中国传统菊花品种‘小林静’再生及转化体系的建立

以中国传统菊花品种‘小林静’叶片为外植体,建立了‘小林静’较好的再生体系及遗传转化体系.结果表明,‘小林静’叶盘最适不定芽分化培养基为MS+2.0 mg/L 6-BA+1.5 mg/L NAA,不定芽分化率为92.76％,平均再生不定芽数为2.3767个；试管苗最佳生根培养基为1/2MS,生根率达100％.移栽采用灭菌的蛭石,再生苗移栽成活率达90％以上.采用根癌农杆菌C58C1介导的叶盘转化法进行‘小林静’的遗传转化试验,农杆菌OD600=0.5-0.6,侵染10 min后,将叶片外植体接种到MS+2.0 mg/L 6-BA+1.5mg/L NAA的培养基中黑暗共培养2d,之后转接到附加10 mg/L硫酸卡那霉素和400 mg/L羧苄青霉素的分化筛选培养基中进行转化细胞的筛选,待长出抗性芽后转接至生根培养基中进行培养,最终建立了菊花品种‘小林静’的遗传转化体系.

Establishment of Regeneration and Transformation Systems in Chrysanthemum × morifolium ‘Xiao Linjing'

This research established leaves of Chrysanthemum × morifolium‘Xiao Linjing’stability of high efficient plant regeneration and transformation systems.The results showed that the best adventitious buds differentiation medium was MS+2.0 mg/L 6-BA+1.5 mg/L NAA and the rate of adventitious buds differentiation was 92.76%.The average adventitious buds per explant were 2.376 7.For the rooting medium,1/2MS was the best and the rate of rooting was 100%.Living tube seedlings transplanting survival rate was over 90% using vermiculite after sterilization.Agrobacterium C58C1 was chosen for the genetic transformation experiment.The OD600ranged from 0.5 to 0.6 and the infection time was 10 minutes.Co-cultured in the dark for two days,then the leaf explants were put in antibiotic selection medium that contain 10 mg/L kanamycin and 400 mg/L carbenicillin.When resistant shoots formed,it was cultured on the rooting medium.