate dioxygenase (HPPD, EC 1.13.11.27) inhibitors has been an active area of research due to their great potential as herbicides for weed control. Starting from the binding mode of known inhibitors of HPPD, a series of HPPD inhibitors with new molecular scaffolds were designed and synthesized by hybridizing 2-benzoylethen-1-ol and isoindoline-1,3-dione fragments. The results of the in vitro tests indicated that the newly synthesized compounds showed good HPPD inhibitory activity with IC50 values against the recombinant Arabidopsis thaliana HPPD (AtHPPD) ranging from 0.0039 µM to over 1 µM. Most promisingly, compound 4ae, 2-benzyl-5-(5-hydroxy-1,3-dimethyl-1H-pyrazole-4- carbonyl)isoindoline-1,3-dione, showed the highest AtHPPD inhibitory activity with a Ki value of 3.92 nM, making it approximately 10 times more potent than pyrasulfotole (Ki = 44 nM) and slightly more potent than mesotrione (Ki = 4.56 nM). In addition, the cocrystal structure of the AtHPPD-4ae complex was successfully resolved at a resolution of 1.8 Å. The X-ray diffraction analysis indicated that the two carbonyl groups of 2-benzoylethen-1-ol formed a bidentate chelating interaction with the metal ion, while the isoindoline-1,3-dione moiety formed pronounced π-π stacking interactions with Phe381 and Phe424. Moreover, water-mediated hydrogen bonding interactions were observed between Asn282 and the nitrogen atoms of the pyrazole ring of 4ae. The above results showed that the pyrazole-isoindoline-1,3-dione hybrid is a promising scaffold for developing HPPD inhibitors.


Assuntos
4-Hidroxifenilpiruvato Dioxigenase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Herbicidas/farmacologia , Isoindóis/farmacologia , Proteínas de Plantas/antagonistas & inibidores , Pirazóis/farmacologia , 4-Hidroxifenilpiruvato Dioxigenase/química , 4-Hidroxifenilpiruvato Dioxigenase/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Herbicidas/síntese química , Herbicidas/química , Isoindóis/química , Cinética , Estrutura Molecular , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Daninhas/efeitos dos fármacos , Plantas Daninhas/crescimento & desenvolvimento , Pirazóis/química , Relação Estrutura-Atividade
18.
Phytopathology ; 109(10): 1732-1740, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31479403

RESUMO

Plant polygalacturonase-inhibiting protein (PGIP) is a structural protein that can specifically recognize and bind to fungal polygalacturonase (PG). PGIP plays an important role in plant antifungal activity. In this study, a maize PGIP gene, namely ZmPGIP3, was cloned and characterized. Agarose diffusion assay suggested that ZmPGIP3 could inhibit the activity of PG. ZmPGIP3 expression was significantly induced by wounding, Rhizoctonia solani infection, jasmonate, and salicylic acid. ZmPGIP3 might be related to disease resistance. The gene encoding ZmPGIP3 was posed under the control of the ubiquitin promoter and constitutively expressed in transgenic rice. In an R. solani infection assay, ZmPGIP3 transgenic rice was more resistant to sheath blight than the wild-type rice regardless of the inoculated plant part (leaves or sheaths). Digital gene expression analysis indicated that the expression of some rice PGIP genes significantly increased in ZmPGIP3 transgenic rice, suggesting that ZmPGIP3 might activate the expression of some rice PGIP genes to resist sheath blight. Our investigation of the agronomic traits of ZmPGIP3 transgenic rice showed that ZmPGIP3 overexpression in rice did not show any detrimental phenotypic or agronomic effect. ZmPGIP3 is a promising candidate gene in the transgenic breeding for sheath blight resistance and crop improvement.


Assuntos
Resistência à Doença/genética , Oryza , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Rhizoctonia , Regulação da Expressão Gênica de Plantas , Oryza/microbiologia , Plantas Geneticamente Modificadas/microbiologia , Rhizoctonia/fisiologia
19.
Phytopathology ; 109(10): 1679-1688, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31479404

RESUMO

Alternative oxidase (AOX) is a ubiquinol terminal oxidase that is involved in fungal mitochondrial oxidative phosphorylation. In this study, we analyzed the roles of AOX in Botrytis cinerea by generating BcAOX deletion mutants. The mutants exhibited defects in mycelial growth, sporulation, spore germination, and virulence. Furthermore, the sensitivity of the mutants to quinone outside inhibitor fungicides and oxidative stress were increased. All phenotypic variations could be restored in the complemented strain. In summary, these results showed that BcAOX is involved in the regulation for vegetative development, adaptation to environmental stress, and virulence of B. cinerea.


Assuntos
Botrytis , Proteínas Mitocondriais , Oxirredutases , Oxigênio , Proteínas de Plantas , Botrytis/enzimologia , Botrytis/crescimento & desenvolvimento , Botrytis/patogenicidade , Proteínas Mitocondriais/metabolismo , Estresse Oxidativo/fisiologia , Oxirredutases/metabolismo , Proteínas de Plantas/metabolismo , Virulência
20.
J Agric Food Chem ; 67(38): 10624-10636, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31483633

RESUMO

The freshness and color quality of postharvest tea leaves can be markedly prolonged and retained by proper preservation measures. Here, we investigated the dynamic changes of chlorophyll and its derivatives in postharvest tea leaves under different low-temperature treatments using natural withering as a control. Chlorophyll decomposition was found closely related with chlorophyllide, pheophorbide, and pheophytin. Low-temperature withering could slow chlorophyll degradation in postharvest tea leaves via significant inhibition on the enzyme activity and gene expression of Mg-dechelatase, chlorophyllase, and pheophorbide a oxygenase. At the initial stage of withering, a significant increase was observed in the chlorophyll content, expression of chlorophyll-synthesis-related enzymes (such as glutamyl-tRNA synthetase, etc.), and chlorophyll synthase activity in newly picked tea leaves. Moreover, an obvious decrease was found in the content of l-glutamate as the foremost precursor substance of chlorophyll synthesis. Hence, our findings revealed that the chlorophyll synthesis reaction was induced by the light-dehydration-stress in the initial withering of tea leaves. This study provides a theoretical basis for exploring preservation technology in actual green tea production.


Assuntos
Camellia sinensis/genética , Camellia sinensis/metabolismo , Clorofila/metabolismo , Manipulação de Alimentos/métodos , Regulação da Expressão Gênica de Plantas , Camellia sinensis/química , Camellia sinensis/crescimento & desenvolvimento , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , Clorofila/química , Cor , Enzimas/genética , Enzimas/metabolismo , Oxigenases/genética , Oxigenases/metabolismo , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Temperatura Ambiente
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