Estimulación de cardenólidos en brotes de Digitalis purpurea L. cultivados in vitro mediante elicitores

Abstract

ResumenDigitalis purpurea L. es una de las principales fuentes de cardenólidos tales como digoxina y digitoxina. Estos fármacos son ampliamente usados en la disfunción cardíaca y para regular las arritmias del corazón. El presente trabajo se realizó con el objetivo de estudiar el efecto de tres elicitores en el cultivo de brotes de Digitalis purpurea var. Roter Berggold para incrementar la producción in vitro de cardenólidos. La elicitación es una estrategia para incrementar la producción de biomasa y metabolitos secundarios en el cultivo in vitro. Los elicitores evaluados fueron ChitoPlant (0,001; 0,01; 0,1 g.L ) y Jasmonato de metilo (60, 80 y 100 µM), descritos por primera vez para el incremento de cardenólidos. Se demostró que la elicitación es una estrategia viable para el incremento de cardenólidos en brotes de D. purpurea. El ChitoPlant ® , redujo la altura sin afectación en el resto de las variables morfológicas evaluadas. Además indujo un incremento significativo en el contenido de cardenólidos. El SilioPlant) no provocó afectaciones en la biomasa e incrementó significativamente la síntesis de cardenólidos en los brotes en 3,6 y 6,9 veces el contenido de digoxina y digitoxina respectivamente. La elicitación con el jasmonato de metilo provocó una reducción de la biomasa. Los contenidos de digoxina y digitoxina se incrementaron ligera y significativamente con 80 y 100 µM de jasmonato de metilo respectivamente. El mejor resultado integral se obtuvo con 0,01 g.L -1 de SilioPlant, el cual indujo la mayor producción neta de cardenólidos por frasco de cultivo (4,72 µg digoxina y 88,27 µg digitoxina).Palabras clave: chitoplant, digitoxina, digoxina, jasmonato de metilo, silioplant. AbstractDigitalis purpurea L. is one of the main sources of cardenolides such as digitoxin and digoxin. These drugs are widely used to strengthen cardiac diffusion and to regulate heart rhythm. The aim of this study was to evaluate the influence of three elicitors on shoots of Digitalis purpurea var. Roter Berggold in semisolid media in order to increase cardenolides biosynthesis. Elicitation is a strategy to increase biomass and secondary metabolites production on in vitro cultures. The elicitors evaluated were ChitoPlant (0,001; 0,01; 0,1 g.L ) and Methyl jasmonate (60, 80, 100 µM), which are reported here to induce cardenolide production for first time. Elicitation resulted an effective strategy to increase cardenolide production on D. purpurea shoot cultures. ChitoPlant induced a decrease in shoots length, but had no effect on the rest of morphological parameters evaluated. As well, ChitoPlant increased cardenolide content. SilioPlant (0,01 g.L -1 ) did not affect biomass production and at the same time, increased in 3,6-fold and 6,9-fold digoxin and digitoxin content respec-* Doctor en Ciencias Agrícolas.

The Foxgloves (Digitalis) Revisited

“…The large amounts of progesterone used in this experiment suggest that this compound is a precursor or an elicitor of cardenolide formation (see below: Biosynthesis) [57]. Elicitation as a method to enhance cardenolide production was used by Pérez-Alonso et al [58], resulting in the production of about 0.1 mg cardenolides/g dry weight (similar to the figures reported by Hagimori et al [6,59]) compared to about 5 to 10 mg/g in dried D. purpurea leaves [6].…”

Enhancement of cardenolide production in transgenic Digitalis purpurea L. by expressing a progesterone-5β-reductase from Arabidopsis thaliana L.

“…But no digitoxin was detected and higher digoxin production was around 40 μg/gDW. Elicitation was also applied to D. purpurea with the same goal (Patil et al, 2013;Pérez-Alonso et al, 2014a). The effect of multiple abiotic and biotic elicitors on plant growth and cardenolide content was evaluated by Patil et al (2013).…”

“…Several biotechnology strategies, like temporary immersion system culture and elicitation, have been developed to minimize these limitations (Pérez-Alonso et al, 2012, 2014aPatil et al, 2013). But unfortunately, none of these led to the production of more cardenolides than in plants cultivated in vivo (Sales et al, 2011;Verma et al, 2016).…”

Effect of Salicylic Acid, 2,4-D and 2i-P on the Production of Secondary Metabolites in Garcinia brasiliensis Mart. Callus

“…Therefore, in vitro plant cell culture has been used for the production of secondary metabolites of interest [18]. One strategy frequently used is the addition of elicitors, such as salicylic acid (SA), in the culture medium of in vitro cell culture [19]. SA acts as a phytohormone [20] and maintains auxin and cytokinin levels in plant tissues, increasing cell division and dry matter [21].…”