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1.
Nat Plants ; 8(4): 346-355, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35347264

RESUMO

Gene duplications are a hallmark of plant genome evolution and a foundation for genetic interactions that shape phenotypic diversity1-5. Compensation is a major form of paralogue interaction6-8 but how compensation relationships change as allelic variation accumulates is unknown. Here we leveraged genomics and genome editing across the Solanaceae family to capture the evolution of compensating paralogues. Mutations in the stem cell regulator CLV3 cause floral organs to overproliferate in many plants9-11. In tomato, this phenotype is partially suppressed by transcriptional upregulation of a closely related paralogue12. Tobacco lost this paralogue, resulting in no compensation and extreme clv3 phenotypes. Strikingly, the paralogues of petunia and groundcherry nearly completely suppress clv3, indicating a potent ancestral state of compensation. Cross-species transgenic complementation analyses show that this potent compensation partially degenerated in tomato due to a single amino acid change in the paralogue and cis-regulatory variation that limits its transcriptional upregulation. Our findings show how genetic interactions are remodelled following duplications and suggest that dynamic paralogue evolution is widespread over short time scales and impacts phenotypic variation from natural and engineered mutations.


Assuntos
Lycopersicon esculentum , Sinais Direcionadores de Proteínas , Regulação da Expressão Gênica de Plantas , Lycopersicon esculentum/genética , Lycopersicon esculentum/metabolismo , Meristema/metabolismo , Peptídeos/metabolismo , Caules de Planta/genética , Caules de Planta/metabolismo
3.
Gene ; 823: 146384, 2022 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-35248661

RESUMO

UV-B radiation is a typical environmental stressor that can promote phytochemical accumulation in plants. Taxus species are highly appreciated due to the existence of bioactive taxoids (especially paclitaxel) and flavonoids. However, the effect of UV-B radiation on taxoid and flavonoid biosynthesis in Taxus cuspidata Sieb. et Zucc. is largely unknown. In the present work, the accumulation of taxoids and flavonoids in T. cuspidata plantlets was significantly induced by 12 and 24 h of UV-B radiation (3 W/m2), and a large number of significantly differentially expressed genes were obtained via transcriptomic analysis. The significant up-regulation of antioxidant enzyme- and flavonoid biosynthesis-related genes (phenylalanine ammonia lyase 1, chalcone synthase 2, flavonol synthase 1, and flavonoid 3', 5'-hydroxylase 2), suggested that UV-B might cause the oxidative stress thus promoting flavonoid accumulation in T. cuspidata. Moreover, the expression of some genes related to jasmonate metabolism and taxoid biosynthesis (taxadiene synthase, baccatin III-3-amino 3-phenylpropanoyltransferase 1, taxadiene-5α-hydroxylase, and ethylene response factors 15) was significantly activated, which indicated that UV-B might initiate jasmonate signaling pathway that contributed to taxoid enhancement in T. cuspidata. Additionally, the identification of some up-regulated genes involved in lignin biosynthesis pathway indicated that the lignification process in T. cuspidata might be stimulated for defense against UV-B radiation. Overall, our findings provided a better understanding of some potential key genes associated with flavonoid and taxoid biosynthesis in T. cuspidata exposed to UV-B radiation.


Assuntos
Vias Biossintéticas , Flavonoides/biossíntese , Perfilação da Expressão Gênica/métodos , Caules de Planta/crescimento & desenvolvimento , Taxoides/metabolismo , Taxus/genética , Cromatografia Líquida de Alta Pressão , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Estresse Oxidativo , Proteínas de Plantas/genética , Caules de Planta/metabolismo , Caules de Planta/efeitos da radiação , RNA-Seq , Espectrometria de Massas em Tandem , Taxus/crescimento & desenvolvimento , Taxus/metabolismo , Taxus/efeitos da radiação , Raios Ultravioleta/efeitos adversos
4.
Sci Rep ; 12(1): 3992, 2022 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-35273205

