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From the Cover: A Ca2+/calmodulin-dependent protein kinase required for symbiotic nodule development: Gene identification by transcript-based cloning

机译:从封面:共生结节发展所需的Ca2 + /钙调蛋白依赖性蛋白激酶:通过基于转录本的克隆进行基因鉴定

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摘要

In the establishment of the legume-rhizobial symbiosis, bacterial lipochitooligosaccharide signaling molecules termed Nod factors activate the formation of a novel root organ, the nodule. Nod factors elicit several responses in plant root hair cells, including oscillations in cytoplasmic calcium levels (termed calcium spiking) and alterations in root hair growth. A number of plant mutants with defects in the Nod factor signaling pathway have been identified. One such Medicago truncatula mutant, dmi3, exhibits calcium spiking and root hair swelling in response to Nod factor, but fails to initiate symbiotic gene expression or cell divisions for nodule formation. On the basis of these data, it is thought that the dmi3 mutant perceives Nod factor but fails to transduce the signal downstream of calcium spiking. Additionally, the dmi3 mutant is defective in the symbiosis with mycorrhizal fungi, indicating the importance of the encoded protein in multiple symbioses. We report the identification of the DMI3 gene, using a gene cloning method based on transcript abundance. We show that transcript-based cloning is a valid approach for cloning genes in barley, indicating the value of this technology in crop plants. DMI3 encodes a calcium/calmodulin-dependent protein kinase. Mutants in pea sym9 have phenotypes similar to dmi3 and have alterations in this gene. The DMI3 class of proteins is well conserved among plants that interact with mycorrhizal fungi, but it is less conserved in Arabidopsis thaliana, which does not participate in the mycorrhizal symbiosis.
机译:在豆科植物-根瘤菌共生的建立中,被称为Nod因子的细菌脂低聚寡糖信号分子激活了新的根器官即结节的形成。点头因子在植物根毛细胞中引起多种反应,包括细胞质钙水平的振荡(称为钙峰值)和根毛生长的变化。已经鉴定出许多在Nod因子信号传导途径中具有缺陷的植物突变体。一种这样的紫花苜蓿截枝突变体dmi3,响应Nod因子而表现出钙尖峰和根毛肿胀,但未能启动共生基因表达或细胞分裂以形成结节。根据这些数据,认为dmi3突变体感知到Nod因子,但是不能转导钙突增的下游信号。此外,dmi3突变体在与菌根真菌共生中存在缺陷,表明编码的蛋白在多种共生酶中的重要性。我们报告使用基于转录本丰度的基因克隆方法鉴定DMI3基因。我们表明基于转录本的克隆是克隆大麦基因的有效方法,表明该技术在农作物中的价值。 DMI3编码钙/钙调蛋白依赖性蛋白激酶。豌豆sym9中的突变体具有类似于dmi3的表型,并且在该基因中具有改变。在与菌根真菌相互作用的植物中,DMI3类蛋白非常保守,但在不参与菌根共生的拟南芥中,其保守性较低。

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