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General information Notification Number B/SE/20/1726 Member State to which the notification was sent Sweden Date of acknowledgement from the Member State Competent Authority 06/02/2020 Title of the Project Potato with altered resistance to pathogens Proposed period of release: 01/04/2020 to 31/12/2024 Name of the Institute(s) or Company(ies) Swedish University of Agricultural Sciences SLU, Department of Plant Protection Biology, P. O Box 102, 230 53, Alnarp.
Is the same GMPt been notified elsewhere by the same notifier? No Has the same GMPt been notified elsewhere by the same notifier? No Genetically modified plant Complete name of the recipient or parental plant(s):

Common Name	Family Name	Genus	Species	Subspecies	Cultivar/breeding line
potato	solanaceae	solanum	solanum tuberosum	tuberosum

Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications: CHL1: Potato in which the CHL1 gene has been genome edited. Neomycin phosphotransferase II (nptII) coding for kanamycin resistance as a marker.

DMR6: Potato in which the DMR6 gene has been genome edited. Neomycin phosphotransferase II (nptII) coding for kanamycin resistance as a marker.

Man_RNAi: down-regulated Mannanase transcript through an RNAi construct. Neomycin phosphotransferase II (nptII) coding for kanamycin resistance as a marker. Genetic modification Type of genetic modification: Insertion;Deletion; In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted: CHL1 T-DNA: From left to right border; left border sequence (LB) from Agrobacterium tumefaciens, polyadenylation sequence from 35S transcript (35S_T), neomycin phosphotransferase II coding sequence (nptII) that can be isolated from different bacteria (marker, NptII), 2x35S promotor from cauliflower mosaic virus 35S gene (2x35S_P), 35S promotor (35S_P) from cauliflower mosaic virus 35S gene, Csy4 ribonuclease gen from Pseudomonas aeruginosa that has been codon-optimized for tomato (Csy4_enz), P2A ribosomal skipping sequence (P2A), Cas 9 gene that has been codon-optimized for Arabidopsis thaliana (Cas9), heat shock protein terminator (HSP_T) from plants, CmYLCV promotor from yellow leaf curling virus (CmYLCV_P), Csy4 cleavage-site (Csy4_k), 20bp long spacer (CRISPR_A), gRNA Scaffold necessary for Cas9 binding (gRNA_Scaffold), Csy4 cleavage-site (Csy4_k), 20bp long spacer (CRISPR _B), gRNA Scaffold necessary for Cas9 binding (gRNA_Scaffold), polyadenylation sequence from cauliflower mosaic virus 35S transcript (35S_T), right border (RB) from Agrobacterium tumefaciens. CRISPR_A = 20bp long spacer sequence from the potato CHL1 gene. CRISPR_B = 20bp long spacer sequence from the potato CHL1 gene.

DMR6 T-DNA: From left to right border; left border sequence (LB) from Agrobacterium tumefaciens, polyadenylation sequence from 35S transcript (35S_T), neomycin phosphotransferase II coding sequence (nptII) that can be isolated from different bacteria (marker, NptII), 2x35S promotor from cauliflower mosaic virus 35S gene (2x35S_P), 35S promotor (35S_P) from cauliflower mosaic virus 35S gene, Csy4 ribonuclease gen from Pseudomonas aeruginosa that has been codon-optimized for tomato (Csy4_enz), P2A ribosomal skipping sequence (P2A), Cas 9 gene that has been codon-optimized for Arabidopsis thaliana (Cas9), heat shock protein terminator (HSP_T) from plants, CmYLCV promotor from yellow leaf curling virus (CmYLCV_P), Csy4 cleavage-site (Csy4_k), 20bp long spacer (CRISPR_A), gRNA Scaffold necessary for Cas9 binding (gRNA_Scaffold), Csy4 cleavage-site (Csy4_k), 20bp long spacer (CRISPR _B), gRNA Scaffold necessary for Cas9 binding (gRNA_Scaffold), polyadenylation sequence from cauliflower mosaic virus 35S transcript (35S_T), right border (RB) from Agrobacterium tumefaciens. CRISPR_A = 20bp long spacer sequence from the potato DMR6 gene. CRISPR_B = 20bp long spacer sequence from the potato DMR6 gene.

