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METHOD OF HIGH FREQUENCY PLANT REGENERATION FROM ZYGOTIC EMBRYO-DERIVED EMBRYOGENIC CELL SUSPENSION CULTURES OF RANUNCULUS KAZUSENSIS
METHOD OF HIGH FREQUENCY PLANT REGENERATION FROM ZYGOTIC EMBRYO-DERIVED EMBRYOGENIC CELL SUSPENSION CULTURES OF RANUNCULUS KAZUSENSIS
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机译：Kazusensis的合子胚性胚细胞悬浮培养高频率植株再生方法
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摘要
A method for regenerating a somatic embryo or a plantlet of Ranunculus kazusensis Makino is provided to preserve the Ranunculus kazusensis Makino effectively as a genetic resource through in vitro plant multiplication, thereby applying the Ranunculus kazusensis Makino to a material for molecular breeding through a useful transduction. A method for regenerating a somatic embryo or a plantlet of Ranunculus kazusensis Makino comprises the steps of: (a) culturing seeds of the Ranunculus kazusensis Makino in a 1/2 SH solid culture medium having a pH of 5.8 at a temperature of 25 deg.C under dark to germinate the seeds; (b) cutting the germinated seeds in a vertical direction and explanting the cut seeds onto the solid culture medium to elongate zygotic embryos; (c) culturing the elongated zygotic embryos in a 1/2 SH solid culture medium where 0.1-1 mg/liter of 2,4-dichlorophenoxyacetic acid is added at a temperature of 25 deg.C under dark to form calluses; (d) culturing the calluses in an SH liquid culture medium where 0.5mg/L of 2,4-D is added or in a V liquid culture medium prepared by adding 0.1mg/liter of indole acetic acid(IAA), 0.1mg/liter of naphthalene acetic acid(NAA), 1.5mg/liter of 2,4-D, and 0.25 mg/liter of kinetin to a B5 culture medium for one week to establish a suspension culturing system and proliferating the calluses; (e) suspension-culturing the proliferated calluses in an SH liquid culture medium or a V liquid culture medium at a temperature of 25 deg.C with the speed of 100 rpm through subculturing; and (f) after subculturing cell mass formed at the step(e) in a 1/2 solid culture medium and culturing plantlets at a temperature of 25 deg.C for 2 weeks under dark, culturing them at a temperature of 25 deg.C under the light cycle of 16/8 hours under light.
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