Mining the genome of Arabidopsis thaliana as a basis for the identification of novel bioactive peptides involved in oxidative stress tolerance
Mining the genome of Arabidopsis thaliana as a basis for the identification of novel bioactive peptides involved in oxidative stress tolerance
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DOI：
10.1093/jxb/ert295
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发表时间：
2013-12-01
影响因子：
6.9
通讯作者：
Cammue, Bruno P. A.
中科院分区：
文献类型：
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作者：
De Coninck, Barbara;Carron, Delphine;Cammue, Bruno P. A.
尽管已经积累了关于植物肽在响应外部条件中的作用的证据，但基因组中肽编码基因的数量仍然被低估。使用平铺阵列，我们确定了176个未注释的转录活性区（TARs）在拟南芥诱导后产生的除草剂百草枯（PQ）的氧化应激。这176个TAR可以翻译成575个推定的氧化应激诱导肽（OSIP）。在真核生物模式生物酿酒酵母（Saccharomyces cerevisiae）中使用高通量功能测定，使我们能够测试增加氧化应激耐受性的生物活性肽。通过这种方式，我们确定了三种OSIP，在酵母中过表达后，导致对过氧化氢（H2O2）的耐受性显着上升。对于这些肽之一，十肽OSIP108，外源性应用于H2O2处理的酵母也导致存活率显着增加。OSIP108包含在假基因中，并在A.通过活性氧诱导剂PQ和坏死营养型真菌病原体灰葡萄孢（Botrytis cinerea），OSIP108在A.拟南芥叶片对PQ处理的耐受性增加。总之，OSIP108的鉴定和表征证实了我们基于A.和酵母中的功能筛选，以鉴定生物活性肽。
Although evidence has accumulated on the role of plant peptides in the response to external conditions, the number of peptide-encoding genes in the genome is still underestimated. Using tiling arrays, we identified 176 unannotated transcriptionally active regions (TARs) in Arabidopsis thaliana that were induced upon oxidative stress generated by the herbicide paraquat (PQ). These 176 TARs could be translated into 575 putative oxidative stress-induced peptides (OSIPs). A high-throughput functional assay was used in the eukaryotic model organism Saccharomyces cerevisiae allowing us to test for bioactive peptides that increase oxidative stress tolerance. In this way, we identified three OSIPs that, upon overexpression in yeast, resulted in a significant rise in tolerance to hydrogen peroxide (H2O2). For one of these peptides, the decapeptide OSIP108, exogenous application to H2O2-treated yeast also resulted in significantly increased survival. OSIP108 is contained within a pseudogene and is induced in A. thaliana leaves by both the reactive oxygen species-inducer PQ and the necrotrophic fungal pathogen Botrytis cinerea. Moreover, infiltration and overexpression of OSIP108 in A. thaliana leaves resulted in increased tolerance to treatment with PQ. In conclusion, the identification and characterization of OSIP108 confirms the validity of our high-throughput approach, based on tiling array analysis in A. thaliana and functional screening in yeast, to identify bioactive peptides.