Somatic embryogenesis and plant regeneration from protoplasts of Cichorium intybus L. x Cichorium endivia L.

Somatic embryos and adult plants were regenerated from mesophyll protoplasts of a clone of chicory 474 (Cichorium intybus L. x Cichorium endivia L.). Embryos were obtained in three different ways:

–	- plating of 7-day-old protoplast cultures on PM-3 solid induction medium with 2.5 M 2-isopentenyladenine (2-iP) and 0.5 M naphthaleneacetic acid (NAA);
–	- transfer of 1–2 mm microcolonies into liquid M-17 induction medium with 2.5 M (2-iP) and 0.1 M (NAA);
–	- transfer of well-established protoplast-derived calluses onto M-17.

Development of embryos was accomplished in agitated liquid Heller medium with 0.15 M gibberellic acid and germination on solid Heller medium without growth regulators. The total time for plantlet (4-leaves stage) recovery, following protoplast isolation, was 13 to 15 weeks. After acclimatization all protoplast-derived plants of Cichorium 474 were phenotypically normal and fertile.