RESUMO

Bio-impedance non-invasive measurement techniques usage is rapidly increasing in the agriculture industry. These measured impedance variations reflect tacit biochemical and biophysical changes of living and non-living tissues. Bio-impedance circuit modeling is an effective solution used in biology and medicine to fit the measured impedance. This paper proposes two new fractional-order bio-impedance plant stem models. These new models are compared with three commonly used bio-impedance fractional-order circuit models in plant modeling (Cole, Double Cole, and Fractional-order Double-shell). The two proposed models represent the characterization of the biological cellular morphology of the plant stem. Experiments are conducted on two samples of three different medical plant species from the family Lamiaceae, and each sample is measured at two inter-electrode spacing distances. Bio-impedance measurements are done using an electrochemical station (SP150) in the range of 100 Hz to 100 kHz. All employed models are compared by fitting the measured data to verify the efficiency of the proposed models in modeling the plant stem tissue. The proposed models give the best results in all inter-electrode spacing distances. Four different metaheuristic optimization algorithms are used in the fitting process to extract all models parameter and find the best optimization algorithm in the bio-impedance problems.


Assuntos
Algoritmos , Biofísica , Impedância Elétrica , Eletrodos , Caules de Planta
5.
Mar Drugs ; 20(3)2022 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-35323487

RESUMO

Catharanthus roseus (L.) G. Don is a plant belonging to the genus Catharanthus of the Apocynaceae family. It contains more than one hundred alkaloids, of which some exhibit significant pharmacological activities. Chitooligosaccharides are the only basic aminooligosaccharides with positively charged cations in nature, which can regulate plant growth and antioxidant properties. In this study, the leaves of Catharanthus roseus were sprayed with chitooligosaccharides of different molecular weights (1 kDa, 2 kDa, 3 kDa) and different concentrations (0.01 µg/mL, 0.1 µg/mL, 1 µg/mL and 10 µg/mL). The fresh weights of its root, stem and leaf were all improved after chitooligosaccharides treatments. More importantly, the chitooligosaccharides elicitor strongly stimulated the accumulation of vindoline and catharanthine in the leaves, especially with the treatment of 0.1 µg/mL 3 kDa chitooligosaccharides, the contents of them were increased by 60.68% and 141.54%, respectively. Furthermore, as the defensive responses, antioxidant enzymes activities (catalase, glutathione reductase, ascorbate peroxidase, peroxidase and superoxide dismutase) were enhanced under chitooligosaccharides treatments. To further elucidate the underlying mechanism, qRT-PCR was used to investigate the genes expression levels of secologanin synthase (SLS), strictosidine synthase (STR), strictosidine glucosidase (SGD), tabersonine 16-hydroxylase (T16H), desacetoxyvindoline-4-hydroxylase (D4H), deacetylvindoline-4-O-acetyltransferase (DAT), peroxidase 1 (PRX1) and octadecanoid-responsive Catharanthus AP2-domain protein 3 (ORCA3). All the genes were significantly up-regulated after chitooligosaccharides treatments, and the transcription abundance of ORCA3, SLS, STR, DAT and PRX1 reached a maximal level with 0.1 µg/mL 3 kDa chitooligosaccharides treatment. All these results suggest that spraying Catharanthus roseus leaves with chitooligosaccharides, especially 0.1 µg/mL of 3 kDa chitooligosaccharides, may effectively improve the pharmaceutical value of Catharanthus roseus.


Assuntos
Catharanthus/efeitos dos fármacos , Quitosana/farmacologia , Oligossacarídeos/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Antioxidantes/metabolismo , Catharanthus/genética , Catharanthus/crescimento & desenvolvimento , Catharanthus/metabolismo , Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oxirredutases/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Caules de Planta/efeitos dos fármacos , Caules de Planta/crescimento & desenvolvimento , Vimblastina/análogos & derivados , Vimblastina/metabolismo , Alcaloides de Vinca/metabolismo
6.
J Agric Food Chem ; 70(10): 3136-3141, 2022 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-35232018

