A HIGS approach targeting the DCL1, CYP51 and CHS genes of the pathogen to control Colletotrichum acutatum infection of strawberry

J.J. Higuera-Sobrino, R. Blanco-Portales, E. Moyano, A. Rodríguez-Franco, J. Muñoz-Blanco, J.L. Caballero
Among the berries, the strawberry (Fragaria × ananassa) represents one of the most valuable cultivated species worldwide. However, this crop suffers significant losses due to the attack by many fungal pathogens including Colletotrichum acutatum, which devalues the fruit in the harvest and postharvest stages. To date, due to the lack of truly effective alternatives, chemical and fungicides are being used to control C. acutatum infection, despite the fact that these procedures are not environmentally friendly enough and are also risky for health. New biotechnological approaches, such as host induced gene silencing (HIGS) applied to this species, emerge as a valuable alternative to control pathogen infection while reducing the use of chemicals and thus, the environmental impact. In this study, using the Colletotrichum databases currently available, several hairpin-based gene cassettes which target five important pathogen genes, i.e., the dcl-1 and dcl-2, the cytochrome p450-51A and 51B (cyp51A/cyp51B) and the chitin synthase class V (chsV) have been designed and their DNA sequences, either individually or linked, have been obtained by synthetic biology and cloned into gateway derivative T-vectors under the control of a 35S promoter. Putative HIGS lines of strawberry (Fragaria × ananassa ‘A1’) have been obtained by Agrobacterium transformation and kanamycin-based selection. Positive HIGS lines have been molecular characterized by PCR analysis. The effectiveness of dsRNAs stably expressed in the HIGS plants is being preliminary tested by inoculating detached leaves from in vitro elongated shoots of strawberry HIGS lines.
Higuera-Sobrino, J.J., Blanco-Portales, R., Moyano, E., Rodríguez-Franco, A., Muñoz-Blanco, J. and Caballero, J.L. (2023). A HIGS approach targeting the DCL1, CYP51 and CHS genes of the pathogen to control Colletotrichum acutatum infection of strawberry. Acta Hortic. 1381, 149-156
DOI: 10.17660/ActaHortic.2023.1381.20
https://doi.org/10.17660/ActaHortic.2023.1381.20
RNA interference, Fragaria × ananassa, Colletotrichum
English

Acta Horticulturae