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PTK1是一种丝裂原活化蛋白激酶基因,它是大麦网斑病菌分生孢子形成、附着胞形成及致病性所必需的。

PTK1, a mitogen-activated-protein kinase gene, is required for conidiation, appressorium formation, and pathogenicity of Pyrenophora teres on barley.

作者信息

Ruiz-Roldán M C, Maier F J, Schäfer W

机构信息

Institute of General Botany, Department of Molecular Phytopathology and Genetics (AMP III), University of Hamburg, Germany.

出版信息

Mol Plant Microbe Interact. 2001 Feb;14(2):116-25. doi: 10.1094/MPMI.2001.14.2.116.

Abstract

Mitogen-activated protein kinases (MAPKs) are a group of protein kinases that execute a wide variety of roles in cellular signal transduction pathways such as osmoregulation, cell wall biosynthesis, growth, and differentiation. A polymerase chain reaction (PCR) with degenerate primers based on conserved regions of known MAPKs was used to clone the MAPK gene PTK1 from the leaf pathogen Pyrenophora teres (anamorph Drechslera teres), the causal agent of net blotch of barley (Hordeum vulgare L.). The predicted amino acid sequence shows high homology with MAPKs from other phytopathogenic fungi. The gene is present in the genome as a single copy. PTK1 is expressed during in vitro growth on complete medium, under conidiation-inducing conditions and during infection of barley leaves, as shown by reverse transcription-PCR studies. In order to assess the role of PTK1 in the life cycle of P. teres, targeted gene disruption was conducted. Mutants carrying an interrupted copy of the gene were deficient in conidiation, did not form appressoria on glass surfaces or on barley leaves, lost their ability to infect barley leaves, and could not colonize host tissues following artificial wounding.

摘要

丝裂原活化蛋白激酶(MAPKs)是一类蛋白激酶,在细胞信号转导途径中发挥多种作用,如渗透调节、细胞壁生物合成、生长和分化。基于已知MAPKs保守区域的简并引物进行聚合酶链反应(PCR),用于从大麦网斑病病原菌——叶部病原菌网斑核腔菌(无性型为圆突状德氏霉)中克隆MAPK基因PTK1。预测的氨基酸序列与其他植物病原真菌的MAPKs具有高度同源性。该基因在基因组中以单拷贝形式存在。逆转录PCR研究表明,PTK1在完全培养基上的体外生长过程中、在分生孢子诱导条件下以及在感染大麦叶片期间均有表达。为了评估PTK1在网斑核腔菌生命周期中的作用,进行了靶向基因敲除。携带该基因中断拷贝的突变体在分生孢子形成方面存在缺陷,在玻璃表面或大麦叶片上不形成附着胞,失去了感染大麦叶片的能力,并且在人工创伤后无法定殖于宿主组织。

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