Articles
Intragenic and synthetic biotechnology for genetic improvement in strawberry
Article number
1454_8
Pages
57 – 64
Language
English
Abstract
Strawberry is one of the most consumed and valued fruits for its nutritional and organoleptic properties, making it one of the most profitable crops in the Rosaceae family worldwide.
However, many pathogens can attack this plant, significantly affecting the production and quality of the fruit during pre- and postharvest stages.
To address this issue, the use of chemicals is common, but it raises public concerns.
However, we can currently turn to genetic improvement targeting key plant genes involved in disease protection such as the NPR gene family, which has been successfully tested in many crops.
Moreover, thanks to new genome sequencing data, bio-informatics tools, and synthetic biology, it is possible to achieve these objectives through intragenic strategies, which are better accepted by consumers, to accelerate genetic gains in strawberries.
Using synthetic biology, our research group has designed four intragenic RNA interference (RNAi) silencing cassettes by combining new strawberry-specific promoters.
Also, using a transient assay in the fruit, we detected the expression of GUS by the two synthetic promoters FvAAT2 and FvDOF2, confirmed by both histochemical assays and qPCR analysis of GUS transcription levels, thus ensuring their capability to drive the expression of the silencing cassettes in this strawberry tissue.
The targets are members of the FaNPR3 clade of Fragaria × ananassa, whose silencing we have shown increases tolerance and significantly reduces fruit tissue damage following infection with Colletotrichum acutatum. Therefore, we can anticipate that members of the FaNPR3 clade in strawberries negatively regulate the defense response to pathogens, like their orthologs AtNPR3/AtNPR4 do in Arabidopsis. Indeed, the ectopic expression of FaNPR3.2 in the Arabidopsis thaliana double mutant npr3npr4, reverses the disease resistance phenotype against Pseudomonas syringae to wild-type levels.
The approaches described here represent valuable new tools for the rapid development of improved strawberry lines through synthetic biology.
Currently, we are moving toward HIGS and SIGS approaches that use RNAi silencing constructs engineered by synthetic technology.
However, many pathogens can attack this plant, significantly affecting the production and quality of the fruit during pre- and postharvest stages.
To address this issue, the use of chemicals is common, but it raises public concerns.
However, we can currently turn to genetic improvement targeting key plant genes involved in disease protection such as the NPR gene family, which has been successfully tested in many crops.
Moreover, thanks to new genome sequencing data, bio-informatics tools, and synthetic biology, it is possible to achieve these objectives through intragenic strategies, which are better accepted by consumers, to accelerate genetic gains in strawberries.
Using synthetic biology, our research group has designed four intragenic RNA interference (RNAi) silencing cassettes by combining new strawberry-specific promoters.
Also, using a transient assay in the fruit, we detected the expression of GUS by the two synthetic promoters FvAAT2 and FvDOF2, confirmed by both histochemical assays and qPCR analysis of GUS transcription levels, thus ensuring their capability to drive the expression of the silencing cassettes in this strawberry tissue.
The targets are members of the FaNPR3 clade of Fragaria × ananassa, whose silencing we have shown increases tolerance and significantly reduces fruit tissue damage following infection with Colletotrichum acutatum. Therefore, we can anticipate that members of the FaNPR3 clade in strawberries negatively regulate the defense response to pathogens, like their orthologs AtNPR3/AtNPR4 do in Arabidopsis. Indeed, the ectopic expression of FaNPR3.2 in the Arabidopsis thaliana double mutant npr3npr4, reverses the disease resistance phenotype against Pseudomonas syringae to wild-type levels.
The approaches described here represent valuable new tools for the rapid development of improved strawberry lines through synthetic biology.
Currently, we are moving toward HIGS and SIGS approaches that use RNAi silencing constructs engineered by synthetic technology.
Authors
V. Súnico, J.J. Higuera, J. Molina Hidalgo, R. Blanco-Portales, E. Moyano, A.M. Maldonado-Alconada, J. Muñoz-Blanco, E. Baraldi, B. Mezzetti, S. Sabbadini, J.L. Caballero
Keywords
intragenic strawberry, synthetic biology, RNA interference, strawberry promoters, plant immunity
Groups involved
- Division Plant Genetic Resources, Breeding and Biotechnology
- Working Group Genetic Transformation and Gene Editing
- Working Group Horticultural Biotechnology and Breeding
- Division Ornamental Plants
- Division Vegetables, Roots and Tubers
- Division Temperate Tree Fruits
- Division Vine and Berry Fruits
- Division Horticulture for Development
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