Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B-napus
Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B-napus
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DOI：
10.1007/s00299-005-0973-5
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发表时间：
2005-10-01
影响因子：
6.2
通讯作者：
Kameya, T
中科院分区：
文献类型：
--
作者：
Park, BJ;Liu, ZC;Kameya, T
采用农杆菌介导的方法，建立了超声波和真空渗透法辅助转化萝卜的技术路线。农杆菌菌株LBA 4404含有双元载体pBI 121-莱亚（晚期胚状体丰富），其携带来自甘蓝型油菜的组3莱亚基因。在6个组合中，农杆菌介导的转化结合5 min超声波处理和5 min真空渗入的转化频率最高。通过PCR、基因组Southern和Western blot分析，证实了转入的莱亚基因在转基因植株中的存在、整合和表达。在渗透胁迫和盐胁迫条件下，转基因萝卜表现出比非转化对照植株更好的生长表现。第3组莱亚蛋白在转基因萝卜营养组织中的积累增强了对水分亏缺和盐胁迫的耐受性。
A protocol for producing transgenic radish (Raphanus sativus) was obtained by using both ultrasonic and vacuum infiltration assisted, Agrobacterium-mediated transformation. The Agrobacterium strain LBA4404 contained the binary vector pBI121-LEA (late embyogenesis abundant), which carried a Group 3 LEA gene, from Brassica napus. Among six combinations, Agrobacterium-mediated transformation assisted by a combination of 5-min sonication with 5-min vacuum infiltration resulted in the highest transformation frequency. The existence, integration and expression of transferred LEA gene in transgenic T-1 plants were confirmed by PCR, genomic Southern and Western blot analysis. Transgenic radish demonstrated better growth performance than non-transformed control plants under osmotic and salt stress conditions. Accumulation of Group 3 LEA protein in the vegetative tissue of transgenic radish conferred increased tolerance to water deficit and salt stress.