Fighting Extinction - Conservation biotechnologies for endangered plants

In Austria, about 35% of higher plants and ferns are found on the Austrian Red List in categories 0 (extinct or missing) and 1- 3 (threatened). The conservation of endangered species in their natural environment is of utmost importance, but may not be sufficient to safeguard survival, in particular, if the habitat itself is under threat. Materials in ex situ conservations such as living plant collections and seed banks provide some degree of backup. However, they suffer from practical limitations and cannot meet the requirements of all species nor all conservation concepts. Alternatively, cryopreservation allows the conservation of valuable germplasm in liquid nitrogen at -196 oC on a long-term basis. Significant progress has been made in recent years and improved vitrification based cryopreservation protocols are now widely applied. However, the development of a protocol for a new species is still slow due to the need to optimise each step of the procedure through trial and error. In this project, fundamental processes underlying the cryostorage of plants will be studied and will provide a rationale for optimising cryopreservation methods for a wider range of species. Pre-conditioning cold treatments stimulate stress response and promote complex changes within plant cells such as altered membrane composition and sugar accumulation. These changes in cold-acclimated and non-acclimated (control) shoot tips will be studied through biochemical and electrolyte leakage analysis. Further, to measure and determine glass transition phases and crystallisation in tissues used for cryopreservation differential scanning calorimetry will be carried out. The findings will be correlated with regrowth after cryostorage, which will elaborate if induced cold stress responses increased cryotolerance. A critical aspect in the cryopreservation process is the regeneration of plants after rewarming. The ultrastructure of cells within the meristematic tissue in shoot tips will be studied before and after cryostorage. It will reveal which parts of the shoot tips survive and eventually contribute to the regeneration process. The final objective is to generate vigorous and true-to-type plants. The project will establish if regenerated plants are free from somaclonal variation and of good physiological health using flow cytometry, phenotypic and biochemical analysis. This project uses Austrian endangered plant species as model species; a group of plants that is under-researched and has never undergone cryopreservation. The project will acquire locally relevant data to provide evidence of the feasibility but also limitations of this methodology to reduce the perceived inherent risks and to overcome adoption barriers leading to application in Austria. As part of the project, a cryopreservation facility will be established in Austria in collaboration with the cryo-genebank IPK, Gatersleben (Germany) and the Botanic Gardens Kew (UK) and Kings Park (Australia).

In Austria, about 35% of higher plants and ferns are found on the Austrian Red List in categories 0 (extinct or missing) and 1- 3 (threatened). The conservation of endangered species in their natural environment is of utmost importance, but may not be sufficient to safeguard survival, in particular, if the habitat itself is under threat. Materials in ex situ conservations such as living plant collections and seed banks provide some degree of backup. However, they suffer from practical limitations and cannot meet the requirements of all species nor all conservation concepts. Alternatively, cryopreservation allows the conservation of valuable germplasm in liquid nitrogen at -196 oC on a long-term basis. We developed cryopreservation protocols for endangered Austrian wild plant species for germplasm conservation. In particular, we looked into the occurrence of latent bacteria and their impact on regeneration. The use of winter-acclimated field material of herbaceous dicots as explant source for cryopreservation has been first described as part of this project. We further performed a genetic population study in Artemisia laciniata and included old herbarium specimen of nowadays extinct European populations. Our work demonstrated proof of concept and the feasibility of using biotechnology in conservation. Numerous outreach activities complemented our research to raise awareness for conservation issues among the public.