Unraveling the genetic basis of amino acid composition in dry Arabidopsis seeds

揭示干燥拟南芥种子氨基酸组成的遗传基础

• 批准号: 1754201
• 负责人: Ruthie Angelovici
• 金额: $ 82.78万
• 依托单位: University of Missouri-Columbia
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1754201&HistoricalAwards=false
• 关键词: Unraveling; genetic; basis; amino; acid

Essential amino acids are necessary for human health but must come from dietary sources, as the human body cannot produce them. A lack of these essential amino acids in a diet leads to malnutrition. Much of the world's human and livestock populations rely on seeds for protein, but most staple crops seeds are deficient in several essential amino acids. Efforts to fortify the amino acid composition of staple crop seeds have had limited success since plants respond to induced protein composition alterations by activating a regulatory mechanism that "resets" it back to an original state. While beneficial to the plant growth and development, this mechanism has been a major hurdle to biofortification efforts. Fortunately, it is also known that different cultivars/varieties display natural variation in their seed amino acid composition, which means that the regulation of these compounds is genetically driven and thus should be amenable to manipulation. Therefore, this project aims to identify the key genes involved in this regulatory mechanism by analyzing hundreds of Arabidopsis strains, which display natural genetic and seed amino acid composition variation. Subsequently the potential role of the identified candidate genes will be tested using classical physiological and genetic approaches. Identifying new genetics targets will help facilitate the breeding of new crop varieties that can support our nation's and world's growing population.It is important to identify the genes that underlie the amino acid composition of dry seeds. Doing so will shed new light on a complex regulatory mechanism in plants as well as facilitate efforts to fortify staple seed crops. While much is known about amino acid metabolic pathways, little is known about their regulation, especially in seeds. So far, it is known that alterations to the composition of seed storage proteins (i.e., the major amino acid sink) lead to proteomic reprogramming and activation of a rebalancing mechanism, a response that suggests tight regulation of this trait. Seed amino acid composition varies substantially across genotypes within the same genus. This observation implies that the genetic architecture responsible for this natural variation includes, at least in part, the genes involved in responding to amino acid sink alteration and activation of the rebalancing mechanism. This project aims to elucidate key components of this genetic architecture in Arabidopsis thaliana using two approaches: (1) a GWAS on amino acid composition measured from dry seeds of an 800-accession association panel and (2) an amino acid/gene expression correlation-based network analysis of wildtype and two storage protein mutants with active rebalancing phenotypes. The candidate genes identified from each approach will be compared, and the top candidate genes will be tested for involvement in amino acid regulation via knockdown and overexpression experiments. The identified genetic regulators will provide new targets for breeding programs.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.

必需的氨基酸对于人类健康是必需的，但必须来自饮食来源，因为人体无法产生它们。饮食中缺乏这些必需氨基酸会导致营养不良。世界上许多人类和牲畜种群都依赖种子来蛋白质，但大多数主食种子在几种必需的氨基酸中都缺乏。固定基本种子种子的氨基酸组成的努力取得了有限的成功，因为植物通过激活将其“重置”回原始状态的调节机制来应对诱导的蛋白质组成改变。虽然对植物的生长和开发有益，但这种机制一直是生物进行努力的主要障碍。幸运的是，众所周知，不同的品种/品种在其种子氨基酸组成中表现出自然变化，这意味着这些化合物的调节是遗传驱动的，因此应适合操纵。因此，该项目旨在通过分析数百种显示自然遗传和种子氨基酸组成变化的拟南芥菌株来鉴定该调节机制所涉及的关键基因。 随后，将使用经典的生理和遗传方法测试已确定的候选基因的潜在作用。确定新的遗传学目标将有助于促进可以支持我们国家和世界增长的新作物品种的繁殖，这对于识别基于干燥种子氨基酸组成的基因很重要。这样做将为植物中复杂的调节机制提供新的启示，并有助于促进固定种子作物的努力。虽然对氨基酸代谢途径知之甚少，但对它们的调节，尤其是在种子中，知之甚少。到目前为止，众所周知，对种子储存蛋白组成（即主要氨基酸下水道）的组成改变会导致蛋白质组学重编程和重新平衡机制的激活，这一反应表明对这种性状的严格调节。种子氨基酸组成在同一属内的基因型之间有很大的变化。该观察结果表明，负责这种自然变异的遗传结构至少部分包括响应氨基酸下水道改变和重新平衡机制的激活所涉及的基因。该项目的目的是使用两种方法阐明这种遗传结构的关键组成部分：（1）从800个分析关联面板的干燥种子和（2）基于氨基酸/基因表达相关的基于Wildtype和两个具有活性反复平衡现象的氨基酸/基因表达相关网络分析的GWA氨基酸组合物和（2）基于氨基酸/基因表达相关的网络分析。将比较从每种方法中鉴定出的候选基因，并将通过敲低和过表达实验测试顶部候选基因以参与氨基酸调节。 已确定的遗传调节器将为育种计划提供新的目标。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛影响的评论标准来评估值得支持的。

项目成果

HAPPI GWAS: Holistic Analysis with Pre- and Post-Integration GWAS

HAPPI GWAS：集成前后 GWAS 的整体分析

• DOI: 10.1093/bioinformatics/btaa589
• 发表时间: 2020
• 期刊: Bioinformatics
• 影响因子: 5.8
• 作者: Slaten, Marianne L;Chan, Yen On;Shrestha, Vivek;Lipka, Alexander E;Angelovici, Ruthie
• 通讯作者: Angelovici, Ruthie

A High-Throughput Absolute-Level Quantification of Protein-Bound Amino Acids in Seeds

种子中蛋白质结合氨基酸的高通量绝对水平定量

• DOI: 10.1002/cppb.20084
• 发表时间: 2018
• 期刊: Current Protocols in Plant Biology
• 作者: Yobi, Abou;Angelovici, Ruthie
• 通讯作者: Angelovici, Ruthie

A High‐Throughput Absolute Quantification of Protein‐Bound Tryptophan from Model and Crop Seeds

对模型和作物种子中的蛋白质结合色氨酸进行高通量绝对定量

• DOI: 10.1002/cpz1.862
• 发表时间: 2023
• 期刊: Current Protocols
• 作者: Ansaf, Huda;Yobi, Abou;Angelovici, Ruthie
• 通讯作者: Angelovici, Ruthie

A High‐Throughput Absolute Quantification of Protein‐Bound Sulfur Amino Acids from Model and Crop Plant Seeds

对模型和作物种子中的蛋白质结合硫氨基酸进行高通量绝对定量

• DOI: 10.1002/cpz1.861
• 发表时间: 2023
• 期刊: Current Protocols
• 作者: Yobi, Abou;Ansaf, Huda;Angelovici, Ruthie
• 通讯作者: Angelovici, Ruthie

A plant balancing act: Meshing new and existing metabolic pathways towards an optimized system

• DOI: 10.1016/j.pbi.2022.102173
• 发表时间: 2022-02-07
• 期刊: CURRENT OPINION IN PLANT BIOLOGY
• 影响因子: 9.5
• 作者: Angelovici, Ruthie;Kliebenstein, Dan
• 通讯作者: Kliebenstein, Dan