RESUMO
A successful regeneration protocol was developed for micropropagation of Lilium akkusianum R. Gämperle, an endemic species of Türkiye, from scale explants. The study also aimed to evaluate the effects of Meta-Topolin (mT) and N6-Benzyladenine (BA) on in vitro regeneration. The Murashige and Skoog medium (MS) supplemented with different levels of α-naphthaleneacetic acid (NAA)/BA and NAA/mT were used for culture initiation in the darkness. The highest callus rates were observed on explants cultured on MS medium with 2.0 mg/L NAA + 0.5 mg/L mT (83.31%), and the highest adventitious bud number per explant was 4.98 in MS medium with 0.5 mg/L NAA + 1.5 mg/L mT. Adventitious buds were excised and cultured in 16/8 h photoperiod conditions. The highest average shoot number per explant was 4.0 in MS medium with 2.0 mg/L mT + 1.0 mg/L NAA. Shoots were rooted with the highest rate (90%) in the medium with the 1.0 mg/L IBA, and the highest survival rate (87.5%) was recorded in rooted shoots in the same medium. The ISSR marker system showed that regenerated plantlets were genetically stable. Besides traditional tissue culture techniques used in the current study, the potential for improving the effectiveness of L. akkusianum propagation protocols by incorporating machine learning methodologies was evaluated. ML techniques enhance lily micropropagation by analyzing complex biological processes, merging with traditional methods. This collaborative approach validates current protocols, allowing ongoing improvements. Embracing machine learning in endemic L. akkusianum studies contributes to sustainable plant propagation, promoting conservation and responsible genetic resource utilization in agriculture.
Assuntos
Lilium , Aprendizado de Máquina , Lilium/crescimento & desenvolvimento , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/efeitos dos fármacos , Regeneração/efeitos dos fármacos , Ácidos Naftalenoacéticos/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Meios de Cultura , Purinas/farmacologiaRESUMO
Novel magnetic covalent organic frameworks (COFs) were prepared by one-pot synthetic strategy and employed as an efficient adsorbent for magnetic solid-phase extraction (MSPE) of naphthaleneacetic acid (NAA) in food samples. Depending on the predesigned the hydrogen bonding, π-π and hydrophobic interactions of magnetic COFs, the efficient and selective extraction process for NAA was achieved within 15 min. The magnetic COFs adsorbent combined with HPLC-UV was devoted to develop a novel quantitative method for NAA in complex food. The method afforded good coefficient in range of 0.002-10.0 µg mL-1 and low limit of detection was 0.0006 µg mL-1. And the newly established method afforded less adsorbent consumption, wider linearity and lower LODs than the reported analytical methods. Ultimately, the method was successfully applied to determine NAA in fresh pear, tomato and peach juice. The magnetic COFs based MSPE coupled with HPLC-UV method provided a simple, efficient and dependable alternative to monitor trace NAA in food samples.
Assuntos
Limite de Detecção , Estruturas Metalorgânicas , Ácidos Naftalenoacéticos , Extração em Fase Sólida , Extração em Fase Sólida/métodos , Cromatografia Líquida de Alta Pressão/métodos , Ácidos Naftalenoacéticos/análise , Ácidos Naftalenoacéticos/química , Estruturas Metalorgânicas/química , Adsorção , Contaminação de Alimentos/análise , Solanum lycopersicum/química , Sucos de Frutas e Vegetais/análiseRESUMO
Callus cultures of the Iranian medicinal plant Salvia atropatana were initiated from three-week-old seedlings on Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA) and various cytokinins. Although all tested hormonal variants of the medium and explant enabled callus induction, the most promising growth was noted for N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU)-induced calli. Three lines obtained on this medium (cotyledon line-CL, hypocotyl line-HL, and root line-RL) were preselected for further studies. Phenolic compounds in the callus tissues were identified using UPLC-MS (ultra-performance liquid chromatography-mass spectrometry) and quantified with HPLC (high-performance liquid chromatography). All lines exhibited intensive growth and contained twelve phenolic acid derivatives, with rosmarinic acid predominating. The cotyledon-derived callus line displayed the highest growth index values and polyphenol content; this was exposed to different light-emitting diodes (LED) for improving biomass accumulation and secondary metabolite yield. Under LED treatments, all callus lines exhibited enhanced RA and total phenolic content compared to fluorescent light, with the highest levels observed for white (48.5-50.2 mg/g dry weight) and blue (51.4-53.9 mg/g dry weight) LEDs. The selected callus demonstrated strong antioxidant potential in vitro based on the 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) tests. Our findings confirm that the S. atropatana callus system is suitable for enhanced rosmarinic acid production; the selected optimized culture provide high-quality plant-derived products.
