The conservation of genetic diversity and its sustainable use are essential to meet a number of challenges facing humanity, from coping with the predicted climate changes to achieving food security despite a still growing world population. European plum (Prunus domestica L.), as the leading fruit species in Serbian agriculture, is facing with climate changes and long-term inoculum pressure of numerous pathogens. Autochthonous plum genotypes of immeasurable genetic and cultural value are endangered and suppressed by newly bred and commercially important cultivars. Valuable plum germplasm is exposed to slight and irreversible disappearance. The strategic objective of the CryoPlum project is to provide methods and tools to face this menace.
On that purpose, the project will:
In the field of plant pathology and cryobiology brought new knowledge by:
- Evaluating health status of the autochthonous plum genotypes with available laboratory techniques (ELISA, PCR and Real-time PCR) for the detection of viruses and phytoplasmas, with the special reference to plum pox virus (PPV);
- Optimizing and elaborating protocols for micropropagation and in vitro cryopreservation using newly developed vitrification-based techniques (V and D cryo-plate techniques) of virus-free and PPV-infected genotypes;
- Assessing into the health status of the recovered plants after cryopreservation, i.e. evaluation of the cryotherapy as a method for plum pox virus elimination.
In the field of agricultural management, help the interested parties (breeders, biologists) to take advantage of the project outcomes by:
- Establishing guidelines for the collection of plum material and its analysis on the presence of pathogens for conservation purpose;
- Delivering protocols for plum germplasm conservation using in vitro cryo-techniques;
- Delivering protocols for plum cryotherapy;
- Opening new possibilities to implement this technique to other fruit species.
The concept of CryoPlum is to combine the latest methodology in plum germplasm conservation, pathogen detection and virus elimination. The project scope covers the entire chain from the collection of autochthonous plum genotypes, analyses of the collected material on the presence of infectious pathogens, in vitro cryopreservation of the material, recovery and continuous laboratory tests of the recovered material. The project addresses virologists, physiologists, cryobiologists, plum breeders and pomologists, with relevant outcomes including autochthonous plum cultivars, sample collection, detection methods and tools for pathogen detection, cryopreservation and cryotherapy. Cryotherapy is a novel application of plant cryopreservation for pathogen (viruses, phytoplasmas and bacteria) elimination from infected plant material. After the establishment of the protocol for the stated genotype, the technique requires only basic equipment available in a tissue culture laboratory.
The major challenge for cryotherapy technique application is significantly different response of the genotypes of the same species to this treatment. Several protocols with different cryotherapy methods have been reported for dozen of species. Their efficiency differ between laboratories (levels of survival, regeneration and pathogen presence). Considering the wide distribution of plum pox virus in Serbia it should be expected that a large portion of the autochthonous genotypes (especially sensitive ones) are in high percentage infected indicating need for virus elimination. Considering the importance of preserving plant genetic resources and plum significance for Serbian agriculture production and tradition, the main idea of the CryoPlum project is to develop and optimize protocols for conservation of the most important autochthonous plum genotypes using new technology-based conservation methods and pathogen detection techniques. The concept is to elaborate methodology for cryopreservation of selected autochthonous plum genotypes, but also to assimilate it with cryotherapy to obtain plants free of viruses from infected starting material.