General informationNotification NumberB/DE/03/152Member State to which the notification was sentGermanyDate of acknowledgement from the Member State Competent Authority14/10/2003Title of the ProjectFungal resistant wheat Germany 2004 (II)Proposed period of release:01/03/2004 to 31/10/2004Name of the Institute(s) or Company(ies)Syngenta Seeds GmbH, Alte Reeser Straße 95
3. Is the same GMPt release planned elsewhere in the Community?NoHas the same GMPt been notified elsewhere by the same notifier?YesIf yes, notification number(s): Genetically modified plantComplete name of the recipient or parental plant(s)
2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:FRG: Gene of fungal origin conferring tolerance to Fusarium pathogens.
|Common Name||Family Name||Genus||Species||Subspecies||Cultivar/breeding line|
|spring wheat||poaceae||triticum||triticum aestivum|
PMI: Phosphomannose Isomerase gene isolated from Escherichia coli, conferring tolerance to mannose.Genetic modification3. Type of genetic modification:Insertion; In case of insertion of genetic material, give the source and intended function of each constituent fragment of the region to be inserted:The plasmids used for the transformation were derived from the pUC19 plasmid described by Yanisch-Perron et al (1985).
pZMLR 14 (6610bp)
Intron: Intron #9pepc (I-9)
Ampicillin resistance gene controlled by a prokaryotic promoter sequence: Amp (bla gene).
pZMLR 69 (6636bp)
Ampicillin resistance gene controlled by a prokaryotic promoter sequence: Amp (bla gene).
Nature and source of the DNA coding sequences within PZMLR 14 and PZMLR 69
- Ubi: promoter of a maize ubiquitin gene together with the 1st exon and the 1st intron of the gene (Christensen et al, 1992).
- PMI: Phosphomannose Isomerase Gene isolated from Escherichia coli (Miles and Guest,1984). This gene is used as a marker for the transformation, and allows positive selection on mannose (Bojsen et al 1994, Joersbo et al 1998, Reed 1999, Negrotto et al 2000).
- Intron #9pepc (I-9): Intron isolated from the maize phosphoenolpyruvate carboxylase gene (pep-c) (Hudspeth and Grula, 1989); its role is to stimulate the expression of the PMI gene.
- 35S: Termination sequence of the cauliflower mosaic virus gene.
- Nos: Termination sequence of the nopaline synthase gene, isolated from Agrobacterium tumefaciens (Depicker et al 1982, Bevan et al 1983).
- FRG: cDNA isolated form a fungal species, coding for an enzyme capable of conferring Fusarium tolerance.
Prior to gene delivery, these plasmids were cut using restriction enzymes in order to eliminate the carrier fragment of the bacterial marker that confers resistance to ampicillin and to achieve transfer to the plant of only the plasmid fragment that carries the necessary genetic material for effective transformation.
Wheat plants with the same modification were evaluated in field trials in the USA and Canada in 2002 and 2003.6. Brief description of the method used for the genetic modification:Biolistics method.7. 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 Release1. Purpose of the release:To compare the agronomic performance (pathogen infestation level and mycotoxin level) of wheat modified to express an enhanced resistance to Fusarium pathogens with existing non-modified varieties, grown under standard agronomic conditions in Germany.
Two different transformation events will be assessed, compared with five conventional genotypes used as controls.2. Geographical location of the site:Location: Baalberge
Federal state: Saxony-Anhalt
Agrargenossenschaft Baalberge eG
Saxony-Anhalt3. Size of the site (m2):- The total trial ground (surface of the trial plus stripped area plus border of dicotyleton plants) will be approx. 8.300 m².
- The area comprising the released GM wheat plus the non GM wheat border rows (no less than 5 m) will be approximately 450 m².
- The trial will be surrounded by a 5m area stripped of all plants and a 30m border of dicotyleton plants.
- The total number of GM seeds to be used in this field trial will not be more than 6.000.4. 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:No adverse effects were noted from wheat plants with the same genetic modification released in the field in the USA and Canada in 2002 and 2003.Environmental Impact and Risk ManagementSummary of the potential environmental impact from the release of the GMPts:This application concerns a small-scale research trial which will take place in Baalberge, Germany. Due to the characteristics of the wheat plants, effects are expected to be limited to the site of the trial, and to be transient in nature.