Assuntos
Polifenóis , Salvia , Polifenóis/metabolismo , Salvia/metabolismo , Salvia/química , Antioxidantes/metabolismo , Antioxidantes/química , Cromatografia Líquida de Alta Pressão , Cinamatos/metabolismo , Cinamatos/química , Ácido Rosmarínico , Depsídeos/metabolismo , Cotilédone/metabolismo , Cotilédone/química , Ácidos Naftalenoacéticos/farmacologia , Ácidos Naftalenoacéticos/química , Ácidos Naftalenoacéticos/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Reguladores de Crescimento de Plantas/metabolismo , Plântula/metabolismo , Plântula/crescimento & desenvolvimento , Plântula/efeitos dos fármacosRESUMO
Auxin is an important phytohormone that regulates diverse biologic processes, including plant growth and immunity. Indole-3-acetic acid (IAA), known as one of the main forms of auxin, is able to activate plant immunity. However, it is unknown whether IAA enhances plant resistance and/or suppresses the growth of the fungal pathogen Magnaporthe oryzae. Here, we found that IAA could induce expression levels of pathogenesis-related genes to enhance disease resistance and could control the development of blast disease through inhibiting M. oryzae infection. Exogenous IAA suppressed mycelial growth and delayed spore germination by inhibiting fungal endogenous IAA biosynthesis and impairing redox homeostasis, respectively. When applied to a field test, two IAA analogues, 1-naphthaleneacetic acid and 2,4-dichlorophenoxy acetic acid, can effectively control rice blast disease. Our study advances the understanding of IAA in controlling rice blast disease through suppressing pathogen growth and enhancing plant resistance.
Assuntos
Resistência à Doença , Ácidos Indolacéticos , Oryza , Doenças das Plantas , Ácidos Indolacéticos/metabolismo , Oryza/microbiologia , Oryza/crescimento & desenvolvimento , Oryza/imunologia , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Doenças das Plantas/prevenção & controle , Resistência à Doença/genética , Resistência à Doença/efeitos dos fármacos , Reguladores de Crescimento de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Ascomicetos/efeitos dos fármacos , Ascomicetos/fisiologia , Ácidos Naftalenoacéticos/farmacologia , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimentoRESUMO
In vitro cell culturing witnessed its applications in scientific research and industrial activities. Attempts to shorten the doubling time of cultured cells have never ceased. In plants, auxin is applied to promote plant growth, the synthetic derivative 1-Naphthaleneacetic acid (NAA) is a good example. Despite the auxin's naturally occurring receptors are not present in mammalian cells, studies suggested they may affect cell culturing. Yet the effects and mechanisms are still unclear. Here, an up to 2-fold increase in the yield of in vitro cultured human cells is observed. Different types of human cell lines and primary cells are tested and found that NAA is effective in all the cells tested. The PI staining followed by FACS suggested that NAA do not affect the cell cycling. Apoptosis-specific dye staining analysis implicated that NAA rescued cell death. Further bulk RNA sequencing is done and it is identified that the lipid metabolism-engaging and anti-apoptosis gene, ANGPTL4, is enhanced in expression upon NAA treatment. Studies on ANGPTL4 knockout cells indicated that ANGPTL4 is required for NAA-mediated response. Thus, the data identified a beneficial role of NAA in human cell culturing and highlighted its potency in in vitro cell culturing.
Assuntos
Ácidos Indolacéticos , Ácidos Naftalenoacéticos , Animais , Humanos , Ácidos Indolacéticos/metabolismo , Ácidos Indolacéticos/farmacologia , Células Cultivadas , Ácidos Naftalenoacéticos/farmacologia , Ácidos Naftalenoacéticos/metabolismo , Apoptose , Mamíferos/metabolismoRESUMO
It is widely accepted that prevéraison application of naphthaleneacetic acid (NAA) can delay the ripening of grapes and improve their quality. However, how NAA impacts grape aroma compound concentrations remains unclear. This study incorporated the analyses of aroma metabolome, phytohormones, and transcriptome of Vitis vinifera L. cv. Cabernet Sauvignon grapes cultivated in continental arid/semiarid regions of western China. The analyses demonstrated that NAA application increased ß-damascenone and 1,1,6-trimethyl-1,2-dihydronaphthalene (TDN) in the harvested grapes by delaying véraison and upregulating VvPSY1 and VvCCD4b expressions. Additionally, NAA treatment decreased 2-isobutyl-3-methoxypyrazine (IBMP) at the same phenological stage. Notably, abscisic acid (ABA) levels increased in NAA-treated grapes during véraison, which triggered further changes in norisoprenoid metabolisms. The ABA-responsive factor VvABF2 was potentially involved in VvPSY1 positive modulation, while the auxin response factor VvARF10 may play a role in VvCCD4b upregulation and VvOMT2 downregulation during NAA induction. VvARF10 possibly acts as a crosstalk node between the ABA and auxin signaling pathways following NAA treatment in regulating aroma biosynthesis.
