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1.
Sheng Wu Gong Cheng Xue Bao ; 36(2): 362-371, 2020 Feb 25.
Artigo em Chinês | MEDLINE | ID: mdl-32148008

RESUMO

Solanum tuberosum Zinc transporter 11 (StZnT11) is very important for maintaining zinc homeostasis in cells. The study on the expression of StZnT11 under abiotic stress and biotic stress laid a foundation for verifying the role of potato StZnT11 in the process of biotic stress of Ralstonia solanacearum species complex. According to the designated EST sequence, the homology of the original sequence was analyzed by using the Blast tool in NCBI, and a homologous object sequence with the highest similarity, coverage and e expectation value was selected. StZnT11 gene is obtained by Silico Cloning. The sequence and coding amino acid composition, physicochemical properties, molecular evolution, phosphorylation site and advanced structure of Solanum tuberosum StZnT11 gene were analyzed by bioinformatics method. The results showed that the cDNA gene is 1 300 bp in length, encoding a protein containing 348 amino acid residues, including 23 phosphorylation sites, one signal peptide and nine transmembrane regions, and is a hydrophobic protein located the plasma membrane. Through amino acid sequence alignment, StZnT11 protein has a high homology with zinc transporter from tobacco, tomato, pepper and other plants. The results of real-time fluorescence quantitative polymerase chain reaction showed that, StZnT11 is up-regulated by different concentrations of exogenous plant hormone abscisic acid (ABA). Tissue localization showed that StZnT11 was mainly expressed in specific tissues (phloem and leaf vascular bundles of stem vascular system). These results provide a theoretical basis for further experimental cloning and functional verification of the gene.


Assuntos
Solanum tuberosum , Sequência de Aminoácidos , Proteínas de Transporte , Clonagem Molecular , Biologia Computacional , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas
2.
Science ; 367(6485): 1482-1485, 2020 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-32217727

RESUMO

The premature abscission of flowers and fruits limits crop yield under environmental stress. Drought-induced flower drop in tomato plants was found to be regulated by phytosulfokine (PSK), a peptide hormone previously known for its growth-promoting and immune-modulating activities. PSK formation in response to drought stress depends on phytaspase 2, a subtilisin-like protease of the phytaspase subtype that generates the peptide hormone by aspartate-specific processing of the PSK precursor in the tomato flower pedicel. The mature peptide acts in the abscission zone where it induces expression of cell wall hydrolases that execute the abscission process. Our results provide insight into the molecular control of abscission as regulated by proteolytic processing to generate a small plant peptide hormone.


Assuntos
Secas , Flores/fisiologia , Lycopersicon esculentum/fisiologia , Hormônios Peptídicos/fisiologia , Proteínas de Plantas/fisiologia , Estresse Fisiológico , Parede Celular/enzimologia , Etilenos , Frutas/fisiologia , Técnicas de Silenciamento de Genes , Hidrolases/fisiologia , Ácidos Indolacéticos , Peptídeos , Plantas Geneticamente Modificadas/fisiologia , Transdução de Sinais
3.
J Agric Food Chem ; 68(10): 3238-3249, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32059106

RESUMO

Enrichment of multiple bioactive components with different characters into one food substrate simultaneously is a challenge. In this study, the hydrophilic epigallocatechin gallate (EGCG) and the hydrophobic quercetin were simultaneously enriched in the cavity of phytoferritin from red bean seed deprived of iron (apoRBF), a cagelike protein. The interactions of apoRBF with EGCG and quercetin were evaluated by UV/visible absorption, fluorescence, and circular dichroism technologies. By combination of the reversible assembly and urea induced approaches, both EGCG and quercetin were successfully coencapsulated in apoRBF to fabricate four kinds of apoRBF-EGCG-quercetin nanocomplexes FEQ (FEQ1, FEQ2, FEQ3, and FEQ4) with good solubility in aqueous solution. All FEQ samples maintained the typically spherical morphology of ferritin cage with a diameter around 12 nm. Among the four FEQ samples, the FEQ1 prepared by involving a pH 2.0/6.7 transition scheme was more effective in encapsulating EGCG and quercetin molecules than that by the urea induced method. Furthermore, all FEQs facilitated the stability of EGCG and quercetin molecules relative to free ones, and simultaneous coencapsulation of EGCG and quercetin could significantly improve the quercetin stability as compared with that of the free one and quercetin-loaded ferritin (p < 0.05), respectively. This work provides a new scheme to design and fabricate the ferritin based carrier for encapsulation of multiple bioactive components, and it is beneficial for the intensification of multifunction in one food substrate.