RESUMO

Maize stalks support leaves and reproductive structures and functionally support water and nutrient transport; besides, their anatomical and biochemical characteristics have been described as a plant defense against stress, also impacting economically important applications. In this study, we evaluated agronomical and stem description traits in a subset of maize inbred lines that showed variability for cell wall composition in the internodes. Overall, a great proportion of lignin subunit G and a low concentration of p-coumaric acid and lignin subunit S are beneficial for greater rind puncture resistance and taller plants, with a greater biomass yield. Also, the greater the proportions of subunit H, the longer the internode. Finally, the lower the total hemicellulose content, the greater the rind puncture resistance. Our results confirmed the effect of the cell wall on agronomic and stalk traits, which would be useful in applied breeding programs focused on biomass yield improvement.


Assuntos
Parede Celular , Melhoramento Vegetal , Biomassa , Parede Celular/química , Lignina/metabolismo , Caules de Planta/metabolismo , Zea mays/química
7.
Artigo em Inglês | MEDLINE | ID: mdl-35130136

RESUMO

A Gram-stain-negative, facultatively anaerobic, motile by gliding, rod-shaped, oxidase- and catalase-positive bacterial strain, designated BB8T, was isolated from the stems of a Korean soybean cultivar (Glycine max L. cv. Gwangan). The strain produced a yellow pigment on tryptic soy agar. Growth of strain BB8T occurred at pH 5.0-8.0 (optimum, pH 7.0), at 10-35 °C (optimum, 25-30 °C) and in the presence of 0-1 % (w/v) NaCl (optimum, 0.5%). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain BB8T formed a lineage within the genus Flavobacterium and was most closely related to Flavobacterium artemisiae SYP-B1015T (96.9 % 16S rRNA gene sequence similarity) and Flavobacterium ustbae T13T (96.8%). The complete genome sequence of strain BB8T was 5 513 159 bp long with a G+C content of 34.1 mol%. The major fatty acids (>10 %) of strain BB8T were iso-C15 : 0 (21 %), summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c, 20.3%) and iso-C16 : 0 3-OH (13.7%). The predominant polar lipids were phosphatidylethanolamine and unidentified aminolipids, and the major respiratory quinone was menaquinone-6. Based on these phenotypic, genotypic and chemotaxonomic characteristics, strain BB8T is considered to represent a novel species of the genus Flavobacterium, for which the name Flavobacterium endoglycinae sp. nov. is proposed. The type strain is BB8T (=KCTC 82167T=CCTCC AB 2020070T).


Assuntos
Flavobacterium , Filogenia , Soja , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Flavobacterium/classificação , Flavobacterium/isolamento & purificação , Fosfolipídeos/química , Caules de Planta/microbiologia , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Soja/microbiologia , Vitamina K 2/análogos & derivados , Vitamina K 2/química
8.
Molecules ; 27(3)2022 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-35164380

RESUMO

Two undescribed ether derivatives of sesquiterpenes, 1-ethoxycaryolane-1, 9ß-diol (1) and 2-ethoxyclovane-2ß, 9α-diol (3), and one new monoterpene glycoside, p-menthane-1α,2α,8-triol-4-O-ß-D-glucoside (5), were obtained, together with eight known compounds from the stems and leaves of I. simonsii. Their structures were elucidated by spectroscopic methods. Compounds 1-11 were evaluated for their potency against Staphylococcus aureus and clinical methicillin-resistant S. aureus (MRSA). Among them, compound 3 was weakly active against S. aureus (MIC = 128 µg/mL), and compounds 6 and 7 exhibited good antibacterial activity against S. aureus and MRSA (MICs = 2-8 µg/mL). A primary mechanism study revealed that compounds 6 and 7 could kill bacteria by destroying bacterial cell membranes. Moreover, compounds 6 and 7 were not susceptible to drug resistance development.


