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RESUMEN La utilización de elicitores es frecuente en la inducción de diferentes respuestas relacionadas con la resistencia sistémica de las plantas; no obstante, no se ha caracterizado el gasto metabólico que generan en el cultivo de papa variedad Diacol Capiro, que es la más utilizada para el procesamiento industrial en Colombia, debido a su calidad y alta producción. En esta investigación, se evaluó el efecto de cuatro elicitores en el crecimiento y desarrollo del cultivo de papa, en la vereda El Toval, del municipio de Tutazá, Boyacá. El diseño experimental fue en parcelas divididas, en donde la parcela principal fue la etapa fenológica, en la que se hizo la aplicación (E1; formación de tallos secundarios), época dos (E2; floración) y época tres (E3; aplicación tanto en formación de tallos secundarios como en floración) y la subparcela, con los elicitores aplicados (glucosamina, ácido γ-amino butírico, fosfito de potasio y ácido salicílico) y el testigo sin aplicación. Los datos fueron tomados a los 60, 80 y 100 días después de la siembra, teniendo como variables: altura de la planta, número de foliolos, clorofilas totales, eficiencia fotosintética, biomasa seca y producción. La glucosamina fue el tratamiento que obtuvo menores resultados en la mayoría de variables evaluadas con respecto al testigo; asimismo, los elicitores tienen efectos diferentes, dependiendo del estado fenológico del cultivo, especialmente, en la época de formación de tallos secundarios. El ácido salicílico (AS) presentó un comportamiento estadísticamente similar al del testigo, mientras que los demás elicitores aplicados, disminuyeron la producción de papa.
ABSTRACT Elicitors are frequently employed for the induction of different responses related to the systemic resistance of plants, however, it has not been fully characterized in the metabolic expenditure generated in the cultivation of the potato variety Diacol Capiro, the most planted for industrial processing in Colombia due to its quality and production. In this research, the effect of four elicitors on the growth and development of the potato crop was evaluated, in the municipality of Tutazá, Boyacá. The experimental design was divided plots, being the main plot the phenological stage in which the application was made (E1; formation of secondary stems), season two (E2; flowering) and season three (E3; application both in formation of secondary stems as in flowering) and the subplot, the applied elicitors (glucosamine, γ-amino butyric acid, potassium phosphite and salicylic acid) and the control without application. Data were taken 60, 80 and 100 days after sowing, having as variables: plant height, number of leaflets, total chlorophylls, photosynthetic efficiency, dry biomass and production. With the use of glucosamine the lowest results were obtained in most of the variables evaluated with respect to the control, likewise, elicitors had different effects depending on the phenological state of the crop, especially at the moment of secondary stem formation. Salicylic acid (AS) presented a statistically similar behavior to that of the control while the other applied elicitors decreased potato production.
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Abstract Thevetia peruviana is a medicinal plant that has valuable secondary terpenoid-type metabolites and phenolic compounds. Some flavonoid compounds of pharmaceutical interest stand out in the latter group. The concentration of these bioactive compounds in natural conditions is limited by environmental; therefore, it has been considered necessary to make in vitro plant cell suspension cultures that admit the use of elicitors to increase the content of active principles. Accordingly, in this study, for the optimization of flavonoid production in cell suspension culture of T. peruviana, different parameters related to elicitation with methyl jasmonate (MeJa), and salicylic acid (SA) were evaluated, at stirred flask scale. Firstly, 3 μM MeJa and 300 μM SA were added separately in cell cultures of T. peruviana, to assess their potential effects. Secondly, several experimental conditions were evaluated, for optimization purpose. In the first part, MeJa and SA increased the total flavonoid content, in 1.07 and 1.3 times, respectively, compared to the control culture; in the second part, total flavonoid content produced in MeJa mediated cell suspension cultures were 4.14 mg QE/g DW (milligrams of quercetin equivalent per gram of dry biomass) with: concentration 0.3 μM, addition time day 5 and harvest time 90 h. On the other hand, total flavonoid content produced in SA mediated cell suspension cultures were 3.75 mg QE/g DW with: concentration 100 μM, addition time day 0 and harvest time 96 h. Elicitation of cell suspension cultures of T. peruviana with MeJa and SA under their ideal parameter values increased flavonoid content.