Assuntos
Catequina/análogos & derivados , Ferritinas/química , Proteínas de Plantas/química , Catequina/química , Portadores de Fármacos/química , Composição de Medicamentos , Estabilidade de Medicamentos , Fabaceae/química , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Sementes/química
4.
J Agric Food Chem ; 68(10): 3071-3078, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32078770

RESUMO

This work reports halogenated 5-(2-hydroxyphenyl)pyrazoles as pseudilin analogues with the potential to target the enzyme IspD in the methylerythritol phosphate (MEP) pathway. Such analogues were designed using the bioisosteric replacement of the pseudilin core structure and synthesized via an efficient three-step route. With AtIspD-based screening and pre- and post-emergence herbicidal tests, these compounds were demonstrated to have considerable activities against AtIspD, with IC50 up to 3.27 µM, and against model plants rape and barnyard grass, with moderate to excellent activities. At a rate of 150 g/ha in the greenhouse test, three compounds exhibited higher or comparable herbicidal activities than pseudilin. Molecular docking of representative compounds into the allosteric site of AtIspD revealed a binding mode similar to that of pseudilin. The established bioisosterism and synthesis method in this work may serve as an important tool for the development of new herbicides and antimicrobials targeting IspD in the MEP pathway.


Assuntos
Echinochloa/enzimologia , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Herbicidas/química , Herbicidas/farmacologia , Proteínas de Plantas/antagonistas & inibidores , Pirazóis/química , Pirazóis/farmacologia , Echinochloa/efeitos dos fármacos , Echinochloa/genética , Echinochloa/metabolismo , Inibidores Enzimáticos/síntese química , Eritritol/metabolismo , Halogenação , Herbicidas/síntese química , Simulação de Acoplamento Molecular , Estrutura Molecular , Fosfatos/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Pirazóis/síntese química , Relação Estrutura-Atividade
5.
J Agric Food Chem ; 68(10): 3214-3219, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32079394

RESUMO

Four terpene synthases for the biosynthesis of volatile terpenoids were identified from the transcriptome of Stellera chamaejasme L. flowers, including SchTPS1, SchTPS2, SchTPS3, and SchTPS4. Their functions were characterized by synthetic biology approaches in Escherichia coli and in vitro enzymatic assays. SchTPS1, SchTPS2, and SchTPS3 are guaiene synthases, while SchTPS4 is an (E,E)-geranyl linalool synthase. Next, SchTPS1 and α-guaiene 2-oxidase VvSTO2 were co-expressed in Saccharomyces cerevisiae to reconstruct the biosynthetic pathway of (-)-rotundone, which is a unique aroma compound in fruits, vegetables, and wines. This is the first report for the construction of a (-)-rotundone-producing microbial platform.


Assuntos
Alquil e Aril Transferases/metabolismo , Azulenos/metabolismo , Proteínas de Plantas/metabolismo , Saccharomyces cerevisiae/metabolismo , Sesquiterpenos de Guaiano/metabolismo , Sesquiterpenos/metabolismo , Thymelaeaceae/enzimologia , Alquil e Aril Transferases/genética , Vias Biossintéticas , Flores/enzimologia , Flores/genética , Expressão Gênica , Proteínas de Plantas/genética , Saccharomyces cerevisiae/genética , Thymelaeaceae/genética
6.
J Agric Food Chem ; 68(10): 3267-3276, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32101430

RESUMO

Cross-talk between various hormones is important in regulating many aspects of plant growth, development, and senescence, including fruit ripening. Here, exogenous ethylene (ETH, 100 µL/L, 12 h) rapidly accelerated 'Hayward' kiwifruit (Actinidia deliciosa) softening and ethylene production and was enhanced by supplementing with continuous treatment with methyl jasmonate (MeJA, 100 µM/L, 12 h) (ETH+MeJA). ETH+MeJA enhanced ACC synthase (ACS) activities and 1-aminocyclopropane-1-carboxylic acid (ACC) accumulation but not ACC oxidase (ACO) activity. Increased transcripts of ACS genes AdACS1 and AdACS2, ACS activity, and ethylene production were positively correlated. The abundance of AdACS1 was about 6-fold higher than AdACS2. RNA-seq identified 6 transcription factors among the 87 differentially expressed unigenes induced by ETH+MeJA. Dual-luciferase and electrophoretic mobility shift assays (EMSA) indicated that AdNAC2/3 physically interacted with and trans-activated the AdACS1 promoter 2.2- and 3.5-fold, respectively. Collectively, our results indicate that MeJA accelerates ethylene production in kiwifruit induced by exogenous ethylene, via a preferential activation of AdACS1 and AdACS2.


