Improvement of Agrobacterium-mediated transformation of cucumber (Cucumis sativus L.) by combination of vacuum infiltration and co-cultivation on filter paper wicks.
Improvement of Agrobacterium-mediated transformation of cucumber (Cucumis sativus L.) by combination of vacuum infiltration and co-cultivation on filter paper wicks.
复制标题
通过对滤纸芯上真空浸润和共培养的结合，改善了农杆菌介导的黄瓜（Cucumis sativus L.）的转化。
DOI：
10.1007/s11816-012-0260-1
复制
发表时间：
2013-07
影响因子：
2.4
通讯作者：
Tabei Y
中科院分区：
文献类型：
--
作者：
Nanasato Y;Konagaya KI;Okuzaki A;Tsuda M;Tabei Y
开发了一种改进的黄瓜遗传转化方法（Cucumis sativus L. cv. Shinhokusei No. 1）。农杆菌悬浮液对子叶外植体的真空浸润提高了外植体近端区域农杆菌感染的效率。滤纸芯上的共培养抑制了外植体的坏死，从而提高了再生效率。通过卡那霉素抗性和绿色荧光蛋白（GFP）荧光筛选假定的转基因植物，并通过基因组聚合酶链式反应（PCR）和Southern印迹证实转基因整合到黄瓜基因组中。这些转基因植物生长正常，并从7个品系中获得T1种子。最后，通过GFP荧光和基因组PCR证实了转基因在T1代中的稳定整合和传递。使用我们的方法生产黄瓜的平均转基因效率为 11.9 ± 3.5%，这是迄今为止使用卡那霉素作为选择剂报道的最高值之一。本文的在线版本 (doi:10.1007/s11816-012-0260-1) 包含补充材料，可供授权用户使用。
An improved method for genetic transformation of cucumber (Cucumis sativus L. cv. Shinhokusei No. 1) was developed. Vacuum infiltration of cotyledonary explants with Agrobacterium suspension enhanced the efficiency of Agrobacterium infection in the proximal regions of explants. Co-cultivation on filter paper wicks suppressed necrosis of explants, leading to increased regeneration efficiency. Putative transgenic plants were screened by kanamycin resistance and green fluorescent protein (GFP) fluorescence, and integration of the transgene into the cucumber genome was confirmed by genomic polymerase chain reaction (PCR) and Southern blotting. These transgenic plants grew normally and T1 seeds were obtained from 7 lines. Finally, stable integration and transmission of the transgene in T1 generations were confirmed by GFP fluorescence and genomic PCR. The average transgenic efficiency for producing cucumbers with our method was 11.9 ± 3.5 %, which is among the highest values reported until date using kanamycin as a selective agent. The online version of this article (doi:10.1007/s11816-012-0260-1) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
3.7
作者：
Ren Y;Zhang Z;Liu J;Staub JE;Han Y;Cheng Z;Li X;Lu J;Miao H;Kang H;Xie B;Gu X;Wang X;Du Y;Jin W;Huang S
通讯作者：
Huang S
影响因子：
5.2
作者：
Lee, YK;Chung, WI;Ezura, H
通讯作者：
Ezura, H
影响因子：
6.2
作者：
Peixoto de Oliveira, Maria Luiza;Febres, Vicente J.;Otoni, Wagner Campos
通讯作者：
Otoni, Wagner Campos
影响因子：
6.2
作者：
He, Zhengquan;Duan, ZhenZhen;Dai, Jianwu
通讯作者：
Dai, Jianwu
影响因子：
5.2
作者：
Ozawa, Kenjirou
通讯作者：
Ozawa, Kenjirou