Assuntos
Antibacterianos/análise , Illicium/química , Monoterpenos/análise , Sesquiterpenos/análise , Antibacterianos/farmacologia , Humanos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Monoterpenos/farmacologia , Folhas de Planta/química , Caules de Planta/química , Sesquiterpenos/farmacologia , Infecções Estafilocócicas/tratamento farmacológico
9.
Int J Mol Sci ; 23(3)2022 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-35163510

RESUMO

In order to separate transformed cells from non-transformed cells, antibiotic selectable marker genes are usually utilized in genetic transformation. After obtaining transgenic plants, it is often necessary to remove the marker gene from the plant genome in order to avoid regulatory issues. However, many marker-free systems are time-consuming and labor-intensive. Homology-directed repair (HDR) is a process of homologous recombination using homologous arms for efficient and precise repair of DNA double-strand breaks (DSBs). The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 (Cas9) system is a powerful genome editing tool that can efficiently cause DSBs. Here, we isolated a rice promoter (Pssi) of a gene that highly expressed in stem, shoot tip and inflorescence, and established a high-efficiency sequence-excision strategy by using this Pssi to drive CRISPR/Cas9-mediated HDR for marker free (PssiCHMF). In our study, PssiCHMF-induced marker gene deletion was detected in 73.3% of T0 plants and 83.2% of T1 plants. A high proportion (55.6%) of homozygous marker-excised plants were obtained in T1 progeny. The recombinant GUS reporter-aided analysis and its sequencing of the recombinant products showed precise deletion and repair mediated by the PssiCHMF method. In conclusion, our CRISPR/Cas9-mediated HDR auto-excision method provides a time-saving and efficient strategy for removing the marker genes from transgenic plants.


Assuntos
Edição de Genes/métodos , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/genética , Regiões Promotoras Genéticas , Sistemas CRISPR-Cas , Embaralhamento de DNA , Flores/genética , Flores/crescimento & desenvolvimento , Recombinação Homóloga , Oryza/genética , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Caules de Planta/genética , Caules de Planta/crescimento & desenvolvimento
10.
Fitoterapia ; 157: 105133, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35114336

RESUMO

Six diterpenoids including three ent-kauranes (1-2, 4) and three cleistanthanes (3, 5-6) were isolated from the roots and stems of Phyllanthus acidus (L.) Skeels. Of them, (16S)-ent-16,17,18-tri-hydroxy-19-nor-kaur-4-en-3-one (1), phyllanthone A (2), and 6-hydroxycleistanthol (3) are new compounds, while the ent-kaurane diterpenoids were reported from the titled plant for the first time. Their structures were elucidated on the basis of the extensive spectroscopic analyses. Compounds 2 and 4-6 displayed cytotoxic potential with IC50 values ranging from 1.96 to 29.15 µM. They also showed moderate anti-inflammatory activities (IC50 = 6.30-12.05 µM). Particularly, the new ent-kaurane 2 displayed cytotoxic potential against HL-60 (IC50 = 2.00 µM) and MCF-7 (IC50 = 3.55 µM) cells, and anti-inflammatory activity (IC50 = 6.47 µM).


Assuntos
Diterpenos do Tipo Caurano/toxicidade , Diterpenos/toxicidade , Phyllanthus/química , Extratos Vegetais/toxicidade , Alcaloides/química , Alcaloides/isolamento & purificação , Alcaloides/toxicidade , Linhagem Celular Tumoral , Diterpenos/química , Diterpenos do Tipo Caurano/química , Humanos , Concentração Inibidora 50 , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Extratos Vegetais/síntese química , Raízes de Plantas/química , Caules de Planta/química
11.
Gene ; 822: 146329, 2022 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-35181500

RESUMO

Chitinases, the chitin-degrading enzymes, have been shown to play important role in defense against the chitin-containing fungal pathogens. In this study, we identified 48 chitinase-coding genes from the woody model plant Populus trichocarpa. Based on phylogenetic analysis, the Populus chitinases were classified into seven groups. Different gene structures and protein domain architectures were found among the seven Populus chitinase groups. Selection pressure analysis indicated that all the seven groups are under purifying selection. Phylogenetic analysis combined with chromosome location analysis showed that Populus chitinase gene family mainly expanded through tandem duplication. The Populus chitinase gene family underwent marked expression divergence and is inducibly expressed in response to treatments, such as chitosan, chitin, salicylic acid and methyl jasmonate. Protein enzymatic activity analysis showed that Populus chitinases had activity towards both chitin and chitosan. By integrating sequence characteristic, phylogenetic, selection pressure, gene expression and protein activity analysis, this study shed light on the evolution and function of chitinase family in poplar.


