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首页> 外文期刊>Biologia Plantarum >Marker-free transgenic cucumber expressing Arabidopsis cbf1 gene confers chilling stress tolerance
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Marker-free transgenic cucumber expressing Arabidopsis cbf1 gene confers chilling stress tolerance

机译：表达拟南芥cbf1基因的无标记转基因黄瓜赋予抗寒性

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摘要

Marker-free transgenic cucumber (Cucumis sativus L.) cv. Poinsett 76 SR plants were produced by Agrobacterium mediated transformation. A transformation efficiency of 1.62 was observed on using Agrobacterium tumefaciens strain LBA4404 harbouring Arabidopsis cbf1 gene driven by the inducible promoter RD29A in a binary vector system pCAMBIA. Transgene integration and single copy insert in transgenic cucumber was confirmed by polymerase chain reaction (PCR) and Southern blot analysis in T0 lines and also confirmed marker-free status in T1 generation. Transgene expression was confirmed by reverse transcription (RT)-PCR in T1 generation transgenic cucumber and advanced to T2 generation. Upon exposure to chilling stress (4 °C), the T2 generation transgenic plants survived up to 36 h; however, wild-type plants could not survive and gradually died. A significant decrease in membrane injury index (MII), increase in activities of antioxidant enzymes (SOD and CAT), free proline content and relative water content (RWC) in the leaves were observed in transgenic cucumber as compared to wild-type under chilling stress. Thus, the transgenic cucumber plants expressing Arabidopsis cbf1 gene conferred protection against chilling stress.
机译：无标记转基因黄瓜（Cucumis sativus L.）cv。通过农杆菌介导的转化产生Poinsett 76 SR植物。在二元载体系统pCAMBIA中，使用含有由诱导型启动子RD29A驱动的拟南芥cbf1基因的根癌农杆菌菌株LBA4404，转化效率为1.62。通过聚合酶链反应（PCR）和Southern印迹分析在T 0 品系中确认了转基因黄瓜的转基因整合和单拷贝插入，并在T 1 中确认了无标记状态代。通过逆转录（RT）-PCR在T 1 世代转基因黄瓜中证实了转基因表达，并进展到T 2 世代。暴露于低温胁迫（4°C）时，T 2 世代转基因植物存活长达36小时；但是，野生型植物无法生存并逐渐死亡。与低温胁迫相比，转基因黄瓜与野生型黄瓜相比，膜损伤指数（MII），抗氧化酶（SOD和CAT）活性增加，叶片中游离脯氨酸含量和相对水分含量（RWC）显着降低。 。因此，表达拟南芥cbf1基因的转基因黄瓜植物具有抗寒胁迫的保护作用。

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