-Plants: Basics-Technology-Legislation-Analysis Dr. F. Waldherr CONGEN Biotechnologie GmbH
In September 2009 unauthorized linseed FP967/CDC Triffid from Canada was detected in europoean raw materials and end products!... plants in this world
What are? Most plants are transgenic organisms that have introduced additional genetic information that cannot be introduced by conventional breeding techniques. Expected benefits concerning: Resistance against herbicides Resistance against plant diseases, pests and insects increased resistance against geologic and climatic challenges higher yields altered composition (nutritional and technical factors) The flavr savr tomato was the first commercially used plant (1994). An introduced antisense RNA gene blocked the expression of a pectin degrading enzyme. It was cultivated and introduced in the USA. Later on there were also products from this tomato in the European market. The concept was not successfull and the tomato was removed from the market (1997).
How transgenic plants are made Promoter Start Gene new variety Terminator Stop marker Genetic transfer construct (contains e.g. target gene, promotor, terminator, selection marker...) Agrobacterium mediated transformation (only for dicotyledonous, not for cereals!) Direct transformation (e.g. biolistic (gene gun), electroporation, PEG-transfer) plant DNA Coincidental integration in plant genome at undefined location Genetic construct plant DNA 5 Promoter Gene Terminator 3 Start new variety Stop marker
Damages by Triffid: Canada exports nearly two thirds of its linseed to Europe When unauthorized flax was detected in Canadian linseed imports, $ the prices dropped drastically and enormous economical damage was caused!...the current situation of plants
Spread of GM-plants Developing countries Industrialized countries total Where GM plants are cultivated Global cultivation area with GM-plants 1996-2009 in Mio. Hektar (Source: www.transgen.de)
Source: David Fischhoff, Ph.D. Vice President, Technology Strategy & Development, Monsanto Company
Legal regulation of in Europe as a (growing) plant: Directive 2001/18/EC of the European Parliament and the Council of 12 March 2001 on the deliberate release into the environment of gentically modified organisms Regulation of application, authorization procedure, risk assessment, handling, trading, labelling (...) of as food and feed: Regulation (EC) 1829/2003 of the European Parliament and the Council of 22 September 2003 on gentically midfied food and feed Regulation (EC) 1830/2003 of the European Parliament and the Council of 22 September 2003 concerningn the traceability and labelling of genetically modified organisms and the traceability fo food and feed products produced from genetically modified organsims and amending Directive 2001/18 And additional regulations...
Labelling of containing food and feed Labelling: Food and Feed that are, contain or are produced from authorized (including food additives!) independant from the detectability of the Exceptions: Food, feed and additives that are produced by the help of (e.g. eggs and meat from animals that are feed with ) Food, feed and additives that contain less than 0.9 % of an authorized (adventitious presence) Not allowed at all: Food and feed containing unauthorized (0 % tolerance)
99.5% maize 0.5% 98.5% 1.5% maize maize maize maize maize No labelling Labelling
Turkish regulation ( in food) There is a list of 31 permitted events (all of the plants have also an authorization in EU) If these events are present (zero tolerance) the respective product must be labelled. If a not authorized is present (zero tolerance), the product may not be introduced to the market in Turkey.
Triffid inside?... the analytical question
General strategy for detection (EU regulation) Food product Detection/Screening Negative Positive Prerequisite: Availability of detailed information on the molecular make-up Identification Authorized? Joint Research Center of the European Commission Quantification Yes No Less than 0.9% Labelling not required More than 0.9% Labelling required
-analysis: DNA-extraction und control DNAextraction The effective extraction of plant DNA from raw material and food samples and the purification from PCR inhibiting substances is the initial step in succesfull -analysis.
family: extraction and control PREP Plant S1002 & PREP Plant X S1006 effective DNA extraction even for processed food samples (Plant X) effective purification from PCR inhibitory substances easy-to-use spinfilter system Plant PLUS S2049 CaMV S2027 control of plant DNA extraction internal inhibition control for all real-time PCR devices ( PLUS System) exclusion of flase positive results due to the presence of naturally occuring Cauliflower Mosaic Virus (CaMV)
-analysis: screening plant DNA Genetic construct plant DNA 5 Promoter Gene Terminator 3 Start new variety Stop marker Not detected Not detected Not detected! All 35S+ NOS yesterday All +FMV today 35S+ NOS All +FMV 35S+ NOS tomorrow A suitable screening system must be able to quickly and easily adapt to future challenges!
screening selection of Events genetically elements P35S CAMV Soy T-NOS FMV BAR GTS 40-3-2 (Roundup Ready) + + - - MON 89788 (Roundup Ready2Yield) - - + - Corn 59122 (Herculex RW) + - - - Bt176 (176; Maximizer) + - - + GA 21 (Roundup Ready) - + - + MIR604 - + - - MON 810 + - - - Cotton MON531/757/1076 (Bollgard) + + - - Canola GT200 (Roundup Ready) - - + - Rice Bt63 - + - -
family: screening 35S S2045 NOS S2046 FMV S2047 BAR S2048... modular screening system for all important genetic elements combination according to users requirements simple to expand to further paramenters available as packages for 35S+NOS Double Screening (S2025) and 35S+NOS+FMV Triple Screening (S2026)
-anaylsis: identification plant DNA Genetic construct plant DNA 5 Promoter Gene Terminator 3 Start new variety Stop marker construct-specific Event-specific The identification of a specific -event may take special experience, analytical knowledge and a selection of the suitable detection systems.
quantification Standard curves with DNA of known concentration Amplification of a plant species specific target (endogenous gene) reference system and a specific target (transgene) system The percentage of genetically modified material (DNA) is determined by the ratio of GM to total (reference) DNA quantities. GM DNA [%] = Q GM system Q ref. system x 100
family: identification and quantification Bt176 Corn S2015 NK603 Corn S2050 LL Canola S2018 Bt11 Corn S2016 GA21 Corn S2054 Bt176 Corn S2015... T25 Corn S2017 RR Soya S2014 MON810 Corn S2019 RR2Y Soya S2029 Kits for identification/quantification of selected, relevant -events quick adaption of product range according to legislation and customers demand Alternative and additional: analysis range of the Service in addition to your own anaylsis (e. g. screening) confirmation or your own results broad range of parameters and testing systems experienced, trained staff comprehensive support and advice short turn-around-times reliable results
Thank you for listening!