Assuntos
Quitinases/genética , Quitinases/metabolismo , Mapeamento Cromossômico/métodos , Populus/enzimologia , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Família Multigênica , Filogenia , Folhas de Planta/enzimologia , Folhas de Planta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/enzimologia , Raízes de Plantas/genética , Caules de Planta/enzimologia , Caules de Planta/genética , Populus/genética , Seleção Genética
12.
Molecules ; 27(4)2022 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-35209099

RESUMO

Natural compounds have been recognized as valuable sources for anticancer drug development. In this work, different parts from Momordica cochinchinensis Spreng were selected to perform cytotoxic screening against human prostate cancer (PC-3) cells. Chromatographic separation and purification were performed for the main constituents of the most effective extract. The content of the fatty acids was determined by Gas Chromatography-Flame Ionization Detector (GC-FID). Chemical structural elucidation was performed by spectroscopic means. For the mechanism of the apoptotic induction of the most effective extract, the characteristics were evaluated by Hoechst 33342 staining, sub-G1 peak analysis, JC-1 staining, and Western blotting. As a result, extracts from different parts of M. cochinchinensis significantly inhibited cancer cell viability. The most effective stem extract induced apoptosis in PC-3 cells by causing nuclear fragmentation, increasing the sub-G1 peak, and changing the mitochondrial membrane potential. Additionally, the stem extract increased the pro-apoptotic (caspase-3 and Noxa) mediators while decreasing the anti-apoptotic (Bcl-xL and Mcl-1) mediators. The main constituents of the stem extract are α-spinasterol and ligballinol, as well as some fatty acids. Our results demonstrated that the stem extract of M. cochinchinensis has cytotoxic and apoptotic effects in PC-3 cells. These results provide basic knowledge for developing antiproliferative agents for prostate cancer in the future.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Momordica/química , Extratos Vegetais/farmacologia , Caules de Planta/química , Antineoplásicos Fitogênicos/química , Apoptose/genética , Biomarcadores Tumorais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Modelos Moleculares , Estrutura Molecular , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/química , Relação Estrutura-Atividade
13.
Gene ; 821: 146276, 2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35176432

RESUMO

BACKGROUND: A total of 74,936 SNPs were employed to carry out population structure and genome-wide association studies and post-GWAS for hairiness character of the fifty-six samples including thirty-six Actinidia chinensis var. deliciosa, eighteen A. chinensis var. chinensis, and two A. polygama in the light of morphological observations. RESULTS: The percentage of heterozygous sites of A. chinensis var. deliciosa is higher than that of A. chinensis var. chinensis, which could be one of the reasons for A. chinensis var. deliciosa high disease resistance. Fifty-six samples were divided into two subgroups, in which the genetic distance, ranged from 0.17 to 0.99, according to their genetic divergence. Analysis of molecular variance shows that the frequency of genetic variations within the population is 83.53% and 16.47% between populations. Fst between the two populations is 0.14, and Nm is 1.60. Set at α ≤ 0.05, a total of 327 SNPs and 260 haplotypes were related to the hairiness character. A total of 246 proteins were annotated using GO and KEGG analyses, which indicated the membrane-related genes and stress-resistant metabolic pathways are related to the hairiness character of leaves, stems, and peels of kiwifruit. Protein interaction analysis showed that DNA-directed RNA polymerase was an important node protein that interacted with many proteins. CONCLUSIONS: The genetic basic in the fifty-six genotypes was rich. The results of clustering and morphological observations are not completely consistent, indicating the hairiness character play an important role in the classification of kiwifruit, in which two A. polygama were clustered together with those of A. chinensis var. chinensis. Phylogeny and haplotype analysis showed that the evolution of A. chinensis var. chinensis is later than that of A. chinensis var. deliciosa in A. chinesis. The loss of hairiness character on leaves, stems and peels of A. chinensis var. chinensis compare with A. chinensis var. deliciosa, which is also the result of its poor resistance.


