Plant regeneration via somatic embryogenesis of Camptotheca acuminata in temporary immersion system (TIS)

Abstract

The present study describes a protocol for plant regeneration via somatic embryogenesis in temporary immersion system (TIS) for Camptotheca acuminata. Somatic embryos were induced by culturing hypocotyl segments from 14-day-old in vitro grown C. acuminata seedlings in TIS. Hypocotyl segments were placed in culture vessels modified with a mechanical device to support the fixation of explants. Cultures were maintained under a 16 h photoperiod with a light intensity of 60 lmol m -2 s -1 PPF at 25 ± 1°C. After 16 weeks of incubation embryogenic calli were formed above the edge of the mechanical device in the basal Murashige and Skoog (MS) medium containing 35 g l -1 sucrose and without hormonal supplementation. For plantlet regeneration, somatic embryos at cotyledonary stage were cultured in three different concentrations of 6-benzylamino-purine (0.5, 1.0 and 1.5 mg l -1 BAP) and in plant growth regulator (PGR) free medium. In general, 0.5 mg l -1 BAP was found to be the most effective concentration for growth and development of Camptotheca embryos in TIS. Conversion of somatic embryos into plantlets was also successfully achieved on sterile substrates moistened with 0.5 mg l -1 BAP. Plantlets derived from cotyledonary embryos were rooted in vitro with 0.5 mg l -1 indole-3-butyric acid (IBA) before transfer to ex vitro conditions.

Morphological and physiological responses of proliferating shoots of teak to temporary immersion and BA treatments

“…Rooting is one of the most difficult steps in the micropropagation of woody species, and forest rooting is usually vitro plant quality and made acclimatization safer for different species (Majada et al, Rodríguez et al 2008;Aragón et al 2010;Yang and Yeh, 2008;Sankar-Thomas et al 2008, Yan et al 2010. The teak plants from 4.44 µM of BA which showed some morpho-physiological disorders during the multiplication stage displayed good development and survived (80%) during acclimatization, suggesting that plantlets were able to revert to a normal morpho-physiological state after the transfer to ex vitro conditions.…”

Camptothecin accumulation in various organ cultures of Camptotheca acuminata Decne grown in different culture systems

“…A rapid and efficient plant regeneration system for C. acuminata was suggested by Sankar-Thomas et al (2008) which could supersede the extraction from plants either from wild populations or from cultivation.…”

Somatic Embryogenesis in Temporary Immersion Bioreactors

“…However, the culturing of explants directly in the container of the bioreactor has been reported in Saccharum spp using leaf disks (Snyman et al 2011), Camptotheca acuminata with hypocotyl segments (Sankar-Thomas et al 2008), and in Agave fourcroydes and Agave tequilana using layers of stem (Monja-Mio and Robert, data no reported), all with the aim of initiating embryogenic culture.…”

“…In Campotheca acuminata, a higher percentage of well-developed plantlets were obtained in the RITA bioreactor than in the twin-flask bioreactor; apparently, the presence of the polyurethane foam in the upper chamber that offers support to the embryos and the RITA container design positively affected development (Sankar-Thomas et al 2008). In B. gasipaes, a better response was achieved using the RITA bioreactor than using the twin-flask bioreactor (modified and unmodified), both in the multiplication of the cluster of somatic embryos and in converting them (Heringer et al 2014).…”

“…24.2) and is the most used bioreactor in the ES of different species. They have been successfully used in species such as C. arabica (Etienne-Barry et al 1999;Albarrán et al 2005;Gatica-Arias et al 2008), C. acuminata (Sankar-Thomas et al 2008), Saccharum spp (Snyman et al 2011), Q. robur (Mallón et al 2012), L. aestivum (Ptak et al 2013), Quercus suber When the pressure is released, the medium flows back into the lower compartment (d). The use of solenoid valve in the air circuit accelerates the return of the medium to the lower compartment.…”