Dissecting heat stress responses in barley

背景：气候条件变化对全球农作物生产的影响。我们需要增加人口来养活世界，但同时要减少对化肥使用的依赖。一种解决方案是开发能够改善土壤资源捕获的品种，从而在次优条件下提高产量。全球农作物产量受到干旱、盐碱和内涝等非生物胁迫的影响。泛素通过PRT6N-Degron途径中的植物半胱氨酸氧化酶(PCO)分支调控参与感受和/或赋予非生物胁迫耐受性的蛋白质的周转，从而控制植物的反应。根皮质衰老(RCS)。皮层衰老对此的重要性，特别是提高产量，同时减少在非生物胁迫下维持皮质组织的代谢负担。RCS是一个非常重要的土壤耐逆性育种目标。对控制这一过程的基因和机制缺乏基本的理解。有趣的是，我们最近观察到PRT6N-degron途径的突变使RCS增加。本项目重点研究新发现的和尚未鉴定的N-Degron途径底物，以阐明其特定的下游应激调节功能，为应激恢复研究提供新的机会。我们之前已经确定了两种这样的底物，BERF1和RAF；这个项目将研究它们作为环境压力关键调节因子的具体作用。最近的一篇论文显示，RAF和BERF1在转录分析中表达上调，这表明它们在根皮质衰老(RCS)的发育过程中发挥了重要作用。因此，在这个DTP项目中，我们的目标是剖析这一点，以提高我们对控制这一过程的基因的基本理解，并识别新的遗传成分/解决方案，以帮助我们开发耐非生物胁迫的品种。工作计划：第1年：绘制突变体和WT中干旱期间根皮质衰老的发展图。确定取样的区域和时间点，对对照和干旱胁迫下的突变体和WT进行RNASEQ，并取样不同的区域，包括RCS形成，以了解基因的表达。根据以前的工作和它在干旱等方面的相关性，优先考虑一些候选基因。今年2-3：开发CRISPR、RNAi、报告系，以验证PRT6 N-degron途径在调控根皮质衰老中的下游靶点。3-4年级。在Hounsfield设施的土柱中进行干旱实验。采集根部进行激光消融断层扫描，看看突变体中RCS的形成是否受到损害，以及它对在突变体与WT中形成RCS的成本效益的影响。使用Hounsfield设施中的PhenoSpex性状探测仪研究地上部的生理参数。第4年：向育种公司部署新的遗传解决方案。项目产出和影响：该项目侧重于新发现的和尚未确定的N-Degron途径底物，以阐明其特定的下游压力调节功能，为压力弹性研究提供新的机会。因此，该项目将基础研究与应用相结合，以了解非生物逆境恢复能力的驱动因素，并应用于培育可恢复能力的大麦用于部署。

Background: Impact of changing climatic conditions on global crop production. We need to increase the population to feed the world but while reducing the reliance on fertiliser application. One solution is to develop varieties that have improved soil resource capture and thus improved yield under suboptimal conditions. Crop yields are globally affected by abiotic stresses such as drought, salinity and waterlogging. Ubiquitin-mediated proteolysis via the Plant Cysteine Oxidase (PCO) branch of the PRT6 N-Degron pathway controls the plant response by regulating the turnover of proteins involved in sensing and/or conferring tolerance to abiotic stresses. Root cortical senescence (RCS). Importance of cortical senescence towards this especially improving yield while reducing metabolic burden of maintaining cortical tissues under abiotic stresses. RCS is a very important breeding target for edaphic stress tolerance. The fundamental understanding about genes and mechanisms controlling this process is missing. Interestingly, we recently observed that mutants in PRT6 N-degron pathway have increased RCS. This project focuses on newly discovered and uncharacterized N-Degron pathway substrates to shed light on their specific downstream stress regulation functions, offering new opportunities for stress resilience research. We have previously identified two such substrates, BERF1 and RAF; this project will study their specific roles as key regulators of environmental stress. A recent paper showed RAF and BERF1 were upregulated in a transcriptional analysis, which indicated their important role of these gene in the development of root cortical senescence (RCS). Thus, in this DTP project we aim to dissect this to improve our fundamental understanding about genes controlling this process and identify novel genetic components/solutions that could help us develop varieties tolerant to abiotic stresses.Work plan: Year 1: Map the development of root cortical senescence during drought in the mutant and WT. Decide zones and timepoints for sampling roots and perform RNAseq of mutant vs WT under control and drought stress and sample different zones, including RCS formation to understand genes expressed. Prioritise some candidates' genes based on previous work and its relevance in drought, etc.Year 2-3: Develop CRISPR, RNAi, reporter lines to validate downstream targets of PRT6 N-degron pathway functioning in regulating root cortical senescence. Year 3-4. Perform drought experiment in soil columns in Hounsfield facility. Harvest roots for Laser ablation tomography and see whether RCS formation is impaired in the mutant and its impact on the cost benefit of forming RCS in the mutant vs WT. Also study shoot physiological parameters using PhenoSpex trait finder in the Hounsfield facility. Year 4: Deploy novel genetic solutions to breeding companies. Project output and impact: This project focuses on newly discovered and uncharacterized N-Degron pathway substrates to shed light on their specific downstream stress regulation functions, offering new opportunities for stress resilience research. This project therefore combines fundamental research into understanding the drivers for abiotic stress resilience, with application to generate resilient barley for deployment.