AFBV-WGG Explanatory Note

Explanatory Note supporting the AFBV-WGG Initiative

Suggestions to enable the development of genome editing in Europe

A. The challenges facing Agriculture:

Global agriculture is facing many challenges, the most important of which are a growing world population (9-10 billion people in 2050) and the scarcity of arable land which will remain at best stable. Hence the need to produce more on the same area while taking into account:
  • Environmental constraints related to climatic variations and the need to reduce inputs (crop protection products, fertilizers, water, etc.);
  • The demands and constraints coming from consumers and the food chain.
B. The need to continue to innovate for crop improvement:

To meet these challenges, all stakeholders must continue to develop innovative and efficient agriculture in France, Germany, Europe and the rest of the world. Among the innovations which are required at all steps from seed to fork, those related to plant genetics play an important role. It is essential that all technologies available for the creation of new plant varieties can be used without exclusion in principle.

C. Genome editing:

Genome editing, one of the techniques referred to as NPBT (New Plant Breeding Techniques), brings together a set of technologies allowing the modification of genetic information by addition, deletion or exchange (replacement) of nucleotides at a targeted site of the genome sequence of a recipient plant. These technologies will become essential tools for quickly obtaining, for example, resistance to biotic stresses, pathogens and aggressors, increased tolerance to abiotic stress such as tolerance to drought or temperature variations; as well as improving sanitary, technological and nutritional qualities of harvested products.

These technologies have shown a significant potential for genetic improvement in research and development. In fact, the first plants derived from these technologies are on the market in North America. Various analyses and evaluations of these technologies in France, Europe (in particular the February 2017 report of the Scientific Advice Mechanism on new biotechnologies in agriculture) and other countries conclude that these new seeds are not different in their effects on health or environment from those obtained from traditional breeding techniques. See SAM (2017) “New Techniques in Agricultural Biotechnology”, https://doi.org/10.2777/17902, and SAM (2018) “A Scientific Perspective on the Regulatory Status of Products Derived from Gene Editing and the Implications for the GMO Directive”, https://doi.org/10.2777/407732.

Given the potential of these technologies, it seems essential for Europe to revise the regulatory framework for plants derived from genome editing techniques. For this purpose, we present below our proposal for a revision of that framework.

D. Basis for our approach:

AFBV and the WGG are aware that a complete revision of Directive 2001/18 / EC regulating GMOs will take a long time, which is difficult to reconcile with the need to maintain the competitiveness of research teams and seed companies. Pending a complete overhaul of the European Directives and Regulations concerning GMOs, as well as a harmonization with international treaties, our organizations propose to quickly introduce in Directive 2001/18/EC and in related GMO Regulations and Directives, new provisions that will allow the use of genome editing techniques.

E. Proposed Additions to Directive 2001/18/EC:

Without affecting the logic and coherence of the whole Directive, we propose additions that take into account up-to-date scientific knowledge and technological progress. While these additions summarized below only concern changes in Directive 2001/18/EC, it is understood that the other GMO-related Directives and Regulations in Europe will have to be amended to incorporate the same changes.

Our proposals have been written with the intention of covering plants. They may be adapted, if necessary and where appropriate, to animals and microorganisms.

We propose to address (1) the conditions of use of technologies grouped under the term “genome editing” and (2) the regulatory status of null segregants, as follows:

   1.  Define genome-editing techniques. Include a definition of genome-editing techniques in the Directive (addition of a new 
point (4) to Annex I A, Part 1).

   2.  Remove from the scope of Directive certain categories of plants derived from genome editing. As genome-editing 
technologies can be used to create a broad range of plants with new traits, going from a change in one nucleotide up to the incorporation of whole genes, we are proposing to establish different categories of plants based on the type of edit that has been obtained. At this stage, we are proposing four categories of plants derived from genome-editing techniques which should be excluded from the Directive. Following confirmation of compliance of a proposed plant with an excluded category, in accordance with a confirmation process described below, such plant would then be regulated in the same way as plants derived from traditional breeding techniques. The four categories will be described in a new Annex I C and would include the following:

         ● Category 1: A plant having a native allele that has been edited[1] to reproduce a functionality associated with a known allele
   present  in its natural gene pool [2].
   Making such a change would be equivalent, for instance, to the transfer of a known allele from a wild counterpart to a  
   cultivated variety of the same species accomplished through traditional breeding. 

