Periodic Reporting for period 1 - SSAR (Future bio-pesticides for EU: unlocking the code of Saponin Structure-Activity Relationships for use in pest management) Reporting period: 2016-03-01 to 2018-02-28 Summary of the context and overall objectives of the project This project aims to elucidate the relationship between saponin chemical structures and their biological activities and evolution. The new knowledge gained will have great industrial application potential as a generic platform for the production of bio-active compounds from plants and their use for bio-pesticides as an alternative to chemically produced pesticides. The new bio-pesticides will both help to meet society’s need for more sustainable food production and to contribute to the development of the knowledge based bio-economy. Identification of genes involved in saponin biosynthetic pathway will also open the possibility to engineer and breed crop with tailored saponin-based pest resistance capabilities. First, the genes involved in the biosynthetic pathway of saponins will be identified. Then desired or novel saponins will be produced in plant tissues through metabolic engineering with the identified genes. Finally, purified saponins will be used in bioassays with a range of pest species, to unlock the code of exactly which saponin chemical structures have a given specific biological activity. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far During this project, we elucidated the biosynthetic pathway of insect-resistance associated saponins through identification of the key biosynthesis gene, CYP72A552. With all the biosynthetic genes at hand, we managed to produce desired saponins in tobacco plants through metabolic engineering. Later we showed that those tobacco plants gained resistance against its herbivore insects because of the accumulation of desired saponins. In the end, we managed to produce desired saponins in yeast, a potential production platform, which is a crucial step for commercial production of this compound in large scale. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) We have identified bio-active saponins and shown the possibility to produce them in a production platform. This will help to commercialize the production of bio-active saponins in industry scale, which will contribute to society’s need for more sustainable food production. We successfully enabled tobacco plants to gain resistance against its herbivore through transiently expressing the whole saponin biosynthesis pathway, which showed the possibility to engineer and breed crop with tailored saponin-based pest resistance capabilities.