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Development of an Efficient CRISPR/Cas9 System in Fusarium verticillioides and Its Application in Reducing Mycotoxin Contamination

Development of an Efficient CRISPR/Cas9 System in Fusarium v...

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中国植物病理学会2024年学术年会

作者： TANG Tingting DING Yi GUO Wei Institute of Food Science and Technology Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport ProcessMinistry of Agriculture and Rural Affairs

Fusarium verticillioides is a globally recognized and highly impactful fungal pathogen of maize,causing yield losses and producing harmful mycotoxins that pose a threat to human and animal ***,the genetictools available for studying this crucial fungus are currently limited in comparison with other important fungal *** address this,an efficient CRISPR/Cas9 genome editing system based on an autonomously replicating plasmid with an AMA1 sequence was established in this ***,gene disruption ofpyrG and pyrE via non-homologous end-joining（NHEJ） pathway was successfully achieved,with efficiency ranging from 66% to 100%.Secondly,precise gene deletions were achieved with remarkable efficiency using a dual sgRNA expression ***,the developed genome editing system can be applied to generate designer chromosomes in ***,as evidenced by the deletion of a crucial 38 kb fragment required for fumonisin ***,the pyrG recycling system has been established and successfully applied in ***,the developed ΔFUM1 and ΔFUM mutants can serve as biocontrol agents to reduce the Fumonisin B（FB） contamination produced by the *** together,these significant advancements in geneticmanipulation and biocontrol strategies provide valuable tools for studying and mitigating the impact of *** on maize crops.

关键词： CRISPR/Cas9 Fusarium verticillioides fumonisin uracil auxotrophic selection system atoxigenic strain

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Study on distribution and characterization of Aspergillus flavus strains to screen potential biocontrol agents from different geographical locations of China

Study on distribution and characterization of Aspergillus fl...

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中国菌物学会2016年学术年会

作者： Firew Tafesse Mamo Bo Shang Jonathan Nimal Selvaraj Yan Wang Yang Liu Institute of Food Science and Technology Chinese Academy of Agricultural Sciences/Key Laboratory of Agroproducts ProcessingMinistry of Agriculture

Aflatoxin contamination of crops like peanut,maize and rice is still a major public health and economic problem for the world,the impact is more significant for developing *** promising method for combating aflatoxin formation is biocontrol using competitive exclusion,currently under intensive study in peanut crops in different parts of *** *** strains as a biocontrol must be unable to produce both aflatoxin and cyclopiazonic acids and also must incapable to convert to *** the current study a total of 724 Aspergillus flavus strains isolated from peanut,maize and rice in 21 geographical locations of China were *** these,229 strains did not produce *** incidence of atoxigenic *** strains range from 28-35% from three *** productions in *** isolates were varied significantly among all sampled locations（X220,0.05= 0.012）.Non toxigenic（non AFB1 and CPA） strains were screened for their potential as biocontrol by examining the 28 aflatoxin biosynthesis genes,using a PCR *** which do not have afl R PCR products were selected for analysis of the other 27 biosynthetic *** four strains showed failure,in the,fas-2 and afl J genes,***（67%） of atoxigenic *** strains failed to show a PCR product in 10 or more ***,18 PCR product patterns were observed,usually common to only one or two isolates,exceptionally six of stains have the same（K） deletion pattern,indicating great diversity in the aflatoxin biosynthesis pathway in *** isolates taken from different crops and *** extensive deletions of 10 or more genes in the aflatoxin gene cluster identified in candidate’s nonaflatoxigenic biocontrol *** isolates serve as a safeguard in preventing adverse genetic reversion,thereby ensuring their safe application to agricultural *** genes dma T and mao A could be used a marker genes for CPA production capacity atoxigenic *** control o

关键词： Aflatoxin Cyclopiazonic acid(CPA) Aspergillus flavus atoxigenic strain toxigenic strains biocontrol

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