Collaborative Research: RUI: RESEARCH-PGR Meeting Future Food Demands: Phosphoproteomics to Unravel Signaling Pathways in Soybean's Response to Phosphate and Iron Deficiency

大豆是美国和世界范围内的一种重要作物，主要用于食用油脂和作为食物来源。大豆作物产量对美国和全球经济至关重要，但铁和磷缺乏是美国许多大豆种植区的常见问题，这可能会严重限制大豆产量。磷酸盐是一种有限的资源，而我们面临着在几个世纪内耗尽可获得的(可开采的)岩石磷肥的风险，这一事实加剧了这个问题。这项拟议研究的目标是了解大豆如何感知并立即对磷和铁缺乏做出反应。我们的方法是基于一种名为定量磷酸蛋白质组学的技术，该技术已成功地应用于识别植物中的其他信号途径，但尚未应用于磷或铁缺乏信号转导。更好地了解对两种主要营养胁迫的早期反应可能有助于培育对化肥需求较少的大豆品系。该项目将通过提供实践研究和传播经验，直接影响科学、技术、工程和数学专业的本科生和硕士学生。这反过来又将帮助学生在当地的生物技术行业就业，他们大多属于代表人数较少的少数群体。避免粮食危机是当今世界面临的最大挑战之一，需要提高作物对养分的吸收和利用。磷和铁的缺乏限制了美国和世界各地的大豆产量。尽管已有大量研究致力于确定植物对磷和铁缺乏的反应，但对植物如何感知和发出这些缺乏的信号知之甚少。这是因为大多数实验方法都集中在基因表达的变化上。然而，信号转导成分，虽然通常没有差异表达，但经常是差异磷酸化。我们将把定量磷蛋白组学应用于大豆，以确定短期磷和铁缺乏反应的差异磷酸化蛋白。然后，我们将通过产生RNAi敲除突变体并分析这些受损的营养信号反应来确认潜在的信号作用。虽然定量磷蛋白质组学已经成功地应用于揭示植物逆境反应中的其他信号转导途径，但尚未应用于磷或铁缺乏信号转导。这项拟议的研究将有助于弥合大豆中与营养耐受性有关的已知基因和基因之间的差距，以及潜在的信号通路。从长远来看，更好地了解信号转导和整合各种营养胁迫反应的网络有助于基于系统的方法来培育或生物工程大豆，从而提高大豆的抗逆性、生产力和减少对化肥的需求。我们的教育目标是培训本科生和研究生，他们大多属于STEM中代表性较低的少数群体，在实践研究和传播方面。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Soybean is an important crop in the US and worldwide, grown primarily for its oil and as a food source. Soybean crop yield is critical for the US and global economy, but iron and phosphate deficiencies are common problems in many US soybean-growing regions, and this can severely limit soybean production. This problem is exacerbated by the fact that phosphate is a finite resource, and we are at risk of running out of accessible (minable) rock phosphate fertilizer within centuries. The objective of the proposed research is to understand how soybean senses and immediately responds to phosphate and iron deficiencies. Our approach is based on a technique called quantitative phosphoproteomics which has been successfully applied to identify other signaling pathways in plants but has not yet been applied to phosphate or iron deficiency signaling. A better understanding of early responses to two major nutrient stresses could help to develop soybean lines with reduced need for fertilizers. The project will directly impact undergraduate and master students in Science, Technology, Engineering, and Mathematics by providing experience in hands-on research and dissemination. This in turn will help students, mostly belonging to underrepresented minority groups, to obtain employment in the local biotech industry. Avoiding a food crisis is one of the greatest challenges facing the world today, requiring crops with improved uptake and utilization of nutrients. Phosphate and iron deficiencies limit soybean production in the US and worldwide. Although significant research has been devoted to identifying plant responses to phosphate and iron deficiency, far less is known about how plants sense and signal these deficiencies. This is because most experimental approaches have focused on changes in gene expression. However, signal transduction components, while usually not differentially expressed, are frequently differentially phosphorylated. We will apply quantitative phosphoproteomics to soybeans to identify differentially phosphorylated proteins in response to short-term phosphate and iron deficiency. We will then confirm potential signaling roles by generating RNAi knockdown mutants and analyzing these for impaired nutrient signaling responses. While quantitative phosphoproteomics has been successfully applied to unravel other signal transduction pathways in plant stress responses, it has not yet been applied to phosphate or iron deficiency signaling. The proposed research will help bridge the gap between known loci and genes involved in nutrient tolerance in soybeans and the underlying signaling pathways. In the long term, a better understanding of signal transduction and networks integrating various nutrient stress responses could aid system-based approaches to breed or bioengineer soybeans with increased stress tolerance, productivity, and reduced need for fertilizers. Our educational objective is to train undergraduate and graduate students, most belonging to underrepresented minority groups in STEM, in hands-on research and dissemination.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.