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Safflower(Carthamus tinctorius), a valuable traditional Chinese medicinal plant, has attracted much attention in recent years. This study established a stable tissue culture system of safflower and analyzed the chromatogram of its secondary metabolites, providing high-quality experimental materials for further research on natural products in safflower. The calluses were established from the safflower seeds germinated in a sterile environment, and then they were differentiated into the aseptic seedlings, or cultured to obtain suspension cells in liquid medium. The ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS), Progenesis QI, and principal component analysis(PCA) were used to detect and analyze the secondary metabolites in the suspension cells before and after induction with different elicitors(methyl jasmonate, silver nitrate, salicylic acid and yeast extract). A total of 23 secondary metabolites including flavonoids, phenylpropanoids, alkaloids, fatty acids and aromatic glycosides were detected in safflower suspension cells. In response to the four elicitors, 11 compounds showed increased or decreased relative content. The results indicate that different elicitors have various effects on the accumulation of secondary metabolites in safflower suspension cells, and yeast extract shows more obvious positive induction. Therefore, different elicitors may play a role in the expression of related genes in the biosynthetic pathway of specific secondary metabolites. The results facilitate the discovery of targeted elicitors and the large-scale production of valuable secondary metabolites in the future.
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Carthamus tinctorius , Chromatography, High Pressure Liquid , Chromatography, Liquid , Flavonoids , Glycosides , Mass SpectrometryABSTRACT
Objective: Fusarium oxysporum is a common pathogenic fungus in ginseng cultivation. Both pathogens and antagonistic fungi have been reported to induce plant resistance responses, thereby promoting the accumulation of secondary metabolites. The purpose of this experiment is to compare the advantages of one of the two fungi, in order to screen out more effective elicitors. The mechanism of fungal elicitor-induced plant resistance response is supplemented. Methods: A gradient dilution and the dural culture were carried out to screen strains. The test strain was identified by morphology and 18 s rDNA. The effect of different concentrations (0, 50, 100, 200, 400 mg/L) of Penicillium sp. YJM-2013 and F. oxysporum on fresh weight and ginsenosides accumulation were tested. Signal molecules transduction, expression of transcription factors and functional genes were investigated to study the induction mechanism of fungal elicitors. Results: Antagonistic fungi of F. oxysporum was identified as Penicillium sp. YJM-2013, which reduced root biomass. The total ginsenosides content of Panax ginseng adventitious roots reached the maximum (48.95 ± 0.97 mg/g) treated with Penicillium sp. YJM-2013 at 200 mg/L, higher than control by 2.59-fold, in which protopanoxadiol-type ginsenosides (PPD) were increased by 4.57 times. Moreover, Penicillium sp. YJM-2013 activated defense signaling molecules, up-regulated the expression of PgWRKY 1, 2, 3, 5, 7, 9 and functional genes in ginsenosides synthesis. Conclusion: Compared with the pathogenic fungi F. oxysporum, antagonistic fungi Penicillium sp. YJM-2013 was more conducive to the accumulation of ginsenosides in P. ginseng adventitious roots. Penicillium sp. YJM-2013 promoted the accumulation of ginsenosides by intensifying the generation of signal molecules, activating the expression of transcription factors and functional genes.
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Aims@#A rare marine-derived actinomycete, Plantactinospora sp. KBS50, has been identified as a potential source of bioactive secondary metabolites compounds. The present study aimed to evaluate the secondary metabolites biosynthetic capability of strain KBS50 using the One Strain Many Compound (OSMAC) fermentation strategy. @*Methodology and results@#Strain KBS50 was fermented in a basal medium (ISP2) supplemented with selected biological and chemical elicitors, as well as cultivation at different pH value and incubation temperature. Statistical analysis revealed that the antimicrobial activities were significantly increased, as compared to the basal medium, ISP2. Similarly, the comparative High Performance Liquid Chromatography (HPLC) analysis showed an increase in secondary metabolites production, as well as the detection of potential new metabolites, particularly from the crude extracts of ISP2 medium supplemented with 1% (w/v) sodium chloride and with the culture filtrate of Aspergillus niger. The bioassay-guided fractionation showed that the extract of strain KBS50 contains multiple compounds with antibacterial activity against the Gram-positive strains. Further fractionation led to the isolation of two semi-pure compounds (compound 3 and 4) with bactericidal properties against Staphylococcus aureus. The Minimum Inhibitory Concentration (MIC) values of compound 3 and 4 were recorded at 7.81 μg/mL and 62.50 μg/mL, respectively. The minimum bactericidal concentration (MBC) for compound 3 was recorded at 15.63 μg/mL while the MBC for compound 4 was recorded as 125.00 μg/mL. @*Conclusion, significance and impact of study@#The OSMAC fermentation strategy used in this study had successfully enhanced the detection of antibiotics and secondary metabolites from Plantactinospora sp. KBS50. The bioassay-guided fractionation further established the capability of strain KBS50 as a source of bioactive secondary metabolite compounds with potent antimicrobial activity.
