Identification of endogenous small peptides involved in rice immunity through transcriptomics- and proteomics-based screening
Identification of endogenous small peptides involved in rice immunity through transcriptomics- and proteomics-based screening
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通过基于转录组学和蛋白质组学的筛选鉴定参与水稻免疫的内源小肽
DOI：
10.1111/pbi.13208
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发表时间：
2019-08-08
影响因子：
13.8
通讯作者：
Kawano, Yoji
中科院分区：
文献类型：
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作者：
Wang, Pingyu;Yao, Shaolun;Kawano, Yoji
由较大蛋白质前体产生的小信号肽是协调各种植物过程（例如发育和免疫反应）的重要组成部分。然而，参与植物免疫的小信号肽仍然很大程度上未知。在这里，我们开发了一个使用基于转录组学和蛋白质组学的筛选的管道，以鉴定小信号肽的推定前体：水稻中的小分泌蛋白（SSP），由稻瘟菌 Magnaporthe oryzae 及其诱导子几丁质诱导。我们鉴定了 236 个 SSP，包括两个已知的小信号肽家族的成员，即快速碱化因子和植物硫因子，以及许多其他已知参与免疫的蛋白质家族，例如蛋白酶抑制剂和发病机制相关蛋白质家族。我们还分离了 52 个未注释的 SSP，其中我们发现了一个基因，我们将其命名为免疫反应肽 (IRP)，它似乎编码调节水稻免疫的小信号肽的前体。在水稻悬浮细胞中，IRP 的表达是由细菌肽聚糖和真菌几丁质诱导的。 IRP 的过度表达增强了防御基因 PAL1 的表达，并诱导水稻悬浮细胞中 MAPK 的激活。此外，几丁质处理后悬浮细胞培养基中IRP蛋白水平增加。总的来说，我们建立了一个简单而有效的流程来发现可能在水稻免疫中发挥重要作用的 SSP 候选者，并鉴定了 52 个未注释的 SSP，它们可能有助于进一步阐明水稻免疫。我们的方法可用于识别不仅涉及免疫而且还涉及其他植物功能的 SSP。
Small signalling peptides, generated from larger protein precursors, are important components to orchestrate various plant processes such as development and immune responses. However, small signalling peptides involved in plant immunity remain largely unknown. Here, we developed a pipeline using transcriptomics- and proteomics-based screening to identify putative precursors of small signalling peptides: small secreted proteins (SSPs) in rice, induced by rice blast fungus Magnaporthe oryzae and its elicitor, chitin. We identified 236 SSPs including members of two known small signalling peptide families, namely rapid alkalinization factors and phytosulfokines, as well as many other protein families that are known to be involved in immunity, such as proteinase inhibitors and pathogenesis-related protein families. We also isolated 52 unannotated SSPs and among them, we found one gene which we named immune response peptide (IRP) that appeared to encode the precursor of a small signalling peptide regulating rice immunity. In rice suspension cells, the expression of IRP was induced by bacterial peptidoglycan and fungal chitin. Overexpression of IRP enhanced the expression of a defence gene, PAL1 and induced the activation of the MAPKs in rice suspension cells. Moreover, the IRP protein level increased in suspension cell medium after chitin treatment. Collectively, we established a simple and efficient pipeline to discover SSP candidates that probably play important roles in rice immunity and identified 52 unannotated SSPs that may be useful for further elucidation of rice immunity. Our method can be applied to identify SSPs that are involved not only in immunity but also in other plant functions.