Alkaloids Analysis of Habranthus cardenasianus (Amaryllidaceae), Anti-Cholinesterase Activity and Biomass Production by Propagation Strategies

Abstract

Plants in the Amaryllidaceae family synthesize a diversity of bioactive alkaloids. Some of these plant species are not abundant and have a low natural multiplication rate. The aims of this work were the alkaloids analysis of a Habranthus cardenasianus bulbs extract, the evaluation of its inhibitory activity against cholinesterases, and to test several propagation strategies for biomass production. Eleven compounds were characterized by GC-MS in the alkaloid extract, which showed a relatively high proportion of tazettine. The known alkaloids tazettine, haemanthamine, and the epimer mixture haemanthidine/6-epi-haemanthidine were isolated and identified by spectroscopic methods. Inhibitory cholinesterases activity was not detected. Three forms of propagation were performed: bulb propagation from seed, cut-induced bulb division, and micropropagated bulbs. Finally, different imbibition and post-collection times were evaluated in seed germination assays. The best propagation method was cut-induced bulb division with longitudinal cuts into quarters (T1) while the best conditions for seed germination were 0-day of post-collection and two days of imbibition. The alkaloids analyses of the H. cardenasianus bulbs showed that they are a source of anti-tumoral alkaloids, especially pretazettine (tazettine) and T1 is a sustainable strategy for its propagation and domestication to produce bioactive alkaloids.

Keywords: Amaryllidaceae; GC-MS; bioactive alkaloids; biomass production; propagation methods.

Figure 1

GC chromatogram of H. cardenasianus extract showing the alkaloids identified by MS. (1) Ismine, (2) 5,6-Dihydrobicolorine, (3) Unknown, (4) Galanthindole, (5) Anhydrolycorine, (6) Unknown, (7) 11,12-dehydro anhydrolycorine, (8) Haemanthamine, (9) Tazettine, (10) Haemanthidine/6-epi-haemanthidine, (11) Lycorine.

Figure 2

Alkaloids from H. cardenasianus.

Figure 3

Effect of the propagation method on survival (A); multiplication rate (B); equatorial diameter of bulb (C); and bulb length (D). Means and standard deviation are reported. Data were analyzed by ANOVA and different letters (a, b, c, d) indicate significant differences by the Tukey test (p < 0.05). The treatments were: T0 (Control): whole bulb, T1: bulb quarters, T2: twin scales, T3: bulb basal discs, T4 micro propagated bulbs and T5: seed propagated bulbs.

Figure 4

Effect of the propagation method on biomass increase (A); root diameter (B); root length (C); and root number (D). Means and standard deviation are reported. Data were analyzed by ANOVA and different letters (a, b, c, d) indicate significant differences by the Tukey test (p < 0.05). The treatments were: T0 (Control): whole bulb, T1: bulb quarters, T2: twin scales, T3: bulb basal discs, T4 micro propagated bulbs and T5: seed propagated bulbs.

Figure 5

Linear correlation analysis: root length-equatorial bulb diameter (A), root length-bulb length (B), root length-bulb dry weight (C), root diameter-equatorial bulb diameter (D), root diameter-bulb length (E), root diameter-bulb dry weight (F), root number-equatorial bulb diameter (G), root number-bulb length (H), root number-bulb dry weight (I). Coefficient of determination (R²) and Pearson’s correlation coefficient (r) are reported. p < 0.05 indicate that the linear correlation is statistically significant.

Figure 5

Linear correlation analysis: root length-equatorial bulb diameter (A), root length-bulb length (B), root length-bulb dry weight (C), root diameter-equatorial bulb diameter (D), root diameter-bulb length (E), root diameter-bulb dry weight (F), root number-equatorial bulb diameter (G), root number-bulb length (H), root number-bulb dry weight (I). Coefficient of determination (R²) and Pearson’s correlation coefficient (r) are reported. p < 0.05 indicate that the linear correlation is statistically significant.