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Aims: To evaluate the effect of salicylic acid (SA) and acibenzolar-S-methyl (ASM) on the minimal concentration of juglone (Cmin) inducing foliar necrosis- and to determine the total phenol content and antioxidant activity of extracts of banana leaves after SA and ASM applications and toxin injection. Methodology: Banana cultivars Orishele and Corne 1 were subjected to root and foliar applications of elicitors, then leaves were injected with juglone a toxin of M. fijiensis to the determination of Cmin. The determination of total phenols extracted from leaves was carried out using Folin Reagent. The antioxidant activity of phenolic crude extracts (PCE) was determined through the DPPH radical scavenging ability. Results: In banana without elicitor applications, the minimum concentration inducing necrosis varied between 12.5 and 25 ppm of juglone but reached 250 ppm of juglone into banana treated with elicitors, particularly with ASM at 50 ppm. The phenol contents were highest 14 days after elicitors application. After this incubation time, with ASM at 50 ppm and AS at 25 ppm, the levels were 16.25 mg GAE/g DW in Orishele and 17.20 mg GAE/g DW in Corne 1, respectively. But the levels were lower 28 days after banana elicitation with 50 ppm of SA into Orishele and Corne 1 respectively 5.60 and 6.79 mg GAE/g DW. Treatments with 50 ppm of ASM showed the highest antioxidant activity between 7 and 28 days after elicitors application on banana leaves. With this treatment, the lowest concentration of phenolic crude extract scavenging 50% of DPPH was 13.09 μg/mL at 21 days after foliar elicitation. Conclusion: The applications of elicitors SA and ASM affect phenol content and antioxidant activity for the detoxification of foliar tissues of banana cultivars Orishele and Corne 1 infiltrated with toxin.
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The aim of the present study was to investigate the influence of tyrosine on the in vitro growth and the production of the betacyanin pigment in Alternanthera philoxeroides and A. tenella. Nodal segments were inoculated in MS medium containing different concentrations of tyrosine (0, 25, 50 and 75 μM), and the number of sprouts and buds, height, root length, fresh matter of shoots and roots and betacyanin content were evaluated. In A. philoxeroides , the highest production of betacyanin (51.30 mg 100 g-1 FM) was in the stems with the addition of approximately 45 μM tyrosine, while the increase in the leaves was proportional to the tyrosine concentration, and the best average was obtained with a tyrosine concentration of 75 μM (15.32 mg 100 g-1 FM). Higher tyrosine concentrations were deleterious to the growth of A. tenella plants, and a concentration of 75 μM was considered toxic. However, a tyrosine concentration of 50 μM benefitted betacyanin production, which reached 36.5 mg 100 g-1 FM in the plant shoots. These results showed the positive effect of tyrosine on the production of betacyanin in both species; however, application at high concentrations hampered the growth of Alternanthera plants.
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Leaves of Withania somnifera contained more withaferin A and withanolide A than roots indicating that these compounds mainly accumulate in leaves. With an increase in age of the plant, withaferin A was enhanced with a corresponding decrease in withanolide A. Hairy root cultures were induced from leaf explants using Agrobacterium rhizogenes and the transgenic nature of hairy roots was confirmed by partial isolation and sequencing of rolB gene, which could not be amplified in untransformed plant parts. In hairy roots, withaferin A accumulated at 2, 3 and 4% but not at 6% sucrose, the highest amount being 1733 mg/g dry weight at 4% level. High and equal amounts of withaferin A and withanolide A accumulated (890 and 886 mg/g dry tissue respectively) only at 3% sucrose. Increasing concentrations of glucose enhanced withaferin A and it peaked at 5% level (3866 mg/g dry tissue). This amount is 2842 and 34% higher compared to untransformed roots and leaves (collected from 210-day-old plants) respectively. Withanolide A was detected at 5% glucose but not at other concentrations. While chitosan and nitric oxide increased withaferin A, jasmonic acid decreased it. Acetyl salicylic acid stimulated accumulation of both withaferin A and withanolide A at higher concentrations. Triadimefon, a fungicide, enhanced withaferin A by 1626 and 3061% (not detected earlier) compared to hairy and intact roots respectively.
