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Analysis of plant gene expression responses to the pathogen and natural genetic engineer Agrobacterium tumefaciens.

机译:分析植物基因表达对病原体和自然遗传工程农杆菌的影响。

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摘要

Agrobacterium tumefaciens genetically transforms its natural plant host and other eukaryotic cells, representing both an intriguing pathogen and a vehicle for genetic engineering. Host responses to this bacterium have not been studied extensively in the past, although this understanding is critical for both practical applications in biotechnology and insights into host-microbe interaction. Here we present a study of plant gene expression responses to Agrobacterium infection. Two different plant species and two different large-scale approaches have been used to identify genes with altered expression during infection. By cDNA-AFLP we identified several transcripts from Ageratum conyzoides cell cultures that were regulated at 24 and 48 hours after infection with Agrobacterium . A number of these transcripts code for plant defense-related proteins which are also regulated by a non-pathogenic bacterium. However, a nodulin-like gene was regulated uniquely by Agrobacterium, suggesting that the closely-related symbiotic Rhizobium and Agrobacterium can provoke similar responses in plants. We observed that an attachment-deficient Agrobacterium mutant hyper-induces a set of plant defense genes and we propose that Agrobacterium can dampen plant defense responses by an attachment-dependent mechanism. The plant defense system is likely important in regulating infection since Ageratum cells with heightened defenses hinder transformation. By using microarrays, we also investigated the transcriptome of Arabidopsis thaliana cell cultures during a time-course of infection by Agrobacterium. Statistically significant alterations in gene expression are observed at 48 hours after infection, but not at earlier time points. The identity of the differentially expressed genes suggests similarities with classes of genes characterized in our previous studies and with other defense-related genes. Future microarray studies comparing infection with other pathogens, symbionts and various Agrobacterium mutants will further advance our understanding of this fascinating plant-bacterium interaction.
机译:根癌农杆菌可遗传转化其天然植物宿主和其他真核细胞,既是一种引人入胜的病原体,又是遗传工程的载体。尽管对生物技术的实际应用和对宿主-微生物相互作用的了解都是至关重要的,但是过去对宿主对这种细菌的反应尚未进行广泛的研究。在这里,我们提出对农杆菌感染的植物基因表达反应的研究。已经使用两种不同的植物物种和两种不同的大规模方法来鉴定在感染过程中表达改变的基因。通过cDNA-AFLP,我们鉴定了从香叶香菇细胞培养物中获得的几种转录本,这些转录本在土壤杆菌感染后24和48小时受到调控。这些转录物中的许多编码植物防御相关蛋白,其也由非病原性细菌调节。然而,农杆菌独特地调节一种结节蛋白样基因,这表明密切相关的共生根瘤菌和农杆菌可以在植物中引起类似的反应。我们观察到,一个附着缺陷的农杆菌突变体过度诱导了一组植物防御基因,并且我们提出土壤杆菌可以通过一个依附依赖性机制来抑制植物防御反应。植物防御系统在调节感染中可能很重要,因为防御能力增强的香叶细胞会阻碍转化。通过使用微阵列,我们还研究了农杆菌感染的过程中拟南芥细胞培养的转录组。感染后48小时观察到基因表达有统计学上的显着变化,但在较早的时间点未观察到。差异表达基因的同一性暗示与我们先前研究中表征的基因类别以及与其他防御相关基因的相似性。将来将感染与其他病原体,共生体和各种农杆菌突变体进行比较的微阵列研究将进一步提高我们对这种迷人的植物-细菌相互作用的理解。

著录项

  • 作者

    Ditt, Renata Fava.;

  • 作者单位

    University of Washington.;

  • 授予单位 University of Washington.;
  • 学科 Biology Botany.; Biology Molecular.; Biology Plant Physiology.
  • 学位 Ph.D.
  • 年度 2004
  • 页码 110 p.
  • 总页数 110
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 植物学;分子遗传学;植物学;
  • 关键词

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