A CRISPR-based lateral flow assay for plant genotyping and pathogen diagnostics,Plant Biotechnology Journal

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A CRISPR-based lateral flow assay for plant genotyping and pathogen diagnostics
Plant Biotechnology Journal ( IF 10.1 ) Pub Date : 2022-09-07 , DOI: 10.1111/pbi.13924
Edith Sánchez ₁ , Zahir Ali ₁ , Tofazzal Islam ₂ , Magdy Mahfouz ₁

Affiliation

Efficient pathogen diagnostics and genotyping methods enable effective disease management and breeding, improve crop productivity and ensure food security. However, current germplasm selection and pathogen detection techniques are laborious, time-consuming, expensive and not easy to mass-scale application in the field. Here, we optimized a field-deployable lateral flow assay, Bio-SCAN, as a highly sensitive tool to precisely identify elite germplasm and detect mutations, transgenes and phytopathogens in <1 h, starting from sample isolation to result output using lateral flow strips. As a proof of concept, we genotyped various wheat germplasms for the Lr34 and Lr67 alleles conferring broad-spectrum resistance to stripe rust, confirmed the presence of synthetically produced herbicide-resistant alleles in the rice genome and screened for the presence of transgenic elements in the genome of transgenic tobacco and rice plants with 100% specificity. We also successfully applied this new assay to the detection of phytopathogens, including viruses and bacterial pathogens in Nicotiana benthamiana, and two destructive fungal pathogens (Puccinia striiformis f. sp. tritici and Magnaporthe oryzae Triticum) in wheat. Our results illustrate the power of Bio-SCAN in crop breeding, genetic engineering and pathogen diagnostics to enhance food security. The high sensitivity, simplicity, versatility and in-field deployability make the Bio-SCAN as an attractive molecular diagnostic tool for diverse applications in agriculture.

中文翻译：

基于 CRISPR 的植物基因分型和病原体诊断侧流分析

高效的病原体诊断和基因分型方法可以实现有效的疾病管理和育种，提高作物生产力并确保粮食安全。然而，目前的种质选择和病原菌检测技术费力、耗时、昂贵，且不易在田间大规模应用。在这里，我们优化了可现场部署的侧向层析检测 Bio-SCAN，作为一种高度灵敏的工具，可精确识别优良种质并检测 <1 id=3>Lr34 和Lr67等位基因中的突变、转基因和植物病原体，从而赋予广谱抗性条锈病，证实了水稻基因组中存在合成产生的除草剂抗性等位基因，并以 100% 特异性筛选转基因烟草和水稻植物基因组中转基因元件的存在。我们还成功地将这种新的检测方法应用于植物病原体的检测，包括烟草中的病毒和细菌病原体，以及小麦中的两种破坏性真菌病原体（小麦条锈菌专化型和稻瘟病菌）。我们的结果说明了 Bio-SCAN 在作物育种、基因工程和病原体诊断方面的力量，可增强粮食安全。高灵敏度、简单性、多功能性和现场可部署性使 Bio-SCAN 成为适合农业多种应用的有吸引力的分子诊断工具。

更新日期：2022-09-07

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