2011•Tianjin Agricultural SciencesRequires access
Protoplast Preparation and Culture of Nitraria sibirica Pall.
Chengjin Guo
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Abstract
The cotyledons of Nitraria sibirica Pall.were used as materials in this work.The effects of combinations of enzyme mixture,the concentration of mannitol,the temperature and the time of enzymolysis on the protoplast preparation were studied by orthogonal design with L9(34).The results showed that the optimal preparing condition was by 1% cellulase(Onozuka R-10)+0.2% Pectolyase Y-23+0.8 mol·L-1 mannitol for 3.5 h,under 28 ℃.However,under the same conditions,more protoplast would be get from the cotyledons which had been treated by tissue culture.Besides,protoplast were cultured in solid-liquid double MS medium containing 2,4-D 1 mg·L-1+6-BA 0.5 mg·L-1+0.7 mol·L-1 mannitol,and then developed into cell cluster.