Assuntos
Vitis , Vinho , Ácido Abscísico/metabolismo , Vitis/genética , Vitis/metabolismo , Ácidos Indolacéticos/metabolismo , Odorantes/análise , Transcriptoma , Frutas/química , Metaboloma , Ácidos Naftalenoacéticos/análise , Vinho/análiseRESUMO
A major obstacle in the treatment of tuberculosis (TB) is to combat the emerging resistant strains of its causing agent i.e. Mycobacterium tuberculosis (MTb). The emergence of multidrug-resistant and extensively drug-resistant -TB strains raise a requirement of new potential anti-tubercular compounds. In this direction, different plant parts of Morus alba were tested against MTb and found to be active with a minimum inhibitory concentration ranging between 125 µg/ml to 31.5 µg/ml. Further to identify the phytochompounds having anti-mycobacterium activity, phytocompounds of the plant were docked against the five MTb proteins (PDB ID: 3HEM, 4OTK, 2QO0, 2AQ1 and 6MNA). Among twenty-two tested phytocompounds, four phytocompounds with effective binding energy (kcal/mol): Petunidin-3-rutinoside (3HEM: -8.2, 4OTK: -6.9, 2QO0: -9.0, 2AQ1: -8.3 and 6MNA:-7.8), Quercetin-3'-glucoside (3HEM:-6.7, 4OTK:-7.6, 2QO0:-7.6, 2AQ1:7.6 and 6MNA:-6.4), Rutin (3HEM:-7.8, 4OTK:-7.5, 2QO0:-9.1, 2AQ1:9.3 and 6MNA:-6.9) and Isoquercitrin (3HEM:-7.3, 4OTK:-6.6, 2QO0:-7.7, 2AQ1:8.3 and 6MNA:-6.6) shows promising activity against all the five target proteins. Further molecular dynamics studies of Petunidin-3-rutinoside with three target proteins 3HEM, 2AQ1 and 2QO0 resulted with low values of average RMSD (3.723 Å, 3.261 Å, and 2.497 Å, respectively) show that the complexes have better conformational stability. The wet lab validation of the current study will pave the new dimensions for the cure of TB patients.Communicated by Ramaswamy H. Sarma.
Assuntos
Morus , Mycobacterium tuberculosis , Ácidos Naftalenoacéticos , Tuberculose , Humanos , Simulação de Dinâmica Molecular , Antituberculosos/química , Tuberculose/microbiologia , Simulação de Acoplamento MolecularRESUMO
A symbiotic relationship is observed between Armillaria and the Chinese herbal medicine Gastrodia elata (G. elata). Armillaria is a nutrient source for the growth of G. elata, and its nutrient metabolism efficiency affects the growth and development of G. elata. Auxin has been reported to stimulate Armillaria species, but the molecular mechanism remains unknown. We found that naphthalene acetic acid (NAA) can also promote the growth of A. gallica. Moreover, we identified a total of 2071 differentially expressed genes (DEGs) by analyzing the transcriptome sequencing data of A. gallica at 5 and 10 hour of NAA treatment. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that these unigenes were significantly enriched in the metabolism pathways of arginine, proline, propanoate, phenylalanine and tryptophan. The expression levels of the general amino acid permease (Gap), ammonium transporter (AMT), glutamate dehydrogenase (GDH), glutamine synthetase (GS), Zn(II) 2Cys6 and C2H2 transcription factor genes were upregulated. Our transcriptome analysis showed that the amino acid and nitrogen metabolism pathways in Armillaria were rapidly induced within hours after NAA treatment. These results provide valuable insights into the molecular mechanisms by which NAA promotes the growth of Armillaria species.
