Transformation of microspore-derived embryos of winter oilseed rape (Brassica napus L.) by using Agrobacterium tumefaciens
Transformation of microspore-derived embryos of winter oilseed rape (Brassica napus L.) by using Agrobacterium tumefaciens
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DOI：
10.1007/bf03195632
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发表时间：
2008-01-01
影响因子：
2.4
通讯作者：
Szala, Laurencja
中科院分区：
文献类型：
--
作者：
Cegielska-Taras, Teresa;Pniewski, Tomasz;Szala, Laurencja
单倍体小孢子衍生胚（MDE）构成了将新性状引入冬季油菜（甘蓝型油菜）的独特材料。 MDE 已使用根癌农杆菌菌株 EHA 105 和 LBA4404 进行转化，两者均携带双元载体 pKG1B，其中包含编码葡萄糖醛酸酶 (GUS) 的 uidA 基因和作为膦丝菌素抗性标记的 bar 基因。经叶子涂漆测试证实，转化的胚胎表达 GUS 并再生出对除草剂 Basta 具有抗性的植物。转化体 T-39 的后代植物都是转基因的，因为它们继承了双单倍体亲本植物的 T-DNA。 Southern 印迹分析证实了 T-DNA 整合并传输到 T 植物中。 MDE的转化有利于获得导入基因纯合的冬季油菜。
Haploid microspore-derived embryos (MDEs) constitute a unique material for the introduction of new traits into winter oilseed rape (Brassica napus). MDEs have been transformed by using Agrobacterium tumefaciens strains EHA 105 and LBA4404, both carrying the binary vector pKG1B containing the uidA gene encoding --glucuronidase (GUS) and the bar gene as a marker of resistance to phosphinotricin. Transformed embryos expressed GUS and regenerated plants that here resistant to herbicide Basta, as confirmed by a leaf-painting test. Progeny plants of the transformant T-39 were all transgenic, as they inherited T-DNA from their doubled haploid parental plant. Southern-blot analysis confirmed the integration and transmission of T-DNA into T, plants. Trans formation of MDEs facilitates the obtaining of winter oilseed rape homozygous for the introduced genes.