In vitro propagation of four saponin producing Maesa species

Abstract

A successful micropropagation system was developed for four different medicinal Maesa species. Multiple shoots were induced through both axillary bud formation and adventitious shoot regeneration from leaf explants. The explants were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA), thidiazuron (TDZ) and/or alpha-naphthalene acetic acid (NAA). The success of regeneration varied for different species and depended on the type and concentration of plant growth regulators. Regenerated shoots spontaneously developed roots within 6 weeks on MS hormone-free medium. The rooted shoots were transferred to the greenhouse with a 100% success rate. Furthermore, flow cytometry analysis indicated that there were no changes in ploidy level of those regenerated shoots as compared with wild type adult plants. Thin layer chromatography (TLC) analysis revealed that common and distinguishing spot of saponins were similarly observed in regenerated shoots compared to the control plants. Therefore, the protocol also provides an effective means for the in vitro conservation of Maesa spp. that produce pharmaceutically interesting saponins

Agroinfiltration of intact leaves as a method for the transient and stable transformation of saponin producing Maesa lanceolata

“…The adventitiously formed shoots spontaneously generated roots in hormone-free MS medium. The rooting success was comparable to the non-transformed adventitious shoots as previously reported (Faizal et al 2011). The plants were transferred to soil and developed into vigorously growing plants under greenhouse conditions.…”

“…Sterilized explants were placed on shoot induction medium supplemented with 500 mg/L cefotaxime and 100 mg/L kanamycin to select the transgenic plants. Multiple shoots were induced through adventitious shoot regeneration as previously described (Faizal et al 2011). …”

“…The dose of antibiotics required for selection was tested by incubating the leaf discs on shoot induction medium containing different concentration of cefotaxime and kanamycin (Table 1). Kanamycin at 25 mg/L completely inhibited shoot regeneration from leaf explants following a protocol previously described (Faizal et al 2011). Because leaf explants showed resistance to higher kanamycin concentrations, we used 100 mg/L.…”

Studies on saponin production in tropical medicinal plants Maesa argentea and Maesa lanceolata

“…lanceolata seeds were collected in Moshi, Tanzania by Frank Mbago (Department of Botany, University of Dar-Es-Salaam). The in vitro plants were maintained as previously described in [4]. glass tube containing solid MS medium supplemented with MS vitamins, 3% (w/v) sucrose and 0.8% (w/v) agar (Lab M plant tissue culture agar MC29, Amersham, UK).…”

“…The in vitro culture and propagation method for Maesa species were successfully established [4]. Using in vitro grown material, we studied the saponin production in more detail and in addition to determined saponin production in greenhouse grown plants.…”