        ● Category 2: A plant having a native allele that has been edited to reproduce a functionality associated with a known allele 
  present in a plant species that is outside the plant’s natural gene pool.
  As the model allele exists in a non-sexually compatible plant species, there is no equivalent in traditional breeding. This 
  category would constitute an extension of Category 1 if the donor plant and the recipient plant were sexually compatible.

● Category 3: A plant having a native allele that has been edited to reproduce a new functionality, of which the sequence
  modifications obtained by genome editing are of the same type as those which be obtained by spontaneous or induced 
  mutagenesis.
  In traditional breeding, such changes would be equivalent to those obtained by selecting a plant having a new allele due to a 
  spontaneous or an induced mutation, which plant is then crossed with a cultivated plant in order to select the mutation of 
  interest.
● Category 4: A plant in which a gene known and present in its natural gene pool1 has been inserted into a targeted site of its 
   genome.
  Amongst genotypes of a species there exists a variation in the number (from zero to N) of copies of certain genes (this may 
  be due, for example, by duplication at the locus, uneven cross-overs or translocation via transposons). In traditional breeding 
  one can select for “copy number” as a criteria. The addition of allelic copies by genome editing reproduces directly this 
  breeding process.

With respect to all of the above categories, it is possible, through genome editing, to have in the same plant several edited alleles (or inserted genes). In such cases each edited allele (or inserted gene) shall be analysed independently according to the above-defined criteria. If all of the edited alleles or inserted genes fall under the same category, the plant belongs to such category. If the edited alleles or inserted genes belong to different categories, the plant must comply with each relevant category in order to be excluded. If a new edit is undertaken upon a different allele of a plant which has previously been determined to be excluded, only confirmation of exclusion for the new allele shall be required of the notifier.

Annex I hereof sets forth examples of plants belonging to the excluded categories based upon scientific publications or regulatory files accessible in public databases.

As scientific knowledge and technical progress evolve, additional new categories can be added to Annex I C (see also point 4 below).

3. Create a new, specific, efficient and predictable regulatory pathway for the above categories of genome-edited 
    plants.

   Confirmation of the exclusion of an edited plant must be obtained by the notifier. The confirmation process is adapted to the 
   exclusion category.

 ●  Procedure for submitting the confirmation request
- The notifier shall file its confirmation request with the competent authority of the Member State in charge of GMO regulations
   (in France, the Ministry of Agriculture, and in Germany the Federal Ministry of Food and Agriculture) who will rely on its 
   existing internal departments capable of evaluating GMOs (in France, ANSES or the HCB, and in Germany the BVL 
   [Bundesamt für Verbraucherschutz und Lebensmittelsicherheit - Federal Office of Consumer Protection and Food Safety]);
- The request for confirmation is made by the notifier whenever it wishes to benefit from the exclusion and remove its plant 
   from the scope of Directive 2001/18 / EC, REGULATIONS (EC) No 1829/2003, No 1830/2003 as well as any other GMO 
   regulations of the European Union.
- The exclusion decision for an edited plant shall be valid for all progeny of such plant containing the same edit and binding 
   upon all Member States;
- Once the confirmation of exclusion is obtained, any variety obtained using the edited plant shall be subject to seed and plant 
  variety regulations applicable to relevant crop species in the same manner as any variety obtained through traditional breeding
  techniques, including registration in the common catalogues of varieties of agricultural plant and vegetable species which can    be marketed in the European Union.

Contents of the confirmation request application

The information requirements to be supplied by the notifier shall be adapted to the plant category:

 ●  Standard requirements for all categories :
(i)    Name of the notifier and contact information ;
(ii)  Taxonomic description of the plant which has been edited or in which a gene has been inserted;
(iii) Technique used and main steps that have been followed, including, if applicable, whether or not an intermediate GMO was produced in the editing process, and the modalities of elimination of any inserted recombinant nucleic acid sequence, and confirmation of the elimination of any such inserted sequence (null segregant);

Requirements that are Category specific 
  ●  For Categories 1 et 2 :
(i)  Taxonomic description of the plant containing the model allele and a description of the model allele ;
(ii) Description of the edit realized in the final plant (addition, deletion or replacement) and confirmation that the resulting 
     edited sequence has been obtained and comparison of the functionality of the model and edited alleles ;

 ●  For Category 3 :
(i)  Description of the new allele and its functionality obtained after genome editing and available background information on
      the reasons that led to editing such allele (research work, for example) ;
(ii) Description of the edit realized in the final plant (addition, deletion or replacement) and confirmation that the resulting 
      edited sequence and its functionality have been obtained;

 ●  For Category 4 :
(i)   Taxonomic description of the donor plant containing the inserted gene and a description of such gene ;
(ii)  Confirmation of the sequence of the inserted gene in comparison to the original gene before insertion ;
(iii) Confirmation that the inserted gene is located at the site targeted by genome editing.

