Dataset Information

A new vector system for targeted integration and overexpression of genes in the crop pathogen Fusarium solani.

ABSTRACT: Background:Besides their ability to produce several interesting bioactive secondary metabolites, members of the Fusarium solani species complex comprise important pathogens of plants and humans. One of the major obstacles in understanding the biology of this species complex is the lack of efficient molecular tools for genetic manipulation. Results:To remove this obstacle we here report the development of a reliable system where the vectors are generated through yeast recombinational cloning and inserted into a specific site in F. solani through Agrobacterium tumefaciens-mediated transformation. As proof-of-concept, the enhanced yellow fluorescent protein (eYFP) was inserted in a non-coding genomic position of F. solani and subsequent analyses showed that the resulting transformants were fluorescent on all tested media. In addition, we cloned and overexpressed the Zn(II)2Cys6 transcriptional factor fsr6 controlling mycelial pigmentation. A transformant displayed deep red/purple pigmentation stemming from bostrycoidin and javanicin. Conclusion:By creating streamlined plasmid construction and fungal transformation systems, we are now able to express genes in the crop pathogen F. solani in a reliable and fast manner. As a case study, we targeted and activated the fusarubin (PKS3: fsr) gene cluster, which is the first case study of secondary metabolites being directly associated with the responsible gene cluster in F. solani via targeted activation. The system provides an approach that in the future can be used by the community to understand the biochemistry and genetics of the Fusarium solani species complex, and is obtainable from Addgene catalog #133094. Graphic abstract:

SUBMITTER: Nielsen MR

PROVIDER: S-EPMC6905090 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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A new vector system for targeted integration and overexpression of genes in the crop pathogen <i>Fusarium solani</i>.

Nielsen Mikkel Rank MR Holzwarth Anna Karolina Rilana AKR Brew Emmett E Chrapkova Natalia N Kaniki Samba Evelyne Kabemba SEK Kastaniegaard Kenneth K Sørensen Trine T Westphal Klaus Ringsborg KR Wimmer Reinhard R Sondergaard Teis Esben TE Sørensen Jens Laurids JL

Fungal biology and biotechnology 20191211

<h4>Background</h4>Besides their ability to produce several interesting bioactive secondary metabolites, members of the <i>Fusarium solani</i> species complex comprise important pathogens of plants and humans. One of the major obstacles in understanding the biology of this species complex is the lack of efficient molecular tools for genetic manipulation.<h4>Results</h4>To remove this obstacle we here report the development of a reliable system where the vectors are generated through yeast recomb ...[more]

PMID: 31890232

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Project description:Soil disinfection using high temperatures via steam is a promising approach to manage plant pathogens, pests, and weeds. Soil steaming is a viable option for growers who are moving away from dependence on chemical soil fumigants, especially in plant nursery or high tunnel environments. However, there are few studies that investigate how soil steaming causes substantial disturbance to the soil by killing both target pathogens and other soil biota. Steaming treatments also change the trajectory of the soil microbiome as it reassembles over time. Growers are interested in the health of soils after using steam-disinfection, especially if a virulent pathogen colonizes the soil and then flourishes in a situation where there are very few microbes to suppress its growth. Should recruitment of a virulent pathogen occur in the soil, this could have devasting effects on seed germination, seedling establishment and survival. Beneficial microbes are often used to prevent the colonization of plant pathogens, especially after a soil-steaming event. Here, we experimentally test how soil fungal communities assemble after steaming disinfection. We introduce to steam-treated soil Fusarium solani, an important fungal pathogen of soybean and Trichoderma harzianum, a known beneficial fungus used for soilborne pathogen suppression. Results show that F. solani significantly affects the relative abundance and diversity of the soil fungal microbiome, however, T. harzianum does not mitigate the amount of F. solani in the steam treated soil. Within the T. harzianum microbial addition, the soil fungal communities were similar to the control (steaming only). This result suggests inoculating the soil with T. harzianum does not drastically alter the assembly trajectory of the soil fungal microbiome. Other soil amendments such as a combination of Trichoderma spp. or other genera could suppress F. solani growth and shift soil microbiome composition and function post-steaming, however, more experimental research is needed.

Dataset Information

A new vector system for targeted integration and overexpression of genes in the crop pathogen Fusarium solani.

Publications

A new vector system for targeted integration and overexpression of genes in the crop pathogen <i>Fusarium solani</i>.

Similar Datasets

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