Conidia of Fusarium proliferatum infected epidermal cells of stigma, hairs and filaments on maize when sprayed on maize filament. Hyphae of F. proliferatum extended along with the parenehyma and vascular cells in the ...Conidia of Fusarium proliferatum infected epidermal cells of stigma, hairs and filaments on maize when sprayed on maize filament. Hyphae of F. proliferatum extended along with the parenehyma and vascular cells in the filaments, and infected toward ovary. Both the parenchyma cells and the epidermal cells were wrinkled, and the filament was deformed. The result showed that F. proliferatum could infect filaments directly and cause maize ear rot through filament channel.展开更多
For some Cas nucleases,trans-cleavage activity triggered by CRISPR/Cas-mediated cis-cleavage upon target nucleic acid recognition has been explored for diagnostic detection.Portable single and multiplex nucleic acid-b...For some Cas nucleases,trans-cleavage activity triggered by CRISPR/Cas-mediated cis-cleavage upon target nucleic acid recognition has been explored for diagnostic detection.Portable single and multiplex nucleic acid-based detection is needed for crop pathogen management in agriculture.Here,we harnessed and characterized RfxCas13d as an additional CRISPR/Cas nucleic acid detection tool.We systematically characterized AsCas12a,LbCas12a,LwaCas13a,and RfxCas13d combined with isothermal amplification to develop a CRISPR/Cas nucleic acid-based tool for single or multiplex pathogen detection.Our data indicated that sufficient detection sensitivity was achieved with just a few copies of DNA/RNA targets as input.Using this tool,we successfully detected DNA from Fusarium graminearum and Fusarium verticillioides and RNA from rice black-streaked dwarf virus in crude extracts prepared in the field.Our method,from sample preparation to result readout,could be rapidly and easily deployed in the field.This system could be extended to other crop pathogens,including those that currently lack a detection method and have metabolite profiles that make detection challenging.This nucleic acid detection system could also be used for single-nucleotide polymorphism genotyping,transgene detection,and qualitative detection of gene expression in the field.展开更多
基金Supported by Science and Technology Innovation Team Support Program for Colleges and Universities in Henan Province(2010JRTSTHN012)Henan Foundation and Frontier Technology Research Program of Henan Science and Technology Department(162300410137)+1 种基金Henan Educational Committee Key Research Projects of Henan Higher Education(16A210034)Nanyang Normal University Special Project(ZX2016002)
文摘Conidia of Fusarium proliferatum infected epidermal cells of stigma, hairs and filaments on maize when sprayed on maize filament. Hyphae of F. proliferatum extended along with the parenehyma and vascular cells in the filaments, and infected toward ovary. Both the parenchyma cells and the epidermal cells were wrinkled, and the filament was deformed. The result showed that F. proliferatum could infect filaments directly and cause maize ear rot through filament channel.
基金supported by the National Natural Science Foundation of China(31771808 and 32001551)the China Postdoctoral Science Foundation(2020M680779)the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences(S2021ZD03)。
文摘For some Cas nucleases,trans-cleavage activity triggered by CRISPR/Cas-mediated cis-cleavage upon target nucleic acid recognition has been explored for diagnostic detection.Portable single and multiplex nucleic acid-based detection is needed for crop pathogen management in agriculture.Here,we harnessed and characterized RfxCas13d as an additional CRISPR/Cas nucleic acid detection tool.We systematically characterized AsCas12a,LbCas12a,LwaCas13a,and RfxCas13d combined with isothermal amplification to develop a CRISPR/Cas nucleic acid-based tool for single or multiplex pathogen detection.Our data indicated that sufficient detection sensitivity was achieved with just a few copies of DNA/RNA targets as input.Using this tool,we successfully detected DNA from Fusarium graminearum and Fusarium verticillioides and RNA from rice black-streaked dwarf virus in crude extracts prepared in the field.Our method,from sample preparation to result readout,could be rapidly and easily deployed in the field.This system could be extended to other crop pathogens,including those that currently lack a detection method and have metabolite profiles that make detection challenging.This nucleic acid detection system could also be used for single-nucleotide polymorphism genotyping,transgene detection,and qualitative detection of gene expression in the field.