EN 12468-1997 en Biotechnology - Modified Organisms for Application in the Environment - Guidance for the Monitoring Strategies for Deliberate Releases of Genetically Modified Plan.pdf

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1、- STDoBSI BS EN 124b8-ENGL 1998 1b24bb9 Ob93009 518 BRITISH STANDARD Biotechnology - Modified organisms for application in the environment - Guidance for the monitoring strategies for deliberate releases of genetically modified plants The European Standard EN 12468:1997 has the status of a British s

2、tandard ICs 07.080 BS EN 12468: 1998 NO COPYING WITHOUT BSI PERMISSION EXCEPT AS PERMITTED BY COPYRIGHT LAW STDmBSI BS EN 124b8-ENGL 1778 m 1b24bb9 Ob93010 23T m BS EN 124681998 National foreword This British Standard is the English language version of EN 12468:1997. The UK participation in its prep

3、aration was entrusted to Technical Committee CII/s8, Biotechnology, which has the responsibility to: - aid enquirers to understand the text; - present to the responsible intedonaEuropean committee any enquiries on the interpretation, or proposals for change, and keep the K interests informed; - moni

4、tor related international and European developments and promulgate them in the UK. A list of organizations represented on this committee can be obtained on request to its secretary. Cross-references The British Standards which implement international or European publications referred to in this docu

5、ment may be found in the BSI Standards Catalogue under the section entitled “Intedonal Standards Correspondence Index“, or by using the “Find“ facility of the BSI Standards Electronic Catalogue. A British Standard does not purport to include ali the necessary provisions of a conira b) validaton of t

6、he monitoring protocol; c) execution of the monitoring protocol; d) appropriate record keeping. Responsibility for these steps should be assigned to a specific authoriw, organization or person. The monitoring strategy should be reviewed regularly, in the light of the field inspections, to ensure its

7、 continuing validity 5.2 Criteria for the design of the monitoring protocol The following key factors should be considered in the initiai design of the protocol to ensure the design correlates with the monitoIing strategy objectives as determined by the person designing the test: a) sampling strateg

8、y appropriate to the objectives and needs of the monitoring strategy as described in EN 12305; NOTE The development of a vad monkring and sampling strategy for deliberate releases of genetically modified plants in the environment is summarized in Figure A. 1. b) extent of monitoring necessasy to ful

9、fil the requirements of the experiment such as timescale, scaie and area of sampiing; c) plant species, predicted behviour of plant, particularly with respect to polnation and poiien dispersal, seed dispersal, dispersal by vegetative means, competitive ability and weediness; d) relevant area for mon

10、itoring in line with the monitoring objectives such as the release site andor the potential dispersal area; e) particular features of the release site to monitor such as adjacent sites, streams, soil movement activities, meteorological parameters, wild relatives; f) appropriate choice of methods use

11、d for monitoring with resped to plant species involved in experiments. 6.3 Validity of the design of the monitoring protocol 5.3.1 General Factors and criteria which should be considered for the determination of the suitabiity and validity of a monitoring protocol are given in 5.3.2 to 6.3.6. These

12、should not be considered as exhaustive lists. 6.3.2 Considemtions with respect tojwuing the overall experimental objective In fulfilment of the overall experimental objective, the aim of individual experiments can be: - detection; - identification; - determination of the molecular stability of the g

13、ene expression; - determination of the effect of the modification; - rapid screening or detailed investigation; - quantitative or suaiitatjive approach Q BSI 1998 * rn * STD.BS1 BS EN 124b8-ENGL 1998 m 1624669 Ob93015 811 m Page 6 EN 1246 - gene expression; - presence of geneticaiiy modified plant;

14、- behaviour of the genetically modified piants in the environment. 5.3.4 Considerations with respect to the release site Factors infiuencing the monitoring sixate with respect to the release site can be: - environmental conditions; - presence of organisms which feed on the plant; - presence of organ

15、isms that can serve as carriers for genetic material; - presence of compatible species within a potential dispersal area 6.3.6 considerations with respect to the validity Qfmethods used for monitotng 6.3.6.1 Key considemt.ions with respect to the validity of methods used for monitoring The following

16、 parameters should be considered for method validation to ensure that the performance characteristics of the method(s) are undemtood, and to demonstrate that the method used for monitoring is scientifically sound and valid under the experimental conditions to which it is applied: - fit for purpose;

17、- selectivi% - specificity; - reproducibilisr; - repeatability; - reliability; - feasibiliw, - limitations; - detection limit; - access to appropriate controls. NOTE A full denition of severai of these parameters is given in Eurachem guidance document nul - 1993 (see annex B 5). The following explan

