转玉米C_4型pepc水稻成熟胚为外植体的水稻悬浮细胞系的优化  

Optiming Cell Suspension Culture of Mature Embryo of Transgenic C_4 Phosphoenolpyruvate Carboxylase(pepc) Rice

作　　者：王满[1] 李霞[1] 石牡丹[1] 钱宝云[1] 魏晓东[1] 方先文[1] 

机构地区：[1]江苏省农业科学院粮食作物研究所,江苏省优质水稻工程技术研究中心,南京210014

出　　处：《分子植物育种》2012年第6期644-654,共11页Molecular Plant Breeding

基　　金：国家自然科学基金项目(30871459);江苏省自主创新课题[CX(11)1020]共同资助

摘　　要：为了研究外源玉米光合作用关键酶C4型磷酸烯醇式丙酮酸羧化酶(phosphoenolpyruvate carboxylase,PEPC)基因在C3植物水稻中提高光合效率的分子机理,本研究以高表达玉米C4型pepc水稻(PC)和未转基因野生型Kitaake(WT)为材料,探索高效且稳定水稻悬浮细胞系建立的方法。结果表明:PC成熟胚愈伤组织的诱导培养基中添加2mg/L2,4-D和1mg/L6-BA,可出诱导出高频的愈伤组织;再通过3次继代培养(2～0.5mg/L2,4-D,2～0.2mg/L6-BA,1mg/LABA),逐步降低2,4-D和6-BA的浓度,可获得了疏松、颗粒状且分散的愈伤组织块;然后转接到水稻悬浮细胞液体培养基中(0.5mg/L2,4-D,0.2mg/L6-BA和1mg/LABA),在28℃下培养,新原液的比例为3:1,经42d,可建立稳定且高效的供试水稻悬浮细胞系。In order to study the molecular mechanism to improve photosynthetic efficiency of the exogenous photosynthesis key enzyme,maize-C4 phosphoenolpyruvate carboxylase（PEPC） gene in the C3 plant rice.In this research,transgenic rice with high expressed maize C4 pepc gene（PC） and untransformed wild-type Kitaake（WT） used as materials to explore the establishment of efficient and stable rice suspension cell lines.The results showed that the PC mature embryo callus induction medium by adding 2mg/L 2,4-D and 1mg/L 6-BA can induce calli with high frequency;subculture three times（2～0.5mg/L 2,4-D,2～0.2mg/L 6-BA and 1mg/L ABA） and gradually reduce the concentration of 2,4-D and 6-BA,can generate loose,granular like and dispersed callus block;then transferred into in liquid medium of rice suspension cells（0.5mg/L 2,4-D,0.2 mg/L 6-BA and 1mg/L ABA） under the culture of 42 days at 28℃ with dilutionratio of 3：1,a stable and high efficient rice suspension cell lines can be established.

关 键 词：水稻 转C4pepc基因水稻 成熟胚 悬浮培养 激素调优 

分 类 号：S511[农业科学—作物学]