STABLE GENETIC-TRANSFORMATION OF ARABIDOPSIS-THALIANA BY AGROBACTERIUM INOCULATION IN PLANTA
STABLE GENETIC-TRANSFORMATION OF ARABIDOPSIS-THALIANA BY AGROBACTERIUM INOCULATION IN PLANTA
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DOI：
10.1046/j.1365-313x.1994.5040551.x
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发表时间：
1994-04-01
期刊：
影响因子：
7.2
通讯作者：
NAM, HG
中科院分区：
文献类型：
--
作者：
CHANG, SS;PARK, SK;NAM, HG
通过简单地接种带有双元载体pBI121的根癌农杆菌菌株LBA4404，实现了拟南芥的稳定遗传转化。我们称之为植物转化的转化过程包括在根部切断顶端芽，在切断部位接种农杆菌，以及从切断部位产生芽的植物。平均而言，5.5% 的新形成芽产生转化后代。这些后代（T-2代）在基因组中含有T-DNA，通过测定T-DNA编码的β-葡萄糖醛酸酶和卡那霉素抗性以及通过基因组Southern印迹分析进行检查，拟南芥基因组中T-DNA的拷贝数是单个（33%）或多个。在 T-3 和 T-4 代或用转化体与野生型植物之间异型杂交的 F-2 后代检查的转化体的遗传行为表明，大多数插入的 T-DNA 以孟德尔方式遗传。该过程为拟南芥的简单有效转化提供了一种新方法，无需从体外组织外植体再生植物。
Stable genetic transformation of Arabidopsis thaliana was achieved by simple in planta inoculation of Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pBI121. The transformation procedure, which we call in planta transformation, involves severing of apical shoots at their bases, inoculation with Agrobacterium at the severed sites, and in planta generation of shoots from the severed sites. On average, 5.5% of the newly formed shoots produced transformed progenies. These progenies (T-2 generation) contained T-DNA in the genome as examined by assaying the T-DNA encoded beta-glucuronidase and kanamycin resistance and by genomic Southern blot analysis, the copy number of the T-DNAs in the Arabidopsis genome being single (33%) or multiple. The genetic behavior of the transformants examined at the T-3 and T-4 generations or with the F-2 progenies of the outcrosses between transformants and wild-type plants showed that most of the inserted T-DNA are inherited in a Mendelian fashion. This procedure provides a new approach for simple and efficient transformation of A. thaliana, obviating the need for plant regeneration from tissue explants in vitro.