STTR Phase II: Novel Analysis Tools for Production of Higher Indican Yielding Plants for Bio-based Indigo

STTR 第二阶段：用于生产高产生物靛蓝植物的新型分析工具

• 批准号: 1831949
• 负责人: Shawn Genung
• 金额: $ 75万
• 依托单位: Stony Creek Colors, Inc
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1831949&HistoricalAwards=false
• 关键词: STTR; Phase; II; Novel; Analysis

This Small Business Technology Transfer (STTR) Phase II project will develop a new tool for the analysis of in situ indican precursor in indigo plants, which when combined with genomic analysis and genetic linkage mapping in selectively bred indigo crops, will lead to high indican yielding breeding parental lines and ultimately competitively price natural indigo dye. Additionally, characterization of genetic markers will accelerate further crop improvement by understanding and harnessing the genes crucial to indican synthesis and other aspects of significance to overall indigo yield. These advancements will benefit customers, denim mills, by leading to a more reliable, lower cost plant-derived indigo supply. The success of this multiphase STTR project will enable a cost-competitive, cleaner, and more sustainable denim dyeing process, while greatly expanding the market for domestically produced natural indigo. Commercialization of a more consistent and higher yielding US-grown indigo plant that produces high purity indigo powder can replace the current standard of synthetic, imported indigo powder. While in demand by the marketplace today, plant-derived indigo is currently only used in premium denim products due to the high cost per pound resulting from low yields per plant per acre. The direct result of this research will be to open new market segments and expand existing market penetration for US-grown and manufactured biobased indigo for the textile industry, an addressable market of $1.86B. The methods and technology developed through this project have a direct path to the commercial marketplace and the industry is ready to support biobased textile dyes such as plant-derived indigo.During this project, reference genome resources will be constructed for indigo feedstock crops, F1 mapping populations will be constructed for P. tinctoria, I. tinctoria, and I. suffruticosa varieties, and design of the handheld rapid assay device will be validated through laboratory analysis. The reference genome for I. suffruticosa will be built using Pacific Biosciences SMRT sequencing and assembly. In addition to this reference genome, whole-genome resequencing will be performed on available I. suffruticosa and I. tinctoria varieties. Nucleotide variation and variant effect prediction will be made between the Indigofera-based indigo varieties. This variation will be used to develop markers to evaluate intervarietal crosses. Controlled greenhouse crosses of three species will be made to create F1 mapping populations for use in constructing a genetic map and linkage mapping, in combination with the resources developed. Intraspecies varieties exhibiting distinct phenotypic traits of commercial interest or notable dye yield differences will be selected for crossing to generate the F1 populations. Results from laboratory-based fluorometry equipment will be evaluated for efficacy and then compared against extractive indigo dye analysis from the leaf biomass. A final prototype will be constructed based on these findings and validated through use in the laboratory and in the field.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.

该小型企业技术转让（STTR）第二阶段项目将开发一种新工具，用于分析靛蓝植物中的原位靛蓝前体，该工具与选择性育种靛蓝作物的基因组分析和遗传连锁图谱相结合，将带来高靛蓝产量培育亲本系并最终使天然靛蓝染料的价格具有竞争力。 此外，通过了解和利用对靛蓝合成至关重要的基因以及对靛蓝总产量具有重要意义的其他方面，遗传标记的表征将加速作物的进一步改良。这些进步将带来更可靠、成本更低的植物靛蓝供应，从而使客户和牛仔布工厂受益。这一多阶段 STTR 项目的成功将实现具有成本竞争力、更清洁、更可持续的牛仔布染色工艺，同时极大地扩大国产天然靛蓝的市场。 生产高纯度靛蓝粉末的美国种植的靛蓝植物的一致性和产量更高，其商业化可以取代目前合成的进口靛蓝粉末的标准。尽管当今市场需求旺盛，但植物源靛蓝目前仅用于优质牛仔布产品，因为每英亩每株植物的产量较低，导致每磅成本较高。这项研究的直接结果将是为纺织行业开辟新的细分市场并扩大美国种植和制造的生物基靛蓝的现有市场渗透率，可寻址市场为 $1.86B。通过该项目开发的方法和技术可以直接进入商业市场，并且该行业已准备好支持生物基纺织染料，例如植物源靛蓝。在该项目期间，将为靛蓝原料作物构建参考基因组资源，F1图谱将为 P.tinctoria、I.tinctoria 和 I.suffruticosa 品种构建种群，手持式快速检测装置的设计将通过实验室分析进行验证。 I. suffruticosa 的参考基因组将使用 Pacific Biosciences SMRT 测序和组装构建。 除了该参考基因组之外，还将对现有的 I. suffruticosa 和 I.tinctoria 品种进行全基因组重测序。 将在以靛蓝为基础的靛蓝品种之间进行核苷酸变异和变异效应预测。 这种变异将用于开发标记来评估品种间杂交。将进行三个物种的受控温室杂交，以创建 F1 作图群体，用于结合开发的资源构建遗传图谱和连锁作图。 将选择具有商业利益的不同表型特征或显着染料产量差异的种内品种进行杂交以产生 F1 群体。将评估基于实验室的荧光测定设备的结果的功效，然后与从叶子生物量中提取的靛蓝染料分析进行比较。 最终原型将根据这些发现构建，并通过在实验室和现场使​​用进行验证。该奖项反映了 NSF 的法定使命，并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，被认为值得支持。