Spyridon Oikonomidis, Anush Nersesyan, Hripsik Kosyan, Sonya Vardanyan, Costas A Thanos
Although asymbiotic orchid seed germination was first achieved in vitro in 1922, the prediction of germination requirements under in vitro conditions still remains complicated. To address this, we developed a machine learning framework to classify the ex situ asymbiotic germination potential of wild orchids into four discrete groups: Low (0-30%), Mid (31-50%), High (51-80%), and Max (81-100%). Models were trained on a dataset of 203 species, utilizing seed morphometrics-specifically, the embryo-to-testa (E:S) length ratio-alongside core ecological traits (subfamily, growth habit, habitat, and climate zone), as well as chemical scarification duration as a proxy of seed permeability. Validation leveraged novel germination and trait data from 26 taxa from Greece (17) and Armenia (9), published here for the first time. To mitigate class imbalance and prevent algorithmic bias toward highly germinating species, we applied inverse frequency weighting during training. Iterative testing of six algorithms revealed that the "Step 4" feature matrix (excluding climate zone and pretreatment duration) yielded the optimal predictive balance. K-Nearest Neighbor (KNN) and Support Vector Machine (SVM) emerged as the superior models, achieving overall accuracies of 44.4% and 61.1%, respectively, with both achieving 100% accuracy for low-germinating species. Finally, we synthesized a novel database compiling new seed morphometrics from Armenia (17 taxa), Greece (52 taxa), and the data from the literature (479 taxa). After filtering previously utilized species, we generated a prediction pool of 361 orchid taxa. Applying our Step 5 KNN and SVM models to forecast their germination behavior revealed distinct variations linked to ecological profiles. This high-accuracy framework, particularly for low-germinability groups, offers a powerful screening tool for ex situ conservation planning. The final trained models are compiled in the publicly available R (v. 4.6.0) package OrchidGermClass.