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General information Notification Number B/SE/24/5457 Member State to which the notification was sent Sweden Date of acknowledgement from the Member State Competent Authority 12/04/2024 Title of the Project Aspen as a model system Proposed period of release: 01/09/2024 to 31/08/2029 Name of the Institute(s) or Company(ies) Swedish University of Agricultural Sciences, SE-90183, UMEÅ, SWEDEN
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
hybrid aspen	salicacee	populus	populus trichocarpa x populus deltoides		T89 and 3015-15

Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications: This application deal with 13 different lines/modifications, described below. 1) pgm1/pgm2 (coding for phosphoglucomutase). These plants have a severly decreased starch synthesis due to mutations (induced using CRISPR) in the two genes coding fo a key enzyme in starch metabolism, phosphoglucomutase (PGM). Hygromycin-resistance as selecatble marker. 2) sex4-1/sex4-2 (coding for "Starch excess 4"). Hygromycin-resistance gene under the CaMV 35S-promoter, Cas9 also under the 35S-promoter and a pair of guide-RNAs tageted against sax4-1 and sex4-2. Intended function to induce deletions in the sex4-1 and sex4-2 genes. 3) sex1-1/sex1-2 (coding for "Starch excess 1"). Hygromycin-resistance gene under the CaMV 35S-promoter, Cas9 also under the 35S-promoter and a pair of guide-RNAs tageted against sax1-1 and sex1-2. Intended function to induce deletions in the sex1-1 and sex1-2 genes. 4) Cao1/Cao2 (coding for chlorophyll a oxygenase). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against cao1 and cao2. Intended function to induce deletions in the cao1 and cao2 genes. 5) PsbS (coding for photosystem II-S protein). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against PsbS. Intended function to induce deletions in the PsbS gene. 6) VDE (coding for violaxanthin de-epoxidase). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against VDE. Intended function to induce deletions in the VDE gene. 7) Stn7 (coding for "state transition 7"). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against Stn7. Intended function to induce deletions in the Stn7 gene. 8) Pgr5 (coding for "proton gradient regulation 5". Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against pgr5. Intended function to induce deletions in the pgr5 gene. 9) ZEP1/ZEP2 (coding for zeaxanthin epoxidase). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pairs of guide-RNAs tageted against ZEP1 and ZEP2. Intended function to induce deletions in the ZEP1 and ZEP2 gene. 10. VPZ overexpressor. Kanamycin-resistance gene under the Nos-promoter, VDE under the Rbcs1a promoter, PsbS under the GAPA-1 promoter and ZEP under the FBA2 promoter. 11) CALS3 overexpressor (coding for callose synthase isoform CALS3. Bar-resistance gene, Cal3S under the IRX8 promoter. 12) FMT overexpressor (coding for feruloyl-coenzyme A monolignol transferase. Kanamycin- and hygromycin-resistance genes, FMT under the CAS8A and under the C4H promoter. 13) F5H overexpressor (coding for ferulate 5-hydroxylase). Kanamycin- and hygromycin resiostance resistance genes. F5H under the CES8A promoter VDE under the Rbcs1a promoter, PsbS under the GAPA-1 promoter and ZEP under the FBA2 promoter. 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: 1) pgm1/pgm2 (coding for phosphoglucomutase). Hygromycin-resistance gene under the CaMV 35S-promoter, Cas9 also under the 35S-promoter and a pair of guide-RNAs tageted against pgm1 and pgm2. Intended function to induce deletions in the pgm1 and pgm2 genes. 2) sex4-1/sex4-2 (coding for "Starch excess 4"). Hygromycin-resistance gene under the CaMV 35S-promoter, Cas9 also under the 35S-promoter and a pair of guide-RNAs tageted against sax4-1 and sex4-2. Intended function to induce deletions in the sex4-1 and sex4-2 genes. 3) sex1-1/sex1-2 (coding for "Starch excess 1"). Hygromycin-resistance gene under the CaMV 35S-promoter, Cas9 also under the 35S-promoter and a pair of guide-RNAs tageted against sax1-1 and sex1-2. Intended function to induce deletions in the sex1-1 and sex1-2 genes. 4) Cao1/Cao2 (coding for chlorophyll a oxygenase). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against cao1 and cao2. Intended function to induce deletions in the cao1 and cao2 genes. 5) PsbS (coding for photosystem II-S protein). