Abstract
Holm oak is a key tree species in Mediterranean ecosystems, whose populations have been increasingly threatened by oak decline syndrome, a disease caused by the combined action of Phytophthora cinnamomi and abiotic stresses. The aim of the present study was to produce holm oak plants that overexpress the Ginkbilobin-2 homologous domain gene (Cast_Gnk2-like) that it is known to possess antifungal properties. Proembryogenic masses (PEMs) isolated from four embryogenic lines (Q8, E2, Q10-16 and E00) were used as target explants. PEMs were co-cultured for 5 days with Agrobacterium EHA105pGnk2 and then cultured on selective medium containing kanamycin (kan) and carbenicillin. After 14 weeks on selective medium, the transformation events were observed in somatic embryos of lines Q8 and E2 and a total of 4 transgenic lines were achieved. The presence of the Cast_Gnk2-like gene on transgenic embryos was verified by PCR, and the number of transgene copies and gene expression was estimated by qPCR. Transgenic plants were obtained from all transgenic lines after cold storage of the somatic embryos for 2 months and subsequent transfer to germination medium. In an in vitro tolerance assay with the pathogen P. cinnamomi, we observed that transgenic plants were able to survive longer than wild type.
Highlights
IntroductionOne of the ecosystems affected is the Spanish dehesas and Portuguese montados (dehesas, hereafter), which are experiencing an unprecedented crisis, fundamentally caused by a complex disease commonly known as oak decline [3]
Introduction iationsIn recent decades, global climate change has greatly exacerbated the effects of many plant diseases, especially fungal diseases, which can severely affect forest ecosystems [1,2].One of the ecosystems affected is the Spanish dehesas and Portuguese montados, which are experiencing an unprecedented crisis, fundamentally caused by a complex disease commonly known as oak decline [3]
This syndrome has drastically affected the sustainability of the dehesa ecosystem, which is unique within Europe, and the situation is of great concern due to the associated high economic, ecological and social losses [4]
Summary
One of the ecosystems affected is the Spanish dehesas and Portuguese montados (dehesas, hereafter), which are experiencing an unprecedented crisis, fundamentally caused by a complex disease commonly known as oak decline [3]. This syndrome has drastically affected the sustainability of the dehesa ecosystem, which is unique within Europe, and the situation is of great concern due to the associated high economic, ecological and social losses [4]. This species contributes to the direct production in dehesas, providing
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