Assuntos
Actinidia/genética , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Análise de Sequência de DNA/métodos , Estudo de Associação Genômica Ampla , Haplótipos , Metagenômica , Filogenia , Folhas de Planta/genética , Proteínas de Plantas/genética , Caules de Planta/genética
14.
Fitoterapia ; 157: 105132, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35074542

RESUMO

BACKGROUND: Increasing drug resistance of Helicobacter pylori has highlighted the search for natural compounds with antiadhesive properties, interrupting the adhesion of H. pylori to stomach epithelia. Basella alba, a plant widely used in Asian traditional medicine, was investigated for its antiadhesive activity against H. pylori. METHODS: B. alba extract FE was prepared by aqueous extraction. Polysaccharides were isolated from FE by ethanol precipitation and arabinogalactan-protein (AGP) was isolated with Yariv reagent. Carbohydrate analyses was performed by standard methods and sequence analysis of the protein part of AGP by LC-MS. In vitro adhesion assay of fluorescent-labelled H. pylori J99 to human AGS cells was performed by flow cytometric analysis. RESULTS: Raw polysaccharides (BA1) were isolated and 9% of BA1 were identified as AGP (53.1% neutral carbohydrates L-arabinose, D-galactose, rhamnose, 5.4% galacturonic acid, 41.5% protein). After deglycosylation of AGP, the protein part (two bands at 15 and 25 kDa in tricine SDS-PAGE) was shown to contain peptides like ribulose-bisphosphate-carboxylase-large-chain. Histological localization within the stem tissue of B. alba revealed that AGP was mainly located at the procambium ring. Functional assays indicated that neither FE nor BA1 had significant influence on viability of AGS cells or on H. pylori. FE inhibited the bacterial adhesion of H. pylori to AGS cells in a dose dependent manner. Best anti-adhesive effect of ~67% was observed with BA1 at 2 mg/mL. CONCLUSION: The data obtained from this study characterize in part the mucilage and isolated polysaccharides of B. alba. As the polysaccharides interact with the bacterial adhesion, a potential uses a supplemental antiadhesive entity against the recurrence of H. pylori after eradication therapy may be discussed.


Assuntos
Caryophyllales/química , Galactanos/química , Helicobacter pylori/efeitos dos fármacos , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Cromatografia em Camada Delgada , Eletroforese em Gel de Poliacrilamida , Humanos , Imunodifusão , Extratos Vegetais/isolamento & purificação , Caules de Planta/química , Polissacarídeos/isolamento & purificação , Ribulose-Bifosfato Carboxilase/química , Espectrometria de Massas em Tandem , Células Tumorais Cultivadas
16.
J Am Soc Mass Spectrom ; 33(2): 265-272, 2022 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-35020389

RESUMO

Spatially resolving the relative distribution of analyte molecules in biological matter holds great promise in the life sciences. Mass spectrometry imaging (MSI) is a technique that can provide such spatial resolution but remains underused in fields such as chemical ecology, as traditional MSI sample preparation is often chemically or morphologically invasive. Laser ablation electrospray ionization (LAESI)-MSI is a variation of MSI particularly well-suited for situations where chemical sample preparation is too invasive but provides new challenges related to the repeatability of measurement outcomes. We assess the repeatability of LAESI-MSI by sampling a droplet of [ring-13C6]l-phenylalanine with known concentration and expressing the resulting variability as a coefficient of variation, cv. In doing so, we entirely eliminate variability caused by surface morphology or underlying true sample gradients. We determine the limit of detection (LOD) for13C6-Phe by sampling from droplets with successively decreasing but known concentration. We assess the influence of source geometry on the LOD and repeatability by performing these experiments using four distinct variations of sources: one commercial and three custom-built ones. Finally, we extend our study to leaf and stem samples Arabidopsis thaliana and Gossypium hirsutum. We overcome the challenges of LAESI associated with three-dimensional surface morphology by relying on work previously published. Our measurements on both controlled standard and realistic samples give strong evidence that LAESI-MSI's repeatability in current implementations is insufficient for MSI in chemical ecology.