Assuntos
Acetatos/farmacologia , Actinidia/efeitos dos fármacos , Coenzima A Ligases/metabolismo , Ciclopentanos/farmacologia , Etilenos/biossíntese , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oxilipinas/farmacologia , Reguladores de Crescimento de Planta/farmacologia , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Actinidia/enzimologia , Actinidia/genética , Actinidia/metabolismo , Frutas/efeitos dos fármacos , Frutas/enzimologia , Frutas/genética , Frutas/metabolismo , Proteínas de Plantas/genética , Fatores de Transcrição/genética
7.
Yi Chuan ; 42(2): 194-211, 2020 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-32102776

RESUMO

As a coastal halophyte, Spartina alterniflora has high salt tolerance. However, the mechanism at the molecular level has not been widely studied due to the absence of a reference genome. The proteins of NAC families are plant-specific transcription factors that regulate the growth, development and stress response in plants. To identify the NAC family and explore the relationship between NAC proteins and the growth, development and stress response of Spatina alterniflora, full-length transcriptome data of Spartina alterniflora by the third generation sequencing technology was used as reference sequences in this study to blast with the NAC protein sequences from Oryza sativa, Arabidopsis thaliana and Zea mays. Finally, 62 SaNAC proteins were found in Spartina alterniflora by deep analysis on conserved domains. Then we analyzed sequence alignment, evolution, motif prediction, homology comparison, subcellular localization, tissue and abiotic stress-induced gene differential expression profile on the NAC family members in Spartina alterniflora. As a result, all SaNAC proteins were found containing a conserved NAM domain and having certain evolutionary similarity with rice; two family proteins, SaNAC9 and SaNAC49, were expressed in the nucleus; moreover, SaNAC genes were identified to have distinct expressional profiles in different tissues and stress response of Spartina alterniflora. These results indicated the SaNAC transcription factor family not only had conserved functional domains but also played important role in the regulation of growth, development and abiotic stress response.


Assuntos
Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Poaceae/genética , Plantas Tolerantes a Sal/genética , Fatores de Transcrição/genética , Filogenia , Estresse Fisiológico
8.
Plant Mol Biol ; 102(6): 589-602, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32026326

RESUMO

Leaf angle is a key factor in plant architecture and crop yield. Brassinosteroids (BRs) regulate many developmental processes, especially the leaf angle in monocots. However, the BR signalling pathway is complex and includes many unknown members. Here, we propose that Oryza sativa BRASSINOSTEROID-RESPONSIVE LEAF ANGLE REGULATOR 1 (OsBLR1) encodes a bHLH transcription factor, and positively regulates BR signalling to increase the leaf angle and grain length in rice (Oryza sativa L.). Lines overexpressing OsBLR1 (blr1-D and BLR1-OE-1/2/3) had similar traits, with increased leaf angle and grain length. Conversely, OsBLR1-knockout mutants (blr1-1/2/3) had erect leaves and shorter grains. Lamina joint inclination, coleoptile elongation, and root elongation assay results indicated that these overexpression lines were more sensitive to BR, while the knockout mutants were less sensitive. There was no significant difference in the endogenous BR contents of blr1-1/2 and wild-type plants. These results suggest that OsBLR1 is involved in BR signal transduction. The blr1-D mutant, with increased cell growth in the lamina joint and smaller leaf midrib, showed significant changes in gene expression related to the cell wall and leaf development compared with wild-type plants; furthermore, the cellulose and protopectin contents in blr1-D were reduced, which resulted in the increased leaf angle and bent leaves. As the potential downstream target gene of OsBLR1, the REGULATOR OF LEAF INCLINATION1 (OsRLI1) gene expression was up-regulated in OsBLR1-overexpression lines and down-regulated in OsBLR1-knockout mutants. Moreover, we screened OsRACK1A as an interaction protein of OsBLR1 using a yeast two-hybrid assay and glutathione-S-transferase pull-down.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Brassinosteroides/metabolismo , Oryza/genética , Oryza/metabolismo , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Transdução de Sinais , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Genes de Plantas , Oryza/crescimento & desenvolvimento , Fenótipo , Reguladores de Crescimento de Planta/metabolismo , Folhas de Planta/citologia , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Transdução de Sinais/genética , Fatores de Transcrição
9.
Plant Mol Biol ; 102(6): 603-614, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32052233

