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.

中文翻译：

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通过真空渗透和滤纸芯上共培养的组合改进农杆菌介导的黄瓜 (Cucumis sativus L.) 转化。

开发了一种改良的黄瓜遗传转化方法（Cucumis sativus L. cv. Shinhokusei No. 1）。用农杆菌悬浮液对子叶外植体进行真空浸润提高了外植体近端区域农杆菌感染的效率。滤纸灯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，T ₁种子来自 7 个品系。最后，通过 GFP 荧光和基因组 PCR 证实了转基因在 T ₁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。