Abstract
Castilleja tenuiflora, a species highly valued for its medicinal properties, is threatened in the wild. We evaluated the effects of six different immersion cycles in a temporary immersion bioreactor on C. tenuiflora shoot growth, proliferation rate, phenolics content, flavonoid content, and antioxidant activity. We also evaluated the regeneration capacity of the shoots. The highest proliferation rate (nine shoots per explant) was obtained using an immersion cycle of 5 min every 12 h, and the longest shoots (38.8 ± 1.9 mm) were obtained using an immersion cycle of 5 min every 24 h. Shoots obtained from immersion cycles of 30 min every 24 h or 5 min every 24 h showed 100% rooting efficiency. Shoots obtained from immersion cycles of 30 min every 3 h or 30 min every 12 h accumulated H2O2, developed abnormal stomata, and showed symptoms of hyperhydricity. These characteristics were associated with a low survival rate (16–80%) when the plants were transferred to potting mix. The shoots from an immersion cycle of 30 min every 24 h showed the highest total phenolics content, which coincided with the highest antioxidant activity in the 2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) free-radical scavenging assay (161.74 ± 10.06 μmol Trolox/g dry weight (DW)). The shoots from an immersion cycle of 5 min every 24 h showed the highest activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical scavenging assay, and those from an immersion cycle of 5 min every 3 h showed the strongest reducing power. These results show that temporary immersion culture represents a reliable and efficient method for in vitro micropropagation of C. tenuiflora.
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Acknowledgments
This work was supported by the Secretaría de Investigación y Posgrado del IPN-México (SIP–IPN Grants 20131786) and by the Fondo Mixto de Fomento a la Investigación Científica y Tecnológica CONACYT–Gobierno del Estado de Morelos (Grant MOR-2007-C01-79409). The funding sources were not involved in the preparation of this paper or in the decision to submit it for publication. RVT is indebted to the CONACYT and PIFI-IPN for the Master in Sciences fellowship awarded. GTT, JLTE, and ARLL are grateful to the Sistema de Becas por Exclusividad (IPN) and Programa de Estímulos al Desempeño de los Investigadores (IPN).
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Editor: Praveen Saxena
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Valdez-Tapia, R., Capataz-Tafur, J., López-Laredo, A.R. et al. Effect of immersion cycles on growth, phenolics content, and antioxidant properties of Castilleja tenuiflora shoots. In Vitro Cell.Dev.Biol.-Plant 50, 471–477 (2014). https://doi.org/10.1007/s11627-014-9621-5
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DOI: https://doi.org/10.1007/s11627-014-9621-5