Any information supplied by the notifier for which it wishes to claim confidentiality must be marked "Confidential".

The processing time by the competent authority of a Member State to determine whether or not an edited plant falls under one of the four Categories for exclusion should be no more than sixty days. 

4. Permit periodic updating of the Directive if justified by advances in scientific knowledge and technical progress. 
    As indicated above, these proposals are based on the current state of scientific knowledge and technical progress achieved based 
    upon that knowledge. As scientific knowledge and technical progress evolve rapidly in this field, we propose that every five years,
    after consulting the relevant stakeholders and in collaboration with the competent authorities of the Member States, the 
    Commission reports to the European Parliament on developments in scientific knowledge and technical technological progress and,
    if necessary, proposes a revision of the annexes.

5. Address the status of null segregants (progeny of a GMO plant from which the GMO feature has been removed). 
    As part of this revision of the Directive, we propose that null segregants be confirmed as being excluded from the scope of the
    Directive. A null segregant that is obtained after genome editing and that is also an edited plant is subject to the confirmation 
    process to confirm exclusion under one of the four Categories above.

    These different proposals are included in a draft amendment which you will find attached hereto.

Frankfurt and Paris, January 2020


[1] The terms ‘Editing’ or ‘edited’ refer to the application of ‘genome editing’ techniques.
[2] The term ‘natural gene pool’ refers to the gene pool of a plant species defined as all of the genes and alleles (i.e., different versions of the same gene)
     obtained from plants which can exchange genes by sexual crossing as well as from distantly related plant species with which genes can be exchanged
     by sexual crosses using traditional breeding techniques

Annex 1

Examples of plants falling under excluded categories,
based upon scientific publications or regulatory files accessible in public databases

These examples are taken from the literature or from regulatory files. We tried to find, from available public information, the origin of the model alleles. Thus, for each example, and when available, the first reference discloses the edited plant and the other references describe the probable origin of the model alleles. Except for the plants already marketed in North America, these examples do not prejudge the fate of these edited plants and their commercial opportunities.

