Santos Cristiane, Nogueira Fábio C S, Domont Gilberto B, Fontes Wagner, Prado Guilherme S, Habibi Peyman, Santos Vanessa O, Oliveira-Neto Osmundo B, Grossi-de-Sá Maria Fatima, Jorrín-Novo Jesus V, Franco Octavio L, Mehta Angela
Embrapa Recursos Genéticos e Biotecnologia, Brasília, Brazil.
Departamento de Biologia, Universidade Federal de Juiz de Fora, Juiz de Fora, Brazil.
Front Plant Sci. 2019 Apr 12;10:414. doi: 10.3389/fpls.2019.00414. eCollection 2019.
Black rot is a severe disease caused by the bacterium pv. (Xcc), which can lead to substantial losses in cruciferous vegetable production worldwide. Although the use of resistant cultivars is the main strategy to control this disease, there are limited sources of resistance. In this study, we used the LC-MS/MS technique to analyze young cabbage leaves and chloroplast-enriched samples at 24 h after infection by Xcc, using both susceptible (Veloce) and resistant (Astrus) cultivars. A comparison between susceptible Xcc-inoculated plants and the control condition, as well as between resistant Xcc-inoculated plants with the control was performed and more than 300 differentially abundant proteins were identified in each comparison. The chloroplast enriched samples contributed with the identification of 600 additional protein species in the resistant interaction and 900 in the susceptible one, which were not detected in total leaf sample. We further determined the expression levels for 30 genes encoding the identified differential proteins by qRT-PCR. gene, encoding an endochitinase showing a high increased abundance in resistant Xcc-inoculated leaves, was selected for functional validation by overexpression in Compared to the wild type (Col-0), transgenic plants were highly resistant to Xcc indicating that gene could be an interesting candidate to be used in genetic breeding programs aiming at black rot resistance.
黑腐病是由野油菜黄单胞菌野油菜致病变种（Xcc）引起的一种严重病害，可导致全球十字花科蔬菜生产遭受重大损失。尽管使用抗病品种是控制这种病害的主要策略，但抗性来源有限。在本研究中，我们使用液相色谱-串联质谱（LC-MS/MS）技术，对接种Xcc 24小时后的幼嫩甘蓝叶片和叶绿体富集样本进行分析，使用了感病品种（Veloce）和抗病品种（Astrus）。对感病的接种Xcc植株与对照条件之间，以及抗病的接种Xcc植株与对照之间进行了比较，在每次比较中鉴定出300多种差异丰富的蛋白质。叶绿体富集样本在抗病互作中额外鉴定出600种蛋白质，在感病互作中额外鉴定出900种蛋白质，这些蛋白质在全叶样本中未检测到。我们通过qRT-PCR进一步测定了30个编码已鉴定差异蛋白质的基因的表达水平。选择了一个在抗病的接种Xcc叶片中丰度显著增加的编码内切几丁质酶的基因，通过在拟南芥中过表达进行功能验证。与野生型（Col-0）相比，转基因植物对Xcc具有高度抗性，表明该基因可能是用于旨在抗黑腐病的遗传育种计划的一个有价值的候选基因。
