Helmut Koblitz

Studies on tissue cultures of the genus Cinchona L. alkaloid production in cell suspension cultures

Initiation and culture of callus and cell suspensions of Cinchona ledgeriana and C. succirubra as well as the successful isolation and selection of a high-yielding alkaloid-forming strain derived from the leaf rachis of a C. succirubra plant are described. Results of feeding experiments with L-tryptophan using two different culture procedures are presented and discussed. Maximum alkaloid yields of up to 0.9% (based on dry weight) or 6.35 mg/l have been obtained.

Experiments on tissue culture in the genus Lycopersicon miller : Mesophyll protoplast regeneration to plants in Lycopersicon esculentum cv. 'Nadja'.

A method is described for the isolation and culture of protoplasts from cotyledon mesophyll cells of the tomato cultivar ‘Nadja’ grown axenically. Tissue cultures were produced from which shoots could be recovered. Rooted plantlets grew to normal plants and finally produced mature fruits.

Studies on tissue cultures of the genus Cinchona L. : In vitro mass propagation through meristem-derived plants.

A procedure for the in vitro clonal mass propagation of shoots and plants of two Cinchona species derived from single shoot tips is described. Special attention is given to rooting problems of the shoots under in vitro conditions.

Shikimate pathway activity in shake and fermenter cultures of Cinchona succirubra.

Cell suspension cultures of Cinchona succirubra were cultivated in shake cultures and for the first time in airlift fermenters. Under both conditions L-tryptophan exerts a stimulatory effect on alkaloid formation. In this context the regulatory pattern of some shikimate pathway enzymes was investigated in non-supplemented and tryptophan supplemented Cinchona cell cultures. A remarkable increase of tryptophan decarboxylase (TDC) activity was observed in Cinchona cells under the influence of tryptophan. Apparently, like in some other indole alkaloid producing cell cultures, a high TDC activity is a prerequisite for alkaloid formation. Growth pattern and some enzyme activities of C. succirubra fermenter cultures at controlled and non-regulated pH levels were followed. Optimum growth and alkaloid formation were recorded under non-regulated (normal) pH conditions.

Experiments on tissue culture in the genus Lycopersicon miller : Shoot formation from protoplasts of tomato long-term cell cultures.

Callus cultures from cotyledon explants were established and maintained in culture for more than two years. After several months callus cultures were transferred into liquid medium and cultured as cell suspensions. Protoplasts were isolated from these cell suspension cultures and cultured in a liquid medium. After formation of new cell walls the cells were further cultured in liquid medium and afterwards transferred to an agar-solidified medium to give a vigorously growing callus culture. In the case of the cultivar ‘Lukullus’ shoots were recovered from callus. All efforts to root these shoots failed and this, in addition to variations in appearence, suggests that the shoots are changed genetically possibly due to the prolonged culture period.