Characterization of mutants impaired in Piriformospora indica-induced cytoplasmic calcium elevation in Arabidopsis roots

印度梨形孢子诱导拟南芥根部细胞质钙含量升高的突变体特征

• 批准号: 195458144
• 负责人: Professor Dr. Ralf Oelmüller
• 金额: --
• 依托单位: Lehrstuhl für Pflanzenphysiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/195458144?language=en
• 关键词: Characterization; mutants; impaired; Piriformospora; indica

A 129 D compound from the root-colonizing beneficial endophyte Piriformospora indica (129Piri), which mimics growth promotion of the fungus, induces cytoplasmic Ca2+ ([Ca2+]cyt) elevation and phosphatidic acid (PA) biosynthesis in Arabidopsis roots. Using transgenic Arabidopsis expressing the Ca2+ sensor aequorin, we isolated EMS mutants impaired in [Ca2+]cyt elevation to 129Piri. The Ca2+ response to flg22 is not impaired. Two classes of mutants are investigated: cam1 (ca2+ mutant1) completely lacks [Ca2+]cyt elevation in response to 129Piri, but also to the protein toxin of Alternaria brassicae (toxAlt). cam1 plants do not respond to P. indica and are highly susceptible to A. brassicae infections. caf (ca2+ friendly mutants) show biphasic [Ca2+]cyt elevation patterns in response to 129Piri, but a wildtype (WT) response to toxAlt. The aims are: (a) Chemical synthesis of 129Piri and analysis in WT and mutants, (b) continuation of map-based cloning of cam1, (c) analysis of 129Piri- and toxAlt-induced responses in cam1 including a comparative microarray analysis, (d) molecular and genetic characterization of three caf mutants, (e) MAP-based cloning of a caf gene. (f) We will test whether PA synthesis can be stimulated in the Ca2+ mutants by 129Piri and toxAlt, (g) and whether shingolipids and plastids (CAS protein) control [Ca2+]cyt elevation.

来自根定殖有益内生真菌印度梨形孢菌（Piriformospora indica，129 Piri）的129 D化合物模拟真菌的生长促进作用，诱导拟南芥根中细胞质Ca 2+（[Ca 2 +]cyt）升高和磷脂酸（PA）生物合成。使用转基因拟南芥表达的Ca 2+传感器水母发光蛋白，我们分离EMS突变体受损的[Ca 2 +]cyt升高到129 Piri。对flg 22的Ca 2+反应未受损。研究了两类突变体：cam 1（ca 2 + mupl 1）完全缺乏响应于129 Piri的[Ca 2 +]cyt升高，但也缺乏响应于芸苔链格孢菌蛋白毒素（toxAlt）的[Ca 2 +]cyt升高。cam 1植株对P. indica没有反应，对A.芸苔属侵染caf（Ca ~（2+）友好突变体）对~（129）Piri的反应显示双相[Ca ~（2+）]cyt升高模式，但对toxAlt的反应为野生型（WT）。其目标是：（a）129 Piri的化学合成以及WT和突变体中的分析，（B）cam 1的基于图的克隆的继续，（c）cam 1中129 Piri-和toxAlt-诱导的应答的分析，包括比较微阵列分析，（d）三种caf突变体的分子和遗传表征，（e）caf基因的基于图的克隆。(f)我们将测试129 Piri和toxAlt（g）是否可以刺激Ca 2+突变体中PA的合成，以及是否脂和质体（CAS蛋白）控制[Ca 2 +]细胞的升高。