Seed regeneration was carried out for all accessions collected as tubers and/or whole plants, as well as for accessions with a low number of seed (less than 6000 seeds). Six thousand seeds were selected to ensure both ex situ institutions (INIA and CIP) maintaining this material, would have ample material for distribution requests and conserving the material into perpetuity. These accessions were grown under greenhouse conditions in Lima (coastal site), Huancayo, and Cusco (higher elevation sites) depending on the origin of the accessions and understanding of the best environmental conditions for regeneration.
In the case of seed collections needing a seed increase, a minimum of 100 seeds were germinated. A set of 25-30 seedlings were transplanted in Jiffys-7 pots for 30 days, and then transferred to 20 cm pots using a Promix 8 substrate (Salas et al., 2008 ). Due to the ISO 17025:2017 accreditation at CIP which regulates workflows to ensure that only virus free germplasm is moved around the globe, each seedling was tested for the following viruses using standard laboratory testing (ELISA, PCR, etc.) and complemented with biological indexing on indicator plants: Arracacha Virus B Oca strain (AVB-O), Alfalfa Mosaic Virus (AMV), Andean Potato Latent Virus (APLV), Potato Yellowing Virus (PYV), Tobacco Mosaic Virus (TMV), Potato Virus T (PVT), and the quarantine viroid Potato Spindle Tuber Viroid (PSTVd), after which only virus-free plants were used for seed regeneration. Any plants that were positive for these viruses were destroyed to prevent dissemination of viruses infecting potato.
When plants were flowering, depending on their mode of reproduction, autogamous or allogamous, open pollination or a combination of sib-crosses and bulk crosses were made, respectively, to obtain seed for conservation. The regeneration of seed was performed between 2017 and 2021. The resulting seed was divided and shared between CIP and INIA for long term conservation.
In the case of seed collections needing a seed increase, a minimum of 100 seeds were germinated. A set of 25-30 seedlings were transplanted in Jiffys-7 pots for 30 days, and then transferred to 20 cm pots using a Promix 8 substrate (Salas et al., 2008 ). Due to the ISO 17025:2017 accreditation at CIP which regulates workflows to ensure that only virus free germplasm is moved around the globe, each seedling was tested for the following viruses using standard laboratory testing (ELISA, PCR, etc.) and complemented with biological indexing on indicator plants: Arracacha Virus B Oca strain (AVB-O), Alfalfa Mosaic Virus (AMV), Andean Potato Latent Virus (APLV), Potato Yellowing Virus (PYV), Tobacco Mosaic Virus (TMV), Potato Virus T (PVT), and the quarantine viroid Potato Spindle Tuber Viroid (PSTVd), after which only virus-free plants were used for seed regeneration. Any plants that were positive for these viruses were destroyed to prevent dissemination of viruses infecting potato.
When plants were flowering, depending on their mode of reproduction, autogamous or allogamous, open pollination or a combination of sib-crosses and bulk crosses were made, respectively, to obtain seed for conservation. The regeneration of seed was performed between 2017 and 2021. The resulting seed was divided and shared between CIP and INIA for long term conservation.
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