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Effect of external and internal factors on the expression of reporter genes driven by the N resistance gene promoter.

机译:外部和内部因素对由N抗性基因启动子驱动的报告基因表达的影响。

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摘要

The role of resistance (R) genes in plant pathogen interaction has been studied extensively due to its economical impact on agriculture. Interaction between tobacco mosaic virus (TMV) and the N protein from tobacco is one of the most widely used models to understand various aspects of pathogen resistance. The transcription activity governed by N gene promoter is one of the least understood elements of the model. In this study, the N gene promoter was cloned and fused with two different reporter genes, one encoding beta -glucuronidase (N::GUS) and another, luciferase (N::LUC). Tobacco plants transformed with the N::GUS or N::LUC reporter constructs were screened for homozygosity and stable expression. Histochemical analysis of N::GUS tobacco plants revealed that the expression is organ specific and developmentally regulated. Whereas two week old plants expressed GUS in midveins only, 6-wk-old plants also expressed GUS in leaf lamella. Roots did not show GUS expression at any time during development. Experiments to address effects of external stress were performed using N::LUC tobacco plants. These experiments showed that N gene promoter expression was suppressed when plants were exposed to high but not low temperatures. Expression was also upregulated in response to TMV, but no changes were observed in plants treated with SA.
机译:由于其对农业的经济影响,已经广泛研究了抗性(R)基因在植物病原体相互作用中的作用。烟草花叶病毒(TMV)与烟草N蛋白之间的相互作用是了解病原体抗性各个方面的最广泛使用的模型之一。由N基因启动子控制的转录活性是该模型中鲜为人知的元素之一。在这项研究中,克隆了N基因启动子并与两个不同的报告基因融合,一个编码β-葡萄糖醛酸苷酶(N :: GUS),另一个编码荧光素酶(N :: LUC)。筛选用N :: GUS或N :: LUC报告基因构建体转化的烟草植物的纯合性和稳定表达。 N :: GUS烟草植物的组织化学分析表明,该表达具有器官特异性并受发育调控。两周龄的植物仅在中脉表达GUS,而6周龄的植物也在叶片中表达GUS。根在发育期间的任何时候均未显示GUS表达。使用N :: LUC烟草植物进行了应对外部压力影响的实验。这些实验表明,当植物暴露于高温而不是低温时,N基因启动子的表达被抑制。响应TMV,表达也被上调,但是在用SA处理的植物中未观察到变化。

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