AGROBACTERIUM-TUMEFACIENS-MEDIATED TRANSFORMATION OF ARABIDOPSIS-THALIANA ROOT EXPLANTS BY USING KANAMYCIN SELECTION
AGROBACTERIUM-TUMEFACIENS-MEDIATED TRANSFORMATION OF ARABIDOPSIS-THALIANA ROOT EXPLANTS BY USING KANAMYCIN SELECTION
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DOI：
10.1073/pnas.85.15.5536
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发表时间：
1988-08-01
影响因子：
11.1
通讯作者：
VANLIJSEBETTENS, M
中科院分区：
文献类型：
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作者：
VALVEKENS, D;VANMONTAGU, M;VANLIJSEBETTENS, M
建立了诱导拟南芥（Arabidopsis thaliana（L.）嘿生根扦插，以快速和100%的效率再生芽。芽在体外或在土壤中生根后产生有活力的种子。建立了一种基于卡那霉素筛选的拟南芥根外植体转化方法。通过使用该再生程序和携带嵌合新霉素磷酸转移酶II基因（neo）的农杆菌肿瘤诱导Ti质粒，在基因转移后3个月内以20%至80%的效率获得转化的产种子植物。这些转化体的F1幼苗表现出卡那霉素抗性性状的孟德尔分离。该转化方法可应用于三种不同的拟南芥生态型。除了neo基因之外，还将赋予除草剂Basta抗性的嵌合bar基因引入拟南芥。通过酶法检测bar基因的表达。
Culture conditions were developed that induce Arabidopsis thaliana (L.) Heynh. root cuttings to regenerate shoots rapidly and at 100% efficiency. The shoots produce viable seeds in vitro or after rooting in soil. A transformation procedure for Arabidopsis root explants based on kanamycin selection was established. By using this regeneration procedure and an Agrobacterium tumor-inducing Ti plasmid carrying a chimeric neomycin phosphotransferase II gene (neo), transformed seed-producing plants were obtained with an efficiency between 20% and 80% within 3 months after gene transfer. F1 seedlings of these transformants showed Mendelian segregation of the kanamycin-resistance trait. The transformation method could be applied to three different Arabidopsis ecotypes. In addition to the neo gene, a chimeric bar gene conferring resistance to the herbicide Basta was introduced into Arabidopsis. The expression of the bar gene was shown by enzymatic assay.