Assuntos
Armillaria , Gastrodia , Armillaria/genética , Ácidos Naftalenoacéticos , Perfilação da Expressão Gênica , Ontologia GenéticaRESUMO
Paradormancy of fruit trees occurs in summer and autumn when signals from adjacent organs stimulate buds to develop slowly. This stage has received less attention that the other stages of dormancy, and the underlying mechanism remains uncharacterized. Early defoliation in late summer and early autumn is usually followed by out-of-season blooming in pear (Pyrus spp.), which substantially decreases the number of buds the following spring and negatively affects fruit production. This early bud flush is an example of paradormancy release. Here, we determined that flower bud auxin content is stable after defoliation; however, polar distribution of the pear (Pyrus pyrifolia) PIN-FORMED auxin efflux carrier 1b (PpyPIN1b) implied that auxin tends to be exported from buds. Transcriptome analysis of floral buds after artificial defoliation revealed changes in auxin metabolism, transport, and signal transduction pathways. Exogenous application of a high concentration of the auxin analog 1-naphthaleneacetic acid (300 mg/L) suppressed PpyPIN1b expression and its protein accumulation in the cell membrane, likely leading to decreased auxin efflux from buds, which hindered flower bud sprouting. Furthermore, carbohydrates and additional hormones also influenced out-of-season flowering. Our results indicate that defoliation-induced auxin efflux from buds accelerates bud paradormancy release. This differs from release of apical-dominance-related lateral bud paradormancy after the apex is removed. Our findings and proposed model further elucidate the mechanism underlying paradormancy and will help researchers to develop methods for inhibiting early defoliation-induced out-of-season bud sprouting.
Assuntos
Pyrus , Pyrus/genética , Ácidos Indolacéticos , Ácidos Naftalenoacéticos/farmacologia , Frutas/genética , Transporte BiológicoRESUMO
Coupling microalgae cultivation with wastewater treatment is a promising environmentally sustainable development strategy. However, toxics such as Bisphenol A (BPA) in wastewater damage microalgae cells and reduces bioresources production. Phytohormone regulation has the potential to solve this issue. However, phytohormone research is still in its infancy. In this work, 0.2 µM naphthyl acetic acid (NAA) significantly enhanced Chlorella vulgaris BPA detoxification by 127.3% and Chlorella biomass production by 46.4%. NAA helps Chlorella convert bisphenol A into small non-toxic intermediates by enhancing the expression of associated enzymes. Simultaneously, NAA promoted carbon fixation and photosynthetic metabolism. Activation of the mitogen-activated protein kinase (MAPK) pathway strengthened the downstream antioxidant system while improving photosynthesis and intracellular starch and lipid synthesis. Carbohydrates, pigment, and lipid production was significantly enhanced by 20.0%, 46.9%, and 21.8%, respectively. A new insight is provided into how phytohormones may increase microalgae in wastewater's bioresource transformation and toxicity resistance.
Assuntos
Chlorella vulgaris , Microalgas , Purificação da Água , Antioxidantes/metabolismo , Compostos Benzidrílicos , Biomassa , Carboidratos , Chlorella vulgaris/metabolismo , Lipídeos , Microalgas/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Ácidos Naftalenoacéticos/metabolismo , Fenóis , Reguladores de Crescimento de Plantas/metabolismo , Amido/metabolismo , Águas ResiduáriasRESUMO
Dragon's head plant (Lallemantia iberica), is a flowering species belongs to the mint family (Lamiaceae). The species contains valuable essential oils, mucilage and oil which are used in pharmaceutical and food industries. Tissue culture is a feasible strategy to attain large-scale production of plantlets with a huge potential to produce plants with superior quality. The objective of this study was to develop a simple and efficient method for regeneration and transformation of L. iberica. To reach this goal, the regeneration ability of various explants including leaf, cotyledonary node, hypocotyl and cotyledon segments was investigated in MS medium supplemented with diverse concentrations of NAA (Naphthalene acetic acid) and BAP (6-Benzyl Amino Purine). According to the results, cotyledonary nodes showed the best regeneration response. The maximum rate of regeneration (and number of induced shoots was achieved in 1 mg l-1 BAP in combination with 0.05 mg l-1 NAA from the cotyledonary nodes. Additionally, through the optimized regeneration technique Agrobacterium-mediated transformation of L. iberica was successfully accomplished. Gene transfer was assessed on leaf samples from regenerated plantlets under a fluorescent microscope to detect the GFP signals. Moreover, transgene integration and its expression were confirmed by PCR and RT-PCR analysis, respectively. The establishment of these efficient regeneration and genetic transformation methods paved the way for further application such as plant improvement, functional analysis and gene editing.
