Solasodine Production bySolanum eleagnifoliumCav.in vitroCultures: Influence of Plant Growth Regulators, Age and Inoculum Size. Large-Scale Production

Abstract

The effect of different auxins ( 2,4-D , 2,4,5-T , NAA and IAA ) on calli maintenance and plant cell suspension initiation as well as the influence of auxins in combination with citoquinines on growth and solasodine productivity by cell suspensions of Solanurn eleagnijolium Cav. was studied . 2,4-D and 2,4,5-T produced a friable and productive calli line and a fine and homogeneous cell suspension cultures . 2,4-D (0.5 pM or 5 pM ) or NAA ( 50 pM) produccd the highest growth index and solasodine productivity with values of about 0.5 mg 1-'day -' to 0.8 mg 1-' day-' Simultaneous administration of auxin and cytokinin increased the solasodine productivity with a diminution of cell growth . The combination of NAA ( 50 pM) with kinetin (0.25 pM) produced an increase of solasodine productivity from 0.8 mg 1-1 day to 2.54 mg 1-I day -I . An inoculum size of 5% V N and 19-day-old was selected to be used in fermentation processcs in order to obtain solasodine production by S. eleagnfoliurn Cav. suspension cultures Processes carried out in erlenmeyer flasks and stirred tank reactors by S. eleagnfoliurn Cav. suspension cultures showed similar specific growth rates ( 0.16 day-land 0.19 day-' respectively ) and maximal solasodine yields ( 0.91 mg g-' to 1.05 mg g-l respectively ) but they presented differences in the maximal biomass value obtained ( 3.9 g DW 1-' in erlenmeyer and 5.2 g DW I-lin bioreactor ) .

The expression of the 14D9 catalytic antibody in suspended cells ofNicotiana tabacumcultures increased by the addition of protein stabilizers and by transference from erlenmeyer flasks to a 2‐L bioreactor

“…Soft, friable calli obtained from the leaves of N. tabacum plantlets expressing the secreted (sec‐Ab) or KDEL‐tagged (Ab‐KDEL) mutant of the monoclonal antibody 14D9 were transferred to 150‐mL Erlenmeyer flasks containing 40 mL of MSRT liquid medium (Murashige Skoog culture medium with Khana and Staba vitamins) with 2.0 mg L −1 naphthalene acetic acid (NAA) and 0.2 mg L −1 kinetin (Kin) as plant growth regulators. Cultures were kept in an orbital shaker at 100 rpm, 24°C ± 2°C and a 16‐h photoperiod (1.8 w m −2 seg −1 ).…”

Co-expressing Turnip Crinkle Virus-coat protein with the serine protease α-thrombin precursor (pFIIa) in Nicotiana benthamiana Domin

“…Each callus was transferred every 3 wk. to fresh medium (López et al 2010, Alvarez et al 1993. After 6 mo of culture, independent callus lines showing kanamycin resistance were established.…”