Man_RNAi T-DNA: : From left to right border; left border sequence (LB) from Agrobacterium tumefaciens, nopaline synthase polyadenylation sequence from Agrobacterium tumefaciens (Nos_T), neomycin phosphotransferase II coding sequence (nptII) that can be isolated from different bacteria (marker, NptII), nopaline synthase promoter (Nos_P) from Agrobacterium tumefaciens, polyadenylation sequence from 35S transcript (35S_T), duplicated and inverted fragment of potato Mannnanase gene, between the duplication is a plant intron (Intron), 35S promotor (35S_P) from cauliflower mosaic virus 35S gene, right border (RB) from Agrobacterium tumefaciens. In case of deletion of genetic material, give information on the function of the deleted sequences: CHL1: The deleted regions are coding (i.e. in exon). Removal of these sequences will give a non-functional Chl1 protein

DMR6: The deleted regions are coding (i.e. in exon). Removal of these sequences will give a non-functional Dmr6 protein

Man-RNAi – not applicable Brief description of the method used for the genetic modification: We have mutated susceptibility factors - proteins that make the plant more susceptible to attack by certain pathogens. Loss of function of a sensitivity factor, for example by mutation, produces recessive resistance. The most well-known example is MLO (Mildew resistance Locus O), which has been used in plant breeding for many years.

For transformation of potato a binary vector system where sequences to be transferred can be found inside the border sequences that form a transfer DNA (T-DNA) was used. The DNA mobilization features are available in a modified Ti plasmid that is not transferred to the plant. For transformation of T-DNA to the potato, Agrobacterium tumefaciens containing the vector has been used. Cut potato leaf tissue was transformed and transgenic shoots were selected on antibiotics. After transformation, the Agrobacterium killed with 400 mg/ml Cefotaxime. If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination: Not applicable Experimental Release Purpose of the release: Under field conditions evaluate agricultural value including resistance properties. Study the stability of the modified features, identify any morphological abnormalities, producing field-grown material for laboratory testing, and produce seed for next year's field trials. The long-term goal is late blight and early blight resistance and to gain knowledge about plant resistance mechanisms. The experiment is only for research purposes. Geographical location of the site: Scania (Kristianstad, Lomma and Kävlinge) Size of the site (m2): Less than 10000 m2 Relevant data regarding previous releases carried out with the same GM-plant, if any, specifically related to the potential environmental and human health impacts from the release: Not applicable Environmental Impact and Risk Management Summary of the potential environmental impact from the release of the GMPts: Potatoes are grown on 1% of agricultural land, but as much as 20% of all fungicides used in Sweden are sprayed on potatoes. Cultivation of resistant potatoes would provide great environmental benefits as the amount of fungicide used could be reduced.

Potato is not known as a colonizer of wild ecosystems (OECD, 1997), and does not occur outside the field. Very rarely does potato waste plants occur in the cultivated landscape since tubers are destroyed during the next year's tillage, use of herbicides and competition of subsequent crops. In theory, resistance to pathogens could have a benefit over non-resistant potato varieties, but there is no evidence that the pathogen resistant potato varieties that are grown today are invasive or give rise to more overwintering tubers. Increased resistance is assumed not to give any change in the viability of the modified potatoes or give an increased risk for health and environment. Brief description of any measures taken for the management of risks: Most modern potato varieties have low pollen fertility and the formation of potato berries and seeds is extremely low. According to the literature, the distance of the spread of potato pollen is maximum of ten meters. A safety distance of 20 meters will be held. All flower buds from the genetically modified plants will be removed. Cleaning of machinery, tools, and transportation vehicles will be done after contact with the modified lines. The field will be checked, post-harvest for growth of surviving tubers. The field will not be used to plant potatoes again until at least one growing season without surviving potatoes is observed. Potential waste tubers will be documented and eliminated.

Transport of harvested potatoes will take place by car or truck in sealed double bags. Transport will not take place with other cultivated potatoes. When transporting, documentation must be included, stating that it is GM and the name and telephone number of a contact person. Harvested potatoes will be stored in SLU's premises licensed for such activities or be destroyed either by autoclaving or by incineration. Analyses of the material will be in SLU's premises and leftover crop residues will be destroyed by autoclaving or incineration. Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release: Not applicable Final report - European Commission administrative Information Consent given by the Member State Competent Authority: Yes
03/30/2020
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