农杆菌T-DNA转化桑树的表型分析  

Molecular Identification and Phenotype Analysis of Transgenic Mulberry Plant by Agrobacterium tumefaciens T-DNA Introduction

作　　者：陆小平[1] 楼程富[1] 沈飞英[1] 小岛峯雄[2] 

机构地区：[1]浙江大学动物科学学院,杭州310029 [2]日本信州大学纤维学部

出　　处：《农业生物技术学报》2005年第2期157-161,共5页Journal of Agricultural Biotechnology

摘　　要：用自然杂交桑种子培育的3月龄幼年桑(MorusalbaL.)作材料,以野生型根癌农杆菌(Agrobacteriumtumefaciens)的突变体M-21介导,将根癌农杆菌T-DNA导入桑树基因组中。将幼年桑摘芯后用细针对腋芽分生组织剌伤感染获得4个形态异常的株系。PCR检测表明:4个株系都有T-DNA插入。将当代(T0)进行扦插繁殖获得T1代,T1表现出与T0不同的性状,显然T0是由正常产生生长素及细胞分裂素的未转化部分(第10叶位以下)和产生高水平细胞分裂素的转化部分共同作用决定的嵌合体表型,而T1是T0新梢的继续发育体,扦插后,它脱离了T0的作用。T0和T1的表型差异可能与T-DNA的插入有关。Mediated by the Agrobacterium tumefaciens mutant M-21, the T-DNA was introduced into the mulberry genome using the three-month-old seedlings (Morus alba.L.) derived from natural crossing as materials. Infection was conducted by directly stabbing the meristematic tissues of the axially buds of the materials after their top buds were removed. As a result, four plants with abnormal morphological traits were obtained. PCR confirmed that all four plants had the T-DNA insert, and we designated them as T0. Cuttings (T1) of the T0 displayed different morphological characters from T0, suggesting that the T0 plants consisted of non-transformed tissues (lower than 10th leaf order) where the auxin and cytokinin were normally produced and transformed tissues where the auxin and cytokinin levels were elevated, while the T1 plants were generated from the shootlets of the T0 plants on which the affect of T0 disappeared after cutting. The differences of the phenotype between the T1 and T0 were suggested to be caused by the insertion of the T-DNA.

关 键 词：DNA转化 表型分析 桑树 T-DNA插入 根癌农杆菌 细胞分裂素 DNA导入 PCR检测 种子培育 形态异常 分生组织 扦插繁殖 共同作用 表型差异 杂交桑 野生型 突变体 基因组 嵌合体 生长素 幼年 株系 月龄 介导 刺伤 腋芽 摘芯 

分 类 号：S572.03[农业科学—烟草工业] TQ464.2[农业科学—作物学]