模式籼稻品种Kasalath转基因胚性悬浮细胞系的建立  

Successful Suspension Culture of the Model Indica Rice Variety Kasalath

作　　者：龙起樟 黄永兰 唐秀英 王会民 芦明 袁林峰 蔡耀辉 万建林 LONG Qi-zhang;HUANG Yong-lan;TANG Xiu-ying;WANG Hui-min;LU Min;YUAN Lin-feng;CAI Yao-hui;WAN Jian-lin(Jiangxi Super-rice Research and DevelopmentCenter,Nanchang 330200,China;National Rice Engineering Laboratory Nanchang,Nanchang 330200,China)

机构地区：[1]江西省超级水稻研究发展中心,江西南昌330200 [2]水稻国家工程实验室(南昌),江西南昌330200

出　　处：《江西农业大学学报》2019年第1期13-18,共6页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家重点研发计划项目(2016YFD0101105);江西省科技计划项目(20143ACF60008);江西现代农业科研协同创新专项(JXXTCX2015006-002)~~

摘　　要：Kasalath是一种公认的组培再生性优异的模式籼稻品种。为建立具有G418抗性的Kasalath悬浮细胞系,通过农杆菌转化法获得转入G418抗性基因和GUS报告基因的Kasalath愈伤组织,然后利用获得的胚性转基因愈伤在NBL培养基中进行初步悬浮培养,后转至AA培养基进行终培养,在继代过程中通过持续的小颗粒愈伤筛选最终建立了Kasalath转基因悬浮细胞系;本研究获得的悬浮细胞系分散性良好,增值快(10 d可增殖约3.8倍),而且可从悬浮细胞中分离到高活性的原生质体,分离效率可达6.3×107个原生质体每毫升自然沉降细胞。本研究展示了Kasalath是一种建立悬浮细胞系的优良材料,为进一步进行栽培稻和野生稻的原生质体融合提供依据,建立的悬浮细胞系亦可用于生理生化研究或供分离原生质体以进行蛋白质亚细胞定位等试验。Kasalath,the model indica rice variety,is known for its outstanding regeneration capability in tissue culture.The purpose of the present study is to prepare suspension culture of Kasalath with transgenic G418 resistance,providing useful materials for protoplast isolation in subsequent somatic fusion between wild rice and cultivated rice.Calli arising form mature seeds were transformed with the NPT II and the GUS gene via agrobacterium‐mediated method and the resultant transgenic calli were used in suspension culture.The suspensions were maintained in NBL medium for early‐stage culture and in AA medium for final suspension culture.At all the stages,continuous selection of small‐size calli was taken when renewing media.Finally,the finely dispersed cell suspension culture of Kasalath was established.The established suspension cultures can propagate 3.8 times within 10 days as calculated in naturally settled volume.The suspension cells were ready for efficient isolation of highly viable protoplasts and the efficiency of such isolation could be as high as 6.3×10^7per milliliter naturally settled cell aggregates.Thus,Kasalath was demonstrated in the present study as a competent variety in establishment of rice suspension culture and the suspension cultures established may be useful in physiological and biochemical research or in isolation of protoplasts for somatic fusion,protein subcellular localization investigation and so on.

关 键 词：籼稻 悬浮细胞系 原生质体 细胞培养 转基因 

分 类 号：S511.035[农业科学—作物学]