Methodology and criteria used:
  • The example must describe an edited plant that has been achieved;
  • For the examples of Categories 1 and 2, a model allele is identified in a plant that is sexually compatible (Category 1) or non-sexually compatible (Category 2);
  • For the examples of Category 3, information is provided on the approaches used to obtain the edited gene, including results in transgenic plants (RNAi experiments for example);
  • For category 4, information is provided on the inserted gene;
  • For the edited plants we tried to use the original publication; for the model alleles we sought to find them in the publications cited by the inventors of the edited plant.
Category 1:
  • An edited, salt-tolerant rice plant, following inactivation of the OsRR22 gene (known allele). Zhang et al., 2019; Takagi et al., 2015.
  • A potato plant edited by inactivating the StGBSSI gene (known allele), leading to the accumulation of amylopectin (waxy starch) in the tuber. Based on the availability of potato mutants rich in amylopectin and on knowledge of the synthesis of amylopectin in cassava, corn and wheat. Veillet et al., 2019; Hovenkamp-Hermelink et al., 1987.
  • A rice plant in which the promoter of three genes coding for sucrose transporters, SWEET11, SWEET13 and SWEET14 has been edited (modification of nucleotides) to no longer be sensitive to the transcription factor produced by Xanthomonas oryzae pv. Oryzae. There are rice mutants for these genes; several have been associated in this edited plant. Oliva et al., 2019; Zaka et al., 2018.
  • A pink-fruited tomato plant following inactivation of the SlMYB12 gene (known allele). Deng et al., 2018; Fernandez-Moreno et al., 2016.
  • A maize plant tolerant to Setosphaeria turcica (Helminthosporium turcicum) following the replacement, by edition, of the sensitive allele of the NLB 18 gene coding for a membrane kinase and responsible for the interaction with the fungus by the resistant allele identified in a corn tolerant to this fungus (known allele). Schmidt 2018; Hurni et al., 2015; Li & Wilson 2006.
  • A maize plant accumulating only amylopectin in the seed following inactivation of the waxy (Wx1) gene coding for the Granule Bound Starch Synthase (GBSS) (known allele). Based upon the waxy maize mutant which has been marketed for many years. Schmidt 2016.
  • A soybean plant with a high oleic acid content following inactivation of two fatty acid desaturase genes (FAD2-1A and FA D2-1B) (known alleles). Haun et al., 2014; Pham et al., 2010.
  • A rapeseed plant edited to be tolerant to imidazolinone and sulfonylurea herbicide families by modifying a single nucleotide of the BnAHAS1 gene in Genome C and a single nucleotide of the BnAHAS3 gene in Genome A of Brassica Napus. https://www.inspection.gc.ca/plant-health/plants-with-novel-traits/approved-under-review/decision-documents/dd-2013-100/eng/1427383332253/1427383674669; https://www.canada.ca/en/health-canada/services/food-nutrition/genetically-modified-foods-other-novel-foods/approved-products/novel-food-information-cibus-canola-event-5715-imidazolinone-sulfonylurea-herbicide-tolerant.html. Many mutants of this enzyme are known to exist in rapeseed, conferring tolerance to sulfonylureas. Magha et al., 1993.
Category 2:
  • A tomato plant whose gene SlJAZ2, orthologue of the AtJAZ2 gene of Arabidopsis, has been edited (modification of the nucleotide sequence) to reproduce the dominant mutant version of Arabidopsis (absence of the C-terminal - jas motif) to obtain the resistance to bacterial spot disease (Pseudomonas syringae pv. tomato (Pto) DC3000). This modified receptor, SlJAZ2Δjas, no longer fixes the coronatine synthesized by the bacteria and as a resultthe stomata do not open. Ortigosa et al., 2019; Gimenez-Ibanez et al., 2017.
  • An edited grape cultivar in which (i) the Mlo gene has been suppressed to obtain powdery mildew resistance and (ii) the VvDMR6 gene has been suppressed based upon knowledge of the suppression of the analogous gene in Arabidopsis thaliana resulting in downy mildew resistance. Giacomelli et al., 2019; van Damme et al., 2008.
  • A cassava plant resistant to potyvirus [Cassava brown streak disease (CBSD)] obtained by editing (modification of the nucleotide sequence) of the gene coding for the translation initiation factor elF4E. Many isoforms of this factor giving potyvirus resistance are known in many plants: chilli, tomato, pea, Arabidopsis mutants. Gomez et al., 2019; Bastet et al. 2019.
  • An edited wheat plant in which the three genes corresponding to the Mildew resistance Locus (Mlo) called TaMlo‐A1, TaMlo‐B1 and TaMlo‐D1, located on chromosomes 5AL, 4BL and 4DL, are simultaneously inactivated to reproduce a phenotype resistant to powdery mildew, based upon the knowledge of Mlo alleles naturally present in barley. Wang et al., 2014; Büschges et al., 1997.

Category 3:
  • An apple cultivar where the MdDIPM4 gene (a kinase receptor) is inactivated by editing to obtain resistance to scab (Erwinia amylovora). By analogy with Arabidopsis mutants and studies of receptor interaction with the bacterium effector (DspA / E) a sequence of MdDIPM4 was deleted in the apple gene. Pompili et al., 2019; Degrave et al., 2013; Borejsza-Wysocka et al., 2004.
  • A petunia plant with prolonged flowering by inactivation of the Ph ACO1 gene which codes for a 1-aminocyclopropane-1-carboxylate oxidase involved in the production of ethylene (reduced quantity in the edited plant). By analogy with the results obtained by expressing antisense in petunia. Xu et al., 2019; Huang et al., 2007.
  • A durum wheat plant that has been edited to inactivate up to 35 of the 45 α-gliadin genes (known alleles) on three chromosomes, causing a reduction in the production of α-gliadins and a drop in immunoreactivity by 85%. Sanchez Leon et al., 2018.
  • A tomato plant of which the promoter of the SlCLV3 allele (new allele) has been edited in order to increase fruit size. Rodriguez-Leal et al., 2017.
  • In several citrus species, the promoter of the CsLOB1 gene (LATERAL ORGAN BOUNDARIES 1) has been edited by deletion of the sequence EBEPthA4 (which fixes the effector produced by the bacteria) conferring resistance to citrus canker [Xanthomonas citri subsp. citri (Xcc)]. Based on knowledge of the interactions between the promoter and the effector of the bacteria and on similar works on rice. Jia et al, 2016a (grapefruit tree); Jia et al., 2016b (lemon tree); Peng et al., 2017 (orange tree). In order for these edited plants to benefit from the exclusion provided by this Category 3, the recombinant DNA used for the editing will need to be removed (null segregants).