RESUMO

The WRKY transcription factor family is involved in responding to biotic and abiotic stresses. Its members contain a typical WRKY domain and can regulate plant physiological responses by binding to W-boxes in the promoter regions of downstream target genes. We identified the sweet sorghum SbWRKY50 (Sb09g005700) gene, which encodes a typical class II of the WRKY family protein that localizes to the nucleus and has transcriptional activation activity. The expression of SbWRKY50 in sweet sorghum was reduced by salt stress, and its ectopic expression reduced the salt tolerance of Arabidopsis thaliana plants. Compared with the wild type, the germination rate, root length, biomass and potassium ion content of SbWRKY50 over-expression plants decreased significantly under salt-stress conditions, while the hydrogen peroxide, superoxide anion and sodium ion contents increased. Real-time PCR results showed that the expression levels of AtSOS1, AtHKT1 and genes related to osmotic and oxidative stresses in over-expression strains decreased under salt-stress conditions. Luciferase complementation imaging and yeast one-hybrid assays confirmed that SbWRKY50 could directly bind to the upstream promoter of the SOS1 gene in A. thaliana. However, in sweet sorghum, SbWRKY50 could directly bind to the upstream promoters of SOS1 and HKT1. These results suggest that the new WRKY transcription factor SbWRKY50 participates in plant salt response by controlling ion homeostasis. However, the regulatory mechanisms are different in sweet sorghum and Arabidopsis, which may explain their different salt tolerance levels. The data provide information that can be applied to genetically modifying salt tolerance in different crop varieties.


Assuntos
Homeostase , Tolerância ao Sal/fisiologia , Sorghum/genética , Sorghum/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Biomassa , Proteínas de Transporte , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Regulação da Expressão Gênica de Plantas , Germinação , Peróxido de Hidrogênio/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas , Potássio/metabolismo , Regiões Promotoras Genéticas , Espécies Reativas de Oxigênio/metabolismo , Sementes , Sódio/metabolismo , Trocadores de Sódio-Hidrogênio/genética , Trocadores de Sódio-Hidrogênio/metabolismo , Estresse Fisiológico , Superóxidos/metabolismo , Simportadores/genética , Simportadores/metabolismo
10.
J Agric Food Chem ; 68(10): 3033-3049, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32052629

RESUMO

The high levels of secondary metabolites in rapeseed play important roles in determining the oil quality and feeding value. Here, we characterized the metabolic profiles in seeds of various yellow- and black-seeded rapeseed accessions. Two hundred and forty-eight features were characterized, including 31 phenolic acids, 54 flavonoids, 24 glucosinolates, 65 lipid compounds, and 74 other polar compounds. The most abundant phenolic acids and various flavonoids (epicatechin, isorhamnetin, kaempferol, quercetin, and their derivatives) were widely detected and showed significant differences in distribution between the yellow- and black-seeded rapeseed. Furthermore, the related genes (e.g., BnTT3, BnTT18, BnTT10, BnTT12, and BnBAN) involved in the proanthocyanidin pathway had lower expression levels in yellow-seeded rapeseed, strongly suggesting that the seed coat color could be mainly determined by the levels of epicatechin and their derivatives. These results improve our understanding of the primary constituents of rapeseed and lay the foundation for breeding novel varieties with a high nutritional value.


Assuntos
Brassica napus/química , Extratos Vegetais/química , Brassica napus/classificação , Brassica napus/genética , Brassica napus/metabolismo , Catequina/química , Cromatografia Líquida de Alta Pressão , Cor , Flavonoides/química , Hidroxibenzoatos/química , Metaboloma , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sementes/química , Espectrometria de Massas em Tandem
11.
J Agric Food Chem ; 68(10): 3017-3025, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32059105

