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The conserved arbuscular mycorrhiza-specific transcription of the secretory lectin MtLec5 is mediated by a short upstream sequence containing specific protein binding sites

机译:分泌型凝集素MtLec5的保守丛枝菌根特异性转录是由一个短的上游序列介导的,该序列包含特定的蛋白质结合位点

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In Medicago truncatula a family of mycorrhiza-specific expressed lectins has been identified recently, but the function and regulation of these lectins during the arbuscular mycorrhiza symbiosis are still unknown. In order to characterize a first member of this protein family, MtLec5 was analyzed concerning its localization and regulation. Confocal laser scanning microscopy showed that MtLec5 is a secretory protein indicating a role as a vegetative storage protein, which is specifically expressed in mycorrhizal root systems. To study the molecular mechanisms leading to the mycorrhiza-specific transcription, deletion studies of pMtLec5 were done using reporter gene fusions. Potential cis-acting elements could be narrowed down to a 150 bp fragment that was located approximately at −300/−150 according to the transcription start, suggesting the binding of positive regulators to this area. Similar expression pattern of the reporter gene was found after transforming roots of the non-legume Nicotiana tabacum with the heterologous promoter–reporter fusions. This indicated that the observed mycorrhiza-specific transcriptional induction is not legume-specific. Electrophoretic mobility shift assays showed that several factors which were exclusively present in mycorrhizal roots bind within the 150 bp promoter area. This strengthens the hypothesis of positive regulators mediating the AM-specific gene expression.
机译:最近在Medi藜苜蓿中鉴定了一个菌根特异性表达的凝集​​素家族,但是在丛枝菌根共生期间这些凝集素的功能和调控仍是未知的。为了表征该蛋白家族的第一个成员,对MtLec5的定位和调控进行了分析。共聚焦激光扫描显微镜显示,MtLec5是一种分泌蛋白,表明它是营养存储蛋白,在菌根系统中特别表达。为了研究导致菌根特异性转录的分子机制,使用报道基因融合体进行了pMtLec5的缺失研究。潜在的顺式作用元件可根据转录起始范围缩小到大约-300 / -150处的150 bp片段,这表明阳性调节子与该区域结合。用异源启动子-报告子融合体转化非豆科植物烟草的根后,发现了报告基因的类似表达模式。这表明观察到的菌根特异性转录诱导不是豆科植物特异性的。电泳迁移率变动分析表明,菌根中仅存在的几种因子在150 bp启动子区域内结合。这加强了介导AM特异性基因表达的正调控因子的假设。

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