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La producción de metabolitos secundarios en cultivos celulares de plantas puede ser de interés para obtener compuestos difíciles de sintetizar o aislar de otras fuentes, lo cual generalmente se relaciona con un alto valor económico, aunque también puede ser útil para ayudar a dilucidar las vías metabólicas involucradas en la síntesis de estos compuestos. En este trabajo se presenta una descripción general de las antocianinas, un grupo de pigmentos de gran importancia para la industria, complementada con la referencia de los trabajos científicos recientes que se han publicado sobre la producción in vitro de las mismas. Con relación a esto último, se hace una descripción del efecto de cambios en las condiciones de cultivo, de la adición de precursores, del uso de reguladores de crecimiento, así como de la utilización de inductores y factores de estrés sobre la producción de estos compuestos. Finalmente, se hace mención al uso de raíces en cabellera, en inglés hairy roots, obtenidas mediante el uso de Agrobacterium rhizogenes, para la producción de estos compuestos.
The production of secondary metabolites in plant cell cultures may be of interest for obtaining compounds that are difficult to synthesize or isolate from other sources, which is usually associated with high economic value of the substances, but may also be useful to help elucidating the metabolic pathways involved in the synthesis of such compounds. This paper presents a general description of anthocyanins, a group of pigments of great importance to the industry, complemented by referring the scientific papers that have been recently published on their in vitro production. Regarding the latter, a description of the effect of changes in growing conditions, of the addition of precursors, of the use of growth regulators, and of the utilization of elicitors and stressors on the production of these compounds, is done. Finally, this review mentions the use of hairy roots obtained by the use of Agrobacterium rhizogenes for the production of these compounds.
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Objetivo: Estudiar la producción del metabolito 4b-hidroxiwithanólido E, mediante el cultivo in vitro de raíces transformadas de uchuva (Physalis peruviana L.) y evaluar el efecto de la influencia de la aplicación de diferentes elicitores sobre la producción de dicho metabolito. Materiales y métodos: Se obtuvieron raíces transformadas de Physalis peruviana L. mediante infección con Agrobacterium rhizogenes C106. Se cultivaron las raíces transformadas en medio líquido Murashige & Skoog, durante cuatro semanas, al cabo de las cuales se aplicaron diferentes concentraciones de sulfato de cobre, ácido salicílico y ácido jasmónico durante 24 horas. Se cuantificó el contenido del metabolito por cromatografía líquida...
Effect of elicitor application on the production of 4-b-hydroxy withanolide E by hairy roots of Physalis peruviana. Objectives: To study the metabolite 4-b-hydroxy withanolide E production by the in vitro culture of golden berry (Physalis peruviana L.) transformed roots, and to evaluate the effect of different elicitors on the metabolite production. Materials and methods: Hairy roots of Physalis peruviana L were obtained through infection with Agrobacterium rhizogenes C106. Hairy roots were cultured on Murashige & Skoog liquid medium for four weeks, before being exposed to different concentrations of copper sulfate, salicylic acid and jasmonic acid during 24 hours. Metabolite contents were quantified using High Performance Liquid Chromatography. Results: The highest amount of 4-bhydroxy withanolide E in hairy root tissues (0.323 mg/g of dry roots) was obtained after exposing the tissues to 10 mM salicylic acid as elicitor. Conclusions: 4-b-hydroxy withanolide E production in hairy roots was improved by using elicitors such as salicylic acid and copper sulphate. The highest concentration of the metabolite in hairy roots treated with elicitors was 1.538 times the control concentration (without elicitor treatment)...
Efeito da aplicação de elicitores sobre a produção de 4b- hidroxiwithanólido E, em raízes transformadas de Physalis peruviana L. Objetivo: Estudar a produção do metabolito 4b-hidroxiwithanólido E em cultivo in vitro de raízes transformadas de uchuva (Physalis peruviana L.) e avaliar o efeito da influência da aplicação de diferentes elicitores na produção deste metabolito. Materiais e métodos: Foramobtidas raízes transformadas de Physalis peruviana L. através da infecção com Agrobacterium rhizogenes C106. Cultivaram-se durantequatro semanas as raízes transformadas em meio líquido Murashige & Skoog, depois se aplicaram diferentes concentrações de sulfato de cobre, ácido salicílico e ácido jasmónico durante 24 horas. Quantificou-se o conteúdo do metabolito por cromatografía líquida. Resultados: Ao quantificar a produção de 4b-hidroxiwithanólido E presente nos tecidos encontrou-se a maior produção (0,323mg/g raízes seca) após aplicação do ácido salicílico a uma concentração de 10 mM como elicitor. Conclusões: Aumentou-se a produção de 4b-hidroxiwithanólido E aplicando elicitores como o ácido salicílico e sulfato de cobre, nas raízes de Physalis peruviana L. transformadas com Agrobacteriumrhizogenes. A maior concentração do metabolito é 1,538 vezes a encontrada nas raízes não tratadas com elicitores...