Wheat is an annual plant that reproduces by means of seed. Seeds that are shed on the soil before or during harvest may overwinter and sprout the following spring. However seed is unlikely to survive longer than 2 years due to lack of dormancy (Field Agronomist, personal communication) and volunteers are easily controlled under standard agricultural conditions.
Wheat flowers display characteristics that do not favour cross-pollination. Under natural circumstances the pollination of wheat relies mainly on self-pollination (average level of 97-99%). Compared to other allogamous grasses, the production of pollen by a wheat ear is very limited, 10% and 2.5% compared to that of rye and maize inflorescence respectively (de Vries, 1971).
Wheat pollen is dispersed by the wind (David & Pham, 1993) but it is fairly heavy, which restricts its dispersal distance (de Vries, 1971 and 1974, Anand and Sharma, 1974).
Wheat pollen is highly sensitive to environmental conditions. Pollen viability for periods of time ranging from 1 min to approximately 30 min in optimal field conditions of 20°C, 60% relative humidity have been quoted (Poehlman 1987).
Whilst wheat species may be present at the release site, they will be at least 50 metres away from the transgenic trial site. These crops are being grown for research and development purposes only. There is potential for wheat, rye and triticale crops to be planted in areas surrounding the field, but at a distance of at least 100 metres from the trial.
There are a small number of Aegilops species that have the potential to hybridise with wheat in Germany under natural conditions. All species of Aegilops are annuals, often growing in dry, open habitats, not on managed agricultural land. The species described are most prevalent in Southern Europe, and are not indigenous in Germany.
A survey of the flora surrounding the field site confirmed that sexually compatible wild species are not present in the experimental fields.
In summary, the self pollinating nature of wheat, the short pollen life, the absence of wild relative species with which hybridisation is possible at the site, the 5m border of non modified wheat and the distance the trial will be located from other wheat crops, indicates the likelihood of pollen flow resulting in successful cross-pollination of the same or sexually compatible species, or from the same or other sexually compatible species to the trial plot is minimal, under the conditions of the trial.
Any ungerminated seed remaining after sowing or produced as a result of the trial may emerge as volunteers. To help with volunteer control, any residual grain will be left on the surface of the trial plot and encouraged to germinate by rain or irrigation. The following year, a crop other than cereal that will allow the clear identification of volunteers will be grown and any wheat volunteers emerging will be removed and incinerated or destroyed by treatment with an appropriate herbicide. The site will be monitored for at least a year after harvest.
The genetic modifications described above are unlikely to confer any invasiveness characteristics to the plant since the intended phenotypic expression of the modification is an increase in Fusarium tolerance. If the plants are host to the Fusarium species targeted by the gene, the development of these species on these plants could be reduced. This selective advantage is only likely to be for the duration of the growing season since wheat does not possess the agronomic qualities to enable it to become a weed and cannot survive without human intervention.
In summary, the likelihood of the modified wheat plants becoming more persistent or invasive than non-modified plants is considered to be negligible.
Population levels of non-target organisms are likely to remain unaffected in this trial because of the specificity of the intended effect of the modification. No direct, indirect, immediate or delayed effects have been noted in previous USA and Canadian trials with the same genetically modified wheat.
Being expressed constitutively both proteins encoded by the fungal resistance gene and the PMI gene may be present in the soil as a result of processes such as root damage. However, the FRG is isolated from a common fungal species which is widely prevalent in nature and is likely to be present in plants that are colonised with the fungal species.
Neither the PMI protein sequence nor the FRG protein sequence show homology to known allergens or toxins when searched against allergen/toxin databases.
The trial is a small-scale research trial, and all the plants used in the release (GM or non-GM) will be destroyed at the end of the trial (except those retained for specific laboratory analysis) and will therefore not be used as human food or animal feed.
Biogeochemical processes are not expected to be affected in any way by the presence of the modified plants.