Assuntos
Lasers , Fenilalanina/química , Folhas de Planta/química , Caules de Planta/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Arabidopsis , Gossypium , Limite de Detecção , Reprodutibilidade dos Testes
17.
Proc Natl Acad Sci U S A ; 119(1)2022 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-34983847

RESUMO

Symplasmicly connected cells called sieve elements form a network of tubes in the phloem of vascular plants. Sieve elements have essential functions as they provide routes for photoassimilate distribution, the exchange of developmental signals, and the coordination of defense responses. Nonetheless, they are the least understood main type of plant cells. They are extremely sensitive, possess a reduced endomembrane system without Golgi apparatus, and lack nuclei and translation machineries, so that transcriptomics and similar techniques cannot be applied. Moreover, the analysis of phloem exudates as a proxy for sieve element composition is marred by methodological problems. We developed a simple protocol for the isolation of sieve elements from leaves and stems of Nicotiana tabacum at sufficient amounts for large-scale proteome analysis. By quantifying the enrichment of individual proteins in purified sieve element relative to bulk phloem preparations, proteins of increased likelyhood to function specifically in sieve elements were identified. To evaluate the validity of this approach, yellow fluorescent protein constructs of genes encoding three of the candidate proteins were expressed in plants. Tagged proteins occurred exclusively in sieve elements. Two of them, a putative cytochrome b561/ferric reductase and a reticulon-like protein, appeared restricted to segments of the endoplasmic reticulum (ER) that were inaccessible to green fluorescent protein dissolved in the ER lumen, suggesting a previously unknown differentiation of the endomembrane system in sieve elements. Evidently, our list of promising candidate proteins ( SI Appendix, Table S1) provides a valuable exploratory tool for sieve element biology.


Assuntos
Retículo Endoplasmático/metabolismo , Células Vegetais/metabolismo , Folhas de Planta/metabolismo , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Proteômica , Tabaco/metabolismo , Retículo Endoplasmático/genética , Folhas de Planta/citologia , Folhas de Planta/genética , Caules de Planta/citologia , Caules de Planta/genética , Plantas Geneticamente Modificadas/citologia , Plantas Geneticamente Modificadas/genética , Tabaco/citologia , Tabaco/genética
18.
Nutrients ; 14(2)2022 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-35057446

RESUMO

Bauhinia scandens L. (Family: Fabaceae) is commonly used to treat cholera, diarrhea, asthma, and diabetes disorder in integrative medicine. This study aimed to screen the presence of phytochemicals (preliminary and UPLC-QTOF-M.S. analysis) and to examine the pharmacological activities of Bauhinia scandens L. stems (MEBS) stem extracts. Besides, in silico study was also implemented to elucidate the binding affinity and drug capability of the selected phytochemicals. In vivo anti diarrheal activity was investigated in mice models. In vitro, antibacterial and antifungal properties of MEBS against several pathogenic strains were evaluated using the disc diffusion method. In addition, in silico study has been employed using Discovery studio 2020, UCFS Chimera, PyRx autodock vina, and online tools. In the anti-diarrheal investigation, MEBS showed a significant dose-dependent inhibition rate in all three methods. The antibacterial and antifungal screening showed a remarkable zone of inhibition, of the diameter 14-26 mm and 12-28 mm, by MEBS. The present study revealed that MEBS has remarkable anti-diarrheal potential and is highly effective in wide-spectrum bacterial and fungal strains. Moreover, the in silico study validated the results of biological screenings. To conclude, MEBS is presumed to be a good source in treating diarrhea, bacterial and fungal infections.