Assuntos
Agrobacterium/genética , Cotilédone/citologia , Lamiaceae/crescimento & desenvolvimento , Brotos de Planta/citologia , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Regeneração , Transformação Genética , Hipocótilo/citologia , Lamiaceae/genética , Ácidos Naftalenoacéticos/farmacologiaRESUMO
Thunbergia coccinea Wall. ex D. Don being a rare, ornamental and medicinal plant of India, is needed to propagate for conserving the germplasm and analyzing its phytochemical compounds in the future. A reliable protocol for direct in vitro propagation using nodal shoot meristem of T. coccinea as explant was standardized. The highest number of shoots per explant (22.17 ± 0.54) with maximum shoot length (2.36 ± 0.28) in cm was obtained in Murashige and Skoog (MS) medium supplemented with 9.70 µM of 6-furfurylaminopurine (Kinetin) and 0.053 µM of α-naphthaleneacetic acid (NAA) combination, among all the different plant growth regulators (PGR's) and concentrations tested. The aforesaid PGR's combination was optimum for axillary shoot bud induction and multiplication in T. coccinea. The best rooting was observed on the half-strength MS medium fortified with 2.68 µM NAA with the highest number of roots per shoot (3.75 ± 0.12) and maximum length (5.22 ± 0.32) in cm. All the in vitro raised plantlets were acclimatized in sterile sand and soil mixture (1:1) with a survival rate of 70% on earthen pots under greenhouse conditions. PCR-based RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeat) molecular markers were employed to determine the genetic homogeneity amongst the plantlets. Twelve (12) RAPD and nine (9) ISSR primers developed a total of 104 and 91 scorable bands, respectively. The band profiles of micropropagated plantlets were monomorphic to the mother, donor in vivo plant, and similarity values varied from 0.9542-1.000. The dendrogram generated through UPGMA (unweighted pair group method with arithmetic mean) showed 99% similarities amongst all tested plants confirming the genetic uniformity of in vitro raised plants.
Assuntos
Acanthaceae/genética , DNA de Plantas/genética , Genes de Plantas , Genoma de Planta , Meristema/genética , Repetições de Microssatélites , Técnica de Amplificação ao Acaso de DNA Polimórfico , Acanthaceae/efeitos dos fármacos , Acanthaceae/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Marcadores Genéticos , Instabilidade Genômica , Genótipo , Cinetina/farmacologia , Meristema/efeitos dos fármacos , Meristema/crescimento & desenvolvimento , Ácidos Naftalenoacéticos/farmacologia , Reguladores de Crescimento de Plantas/farmacologiaRESUMO
Auxin plays a dual role in growth regulation and, depending on the tissue and concentration of the hormone, it can either promote or inhibit division and expansion processes in plants. Recent studies have revealed that, beyond transcriptional reprogramming, alternative auxin-controlled mechanisms regulate root growth. Here, we explored the impact of different concentrations of the synthetic auxin NAA that establish growth-promoting and -repressing conditions on the root tip proteome and phosphoproteome, generating a unique resource. From the phosphoproteome data, we pinpointed (novel) growth regulators, such as the RALF34-THE1 module. Our results, together with previously published studies, suggest that auxin, H+-ATPases, cell wall modifications and cell wall sensing receptor-like kinases are tightly embedded in a pathway regulating cell elongation. Furthermore, our study assigned a novel role to MKK2 as a regulator of primary root growth and a (potential) regulator of auxin biosynthesis and signalling, and suggests the importance of the MKK2 Thr31 phosphorylation site for growth regulation in the Arabidopsis root tip.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Hormônios Peptídicos/genética , Fosfoproteínas/genética , Reguladores de Crescimento de Plantas/farmacologia , Raízes de Plantas/genética , Proteínas Quinases/genética , Receptores de Superfície Celular/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Ácidos Indolacéticos/farmacologia , Meristema/genética , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Ácidos Naftalenoacéticos/síntese química , Ácidos Naftalenoacéticos/farmacologia , Hormônios Peptídicos/metabolismo , Fosfoproteínas/classificação , Fosfoproteínas/metabolismo , Fosforilação , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Proteínas Quinases/metabolismo , Processamento de Proteína Pós-Traducional , Proteoma/classificação , Proteoma/genética , Proteoma/metabolismo , ATPases Translocadoras de Prótons/genética , ATPases Translocadoras de Prótons/metabolismo , Receptores de Superfície Celular/metabolismo , Transdução de SinaisRESUMO
