Abstract
Under climate change, the spread of pests and pathogens into new environments has a dramatic effect on crop protection control. Strawberry (Fragaria spp.) is one the most profitable crops of the Rosaceae family worldwide, but more than 50 different genera of pathogens affect this species. Therefore, accelerating the improvement of fruit quality and pathogen resistance in strawberry represents an important objective for breeding and reducing the usage of pesticides. New genome sequencing data and bioinformatics tools has provided important resources to expand the use of synthetic biology-assisted intragenesis strategies as a powerful tool to accelerate genetic gains in strawberry. In this paper, we took advantage of these innovative approaches to create four RNAi intragenic silencing cassettes by combining specific strawberry new promoters and pathogen defense-related candidate DNA sequences to increase strawberry fruit quality and resistance by silencing their corresponding endogenous genes, mainly during fruit ripening stages, thus avoiding any unwanted effect on plant growth and development. Using a fruit transient assay, GUS expression was detected by the two synthetic FvAAT2 and FvDOF2 promoters, both by histochemical assay and qPCR analysis of GUS transcript levels, thus ensuring the ability of the same to drive the expression of the silencing cassettes in this strawberry tissue. The approaches described here represent valuable new tools for the rapid development of improved strawberry lines.
Highlights
Strawberry fruit (Fragaria spp.) is highly appreciated by consumers around the world and represents one of the most profitable crops of the Rosaceae family, whether in a fresh or processed form [1,2]
Each of them carries all the strawberry DNA sequences needed for a fruit ripening-related expression of an intragenic dsRNAi-inducing unit aimed to silence either of two relevant specific pathogen defense-related endogenous genes, FaWRKY1 or FaNPR3.1
Intragenesis associated with the synthetic biology has emerged as a powerful approach to overcome the limitations and barriers of the traditional methods and speed up the improvement of strawberry
Summary
Strawberry fruit (Fragaria spp.) is highly appreciated by consumers around the world and represents one of the most profitable crops of the Rosaceae family, whether in a fresh or processed form [1,2] (http://www.fao.org/faostat/en/#search/strawberries, accessed on 7 May 2021). In addition to sensorial attributes, such as colour, texture, aroma, and taste, which make this fruit very acceptable for human consumption, strawberries, like other berries, provide substantial benefits for health and blood sugar control, being an important source of manganese, potassium, folate (vitamin B9), vitamin C, and bioactive compounds, with high antioxidant capacity and potential cancer prevention effects [3,4,5,6] Characteristics such as firmness and vulnerability to pathogens significantly affect the yield and quality of the strawberry fruit, reducing its market value and consumption and are considered of great importance in breeding programs that seek to produce elite varieties with improved traits [7,8,9,10]. It is of great interest to accelerate genetic resistance in this crop, since management of strawberry is expected to become more difficult under the influence of climate change and globalization
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