Callus culture and somatic embryogenesis in wild colchicum hierosolymitanum feib

Callus was induced from seeds of Colchicum hierosolymitanum Feib inoculated on the surface of MS media supplemented with 0.45 μM 2, 4-D under dark conditions. Callus was subcultured on different levels (0.0, 0.45, 1.40, 2.30, 4.25 μM) of 2, 4-D or different levels (0.0, 0.54, 1.61, 2.70, 5.40 μM) of NAA under dark and light conditions. Increased 2, 4-D up to 4.52 μM significantly increased callus growth and the number of embryos regenerated (7.20 / explant) under dark conditions. However, the use of NAA at 1.61, 2.70, or 5.40 μM caused significant decrease in callus growth under darkness. Callus growth and maintenance was also evaluated using IBA at 0.0, 0.49 and 2.5 μM in combination with kinetin (0.46, 1.40 2.32, 4.70 μM) ; 2iP (2.50, 3.44, 5.0, 10.0 μM); Zeatin (0.46, 1.4, 2.30, 3.20, 4.60 μM) or TDZ (0.45, 1.40, 2.30, 4.50 μM). Calli induced in all treatments that contained the following plant growth regulators IBA and kinetin or 2iP or zeatin or TDZ were healthy, hard, pale yellow and friable. Phenolic browning occurred after three weeks of incubation on media supplemented with IBA at 2.50 in combination with 2.5, 3.44 or 5.0 μM of 2iP. Callus maintenance was tested on different levels (0.0, 0.1, 0.2, 0.3 M) of Sucrose, Mannitol, or Sorbitol as the source of carbon. Sucrose was the best carbon source among all sugars tested and 0.2 M had positive effect on callus growth and embryo regeneration. Different ratios of NH4+ : NO-3 (0 :0, 0 : 30, 10 : 20, 20 : 10, 30 : 0, 0 : 60, 20 : 40, 40 : 20, 60 : 0) were used to evaluate the growth of callus. Maximum callus growth was obtained at 30:0 mM total nitrogen. In addition, the highest number of regenerated embryos was obtained when the ratio between NH4+ NO-3 was 10 : 20 mM. تم استخدام الكالوس من بذور نبات اللحلاح (Colchicum hierosolymitanum Feib) بزراعتها على وسط موارشيج و سكوغ (MS) المضاف إليها منظم النمو (0.45 μM) 2,4-D تحت ظروف الظلام. و قد تمت زراعة الكالوس على مستويات مختلفة من منظم النمو 2,4-D(0.0, 0.45, 1.40, 2.30, 4.52 μM) أو منظم النمو (0.0, 0.45, 1.61, 2.70, 5.40 μM) NAA تحت ظروف الظلام و الإضاءة. و لوحظ أن زيادة تركيز منظم النمو 2,4-D إلى مستوى 4.52 ميكرومولار زاد عدد الأجنة المستنبتة من الكالوس (7.2) و زاد نمو الكالوس تحت ظروف الظلام. إلا أن تركيز منظم النمو NAA عند (1.61, 2.70, 5.40 μM) سبب انخفاضا في نمو الكالوس تحت ظروف الظلام و بفروق معنوية. و تم أيضا تقييم نمو و بقاء الكالوس باستخدام تراكيز مختلفة من منظم النمو (μM 0.0, 0.49, 2.5) IBA بالإضافة إلى التراكيز المختلفة من منظمات النمو التالية :