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.

这个小企业技术转让（STTR）第二阶段项目将开发一种新的工具，用于分析靛蓝植物中的原位靛蓝前体，当与选择性培育的靛蓝作物的基因组分析和遗传连锁图谱相结合时，将导致高靛蓝产量育种亲本系，并最终使天然靛蓝染料的价格具有竞争力。 此外，遗传标记的表征将通过理解和利用对靛蓝合成至关重要的基因以及对总体靛蓝产量具有重要意义的其他方面来加速进一步的作物改良。这些进步将有利于客户，牛仔布米尔斯厂，导致一个更可靠，更低成本的植物来源的靛蓝供应。这个多阶段STTR项目的成功将实现具有成本竞争力、更清洁和更可持续的牛仔布染色工艺，同时大大扩大国产天然靛蓝的市场。 一种更稳定、产量更高的美国种植的靛蓝植物的商业化可以生产高纯度的靛蓝粉末，从而取代目前标准的合成进口靛蓝粉末。虽然在市场需求的今天，植物来源的靛蓝目前只用于优质牛仔产品，由于每磅成本高，导致每英亩每株产量低。这项研究的直接结果将是开辟新的细分市场，并扩大美国种植和制造的纺织业生物基靛蓝的现有市场渗透率，这是一个18.6亿美元的潜在市场。通过该项目开发的方法和技术可以直接进入商业市场，行业已经准备好支持生物基纺织染料，如植物来源的靛蓝。在该项目中，将构建靛蓝原料作物的参考基因组资源，将构建P.tinctoria，I.着色，我。将通过实验室分析验证对木桑品种的鉴定和手持式快速测定装置的设计。参考基因组I.将使用Pacific Biosciences SMRT测序和组装构建suffruticosa。 除此参考基因组外，还将对可用的I. suffruticosa和I.着色剂品种 对靛蓝类品种间的核苷酸变异和变异效应进行预测。 该变异将用于开发标记以评价品种间杂交。将进行三个物种的受控温室杂交，以建立F1作图群体，用于构建遗传图谱和连锁图谱，并结合开发的资源。 将选择表现出具有商业利益的不同表型性状或显着染料产量差异的种内品种进行杂交以产生F1群体。将评价基于实验室的荧光测定设备的结果的有效性，然后将其与来自叶生物量的提取靛蓝染料分析进行比较。 最终的原型将根据这些研究结果构建，并通过在实验室和现场的使用进行验证。该奖项反映了NSF的法定使命，并被认为是值得通过使用基金会的知识价值和更广泛的影响审查标准进行评估的支持。