农杆菌毒力的定量解析以构建合成Ti质粒

摘要(中文译文)

它不仅是代价高昂的植物病原体，也是植物转化的重要工具。尽管人们对农杆菌介导的转化（AMT）已进行了大量研究，但其多基因特性及复杂的转录调控使得通过传统遗传学和生物信息学方法难以确定转化效率的遗传基础。在此，我们采用自下而上的合成方法，对携带致瘤质粒（pTi）进行系统性工程改造，其中编码了大部分毒力机制。利用一套经过验证的工具包来控制基因表达，我们对AMT进行了定量解析，以研究关键基因在不同表达水平下的贡献。我们构建了一个能够实现植物瞬时转化和真菌稳定转化的合成pTi，并刻画了复杂多基因合成pTi设计中的瓶颈及解决方案。我们的还原论方法展示了如何利用自下而上的工程手段来解析和阐明复杂生物学性状的遗传基础，为未来工程化改造以实现对AMT这一关键过程的完全合成控制奠定了基础。

Abstract

is not only a costly plant pathogen but is also an essential tool for plant transformation. Though -mediated transformation (AMT) has been heavily studied, its polygenic nature and complex transcriptional regulation make identification of the genetic basis of transformational efficiency difficult through traditional genetic and bioinformatic approaches. Here, we use a bottom-up synthetic approach to systematically engineer the tumor-inducing plasmid (pTi), wherein the majority of virulence machinery is encoded. Using a validated toolkit to control gene expression , we perform a quantitative dissection of AMT to investigate the contributions of critical -genes at different expression levels. We construct a synthetic pTi capable of transient plant and stable fungal transformation and characterize bottlenecks and solutions for complex polygenic synthetic pTi designs. Our reductionist approach demonstrates how bottom-up engineering can be used to dissect and elucidate the genetic underpinnings of complex biological traits, laying the foundation for future engineering to establish full synthetic control over the critical process of AMT.

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