Assuntos
Antibacterianos/farmacologia , Antidiarreicos/farmacologia , Antifúngicos/farmacologia , Bauhinia/química , Diarreia , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia , Animais , Bactérias/efeitos dos fármacos , Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/microbiologia , Diarreia/induzido quimicamente , Diarreia/tratamento farmacológico , Diarreia/microbiologia , Modelos Animais de Doenças , Fungos/efeitos dos fármacos , Camundongos , Micoses/tratamento farmacológico , Micoses/microbiologia , Fitoterapia , Extratos Vegetais/química , Caules de Planta/química
19.
Nutrients ; 14(2)2022 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-35057448

RESUMO

Diabetes mellitus is a major predisposing factor for cardiovascular disease and mortality. α-Amylase and α-glucosidase enzymes are the rate-limiting steps for carbohydrate digestion. The inhibition of these two enzymes is clinically used for the treatment of diabetes mellitus. Here, in vitro study and machine learning models were employed for the chemical screening of inhibiting the activity of 31 plant samples on α-amylase and α-glucosidase enzymes. The results showed that the ethanolic twig extract of Pinus kesiya had the highest inhibitory activity against the α-amylase enzyme. The respective ethanolic extract of Croton oblongifolius stem, Parinari anamense twig, and Polyalthia evecta leaf showed high inhibitory activity against the α-glucosidase enzyme. The classification analysis revealed that the α-glucosidase inhibitory activity of Thai indigenous plants was more predictive based on phytochemical constituents, compared with the α-amylase inhibitory activity (1.00 versus 0.97 accuracy score). The correlation loading plot revealed that flavonoids and alkaloids contributed to the α-amylase inhibitory activity, while flavonoids, tannins, and reducing sugars contributed to the α-glucosidase inhibitory activity. In conclusion, the ethanolic extracts of P. kesiya, C. oblongifolius, P. anamense, and P. evecta have the potential for further chemical characterization and the development of anti-diabetic recipes.


Assuntos
Diabetes Mellitus Tipo 2/enzimologia , Hipoglicemiantes/farmacologia , Magnoliopsida/química , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/farmacologia , alfa-Amilases/metabolismo , alfa-Glucosidases/metabolismo , Diabetes Mellitus Tipo 2/tratamento farmacológico , Descoberta de Drogas/métodos , Inibidores Enzimáticos/farmacologia , Inibidores de Glicosídeo Hidrolases/farmacologia , Humanos , Aprendizado de Máquina , Fitoterapia , Folhas de Planta/química , Caules de Planta/química , Tailândia
20.
J Nat Prod ; 85(1): 70-82, 2022 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-35040315

RESUMO

The cumulative effects of cell damage result in aging, which gradually decreases human function in various aspects and leads to multiple age-related chronic diseases. To overcome the adverse effects of aging, silent mating type information regulation 2 homologue (SIRT1) activators are promising bioactive compounds that mimic calorie restriction to improve quality of life and prevent aging. In this study, 11 new flavonostilbenes (1-11) and three known compounds (12-14) were purified from stems of Rhamnoneuron balansae. The structures of the new compounds were determined using extensive data from spectroscopic methods, including NMR and HRESIMS. Their absolute configurations were deduced by ECD calculations with coupling constant analysis. All of the isolated new compounds (1-11) were evaluated for their effects on SIRT1 deacetylase activity, the NAD+/NADH ratio, and the AMP-activated protein kinase activation level in cell-based assays. The results showed that rhamnoneuronal D (1) exhibits promising biological activity in several in vitro models related to SIRT1 and suggest it is a potential natural-product-based antiaging agent.


Assuntos
Caules de Planta/química , Sirtuína 1/efeitos dos fármacos , Estilbenos/isolamento & purificação , Adenilato Quinase/metabolismo , Animais , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Ativação Enzimática , Humanos , NAD/metabolismo , Espectroscopia de Prótons por Ressonância Magnética , Espectrometria de Massas por Ionização por Electrospray , Estilbenos/farmacologia , Thymelaeaceae/química
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