RESUMO

Chlorsulfuron has been applied in wheat fields as a recognized herbicide worldwide, yet it was officially banned in China since 2014 for its soil persistence problem. On the basis of our previous research that 5-dimethylamino distinctively accelerated degradation rate in soils, a modified amino moiety (Ia-c) and monosubstituted amino group (Id-e) were introduced onto the fifth position of the benzene ring in sulfonylurea structures, as well as heterocyclic amino substituents (If-g) to seek a suitable soil degradation rate during such an in situ crop rotation system. Referring to the biological data and ScAHAS inhibition and ScAHAS docking results, they turned out to be AHAS inhibitors with high potent herbicidal activities. The various influence on soil degradation rate along with crop safety indicated that different substituents on the fifth position have exerted an apparent impact. Their united study of structure-activity-safety-degradation relationship has great potential to provide valuable information for further development of eco-friendly agrochemicals.


Assuntos
Acetolactato Sintase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Herbicidas/farmacologia , Proteínas de Plantas/antagonistas & inibidores , Poluentes do Solo/química , Compostos de Sulfonilureia/farmacologia , Acetolactato Sintase/metabolismo , Amaranthus/efeitos dos fármacos , Amaranthus/enzimologia , Brassica/efeitos dos fármacos , Brassica/enzimologia , Inibidores Enzimáticos/química , Herbicidas/química , Cinética , Modelos Moleculares , Proteínas de Plantas/metabolismo , Poluentes do Solo/farmacologia , Relação Estrutura-Atividade , Compostos de Sulfonilureia/química
12.
Gene ; 736: 144422, 2020 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-32007584

RESUMO

Late embryogenesis abundant (LEA) proteins are involved in plant stress responses and osmotic regulation, and they are accumulated in the late embryonic stage. There have been no previous genome-wide analyses of the LEA gene family members in wheat and its close relatives. In this study, 281, 53, 151, 89, 99, and 99 LEA genes were identified in wheat (Triticum aestivum), Triticum urartu, Triticum dicoccoides, Aegilops tauschii, barley, and Brachypodium distachyon, respectively. The wheat LEA gene family (TaLEA genes) was divided into eight subfamilies according to the conserved domains. All TaLEA genes contain very few introns (<3) and they are unevenly distributed on the 21 chromosomes. We identified 39 pairs of tandem duplication genes and 9 pairs of segmental duplication genes in the wheat LEA gene family. This proved that the tandem duplication and segmental duplication played an important role in the expansion of the TaLEA gene family. According to published transcriptome data and qRT-PCR analysis, the TaLEA genes exhibit different tissue expression patterns and they are regulated by various abiotic stresses, especially salt and cold stress. This study provides a comprehensive understanding of the wheat LEA gene family.


Assuntos
Regulação da Expressão Gênica de Plantas/genética , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Triticum/genética , Mapeamento Cromossômico/métodos , Cromossomos de Plantas/genética , Perfilação da Expressão Gênica/métodos , Genoma de Planta/genética , Estudo de Associação Genômica Ampla/métodos , Filogenia , Transcriptoma/genética
13.
J Agric Food Chem ; 68(8): 2528-2538, 2020 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-32011878

RESUMO

Albino became a novel kind of tea cultivar in China recently. In this study, transcriptome and whole-genome bisulfite sequencing (WGBS) were employed to investigate the shading effects on leaf color conversion and biosynthesis of three major secondary metabolites in the albino tea cultivar "Yujinxiang". The increased leaf chlorophyll level was likely the major cause for shaded leaf greening from young pale or yellow leaf. In comparison with the control, the total catechin level of the shading group was significantly decreased and the abundance of caffeine was markedly increased, while the theanine level was nearly not influenced. Meanwhile, differentially expressed genes (DEGs) enriched in some biological processes and pathways were identified by transcriptome analysis. Furthermore, whole-genome DNA methylation analysis revealed that the global genomic DNA methylation patterns of the shading period were remarkably altered in comparison with the control. In addition, differentially methylated regions (DMRs) and the DMR-related DEG analysis indicated that the DMR-related DEGs were the critical participants in biosynthesis of the major secondary metabolites. These findings suggest that DNA methylation is probably responsible for changes in the contents of the major secondary metabolites in Yujinxiang.