In vitro cultures of scarlet flax (Linum grandiflorum L.), an important ornamental flax, have been established as a new possible valuable resource of lignans and neolignans for antioxidant and anti-inflammatory applications. The callogenic potential at different concentrations of α-naphthalene acetic acid (NAA) and thidiazuron (TDZ), alone or in combinations, was evaluated using both L. grandiflorum hypocotyl and cotyledon explants. A higher callus induction frequency was observed on NAA than TDZ, especially for hypocotyl explants, with a maximum frequency (i.e., 95.2%) on 1.0 mg/L of NAA. The presence of NAA (1.0 mg/L) in conjunction with TDZ tended to increase the frequency of callogenesis relative to TDZ alone, but never reached the values observed with NAA alone, thereby indicating the lack of synergy between these two plant growth regulators (PGRs). Similarly, in terms of biomass, NAA was more effective than TDZ, with a maximum accumulation of biomass registered for medium supplemented with 1.0 mg/L of NAA using hypocotyls as initial explants (DW: 13.1 g). However, for biomass, a synergy between the two PGRs was observed, particularly for cotyledon-derived explants and for the lowest concentrations of TDZ. The influence of these two PGRs on callogenesis and biomass is discussed. The HPLC analysis confirmed the presence of lignans (secoisolariciresinol (SECO) and lariciresinol (LARI) and neolignan (dehydrodiconiferyl alcohol [DCA]) naturally accumulated in their glycoside forms. Furthermore, the antioxidant activities performed for both hypocotyl- and cotyledon-derived cultures were also found maximal (DPPH: 89.5%, FRAP 866: µM TEAC, ABTS: 456 µM TEAC) in hypocotyl-derived callus cultures as compared with callus obtained from cotyledon explants. Moreover, the anti-inflammatory activities revealed high inhibition (COX-1: 47.4% and COX-2: 51.1%) for extract of hypocotyl-derived callus cultures at 2.5 mg/L TDZ. The anti-inflammatory action against COX-1 and COX-2 was supported by the IC50 values. This report provides a viable approach for enhanced biomass accumulation and efficient production of (neo)lignans in L. grandiflorum callus cultures.
Assuntos
Anti-Inflamatórios/análise , Antioxidantes/análise , Butileno Glicóis/análise , Cotilédone/química , Linho/química , Furanos/análise , Hipocótilo/química , Lignanas/análise , Extratos Vegetais/análise , Biomassa , Cromatografia Líquida de Alta Pressão/métodos , Cotilédone/metabolismo , Meios de Cultura/química , Técnicas de Cultura/métodos , Linho/metabolismo , Hipocótilo/metabolismo , Ácidos Naftalenoacéticos/farmacologia , Fenóis/análise , Compostos de Fenilureia/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Tiadiazóis/farmacologiaRESUMO
Nitrate (NO3-) plays a pivotal role in stimulating lateral root (LR) formation and growth in plants. However, the role of NO3- in modulating rice LR formation and the signalling pathways involved in this process remain unclear. Phenotypic and genetic analyses of rice were used to explore the role of strigolactones (SLs) and auxin in NO3--modulated LR formation in rice. Compared with ammonium (NH4+), NO3- stimulated LR initiation due to higher short-term root IAA levels. However, this stimulation vanished after 7 d, and the LR density was reduced, in parallel with the auxin levels. Application of the exogenous auxin α-naphthylacetic acid to NH4+-treated rice plants promoted LR initiation to levels similar to those under NO3- at 7 d; conversely, the application of the SL analogue GR24 to NH4+-treated rice inhibited LR initiation to levels similar to those under NO3- supply by reducing the root auxin levels at 10 d. D10 and D14 mutations caused loss of sensitivity of the LR formation response to NO3-. The application of NO3- and GR24 downregulated the transcription of PIN-FORMED 2(PIN2), an auxin efflux carrier in roots. LR number and density in pin2 mutant lines were insensitive to NO3- treatment. These results indicate that NO3- modulates LR formation by affecting the auxin response and transport in rice, with the involvement of SLs.