Category 4:
We did not find any plants that met the criteria for this category. There are many examples of plants containing one or more cisgenes (see two examples below), but none are the result of insertion at a site and homologous recombination. The cisgenes introduced into the plants described below were obtained by transgenesis. With genome editing, a cisgene may be inserted in a chosen site by double homologous recombination, without any residual vector sequence.

  • A potato plant in which several mildew resistant genes identified exclusively in wild potato species have been inserted using Agrobacterium tumefaciens, selected on the criteria that (i) all R genes are expressed and (ii) conformity to the varietal type is maintained. Haverkort et al., 2016.
  • An apple cultivar made resistant to scab by inserting the cisgene FB_MR5 from the wild variety Malus × robusta 5 (Mr5) in chromosome 16. Kost et al., 2015.

Examples of edited plants having alleles in different categories:

As indicated earlier in this Explanatory Note, the same edited plant may contain alleles which correspond to different categories. Two examples are presented below.

  • A tomato plant that has been edited by inactivating (1) the SIER gene (which regulates tomato stem length), (2) the SP5G gene (linked to rapid flowering) and (3) the SP gene (linked to precocious growth termination), all three genes having known mutant alleles, to make it compact and early yielding, suitable for urban agriculture. This plant contains edited genes corresponding to Category 1 for the alleles of the SlER and SP genes and to Category 3 for the allele of the SP5G gene. Kwon et al. 2019; Xu et al., 2015; Soyk et al., 2017, and Menda et al., 2004.
  • An edited cassava plant accumulating amylopectin (waxy starch) instead of amylose following inactivation of the PTST1 gene encoding the Protein Targeting to STarch and the GBSS1 gene encoding the Granule Bound Starch Synthase. Based on the availability of cassava mutants rich in amylopectin and knowledge of the synthesis of amylopectin in potatoes, corn and wheat. This plant contains two edited genes, the allele of the GBSS1 gene corresponds to Category 1 and the allele of the PTST1 gene to Category 3. Bull et al., 2018; Morante et al., 2016

