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马铃薯钙依赖蛋白激酶家族分析及致病疫霉感染诱导的成员StCDPK7的特性研究

Analysis of the potato calcium-dependent protein kinase family and characterization of StCDPK7, a member induced upon infection with Phytophthora infestans.

作者信息

Fantino Elisa, Segretin María Eugenia, Santin Franco, Mirkin Federico Gabriel, Ulloa Rita M

机构信息

Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor Torres" (INGEBI-CONICET) Vuelta de Obligado 2490, 2do piso, Ciudad Autónoma de Buenos Aires, Argentina.

Departamento de Fisiología, Biología Molecular y Celular Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina.

出版信息

Plant Cell Rep. 2017 Jul;36(7):1137-1157. doi: 10.1007/s00299-017-2144-x. Epub 2017 Apr 27.

DOI:10.1007/s00299-017-2144-x
PMID:28451820
Abstract

We describe the potato CDPK family and place StCDPK7 as a player in potato response to Phytophthora infestans infection, identifying phenylalanine ammonia lyase as its specific phosphorylation target in vitro. Calcium-dependent protein kinases (CDPKs) decode calcium (Ca) signals and activate different signaling pathways involved in hormone signaling, plant growth, development, and both abiotic and biotic stress responses. In this study, we describe the potato CDPK/CRK multigene family; bioinformatic analysis allowed us to identify 20 new CDPK isoforms, three CDPK-related kinases (CRKs), and a CDPK-like kinase. Phylogenetic analysis indicated that 26 StCDPKs can be classified into four groups, whose members are predicted to undergo different acylation patterns and exhibited diverse expression levels in different tissues and in response to various stimuli. With the aim of characterizing those members that are particularly involved in plant-pathogen interaction, we focused on StCDPK7. Tissue expression profile revealed that StCDPK7 transcript levels are high in swollen stolons, roots, and mini tubers. Moreover, its expression is induced upon Phytophthora infestans infection in systemic leaves. Transient expression assays showed that StCDPK7 displays a cytosolic/nuclear localization in spite of having a predicted chloroplast transit peptide. The recombinant protein, StCDPK7:6xHis, is an active Ca-dependent protein kinase that can phosphorylate phenylalanine ammonia lyase, an enzyme involved in plant defense response. The analysis of the potato CDPK family provides the first step towards the identification of CDPK isoforms involved in biotic stress. StCDPK7 emerges as a relevant player that could be manipulated to deploy disease resistance in potato crops.

摘要

我们描述了马铃薯钙依赖蛋白激酶(CDPK)家族,并确定StCDPK7在马铃薯对致病疫霉感染的反应中发挥作用,在体外鉴定出苯丙氨酸解氨酶是其特定的磷酸化靶点。钙依赖蛋白激酶(CDPKs)解码钙(Ca)信号,并激活参与激素信号传导、植物生长、发育以及非生物和生物胁迫反应的不同信号通路。在本研究中,我们描述了马铃薯CDPK/CRK多基因家族;生物信息学分析使我们能够鉴定出20种新的CDPK异构体、三种CDPK相关激酶(CRKs)和一种CDPK样激酶。系统发育分析表明,26个StCDPKs可分为四组,预计其成员会经历不同的酰化模式,并在不同组织中以及对各种刺激作出反应时表现出不同的表达水平。为了表征那些特别参与植物-病原体相互作用的成员,我们重点研究了StCDPK7。组织表达谱显示,StCDPK7转录本水平在膨大的匍匐茎、根和微型块茎中较高。此外,在系统叶中,致病疫霉感染会诱导其表达。瞬时表达分析表明,尽管StCDPK7具有预测的叶绿体转运肽,但它显示出胞质/核定位。重组蛋白StCDPK7:6xHis是一种活性钙依赖蛋白激酶,可磷酸化参与植物防御反应的苯丙氨酸解氨酶。对马铃薯CDPK家族的分析为鉴定参与生物胁迫的CDPK异构体迈出了第一步。StCDPK7成为一个相关的作用因子,可对其进行调控以在马铃薯作物中部署抗病性。

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