Assuntos
Ácidos Indolacéticos/metabolismo , Nitratos/farmacologia , Oryza/metabolismo , Compostos de Amônio/farmacologia , Mutação , Ácidos Naftalenoacéticos/farmacologia , Nitratos/metabolismo , Oryza/efeitos dos fármacos , Oryza/genética , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismoRESUMO
The phytohormone auxin and its directional transport through tissues are intensively studied. However, a mechanistic understanding of auxin-mediated feedback on endocytosis and polar distribution of PIN auxin transporters remains limited due to contradictory observations and interpretations. Here, we used state-of-the-art methods to reexamine the auxin effects on PIN endocytic trafficking. We used high auxin concentrations or longer treatments versus lower concentrations and shorter treatments of natural indole-3-acetic acid (IAA) and synthetic naphthalene acetic acid (NAA) auxins to distinguish between specific and nonspecific effects. Longer treatments of both auxins interfere with Brefeldin A-mediated intracellular PIN2 accumulation and also with general aggregation of endomembrane compartments. NAA treatment decreased the internalization of the endocytic tracer dye, FM4-64; however, NAA treatment also affected the number, distribution, and compartment identity of the early endosome/trans-Golgi network, rendering the FM4-64 endocytic assays at high NAA concentrations unreliable. To circumvent these nonspecific effects of NAA and IAA affecting the endomembrane system, we opted for alternative approaches visualizing the endocytic events directly at the plasma membrane (PM). Using total internal reflection fluorescence microscopy, we saw no significant effects of IAA or NAA treatments on the incidence and dynamics of clathrin foci, implying that these treatments do not affect the overall endocytosis rate. However, both NAA and IAA at low concentrations rapidly and specifically promoted endocytosis of photo-converted PIN2 from the PM. These analyses identify a specific effect of NAA and IAA on PIN2 endocytosis, thus, contributing to its polarity maintenance and furthermore illustrate that high auxin levels have nonspecific effects on trafficking and endomembrane compartments.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Endocitose/efeitos dos fármacos , Ácidos Indolacéticos/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Arabidopsis/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Ácidos Naftalenoacéticos/farmacologia , Transporte Proteico , Rede trans-Golgi/efeitos dos fármacosRESUMO
The abscisic acid (ABA) increase and auxin decline are both indicators of ripening initiation in grape berry, and norisoprenoid accumulation also starts at around the onset of ripening. However, the relationship between ABA, auxin, and norisoprenoids remains largely unknown, especially at the transcriptome level. To investigate the transcriptional and posttranscriptional regulation of the ABA and synthetic auxin 1-naphthaleneacetic acid (NAA) on norisoprenoid production, we performed time-series GC-MS and RNA-seq analyses on Vitis vinifera L. cv. Cabernet Sauvignon grape berries from pre-veraison to ripening. Higher levels of free norisoprenoids were found in ABA-treated mature berries in two consecutive seasons, and both free and total norisoprenoids were significantly increased by NAA in one season. The expression pattern of known norisoprenoid-associated genes in all samples and the up-regulation of specific alternative splicing isoforms of VviDXS and VviCRTISO in NAA-treated berries were predicted to contribute to the norisoprenoid accumulation in ABA and NAA-treated berries. Combined weighted gene co-expression network analysis (WGCNA) and DNA affinity purification sequencing (DAP-seq) analysis suggested that VviGATA26, and the previously identified switch genes of myb RADIALIS (VIT_207s0005g02730) and MAD-box (VIT_213s0158g00100) could be potential regulators of norisoprenoid accumulation. The positive effects of ABA on free norisoprenoids and NAA on total norisoprenoid accumulation were revealed in the commercially ripening berries. Since the endogenous ABA and auxin are sensitive to environmental factors, this finding provides new insights to develop viticultural practices for managing norisoprenoids in vineyards in response to changing climates.
Assuntos
Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Norisoprenoides/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Transcriptoma/efeitos dos fármacos , Vitis/genética , Ácido Abscísico/metabolismo , Processamento Alternativo , Frutas/genética , Frutas/crescimento & desenvolvimento , Frutas/metabolismo , Perfilação da Expressão Gênica , Ácidos Indolacéticos/metabolismo , Metabolômica , Ácidos Naftalenoacéticos/metabolismo , Vitis/crescimento & desenvolvimento , Vitis/metabolismoRESUMO
Nabumetone (NAB) is a non-steroidal anti-inflammatory drug used clinically, and its biotransformation includes the major active metabolite 6-methoxy-2-naphthylacetic acid (6-MNA). One of the key intermediates between NAB and 6-MNA may be 3-hydroxy nabumetone (3-OH-NAB). The aim of the present study was to investigate the role of flavin-containing monooxygenase (FMO) isoform 5 in the formation of 6-MNA from 3-OH-NAB. To elucidate the biotransformation of 3-OH-NAB to 6-MNA, an authentic standard of 3-OH-NAB was synthesised and used as a substrate in an incubation with human liver samples or recombinant enzymes. The formation of 3-OH-NAB was observed after the incubation of NAB with various cytochrome P450 (CYP) isoforms. However, 6-MNA itself was rarely detected from NAB and 3-OH-NAB. Further experiments revealed a 6-MNA peak derived from 3-OH-NAB in human hepatocytes. 6-MNA was also detected in the extract obtained from 3-OH-NAB by a combined incubation of recombinant human FMO5 and human liver S9. We herein demonstrated that the reaction involves carbon-carbon cleavage catalyzed by the Baeyer-Villiger oxidation (BVO) of a carbonyl compound, the BVO substrate, such as a ketol, by FMO5. Further in vitro inhibition experiments showed that multiple non-CYP enzymes are involved in the formation of 6-MNA from 3-OH-NAB.