References cited in the above examples:
Bastet et al. 2019. Mimicking natural polymorphism in eIF4E by CRISPR-Cas9 base editing is associated with resistance to potyviruses. 
Plant Biotechnology Journal 17: 1736–1750- doi: 10.1111/pbi.13096
Borejsza-Wysocka et al., 2004. Silencing of apple proteins that interact with DspE, a pathogenicity effector from Erwinia amylovora, as a strategy to 
increase resistance to fire blight. Acta Horticulturae 663: 469–474 - doi:10.17660/ActaHortic.2004.663.81
Bull et al., 2018. Accelerated ex situ breeding of GBSS- and PTST1-edited cassava for modified starch. 
Science Advances 4:eaat6086 - doi.org/10.1126/sciad v.aat60 86
Büschges, R. et al., 1997. The barley Mlo gene: A novel control element of plant pathogen resistance. 
Cell 88: 695–705.
Degrave et al., 2013. The bacterial effector DspA/E is toxic in Arabidopsis thaliana and is required for multiplication and survival of fire blight pathogen. 
Molecular Plant Pathology 14: 506–517 - DOI: 10.1111/mpp.12022.
Deng et al., 2018. Efficient generation of pink-fruited tomatoes using CRISPR/Cas9 system. 
Journal of Genetics and Genomics 45: 51-54 - doi.org/10.1016/j.jgg.2017.10.002.
Fernandez-Moreno et al. 2016. Characterization of a new pink-fruited tomato mutant results in the identification of a null allele of the SlMYB12. 
Plant Physiology 171: 1821-1826.
Giacomelli et al., 2019. Generation of mildew-resistant grapevine clones via genome editing, 
ISHS Acta Horticulturae 1248: XII International Conference on Grapevine Breeding and Genetics - DOI: 10.17660/ActaHortic.2019.1248.28.
Ibanez et al., 2017. JAZ2 controls stomata dynamics during bacterial invasion. 
New Phytologist 213: 1378–1392 - doi: 10.1111/nph.14354.
Gomez et al., 2019. Simultaneous CRISPR/Cas9-mediated editing of cassava eIF4E isoforms nCBP-1 and nCBP-2 reduces cassava 
brown streak disease symptom severity and incidence. Plant Biotechnology Journal 17: 421–434 - doi: 10.1111/pbi.1298.
Haun et al., 2014. Improved soybean oil quality by targeted mutagenesis of the fatty acid desaturase 2 gene family. 
Plant Biotechnology Journal 12: 934–940 - doi: 10.1111/pbi.12201.
Haverkort et al., 2016. Durable Late Blight Resistance in Potato Through Dynamic Varieties Obtained by Cisgenesis: Scientific and 
Societal Advances in the DuRPh Project. Potato Research - DOI 10.1007/s11540-015-9312-6.
Hovenkamp-Hermelink et al., 1987. Isolation of an amylose-free starch mutant of the potato (Solanum tuberosum L.). 
Theoretical Applied Genetics 75: 217–221 - https ://doi.org/10.1007/bf002 49167.
Huang et al., 2007. Delayed flower senescence of Petunia hybrida plants transformed with antisense broccoli ACC synthase and
 ACC oxidase genes. Postharvest Biol. Technol. 46: 47–53.
Hurni et al., 2015. The maize disease resistance gene Htn1 against northern corn leaf blight encodes a wall associated receptor-like 
kinase. Proceedings of the National Academy of Sciences 112: 8780-8785 - doi/10.1073/pnas.1502522112.
Jia et al., 2016a. Modification of the PthA4 effector binding elements in Type I CsLOB1 promoter using Cas9/sgRNA to produce 
transgenic Duncan grapefruit alleviating XccDpthA4:dCsLOB1.3 infection. Plant Biotechnol. J. 14, 1291–1301.
Jia et al., 2016b. Genome editing of the disease susceptibility gene CsLOB1 in citrus confers resistance to citrus canker.
 Plant Biotechnol. J., doi.org/10.1111/pbi.12677.
Kwon et al., 2019. Rapid customization of Solanaceae fruits crops for urban agriculture. 
Nature Biotechnology - doi.org/10.1038/s41587-019-0361-2.
Kost et al., 2015. Development of the first cisgenic apple with Increased Resistance to Fire Blight.
 PLoS ONE, 10, e0143980 - DOI:10.1371/journal.pone.0143980.
Li & Wilson, 2006. Composition and methods for enhancing resistance to northern leaf blight in maize. 
World Intellectual Property Organization, Application No. PCT/US2011/041822.
Magha et al., 1993. Characterization of a spontaneous rapeseed mutant tolerant to sulfonylurea and imidazolinone herbicides. 
Plant Breeding 111: 131-141.
Menda et al., 2004. In silico screening of a saturated mutation library of tomato. 
Plant Journal 38: 861–872.
Morante et al., 2016. Discovery of new spontaneous sources of amylose-free cassava starch and analysis of their structure and 
techno-functional properties. Food Colloids 56: 303-395 - doi.org/10.1016/j.foodhyd.2015.12.025.
Oliva et al., 2019. Broad-spectrum resistance to bacterial blight in rice using genome editing. 
Nature Biotechnology 37: 1344-1350.
Ortigosa et al., 2019. Design of a bacterial speck resistant tomato by CRISPR/Cas9-mediated editing of SlJAZ2.
 Plant Biotechnology Journal 17: 665–673 - doi: 10.1111/pbi.13006.
Peng et al., 2017. Engineering canker-resistant plants through CRISPR/Cas9-targeted editing of the susceptibility gene CsLOB1 
promoter in citrus, Plant Biotechnology Journal 15: 1509–1519 - doi: 10.1111/pbi.12733.
Pham et al., 2010. Mutant alleles of FAD2-1A and FAD-1B combine to produce soybeans with the high oleic acid seed oil trait. 
BMC Plant Biology 10: 195-206 - biomedcentral.com/1471-2229/10/195.
Pompili et al., 2019. Reduced fire blight susceptibility in apple cultivars using a high-efficiency CRISPR/Cas9-FLP/FRT-based gene 
editing system. Plant Biotechnology Journal - doi: 10.1111/pbi.13253.
Rodriguez-Leal et al., 2017. Engineering Quantitative Trait Variation for Crop Improvement by Genome Editing,
 Cell 171, 470–480, http://dx.doi.org/10.1016/j.cell.2017.08.030.
Sanchez Leon et al., 2018. Low-gluten, non-transgenic wheat engineered with CRISPR-Cas9. 
Plant Biotechnology Journal 16: 902–910 - doi: 10.1111/pbi.12837.
Schmidt 2016. Corn with high content of amylopectin developed by CRISPR/Cas technology. 15-352-01_air_inquiry_cbidel Pioneer. 
https://www.aphis.usda.gov/aphis/ourfocus/biotechnology/am-i-regulated/regulated_article_letters_of_inquiry/regulated_article_letters_of_inquiry.
Schmidt 2018. Corn with Improved Resistance to Northern Leaf Blight developed by CRISPR-Cas technology. 17-076-
018_air_inquiry_a1_cbidel revised Pioneer, https://www.aphis.usda.gov/aphis/ourfocus/biotechnology/am-i-regulated/regulated_article_letters_of_inquiry/regulated_article_letters_of_inquiry.
Soyk et al., 2017. Variation in the flowering gene SELF PRUNING 5G promotes day-neutrality and early yield in tomato.
Nature Genetics 49: 162–168.
Takagi et al., 2015. MutMap accelerates breeding of a salt-tolerant rice cultivar. 
Nature Biotechnology 33: 445–449.
van Damme et al., 2008. Arabidopsis DMR6 encodes a putative 2OG-Fe(II) oxygenase that is defense-associated but required for 
susceptibility to downy mildew. The Plant Journal 54:785-793 -. doi: 10.1111/j.1365-313X.2008.03427.x.
Veillet et al., 2019. The Solanum tuberosum GBSSI gene: a target for assessing gene and base editing in tetraploid potato,
 Plant Cell Reports 38:1065–1080, https://doi.org/10.1007/s00299-019-02426-w
Wang et al., 2014. Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery 
mildew. Nature Biotechnology 32: 947-952 - doi:10.1038/nbt.2969.
Xu et al., 2020. CRISPR/Cas9-mediated editing of 1-aminocyclopropane-1-carboxylate oxidase1 enhances Petunia flower longevity. 
Plant Biotechnology Journal 18: 287-297 - doi: 10.1111/pbi.13197.
Xu et al., 2015. A cascade of arabinosyltransferases controls shoot meristem size in tomato.
 Nature Genet. 47, 784–792.
Zaka et al., 2018. Natural variations in the promoter of OsSWEET13 and OsSWEET14 expand the range of resistance against
 Xanthomonas oryzae pv. PLoS ONE 13(9): e0203711 - doi.org/10.1371/journal.pone.0203711.
Zhang et al., 2019. Enhanced rice salinity tolerance via CRISPR/Cas9-targeted mutagenesis of the OsRR22 gene. 
Mol Breeding 39: 47-56 - doi.org/10.1007/s11032-019-0954-y.

