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首页> 外文期刊>Biotechnology Progress >Attachment of Agrobacterium tumefaciens to Leaf Tissue in Response to Infiltration Conditions
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Attachment of Agrobacterium tumefaciens to Leaf Tissue in Response to Infiltration Conditions

机译：根癌农杆菌对叶组织的渗透条件的响应。

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摘要

Transient expression of recombinant proteins in plant tissues following Agrobacterium-medi-ated gene transfer is a promising technique for rapid protein production. However, transformation rates and transient expression levels can be sub-optimal depending on process conditions. Attachment of Agrobacterium tumefaciens to plant cells is an early, critical step in the gene transfer pathway. Bacterial attachment levels and patterns may influence transformation and, by extension, transient expression. In this study, attachment of A., tumefaciens to lettuce leaf tissue was investigated in response to varying infiltration conditions, including bacterial density, surfactant concentration, and applied vacuum level. Bacterial density was found to most influence attachment levels for the levels tested (10~8, 10~9, and 10~(10) CFU/mL), with the relationship between bacterial density and attachment levels following a saturation trend. Surfactant levels tested (Break-Thru S240: 1, 10, 100, and 1,000 μL/L) also had a significant positive effect on bacterial attachment while vacuum level (5,25, and 45 kPa) did not significantly affect attachment in areas exposed to bacteria. Tn planta transgene transient expression levels were measured following infiltration with 10~8, 10~9, and 10~(10) CFU/mL bacterial suspension. Notably, the highest attachment level tested led to a decrease in transient expression, suggesting a potential link between bacterial attachment levels and downstream phenomena that may induce gene silencing. These results illustrate that attachment can be controlled by adjusting infiltration conditions and that attachment levels can impact transgene transient expression in leaf tissue.
机译：农杆菌介导的基因转移后植物组织中重组蛋白的瞬时表达是快速生产蛋白质的有希望的技术。然而，取决于工艺条件，转化率和瞬时表达水平可能不是最佳的。根癌农杆菌附着于植物细胞是基因转移途径中的早期关键步骤。细菌附着水平和模式可能会影响转化，进而影响瞬时表达。在这项研究中，研究了根癌农杆菌对生菜叶片组织的附着，以响应各种渗透条件，包括细菌密度，表面活性剂浓度和施加的真空度。发现细菌密度对测试水平（10〜8、10〜9和10〜（10）CFU / mL）的附着水平影响最大，细菌密度与附着水平之间呈饱和趋势之间的关系。测试的表面活性剂水平（Break-Thru S240：1、10、100和1,000μL/ L）对细菌附着也有明显的积极影响，而真空水平（5,25和45 kPa）对暴露区域的附着没有显着影响去细菌。在用10〜8、10〜9和10〜（10）CFU / mL细菌悬液浸润后，测量了植物Tn转基因瞬时表达水平。值得注意的是，测试的最高附着水平导致瞬时表达降低，这表明细菌附着水平与可能导致基因沉默的下游现象之间存在潜在的联系。这些结果表明，可以通过调节渗透条件来控制附着，并且附着水平可以影响叶组织中转基因的瞬时表达。

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