SBIR Phase II: Generating HLB and canker resistant citrus trees using CRISPR/Cas9

SBIR 第二阶段：使用 CRISPR/Cas9 生成抗 HLB 和溃疡病的柑橘树

• 批准号: 2126741
• 负责人: Dusica Coltrane
• 金额: $ 99万
• 依托单位: Soil Culture Solutions, LLC
• 依托单位国家: 美国
• 项目类别: Cooperative Agreement
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2024-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2126741&HistoricalAwards=false
• 关键词: SBIR; Phase; II; Generating; HLB

This award is funded in whole or in part under the American Rescue Plan Act of 2021 (Public Law 117-2). The broader impact of this Small Business Innovation Research (SBIR) Phase II project is to help the US citrus industry mitigate the devastating effects of two diseases: Huanglongbing (HLB) and citrus canker, both of which have killed millions of citrus trees in Florida, cost the state billions in lost revenue, and caused the loss of thousands of jobs. HLB has also spread to other states including California, Texas, Arizona, and Georgia. This Phase II project will produce non-GMO, disease-resistant citrus trees using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) precision breeding technology. These new varieties will boost the US citrus industry, especially in Florida where HLB and canker have led to a 79% production decline. The proposed project aims to combine effector-interactome and CRISPR to create a commercially valuable new citrus variety for growers. This project leverages research that identified citrus genes that interact with bacterial effector proteins causing disease development. The project explores disruption of the pathogen-citrus interactions by a multiplex CRISPR system and the associated potential for disease resistance in commercially valuable citrus varieties. This project will generate citrus plants with edits to promising target genes and test those new varieties for resistance and other agronomic traits in greenhouse and field trials. The methods and approaches are multiplex CRISPR plasmid constructions, and citrus protoplast transformation and regeneration to create non-transgenic resistant citrus varieties. The testing protocol calls for grafting to inoculate the new varieties with HLB to confirm resistance and clonal micropropagation for large-scale experiments and field trials.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.

该奖项全部或部分根据2021年美国救援计划法案（公法117-2）资助。 小企业创新研究（SBIR）第二阶段项目的更广泛影响是帮助美国柑橘产业减轻黄龙病（HLB）和柑橘溃疡病这两种疾病的破坏性影响，这两种疾病都导致佛罗里达数百万柑橘树死亡，使该州损失数十亿美元的收入，并导致数千人失业。HLB还扩展到其他州，包括加州、德克萨斯、亚利桑那和格鲁吉亚。该第二阶段项目将使用成簇定期间隔短回文重复序列（CRISPR）精准育种技术生产非转基因抗病柑橘树。这些新品种将提振美国柑橘产业，特别是在佛罗里达，HLB和溃疡病导致产量下降79%。 拟议的项目旨在将联合收割机效应器相互作用组和CRISPR结合起来，为种植者创造一种具有商业价值的新柑橘品种。该项目利用了鉴定柑橘基因的研究，这些基因与导致疾病发展的细菌效应蛋白相互作用。该项目探索了通过多重CRISPR系统破坏病原体-柑橘相互作用，以及在具有商业价值的柑橘品种中获得抗病性的相关潜力。该项目将产生柑橘植物，对有希望的目标基因进行编辑，并在温室和田间试验中测试这些新品种的抗性和其他农艺性状。所述方法和途径是多重CRISPR质粒构建，以及柑橘原生质体转化和再生以产生非转基因抗性柑橘品种。该测试协议要求嫁接，以HLB的新品种，以确认大规模实验和田间试验的抗性和克隆微繁殖。该奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。