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General information Notification Number B/SE/21/22027 Member State to which the notification was sent Sweden Date of acknowledgement from the Member State Competent Authority 10/12/2021 Title of the Project Changes in lignin with directed mutagenesis Proposed period of release: 01/05/2022 to 31/12/2026 Name of the Institute(s) or Company(ies) SweTree Technologies AB
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
grey poplar salicacee populus populus x canescens INRA 717-1B4
Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications: The trees contain a mutation in each allele in the coding region of the gene encoding the enzyme cinnamoyl-CoA reductase 2 (CCR2). The poplar species P. tremula × alba is diploid and the gene CCR2 is thus found on two alleles in the genome.Cinnamoyl-CoA reductase (CCR) catalyzes hydroxycinnamoyl-CoA esters to their corresponding hydroxycinnamic aldehydes (mainly feruloyl-CoA to coniferaldehyde), which in turn are used in the production of lignin in the plant. Lignin is a polymer in plant cell walls that helps to give it mechanical strength, where lignin together with cellulose gives both strength and flexibility. At the same time, lignin complicates enzymatic processing of biomass and is the most important factor limiting the conversion of plant biomass to fermentable sugars, and in pulp production, lignin has a negative impact on paper quality.The specific combination of mutations that have been generated in these plants, with a null allele and a haploinsufficient CCR2 allele, has resulted in the plant having a 10% lower lignin content when growing normally after all and saccharification efficiency has increased markedly. Furthermore, the plant exhibits a slightly modified lignin composition, which may further contribute to the increased saccharification efficiency. Genetic modification Type of genetic modification: Other; Other

Introduction of genetic material containing CRISPR/Cas9 modules, used to make a targeted gene edit.
Brief description of the method used for the genetic modification: The transgenic trees were obtained by the well-known and established method of Agrobacterium-mediated genetic modification.The purpose of the introduction of the transgene is to supply the plant with a CRISPR/Cas9 complex which in turn, with the help of the enzyme Cas9 and guide RNA (gRNA), makes a targeted gene edit at a pre-specified site in the plant's genome and thus inactivates or changes the function of a gene. This method is also well known and established If the recipient or parental plant is a forest tree species, describe ways and extent of dissemination and specific factors affecting dissemination: Gray poplar is wind-pollinated and produce wind-dispersed seeds. The clone in the experiment is a female clone. Gray poplar can also propagate from suckers. Experimental Release Purpose of the release: The aim of this experiment is to field test plants with CRISPR/Cas9-generated mutations that give a reduced level of lignin while maintaining normal growth of the plant, which have been shown in greenhouse experiments. This to provide a substantially increased fermentable sugar yield from wood biomass, which can lead to environmental benefits in the transition from a fossil-based to bio-based economy. Geographical location of the site: Våxtorp in Laholms municipial Size of the site (m2): Maximum 5000 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: The modifications aim to reduce the lignin content in the wood, with maintained growth.Altered lignin content does not bring any known selective benefits to the trees in a natural environment. The trees are expected to grow as wild-type plants.Reduced lignin content that increases fermentable sugar yield from the wood biomass can mean significant environmental benefits in the transition from fossil-based to bio-based economy.Antibiotic resistance genes, nptII and hph, are expressed in the plant. These resistance genes can only give the trees an increased selective advantage in the presence of high levels of these substances, which does not occur in nature. Therefore, we assess that there is no increased environmental risk with these. Brief description of any measures taken for the management of risks: Regular inspection to detect if any trees form flower buds. The clone used is a female clone that does not spread pollen. It is possible to identify flower buds long before they start flowering. Should any tree, in contrary to expectations, form flower buds, all branches on that tree will be taken down and destroyed. The field site will be fenced and a surrounding safety zone will prevent suckers from spreading from the experiment. After the end of the experiment, all experimental plants will be terminated and the site will continue to be inspected annually for at least 3 years.  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
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