Assuntos
Anti-Inflamatórios não Esteroides/metabolismo , Nabumetona/metabolismo , Ácidos Naftalenoacéticos/metabolismo , Biotransformação , Sistema Enzimático do Citocromo P-450/metabolismo , Humanos , Microssomos Hepáticos/metabolismo , Oxirredução , OxigenasesRESUMO
In vitro multiplication is an important tissue culture technique that is capable of efficiently producing seedlings at any scale. It is a propagation method based on the aseptic culture of small propagules in a suitable culture medium to enable plant regeneration. Multiplication experiments conducted in vitro to set protocols adapted to wild Manihot species have used modified mineral salts and MS vitamins as basic culture medium. Here, 25 treatments based on combinations of the regulators benzylaminopurine (BAP) and naphthaleneacetic acid (NAA) at 0, 0.025, 0.05, 0.075, and 0.1 mg L-1 were used for in vitro multiplication of three genotypes of wild Manihot species (M. violaceae Pohl Müll. Arg., M. pseudoglaziovii Pax & Hoff., and M. flabellifolia Pohl). Plant height and the number of 1 cm minicuttings, number of roots, shoots, green leaves and senescent leaves were recorded 120 days after explant inoculation. M. violaceae Pohl. Müll. Arg. and M. flabellifolia Pohl. presented favorable results with 0.05 and 0.025 mg L-1 NAA, respectively. Culture medium lacking NAA and BAP favored the in vitro growth of M. pseudoglaziovii Pax & Hoff.
Assuntos
Manihot/crescimento & desenvolvimento , Manihot/química , Técnicas In Vitro , Ácidos Naftalenoacéticos/análiseRESUMO
BACKGROUND: Moringa stenopetala belongs to the flowering family Moringaceae and genus Moringa. It is often referred to as the East African Moringa tree because it is native only to southern Ethiopia and northern Kenya. The expansion of its cultivation and utilization throughout the world especially in Africa is becoming important. For such expansion, the existing propagation method is limiting, so it needs a good propagation system to supply enough planting material with a uniform genotype. Therefore, the main objective of this study was to optimize an in vitro shoot multiplication protocol for M. stenopetala by using shoot tip as explants. RESULTS: Shoots were sterilized and cultured on Muraghige and Skoog (MS) medium for in vitro shoot initiation. For multiple shoot induction, the explants were cultured on MS medium supplemented with different concentrations of kinetin (0.5, 1.0, 1.5, 2.0, 2.5 mg/L) with Indole-3- butyric acid (IBA) or α -naphthalene acetic acid (NAA) (0.01, 0.1, 0.5 mg/L) and maintained at 25 ± 2 °C for four weeks. Rooting was achieved by culturing well developed shoots in half-strength MS medium containing IBA (0.1, 0.5, 1.0, 1.5, 2.0 mg/L), NAA (0.1, 0.5, 1.0, 1.5, 2.0 mg/L), and 0.5 mg/L IBA with NAA (0.1, 0.5, 1.0, 1.5, 2.0 mg/L). Statistical analysis revealed that there was a significant difference among all treatments applied in both shoot multiplication and rooting experiments. The maximum number of shoots per explant (3.43 ± 1.41) and 7.97 ± 4.18 leaves per explant were obtained on MS medium containing 0.5 mg/L kinetin with 0.01 mg/LNAA. The highest mean number of roots per shoot (1.63 ± 1.03) and mean root length (0.87 ± 1.22 cm) were obtained on MS medium containing 1.0 mg/LNAA and 0.1 mg/LIBA alone respectively. After acclimatization, 76% of plants were survived in the greenhouse. CONCLUSION: In general, using NAA with kinetin for shoot multiplication was effective than kinetin with IBA. On the other hand, the application of 1.0 mg/L NAA alone and 1.0 mg/L NAA with 0.5 mg/L IBA were more effective for root induction.