A few published reviews - For additional information on the production of plants by genome editing:

Chen et al., 2019. CRISPR/Cas genome editing and precision plant breeding in Agriculture. 
Annual Review of Plant Biology 70: 28.1-28.31 – doi.org/10.1146/annurev-arplant-050718-100049.
Jaganathan et al., 2018. CRISPR for crop improvement. An update review. 
Frontiers in Plant Science - doi:10.3389/fpls.2018.00985
Metje et al., 2020. Genome edited plants in the field. 
Current Opinion in Biotechnology 61: 1-6 - doi.org/10.1016/j.copbio.2019.08.007.
Modrzejewski et al., 2019. Environmental Evidence - What is available evidence for the range of application of genome-editing as a 
new tool? Environmental Evidence 8- https://doi.org/10.1186/s13750-019-0171-5.
Sharma et al., 2019. Recent advances in developing disease resistance in plants, 
F1000Research, 8(F1000 Faculty Rev):1934 Last updated: 19 NOV 2019, doi.org/10.12688/f1000research.20179.1.
Soda et al., 2018. CRISPR-Cas9 based plant genome editing: significance, opportunities and recent advances. 
Plant Physiology and Biochemistry 131: 2-11 – dx.doi.org/10.1016/j.plaphy.2017.10.024.
Zhang et al., 2018. Applications and potential of genome editing in crop improvement. 
Genome Biology 19: 210-XXX – doi.org/10.1186/s13059-018-1586-y.

pdf-file: Explanatory Note supporting the AFBV-WGG Initiative


AFBV-WGG


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