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Notification report


General information

Notification Number
B/NL/15/L01

Member State to which the notification was sent
Netherlands

Date of acknowledgement from the Member State Competent Authority
29/07/2015

Title of the Project
Cisgenic apples with elevated anthocyanin levels

Proposed period of release:
01/01/2016 to 01/01/2026

Name of the Institute(s) or Company(ies)
Stichting Dienst Landbouwkundig Onderzoek (DLO), Stichting Dienst Landbouwkundig Onderzoek (DLO) in particular Praktijkonderzoek Plant & Omgeving / Plant Research International (PPO/PRI);


3. Is the same GMPt release planned elsewhere in the Community?
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 NameFamily NameGenusSpeciesSubspeciesCultivar/breeding line
applerosaceaemalusmalus domesticaGala & Junami

2. Description of the traits and characteristics which have been introduced or modified, including marker genes and previous modifications:
A gene coding for a transcription factor involved in regulating anthocyanin biosynthesis derived from a red-fleshed apple variety has been introduced into two elite apple varieties, cvs. ‘Gala’ and ‘Junami’. The promoter of this particular apple gene has a natural mutation leading to increased expression and elevated anthocyanin levels throughout the plant including flowers and fruits. No further marker or other genes are introduced and are therefore not present.

Genetic modification

3. 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 apple transcription factor gene engaged in regulating anthocyanin biosynthesis is called MdMYB10. A naturally occurring mutant has been identified in the cv. ‘Red Field’, in which the promoter carries a six-fold repeat of a 23 bp DNA binding domain (R6). The wild-type form only has one copy of this domain (R1). This mutation has been found in nearly all naturally occurring red-fleshed apples. In apple, red-fleshed varieties are not considered to be very tasteful. In order to combine elevated anthocyanin levels with potential health-promoting antioxidant effect with high-quality of taste and other characteristics the R6 form of MdMYB10 was introduced into elite varieties. Due to the long generation time and self-incompatibility of apple this cannot be achieved by classical breeding. The gene including its R6 promoter and its own terminator was the only genetic element present on the region to be inserted. On either side of the gene within the T-DNA a 11 bp stopper sequence was placed to avoid undesired readthrough.

6. Brief description of the method used for the genetic modification:
Agrobacterium-mediated transformation of young, in vitro apple leaf explants.

Experimental Release

1. Purpose of the release:
Testing in a field situation, comparable to a production apple orchard, the appearance (phenotype) of the cisgenic apple lines, one of cv. Gala and two of cv. Junami. The occurrence of red colour (anthocyanin levels) will be monitored in all plants parts including flowers and fruits. This will be done throughout the year to monitor any changes related to season or juvenility (age). Stability of the trait will be monitored over several years. The trees can be used to generate budwood for further multiplication.

2. Geographical location of the site:
Municipality of Wageningen, The Netherlands

3. Size of the site (m2):
1750

Environmental Impact and Risk Management

Summary of the potential environmental impact from the release of the GMPts:
No specific environmental impact is expected. The trait (higher anthocyanin level) and the gene that is introduced are already present within the apple germplasm. High anthocynanin levels are not known to be involved in increased persistence or invasiveness, or in adverse effects on organisms, including insects, humans or animals. Anthocyanins are naturally present in high levels in many fruits and are also present in the skin of appels, without causing any negative effects. Benchmarks are provided by the red-fleshed cultivars present e.g. in the Wageningen Arboretum carrying the same gene and commercially available from breeders. Such non-GM apple cultivars with this particular gene are being cultivated and commercialized, however, most still only as ornamental trees. No positive or negative effects on the environment have been observed in cultivation of such varieties up till now. Therefore, no direct or indirect selective advantages in natural environments are to be expected for the GMPts carrying this gene.

Brief description of any measures taken for the management of risks:
No pollinators will be used, only hand pollination. Monitoring will take place with a minimum of four times per month during the growing season. The location is fenced and bird nets will be placed to avoid damage to the fruits and dispersal of fruits and seeds. Apples will not enter the food chain and will be collected, scored for anthocyanin levels and destroyed.

Summary of foreseen field trial studies focused to gain new data on environmental and human health impact from the release:
A secondary goal of this field trial is to gain data on environmental and other aspects required by the Dutch government to be able to prepare for so-called next level field trials with these cisgenic apple lines. Multiplication by generating budwood and preparing for taste panel trials in due time once sufficient appropriate data have been collected in this field trial are additional secondary goals.

Final report
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European Commission administrative information

Consent given by the Member State Competent Authority:
Not known