Microprojectile bombardment-mediated transformation of Lilium longiflorum
Microprojectile bombardment-mediated transformation of Lilium longiflorum
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DOI：
10.1007/s002990050389
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发表时间：
1998-02
影响因子：
6.2
通讯作者：
A. Watad;Dae-Jin Yun;Tracie K. Matsumoto;X. Niu;Y. Wu;A. Kononowicz;R. Bressan;P. Hasegawa
中科院分区：
文献类型：
--
作者：
A. Watad;Dae-Jin Yun;Tracie K. Matsumoto;X. Niu;Y. Wu;A. Kononowicz;R. Bressan;P. Hasegawa
我们已经获得了转基因百合（百合liliumlongiflorum）植株后，使用Biolistics PDS 1000/He系统，形态发生愈伤组织来自小鳞茎鳞片，随后双丙氨磷选择的基因枪轰击。使用的参数，得到最高的transientuidA表达：轰击压力为1100 psi，6厘米的目标距离和48小时的预培养在培养基上与3%的蔗糖。共1800个形态发生愈伤组织与质粒共轰击含有uidA报告或PAT选择标记基因。轰击后，将愈伤组织暴露于2 mg/l双丙氨膦。只有72个芽形成愈伤组织（4%）存活。在含有0.5mg/l双丙氨膦的伸长培养基上，72个芽簇产生342个芽。只有55株幼苗在2.0 mg/l双丙氨磷的后续暴露中存活。PCR分析表明，其中19株含有PAT基因。对其中3株植物的Southern分析表明，它们都含有PAT基因。
We have obtained transgenic lily (Lilium longiflorum) plants after microprojectile bombardment, using the Biolistics PDS 1000/He system, of morphogenic calli derived from bulblet scales, followed by bialaphos selection. Parameters which gave the highest transientuidAexpression were used: a bombardment pressure of 1100 psi, a target distance of 6 cm and a 48-h preculture on medium with 3% sucrose. A total of 1800 morphogenic calli were co-bombarded with plasmids containing either theuidAreporter orPATselectable marker genes. After bombardment, the calli were exposed to 2 mg/l bialaphos. Only 72 of the shoot-forming calli (4%) survived. The 72 shoot clusters produced 342 shoots on elongation medium containing 0.5 mg/l bialaphos. Only 55 plantlets survived subsequent exposure to 2.0 mg/l bialaphos. PCR analysis indicated that 19 of these plantlets contained thePATtransgene. Southern analysis of 3 of the plants indicated that all contained thePATgene.