Assuntos
Camellia sinensis/metabolismo , Folhas de Planta/efeitos da radiação , Metabolismo Secundário/efeitos da radiação , Camellia sinensis/genética , Camellia sinensis/efeitos da radiação , Clorofila/biossíntese , Cor , Regulação da Expressão Gênica de Plantas , Luz , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
14.
J Agric Food Chem ; 68(8): 2366-2372, 2020 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-32017555

RESUMO

Spermidine possesses multiple healthy functions, and soybeans contain the most abundant spermidine. In this study, spermidine contents of soybeans from different varieties and production regions in China were evaluated, and a spermidine synthase gene (speE) was identified by recombinant expression, transcriptional verification, and sequence analysis. Spermidine contents of soybean samples from 18 varieties ranged 72.38-228.82 mg/kg, and those from 19 production regions ranged 134.64-242.32 mg/kg. The highest-spermidine sample GZ was used to clone four predicted speE genes. Expressing the gene speE5 improved the spermidine titer by 54% in Bacillus amyloliquefaciens, confirming that speE5 was involved in spermidine synthesis. Transcriptional verification was performed through a soybean germination model. Germination for 48 h led to a onefold increase of spermidine in samples SHX and HB, and corresponding speE5 transcriptional levels were improved by 26-fold and 18-fold, respectively, further verifying the function of speE5. Finally, the sequences of the speE5 gene and deduced amino acids were analyzed, and the conserved sites and catalysis mechanisms were presented. This study identified an active spermidine synthase gene from soybean for the first time, which provided an important gene resource for genetic breeding of spermidine-rich soybean or microbial cell factory.


Assuntos
Proteínas de Plantas/genética , Soja/enzimologia , Espermidina Sintase/genética , Sequência de Aminoácidos , Bacillus amyloliquefaciens/genética , Bacillus amyloliquefaciens/metabolismo , Germinação , Dados de Sequência Molecular , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Sementes/enzimologia , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Alinhamento de Sequência , Soja/genética , Soja/crescimento & desenvolvimento , Soja/metabolismo , Espermidina/metabolismo , Espermidina Sintase/química , Espermidina Sintase/metabolismo , Transcrição Genética
15.
J Agric Food Chem ; 68(8): 2467-2476, 2020 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-32031791

RESUMO

Enzymatic browning is a major issue affecting the quality of processed potato (Solanum tuberosum L.). To understand the molecular mechanism of browning, transcriptional analyses were performed by employing potatoes that differed in browning. Coexpression analysis indicated that 9 out of 15 upregulated genes in browning-less groups encoded for potato protease inhibitors (StPIs). In addition, gene otology analysis showed that the enriched terms were mainly involved in protease inhibitors. Overexpression of cysteine StPI 143 and StPI 146 individually reduced browning and lowered protease activities and tyrosine and total free amino acid (FAA) contents, but they could not decrease polyphenol oxidase activity. Moreover, supplementing exogenous tyrosine or total FAAs into transgenic potato mash to wild-type amounts promoted mash browning, browning with total FAAs, more than with tyrosine, resembling wild-type levels. These results implied that cysteine StPIs reduced browning via lowering the accumulation of FAAs in addition to tyrosine. Our findings have enriched the knowledge about the roles and mechanisms of protease inhibitors in regulating enzymatic browning of potato, which provide new ways for controlling potato browning.


Assuntos
Aminoácidos/metabolismo , Proteínas de Plantas/metabolismo , Inibidores de Proteases/metabolismo , Solanum tuberosum/metabolismo , Catecol Oxidase/antagonistas & inibidores , Catecol Oxidase/genética , Catecol Oxidase/metabolismo , Cor , Cisteína Proteases/genética , Cisteína Proteases/metabolismo , Proteínas de Plantas/genética , Solanum tuberosum/enzimologia , Solanum tuberosum/genética
16.
J Agric Food Chem ; 68(7): 1958-1965, 2020 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-32003983

RESUMO

Heavy metal stress is a major growth- and yield-limiting factor for plants. Heavy metals include essential metals (copper, iron, zinc, and manganese) and non-essential metals (cadmium, mercury, aluminum, arsenic, and lead). Plants use complex mechanisms of gene regulation under heavy metal stress. MicroRNAs are 21-nucleotide non-coding small RNAs as important modulators of gene expression post-transcriptionally. Recently, high-throughput sequencing has led to the identification of an increasing number of heavy-metal-responsive microRNAs in plants. Metal-regulated microRNAs and their target genes are part of a complex regulatory network that controls various biological processes, including heavy metal uptake and transport, protein folding and assembly, metal chelation, scavenging of reactive oxygen species, hormone signaling, and microRNA biogenesis. In this review, we summarize the recent molecular studies that identify heavy-metal-regulated microRNAs and their roles in the regulation of target genes as part of the microRNA-associated regulatory network in response to heavy metal stress in plants.


