Efficient callus formation and plant regeneration of goosegrass [Eleusine indica (L.) Gaertn.].
Efficient methods in totipotent callus formation, cell suspension culture establishment and whole-plant regeneration have been developed for the goosegrass [ Eleusine indica (L.) Gaertn.] and its dinitroaniline-resistant biotypes. The optimum medium for inducing morphogenic calli consisted of N6 basal salts and B5 vitamins supplemented with 1-2 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg l(-1) glycine, 100 mg l(-1) asparagine, 100 mg l(-1) casein hydrolysate, 30 g l(-1) sucrose and 0.6% agar, pH 5.7. The presence of organogenic and embryogenic structures in these calli was histologically documented. Cell suspension cultures derived from young calli were established in a liquid medium with the same composition. Morphogenic structures of direct shoots and somatic embryos were grown into rooted plantlets on medium containing MS basal salts, B5 vitamins, 1 mg l(-1) kinetin (Kn) and 0.1 mg l(-1) indole-3-acetic acid (IAA), 3% sucrose, 0.6% agar, pH 5.7. Calli derived from the R-biotype of E. indica possessed a high resistance to trifluralin (dinitroaniline herbicide) and cross-resistance to a structurally non-related herbicide, amiprophosmethyl (phosphorothioamidate herbicide), as did the original resistant plants. Embryogenic cell suspension culture was a better source of E. indica protoplasts than callus or mesophyll tissue. The enzyme solution containing 1.5% cellulase Onozuka R-10, 0.5% driselase, 1% pectolyase Y-23, 0.5% hemicellulase and N(6) mineral salts with an additional 0.2 M KCl and 0.1 M CaCl(2) (pH 5.4-5.5) was used for protoplast isolation. The purified protoplasts were cultivated in KM8p liquid medium supplemented with 2 mg l(-1) 2,4-D and 0.2 mg l(-1) Kn.
Yemets
A I
AI
Institute of Cell Biology and Genetic Engineering, National Academy of Sciences of Ukraine, Acad. Zabolotny str. 148, 03143 Kiev, Ukraine. alla@cellbio.freenet.viaduk.net
Klimkina
L A
LA
Tarassenko
L V
LV
Blume
Y B
YB
D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002478:Cells, Cultured; D046508:Culture Techniques; D031748:Eleusine / Q000166:cytology / Q000196:embryology / Q000502:physiology*; D006540:Herbicides / Q000494:pharmacology; D007701:Kinetin; D018613:Microscopy, Confocal; D009024:Morphogenesis / Q000502:physiology; D010937:Plant Growth Regulators / Q000494:pharmacology; D018520:Plant Shoots / Q000187:drug effects / Q000196:embryology / Q000502:physiology*; D011523:Protoplasts / Q000502:physiology; D012038:Regeneration / Q000187:drug effects; D014274:Trifluralin / Q000494:pharmacology
D006540:Herbicides; D010937:Plant Growth Regulators; D015084:2,4-Dichlorophenoxyacetic Acid; D014274:Trifluralin; D000225:Adenine; D007701:Kinetin