Identification of genes required for Cf-dependent hypersensitive cell death by combined proteomic and RNA interfering analyses.
Identification of genes required for Cf-dependent hypersensitive cell death by combined proteomic and RNA interfering analyses.
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DOI：
10.1093/jxb/err397
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发表时间：
2012-04
影响因子：
6.9
通讯作者：
Cai XZ
中科院分区：
文献类型：
--
作者：
Xu QF;Cheng WS;Li SS;Li W;Zhang ZX;Xu YP;Zhou XP;Cai XZ
过敏性细胞死亡（HCD）调节因子的鉴定是剖析植物抗病性分子机制的基础。在这项研究中，结合蛋白质组学和RNA干扰（RNAi）分析，以确定所需的基因HCD赋予番茄抗性基因CF-4和叶霉病无毒基因Avr4。通过蛋白质组学分析，确定了49个差异表达的蛋白质在番茄幼苗安装和那些不安装Cf-4/Avr4依赖HCD。其中包括多种防御相关蛋白，包括半胱氨酸蛋白酶Pip1，它是另一种C. fulvum effector，Avr2.此外，谷胱甘肽介导的抗氧化作用是对Cf-4/Avr4依赖性HCD的主要反应。通过烟草脆裂病毒诱导的基因沉默和瞬时RNAi试验对所选的16个差异表达蛋白进行功能分析，发现编码Pip1同源物NbPip1、SIPK型MAP激酶Nbf 4、天冬酰胺合成酶NbAsn、胰蛋白酶抑制剂LeMir样蛋白NbMir、小GTP结合蛋白、晚期胚胎发生样蛋白和ASR 4样蛋白的7个基因，是Cf-4/Avr4依赖性HCD所必需的。此外，前四个基因是Cf-9/Avr9依赖的HCD所必需的; NbPip1，NbAsn和NbMir，但不是Nbf4，影响非适应性细菌病原体黄单胞菌HCD。水稻诱导的本氏烟草HCD。这些数据表明，Pip1和LeMir可能在HCD和植物免疫中发挥一般作用，并且组合蛋白质组学和RNA干扰分析的应用是鉴定HCD、抗病性以及植物中可能的其他生物过程所需的基因的有效策略。
Identification of hypersensitive cell death (HCD) regulators is essential to dissect the molecular mechanisms underlying plant disease resistance. In this study, combined proteomic and RNA interfering (RNAi) analyses were employed to identify genes required for the HCD conferred by the tomato resistance gene Cf-4 and the Cladosporium fulvum avirulence gene Avr4. Forty-nine proteins differentially expressed in the tomato seedlings mounting and those not mounting Cf-4/Avr4-dependent HCD were identified through proteomic analysis. Among them were a variety of defence-related proteins including a cysteine protease, Pip1, an operative target of another C. fulvum effector, Avr2. Additionally, glutathione-mediated antioxidation is a major response to Cf-4/Avr4-dependent HCD. Functional analysis through tobacco rattle virus-induced gene silencing and transient RNAi assays of the chosen 16 differentially expressed proteins revealed that seven genes, which encode Pip1 homologue NbPip1, a SIPK type MAP kinase Nbf4, an asparagine synthetase NbAsn, a trypsin inhibitor LeMir-like protein NbMir, a small GTP-binding protein, a late embryogenesis-like protein, and an ASR4-like protein, were required for Cf-4/Avr4-dependent HCD. Furthermore, the former four genes were essential for Cf-9/Avr9-dependent HCD; NbPip1, NbAsn, and NbMir, but not Nbf4, affected a nonadaptive bacterial pathogen Xanthomonas oryzae pv. oryzae-induced HCD in Nicotiana benthamiana. These data demonstrate that Pip1 and LeMir may play a general role in HCD and plant immunity and that the application of combined proteomic and RNA interfering analyses is an efficient strategy to identify genes required for HCD, disease resistance, and probably other biological processes in plants.
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影响因子：
7.4
作者：
Brenner, ED;Lambert, KN;Williamson, VM
通讯作者：
Williamson, VM
影响因子：
4.9
作者：
Olea, F;Pérez-García, A;de Vicente, A
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de Vicente, A
影响因子：
5.1
作者：
Karrer, EE;Beachy, RN;Holt, CA
通讯作者：
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影响因子：
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作者：
JOOSTEN, MHAJ;COZIJNSEN, TJ;DEWIT, PJGM
通讯作者：
DEWIT, PJGM
影响因子：
11.6
作者：
Durrant, WE;Rowland, O;Jones, JDG
通讯作者：
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