Assuntos
Metais Pesados/metabolismo , MicroRNAs/metabolismo , Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Homeostase , MicroRNAs/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas/genética
17.
J Agric Food Chem ; 68(9): 2623-2630, 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32058714

RESUMO

Eleusine indica is a typical xerophytic weed species with a cosmopolitan distribution. It is invasive and highly adaptable to diverse habitats and crops. Due to rice cropping-pattern changes, E indica has become one of the main dominant grass weeds infecting direct-seeding paddy fields. A Chinese E. indica population has evolved multiple-herbicide resistance to cyhalofop-butyl and glyphosate. In this study, the multiple-resistance profile of E. indica to these two different types of herbicides and their resistance mechanisms were investigated. Whole-plant dose-response assays indicated that the multiple-herbicide-resistant (MHR) population exhibited 10.8-fold resistance to cyhalofop-butyl and 3.1-fold resistance to glyphosate compared with the susceptible (S) population. ACCase sequencing revealed that the Asp-2078-Gly mutation was strongly associated with E. indica resistance to cyhalofop-butyl. The MHR plants accumulated less shikimic acid than S plants at 4, 6, and 8 days after glyphosate treatment. In addition, no amino acid substitution in the EPSPS gene was found in MHR plants. Further analysis revealed that the relative expression level of EPSPS in MHR plants was 6-10-fold higher than that in S plants following glyphosate treatment, indicating that EPSPS overexpression may contribute to the glyphosate resistance. Furthermore, the effectiveness of nine post-emergence herbicides against E. indica were evaluated, and one PPO inhibitor pyraclonil was identified as highly effective in controlling the S and MHR E. indica populations.


Assuntos
Butanos/farmacologia , Eleusine/efeitos dos fármacos , Glicina/análogos & derivados , Resistência a Herbicidas , Herbicidas/farmacologia , Nitrilos/farmacologia , Eleusine/genética , Eleusine/metabolismo , Regulação da Expressão Gênica de Plantas , Glicina/farmacologia , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Daninhas/efeitos dos fármacos , Plantas Daninhas/genética , Plantas Daninhas/metabolismo
18.
Phytochemistry ; 173: 112300, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32087435

RESUMO

Although dehydrins show cryoprotective activities for freeze-sensitive enzymes, the underlying mechanism is still under investigation. Here, we report that F-segments conserved in some dehydrins cryoprotected lactate dehydrogenase (LDH) as well as K-segments, which were previously identified as cryoprotective segments of dehydrins. The cryoprotective activity levels of four F-segments of Arabidopsis dehydrins were similar to that of a typical K-segment. Amino acid substitution experiments indicated that the activity of the F-segment of Arabidopsis COR47 (designated as Fseg) depended on the hydrophobic residues (L, F, and V). Intriguingly, when all the amino acids other than the hydrophobic residues were changed to glycine, the cryoprotective activity did not change, suggesting that the hydrophobic amino acids were sufficient for Fseg activity. Circular dichroism analysis indicated that Fseg was mainly disordered in aqueous solution as well as Fseg_Φ/T, in which the hydrophobic residues of Fseg were changed to T. This suggested that the hydrophobic interaction might be related to the cryoprotective activities of Fseg.


Assuntos
Arabidopsis , Sequência de Aminoácidos , Interações Hidrofóbicas e Hidrofílicas , L-Lactato Desidrogenase , Proteínas de Plantas
19.
Plant Mol Biol ; 102(6): 571-588, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31927659