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against PsbS. Intended function to induce deletions in the PsbS gene. 6) VDE (coding for violaxanthin de-epoxidase). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against VDE. Intended function to induce deletions in the VDE gene. 7) Stn7 (coding for "state transition 7"). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against Stn7. Intended function to induce deletions in the Stn7 gene. 8) Pgr5 (coding for "proton gradient regulation 5". Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pair of guide-RNAs tageted against pgr5. Intended function to induce deletions in the pgr5 gene. 9) ZEP1/ZEP2 (coding for zeaxanthin epoxidase). Kanamycin-resistance gene under the Nos-promoter, Cas9 under the 35S-promoter and a pairs of guide-RNAs tageted against ZEP1 and ZEP2. Intended function to induce deletions in the ZEP1 and ZEP2 gene. 10. VPZ overexpressor. Kanamycin-resistance gene under the Nos-promoter, VDE under the Rbcs1a promoter, PsbS under the GAPA-1 promoter and ZEP under the FBA2 promoter. 11) CALS3 overexpressor (coding for callose synthase isoform CALS3. Bar-resistance gene, Cal3S under the IRX8 promoter. 12) FMT overexpressor (coding for feruloyl-coenzyme A monolignol transferase. Kanamycin- and hygromycin-resistance genes, FMT under the CAS8A and under the C4H promoter. 13) F5H overexpressor (coding for ferulate 5-hydroxylase). Kanamycin- and hygromycin resiostance resistance genes. F5H under the CES8A promoter VDE under the Rbcs1a promoter, PsbS under the GAPA-1 promoter and ZEP under the FBA2 promoter. In case of deletion of genetic material, give information on the function of the deleted sequences: 1) pgm1/pgm2. Deletions in the pgm1 and pgm2 genes.2) sex4-1/sex4-2. Deletions in the sex4-1 and sex4-2 genes.3) sex1-1/sex1-2. Deletions in the sex1-1 and sex1-2 genes.4) cao1/cao2. Deletions in the cao1 and cao2 genes.5) PsbS. Deletions in the PsbS gene.6) VDE. Deletions in the vde gene.7) Stn7. Deletions in the Stn7 gene.8) ZEP1/ZEP2. Deletions in the ZER1 and ZEP2 genes.9-13) None Brief description of the method used for the genetic modification: Agrobacterium-mediated transformation If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination: Populus sp. seeds are distributed by wind, and new shoots are also generated from roots. Experimental Release Purpose of the release: Basic research on tree biology Geographical location of the site: Alnarp, Skåne and Umeå university, Västerbotten, both Sweden Size of the site (m2): ca. 1 ha i Alnarp, ca 10 m2 (in pots) in Umeå 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: Note especially if the introduced traits could directly or indirectly confer an increased selective advantage in natural environments; also explain any signifant expected environmental benefitsThis is mainly basic research, in most cases deletions in genes important for starch metabolism and regulation of photosynthesis. Those modifications will most likely only have selective disadvantages, plants will grow less well or as good as wild type plants and it is hard to imagine any impact on the environment. In a few cases (like VPZ) photosynthesis may increase, it would be advantageous with trees that would grow better under natural conditions but it is hard to imagine any negative impact on the environment of this. Some trees could get modified cell wall composition which may improve wood quality for processing, also in this case it is hard to imagine any negative consequences on the environment. It could be added that we have performed several similar experiments in the past and no negative impact on the envorinment has been found. Of course, increased productivity and/or better processability of the wood could in the long run give substantial environmental benefits. Brief description of any measures taken for the management of risks: Alnarp: The trees will be cut down at the end of the experiment. Stumps and roots are eliminated mechanically (digging,harrowing, milling), or chemically if necessary. The experimental site will be plowed and the trial area monitored for at least 3 years after the experiment or after the last residual tree sapling was found. Regular inspections to find trees that may go into flowering (those would be destructed), suckers will be removed mechanically and the site is fenced. Umeå: Trees and soil in the pots will be destructed 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: Not known