Recombinant cystatins in plants

植物中的重组半胱氨酸蛋白酶抑制剂

• 批准号: RGPIN-2022-03997
• 负责人: Michaud, Dominique
• 金额: $ 3.42万
• 依托单位: Université Laval
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=763524
• 关键词: Recombinant; cystatins; plants

Protease inhibitors have long been recognized as potential biopesticides for the development of pest-resistant crops. In recent years, studies have also described indirect effects for these proteins on the plant's own physiology, potentially useful to introduce such new traits of agronomical value as pathogen, cold or drought tolerance. Taking advantage of recent developments in functional proteomics and protein engineering, this project aims at further understanding the effects of recombinant protease inhibitors in plants, and at expanding their usefulness to protect plants in drought conditions. Protease inhibitors of the plant cystatin protein family will be used as model for the project, given the number of recent reports suggesting their positive impact on plants under stress conditions. Candidate cystatins with different affinities for plant proteases will first be isolated among a collection of cystatin variants designed based on a novel protein design approach developed during the last funding cycle of this Discovery program. The selected variants will then be used for expression studies to test the feasibility of triggering variable effects in the plant depending on each variant's target proteases. Finally, the most promising variants will be used to implement drought tolerance in potato selected as a drought-susceptible crop model of major agronomic significance. A first output of this project will be a novel way to implement drought stress tolerance in crop plants, in the current climate change context where an increased incidence of drought episodes is likely to affect several major crops worldwide, including potato. Another output will be a proof-of-concept of our ability to rationally controlling the fine balance between intended and unintended effects of foreign proteins in genetically modified crops using the new tools of protein design and functional proteomics.

蛋白酶抑制剂一直被认为是开发抗虫作物的潜在生物农药。近年来，研究还描述了这些蛋白质对植物自身生理的间接影响，可能有助于引入具有农学价值的新性状，如病原体，耐寒性或耐旱性。利用功能蛋白质组学和蛋白质工程的最新发展，本项目旨在进一步了解重组蛋白酶抑制剂在植物中的作用，并扩大其在干旱条件下保护植物的用途。植物半胱氨酸蛋白酶抑制剂蛋白家族的蛋白酶抑制剂将被用作该项目的模型，因为最近的一些报告表明它们对植物在胁迫条件下的积极影响。对植物蛋白酶具有不同亲和力的候选半胱氨酸蛋白酶抑制剂将首先从一系列半胱氨酸蛋白酶抑制剂变体中分离出来，这些变体是基于本发现计划最后一个资助周期期间开发的新型蛋白质设计方法设计的。然后将选择的变体用于表达研究，以测试根据每个变体的靶蛋白酶在植物中触发可变效应的可行性。最后，最有前途的变种将被用来实现耐旱性在马铃薯中选择作为一个干旱敏感的作物模型的主要农艺意义。该项目的第一个产出将是在当前气候变化的背景下，在作物中实施干旱胁迫耐受性的新方法，在这种背景下，干旱事件的发生率增加可能会影响全球几种主要作物，包括马铃薯。另一个产出将是我们使用蛋白质设计和功能蛋白质组学的新工具合理控制转基因作物中外源蛋白质的预期和非预期影响之间的精细平衡的能力的概念验证。