RESUMO

KEY MESSAGE: A family of repetitive proline-rich proteins interact with acidic pectins and play distinct roles in legume root cell walls affecting cortical and vascular structure. A proline-rich protein (PRP) family, composed of tandemly repeated Pro-Hyp-Val-X-Lys pentapeptide motifs, is found primarily in the Leguminosae. Four distinct size classes within this family are encoded by seven tightly linked genes: MtPRP1, MtPRP2 and MtPRP3, and four nearly identical MtPRP4 genes. Promoter fusions to ß-glucuronidase showed strong expression in the stele of hairy roots for all 4 PRP genes tested, with additional expression in the cortex for PRP1, PRP2 and PRP4. All except MtPRP4 are strongly expressed in non-tumorous roots, and secreted and ionically bound to root cell walls. These PRPs are absent from root epidermal cell walls, and PRP accumulation is highly localized within the walls of root cortical and vascular tissues. Within xylem tissue, PRPs are deposited in secondary thickenings where it is spatially exclusive to lignin. In newly differentiating xylem, PRPs are deposited in the regularly spaced paired-pits and pit membranes that hydraulically connect neighboring xylem elements. Hairpin-RNA knock-down constructs reducing PRP expression in Medicago truncatula hairy root tumors disrupted cortical and vascular patterning. Immunoblots showed that the knockdown tumors had potentially compensating increases in the non-targeted PRPs, all of which cross-react with the anti-PRP antibodies. However, PRP3 knockdown differed from knockdown of PRP1 and PRP2 in that it greatly reduced viability of hairy root tumors. We hypothesize that repetitive PRPs interact with acidic pectins to form block-copolymer gels that can play distinct roles in legume root cell walls.


Assuntos
Medicago truncatula/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Domínios Proteicos Ricos em Prolina/genética , Parede Celular/metabolismo , Regulação da Expressão Gênica de Plantas , Técnicas de Silenciamento de Genes , Vetores Genéticos , Glucuronidase , Medicago truncatula/genética , Raízes de Plantas/citologia , Raízes de Plantas/genética , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Proteínas Salivares Ricas em Prolina , Xilema/metabolismo
20.
Plant Mol Biol ; 102(6): 625-644, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31965448

RESUMO

KEY MESSAGE: Comprehensive transcriptome analysis suggested that the primary metabolism is modulated to augment the supply of substrates towards secondary metabolism operating in the glandular trichomes of Nicotiana tabacum. The comparative gene expression and co-expression network analysis revealed that certain members of transcription factor genes belonging to the MYB, HD-ZIP, ERF, TCP, SRS, WRKY and DOF families may be involved in the regulation of metabolism and/other aspects in the glandular trichomes of N. tabacum The glandular trichomes of Nicotiana tabacum are highly productive in terms of secondary metabolites and therefore have been projected to be used as a prognostic platform for metabolic engineering of valuable natural products. For obvious reasons, detailed studies pertaining to the metabolic and gene regulatory networks operating in the glandular trichomes of N. tabacum are of pivotal significance to be undertaken. We have carried out next-generation sequencing of glandular trichomes of N. tabcaum and investigated differential gene expression among different tissues, including trichome-free leaves. We identified a total of 37,269 and 37,371 genes, expressing in trichome free leaf and glandular trichomes, respectively, at a cutoff of FPKM ≥ 1. The analysis revealed that different pathways involved with the primary metabolism are modulated in glandular trichomes of N. tabacum, providing a plausible explanation for the enhanced biosynthesis of secondary metabolism in the glandular trichomes. Further, comparative gene expression analysis revealed several genes, which display preferential expression in the glandular trichomes and thereby seem to be potential candidate genes for future studies in connection to the discovery of novel trichome specific promoters. The present study also led to the comprehensive identification of 1750 transcription factor genes expressing at a cutoff of FPKM ≥ 1 in the glandular trichomes of N. tabacum. The clustering and co-expression analysis suggested that transcription factor genes belonging to HD-ZIP, ERF, WRKY, MYB, TCP, SRS and DOF families may be the major players in the regulation of gene expression in the glandular trichomes of N. tabacum. To the best of our knowledge, the present work is the first effort towards detailed identification of genes, especially regulatory genes expressing in the glandular trichomes of N. tabacum. The data resource and the empirical findings from present work in all probability must, therefore, provide a reference and background context for future work aiming at deciphering molecular mechanism of regulation of secondary metabolism and gene expression in the glandular trichomes of N. tabacum.


Assuntos
Perfilação da Expressão Gênica/métodos , Redes Reguladoras de Genes/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Tabaco/genética , Tricomas/genética , Aminoácidos/biossíntese , Aminoácidos/genética , Parede Celular , Regulação da Expressão Gênica de Plantas , Glicólise/genética , Metabolismo dos Lipídeos/genética , Engenharia Metabólica , Redes e Vias Metabólicas/genética , Folhas de Planta/genética , Proteínas de Plantas/metabolismo , Regiões Promotoras Genéticas , Metabolismo Secundário/genética , Tabaco/metabolismo , Fatores de Transcrição/genética , Transcriptoma
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