Development of new knowledge on the biological targets of 2-hexenal with a view to its use as a bioherbicide

气候变化给农业系统带来了压力，要求它们开发更环保的方法，以减轻粮食生产中植物病原体和害虫的威胁。满足这一需求的一种方法是了解植物天然武器的作用方式和目标，并利用这些信息进行害虫和杂草的生物控制。为此，本研究将研究植物在虫害发生时释放的生物源分子2-己烯醛（2-Hex）所靶向的蛋白质，这些蛋白质可能具有化感作用，因为它被发现可以抑制杂草拟南芥（Arabidopsis thaliana）的根生长。我们推测2-Hex抑制了琥珀酸半醛脱氢酶的酶活性，这种突变严重损害了植物的生长。为了验证这一假设，我们希望达到以下三个具体目标：1)表征2-己烯六对琥珀酸半醛脱氢酶活性的影响；2)在基因组尺度上生成2- hexx反应蛋白的排序列表。这是为了验证我们的另一种假设，即2-Hex针对的是琥珀酸半醛脱氢酶以外的蛋白质；3)验证2-Hex与细胞和整个植物中选定的靶蛋白相互作用的生物学重要性。为了实现这些目标，我们将主要使用分子生物学工具和基于液相色谱和串联质谱的化学蛋白质组学方法。这些技术将被教授给新一代的研究人员，即2名博士生和3名本科生。我们将在五个物种中测试我们的发现，其中包括加拿大的三种主要栽培植物：油菜、番茄和玉米。我们的目标也是促进知识向生产者和农业转移。这就是为什么我们计划与利益相关者举行会议，包括魁北克省和安大略省的两个主要聚合商和生产者网络，以传播这项研究的结果。总的来说，该项目将允许1)识别与2-己烯醛结合的蛋白质，作为绿叶挥发性醛释放，2)设计新的生物控制杂草或综合虫害管理策略。

The climate changes put pressure on the agriculture system to develop more environmental-friendly approaches to mitigate the threats of plant pathogens and pests in food production. One way to fulfill this need is to understand the mode of action and targets of the plants' natural weapons and exploit this information for the biological control of pests and weeds. To this end, the proposed research will investigate the proteins targeted by the biogenic molecule 2-Hexenal (2-Hex) emitted by plants in the event of pest attack and which may have allelopathic properties as it was found to inhibit the root growth of the weed Arabidopsis thaliana. We hypothesize that 2-Hex inhibits the enzyme activity of the succinate semialdehyde dehydrogenase enzyme which the mutation severely impairs plant growth. To test this hypothesis, we want to achieve the following three specific aims: 1) To characterize the effect of 2-Hex on succinate semialdehyde dehydrogenase activity; 2) To generate a ranked list of 2-Hex-reactive proteins at the genome scale. This is to test our alternative hypothesis that 2-Hex targets proteins other than succinate semialdehyde dehydrogenase; 3) To validate the biological importance of the interaction of 2-Hex with the selected target proteins in the cells and the whole plant. To achieve these objectives, we will mainly use molecular biology tools and chemical proteomics approaches based on liquid chromatography coupled with tandem mass spectrometry. These techniques will be taught to a new generation of researchers, namely 2 Ph.D. students and 3 undergraduates. We will test our findings in five species including three major cultivated plants in Canada: canola, tomato, and maize. Our goal is also to facilitate knowledge transfer to the producers and the agriculture industry. That is why we plan meetings with stakeholders including two major aggregators and producers' networks in Quebec and Ontario to disseminate the outcome of this research. Overall, the project will allow 1) to identify the proteins that bind to 2-Hexenal emitted as a green leaf volatile aldehyde, and 2) to devise new strategies for biological control of weeds or integrated pest management.