There is, at the present time, a great demand for chestnut rootstocks with improved resistance to Phytophthora cinnamomi Rands in the nurseries. New genotypes are emerging from European chestnut breeding programs and the production of thriving plants to restore old orchards with low yields due to a high incidence of diseases, namely root rot, is necessary. Micropropagation is a useful technique for clonal propagation. Nevertheless, in vitro culture propagation is genotype-dependent. Consequently, the existing protocols may demonstrate poor reproducibility and low efficacy. Thus, the need to contribute to the development of new micropropagation protocols suitable for large production of emerging genotypes. As a contribution to fill this gap, a three-step protocol was developed by using new combinations of Murashige & Skoog, Woody Plant, and adapted modified Melin-Norkrans media in different stages of the propagation process. About 90% of shoots were rooted, and after three months of acclimatization, 85% of these plants survived and were capable of continuous growth in the field. Currently, this protocol is being used in the production of several hybrid genotypes (with improved resistance to P. cinnamomi), selected from our ongoing breeding program and also in Castanea sativa Mill. and Castanea crenata Siebold and Zucc. species.
American chestnut (Castanea dentata (Marsh.) Borkh) was almost completely wiped out by the fungal pathogen, Cryphonectria parasitica (Murrill) M.E. Barr. Another invasive pathogen, Phytophthora cinnamomi Rands, is devastating American chestnuts in the southern region of the United States. An alternative approach for controlling these pathogens is to use genetic engineering or gene editing. We successfully transformed American chestnut with a detoxifying enzyme, oxalate oxidase, to enhance blight tolerance and more recently with the Cast_Gnk2-like gene, which encodes for an antifungal protein, to be tested for P. cinnamomi putative tolerance. Eight somatic embryo lines were transformed using three methods of selection: semisolid medium in Petri plates, liquid medium in RITA® temporary immersion bioreactors, or liquid medium in We Vitro containers. No significant differences were found between the treatments. These methods will allow for further testing of transgenes and the development of enhanced pathogen resistance in chestnut. It can serve as a model for other tree species threatened by invasive pests and pathogens.
Chestnuts are multipurpose trees significant for the economy and wildlife. These trees are currently found around the globe, demonstrating their genetic adaptation to different environmental conditions. Several biotic and abiotic stresses have challenged these species, contributing to the decline of European chestnut production and the functional extinction of the American chestnut. Several efforts started over the last century to understand the cellular, molecular, and genetic interactions behind all chestnut biotic and abiotic interactions. Most efforts have been toward breeding for the primary diseases, chestnut blight and ink disease caused by the pathogens, Cryphonectria parasitica and Phytophthora cinnamomi, respectively. In Europe and North America, researchers have been using the Asian chestnut species, which co-evolved with the pathogens, to introgress resistance genes into the susceptible species. Breeding woody trees has several limitations which can be mostly related to the long life cycles of these species and the big genome landscapes. Consequently, it takes decades to improve traits of interest, such as resistance to pathogens. Currently, the availability of genome sequences and next-generation sequencing techniques may provide new tools to help overcome most of the problems tree breeding is still facing. This review summarizes European and American chestnut’s main biotic stresses and discusses breeding and biotechnological efforts developed over the last decades, having ink disease and chestnut blight as the main focus. Climate change is a rising concern, and in this context, the adaptation of chestnuts to adverse environmental conditions is of extreme importance for chestnut production. Therefore, we also discuss the abiotic challenges on European chestnuts, where the response to abiotic stress at the genetic and molecular level has been explored.
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