Wheat-associated bacterial and fungal communities during the interaction of Fusarium graminearum and Trichoderma gamsii T6085 and infection of a CRISPR/Cas9 derived green-fluorescent protein labeled Leptosphaeria maculans strain on Brassica napus
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Date
2022-03-10
Authors
Alukumbura, Dhathuge
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Abstract
The microbial diseases in agriculturally important crops have become a critical problem in countries all around the world. The plant diseases caused by various pathogens are unique and require different management strategies to prevent spreading. The Fusarium head blight (FHB) disease in wheat and the blackleg disease in canola are two major diseases that have been reported all around the world. While the primary causative agents of FHB are Fusarium graminearum and Fusarium culmorum, the primary causative agent of the blackleg disease is the Leptosphaeria maculans. All these pathogens are Ascomycetes and feed on decaying plant material. They can survive throughout the winter within dead plant material and initiate infections during spring and summer. The first objective of the current work was to analyze the microbiome associated with wheat plants and to evaluate the effectiveness of the biocontrol method for FHB management in wheat. This work examined the impact of inoculation with Trichoderma gamsii strain T6085 as a biocontrol agent on the natural bacterial and fungal community associated with wheat plants. The effect of T. gamsii T6085 on the microbial communities in wheat tissues was minimal. However, the current T. gamsii T6085 application method needs to be improved to have a good inhibition in the growth of F. graminearum. The second objective of this work was to understand the behaviour of a mutant L. maculans isolate generated through the CRISPR/Cas9 gene engineering technique. The mutant and its wild-type progenitor were tagged with the egfp gene, which expressed the green fluorescent reporter protein. The transformation of L. maculans with the egfp gene led to a decrease in their pathogenicity. According to the results, the mutant L. maculans isolate can colonize the plant tissues, but the plant defence responses inhibit its growth through a hypersensitive reaction.
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Keywords
Fusarium head blight, Wheat, Biological control, Trichoderma gamsii, Blackleg, Canola, Green fluorescent protein