The cultivation and management techniques used for the modified wheat are the same as those used in good agricultural practice.Brief description of any measures taken for the management of risks:During the trial, the plot will be monitored on a regular basis for volunteer plants. Any found will be destroyed. After termination of the trial, the plot will be monitored for at least a year. Any volunteers detected during the monitoring period will be destroyed. If during the monitoring procedure, concerns arise as to the persistency of the plants the monitoring period will be extended. A crop other than cereal that will allow the clear identification of volunteers will be grown in the following season and any wheat volunteers emerging will be removed and incinerated or destroyed by treatment with an appropriate herbicide.
If necessary the wheat plants can be easily destroyed using a herbicide. In the event of small-scale vandalism, individual plants uprooted or damaged will be removed and destroyed by incineration.
If practicable, without compromising the data that would have been obtained, the plants may be replaced and /or replanted.
In the event of large-scale vandalism where it is deemed the trial cannot continue, the trial will be terminated by the application of a herbicide and the plant material will be disposed of as described above.Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:The planned trial is a research field trial which aims to compare the agronomic performance of wheat modified to express an enhanced resistance to Fusarium pathogens with existing non-modified varieties, grown under standard agronomic conditions in Germany. It is not specifically designed to gain new data on the environmental and human health impact of the release.
- Anand S C and Sharma S C (1974). Pollen dispersal in wheat (Triticum aestivum). Wheat Information Service, 1974, 38: 27-28. http://www.grs.nig.ac.jp/wheat/wis/No38/p27/1.html
- Bevan M W, Barnes W M, Chilton M D (1983). Structure and transposition of the nopaline synthase gene region of T-DNA. Nucl. Acids Research 11 369-385
- Bojsen K I, Donaldson A, Haldrup M, Joersboe J, Kreiberg J, Nielsen F, Okkels and Petersen S (1994). Mannose or Xylose based positive selection Patent WO 94/20627 1994.
- Christensen, AH Sharrock, RA and Quail, PH (1992). Maize polyubiquitin genes - structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Molecular Biology 18 675-689.
- David J L & Pham J L (1993). Rapid changes in pollen production in experimental outcrossing populations of wheat. J. Evol. Biol. 6 659-676
- Depicker A et al. (1982) "Nopaline synthase: transcript mapping and DNA sequence". J. Mol. Appl. Genet. 1 561-573
- de Vries A P (1971). Flowering biology of wheat, particularly in view of hybrid seed production - a review. Euphytica 20 152-170
- de Vries A P (1974). Some aspect of cross-pollination in wheat (Triticum aestivum L;) 4. Seed set on male sterile plants as influenced by distance from the pollen source, pollinator : male sterile ratio and width of the male sterile strip. Euphytica 23: 601-622
- Hudspeth R L and Grula J W (1989). Structure and expression of the maize gene encoding the phosphoenolpyruvate carboxylase isozyme involved in C4 photosynthesis. Plant Molecular Biology 12 579-589
- Joersbo M, Donaldson I, Kreiberg J, Petersen S G, Brunstedt J, and Okkels F T, (1998). Analysis of mannose selection used for transformation of sugarbeet. Molecular Breeding 4 111-117.
- Miles J S and Guest J R (1984). Nucleotide sequence and transcriptional start point of the phosphomannose isomerase gene (man A) of Escherichia coli. Gene 32 41-48.
- Negrotto D, Jolley M, Beer S, Wenck A R, and Hansen G (2000). The use of phosphomannose-isomerase as a selectable marker to recover transgenic maize plants (Zea mays L.) via Agrobacterium transformation. Plant Cell Reports 19 798-803.
- Poehlman J M (1987). Breeding Wheat and Triticale, p. 220-239, In J. M. Poehlman, ed. Breeding Field Crop. AVI Publishing Company Inc., Westport.
- Reed J N, Chang Y F, McNamara D D, Beer S and Miles P J (1999). High frequency transformation of wheat with the selectable marker mannose-6-phosphate isomerase (PMI). In Vitro 35 57-A abstract P-1079.
- Yanisch-Perron C, Viera J and Messing J (1985). Improved M13 phage cloning vectors and host strains; nucleotide sequence of M13mp18 and pUC19 vectors. Gene 33 103-109.Final report-European Commission administrative informationConsent given by the Member State Competent Authority:Yes01/04/2004 00:00:00Remarks: