A CRISPR-based lateral flow assay for plant genotyping and pathogen diagnostics
Plant Biotechnology Journal - 2022 - S nchez - A CRISPR‐based lateral flow assay for plant genotyping and pathogen.pdf
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KAUST DepartmentBioengineering Program
Biological and Environmental Science and Engineering (BESE) Division
Center for Desert Agriculture
Laboratory for Genome Engineering
Laboratory for Genome Engineering and Synthetic Biology, Division of Biological Sciences 4700 King Abdullah University of Science and Technology Thuwal -23955-6900 Saudi Arabia
KAUST Grant NumberBAS/1/1035-01-01
Permanent link to this recordhttp://hdl.handle.net/10754/681129
MetadataShow full item record
AbstractEfficient 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 less than 1 hour, 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.
CitationSánchez, E., Ali, Z., Islam, T., & Mahfouz, M. (2022). A CRISPR-based lateral flow assay for plant genotyping and pathogen diagnostics. Plant Biotechnology Journal. Portico. https://doi.org/10.1111/pbi.13924
SponsorsThis work was supported, in part, by BAS/1/1035-01-01 grant from the KAUST to MM. TI received partial funding through the OFANS project of the K rishi Gobeshona Foundation of Bangladesh. We would like to thank Professor Simon rattinger, and his team at the Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia, for providing the Lr34 referenced wheat germplasm. We would like to thank NurUddin Mahmud of the Institute of Biotechnology and Genetic Engineering (IBGE), Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh, for assistance in extracting DNA from wheat blast fungus, Magnaporthe oryzaeTriticum isolates. We would like to thank Dr. Haroon Butt at the Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia, for providing the SGR3, SGR5 and OsmALS rice lines. We would like to thank Dr Nasir Saeed, Wheat Biotechnology Lab, NIBGE, Pakistan for providing wheat landraces and genomic DNA samples. We also thank members of the genome engineering and synthetic biology laboratory for insightful discussions and technical support.
JournalPlant Biotechnology Journal
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