18、ations are intended as a guide for the application of this European Standard. a) Fit for purpose should address or mer the specific experimental needs and is relevant to the aim, scope and available resources of the experiment. b) Selectivity of a method refers to the extent to which it can determin

19、e particUiar anaiyte(s) in a complex mixture without interference from the other components in the mixture. c) Specificity quaiies a method which is perfectly selective for an anaiyte or group of anaiytes of interest. This method is said to be specific. d) Reproducibility is the closeness of the agr

20、eement between the results of successive measurements carried out independently using an identical experimental protocol but under changed conditions. The changed conditions can include operator, location and time. e) Repeafability is the closeness of the agreement between the results of successive

21、measurements carried out under the same conditions. Repeatability conditions include the same operator, the same location and repetition over a short period of time. f) Reliability pualifies a dependable method which gives both repeatible and reproducible results. g) Feasibility indicates how workab

22、le the method is in practice. h) Limitations are the shortcomings of a method and any restrictions to use. i) Detection limit is the sdest amount of anaiyte that can be reliabiy deteded. This issue impiies the concept of sensitivity. 5.3.6.2 Frthm factors influenciw ULe choice of valid mdhoaS used f

23、or monitoring The following considerations can also be relevant to the choice of valid methods used for monitoring, depending on the nature and objectives of the monitoring experiment Not aii the considerations are applicable for each experimental situation. a) Many methods can be used to monitor ge

24、netically modified plants released into the field. These vary from simple, traditional methods to the most modern and complex, and can be based on observations or physid or biological testing. b) The methods used for monitoring can vary between experiments depending primarily upon the monitoring obj

25、ectives and should be identified in accordance with the validity criteria identified in 6.3. c) Methods used for monitoring shotd be reliable, consistent between release sites and seasons, cost effective and feasible. Methods of appropriate sensitivity for the experiment should be selected. Experime

26、nte= should aim to reach a baiance between sensitivity and practicability to ensure the suitability for the monitoring experiment. d) The validity of any one method, or combination of methods, depends partially upon the ease and acumcy of ientification of the introduced genetically modified plants a

27、nd their propagules or Poilen. Identification should ideaily be by means of easily recognisable phenotypic or genotypic characteristics (see annex B 3, 4). e) Direct observation of the behaviour of the genetidy modified piant at the release site forms the basis of all monitoring methods. Appropriate

28、 and methodical inspection of the experimental field and data recording can provide usehi monitoring information. The frequency of inspection of the experimental field depends on the requirements of the experiment For monitoring by direct O BSI 1998 STD.BS1 BS EN 12468-ENGL L998 Lb24b69 Ob930Lb 758

29、Page 6 EN 12468:1997 observation alone, the released geneticaily modified plant should, where possible, be easily and unequivdy identifiable. Any idenmg character should be stably inherited and expressed, and clearly Werent from the equivaent character dispiayed by local crops and uncultivated popul

30、ations of the same species. f) other traits dependent on the genetic modification of interest that can be suitable for monitoring purposes include herbicide tolerance, pest and disease mistance, biochemical characteristics or end-products synthesized from the gene of interest (for example, allozyme

31、analysis, carbohydrate analysis), and DNA characteristics, including molecular mapping, DNARNA hybridization analysis and RNA/DNA amplification. g) The choice of method used for monitoring should enable detection of events under study. Spread of one or more of the inserted genes to either nearby cro

32、p plants, volunteer plants, or pollencompatible weedy andor wild relatives can need to be monito- in these cases the monitoring design should focus on the specific detection of the genetic modification of interest (see annex B 3). h) Detection of gene transfer into a non-modifed plant can be by mean

33、s of various biological methods. One such method could be to assess the presence of a gene by examining potential non-modified plants for phenotypic traits, for example, herbicide tolerance. An example of another method would be to examine non-modified plants for morphological characteristics specif

34、ic to the geneticaily modified plant such as hybrid vigour, flower colour, leaf morphology, seed shape and colow. Such events can be interpreted to presume flow of the inserted gene. i) Id-, marker characterisics that are cheap and easy to identify would be the most suitable for assessing the spread

35、 of the geneticaily modied plant or introgression of genetic markem. j) “kiphait plants” of the same species with identicai flowering period as the genetically modified plant to be released can be used to detect the sprea of pollen hm the experimenta genetically modined plants. Bander can be inferre

36、d from analysis of seeds or progeny of the traphait plants. Maie-stede lines can be particularly useful for this purpose to elimlliate the possibiiity of self poihation in the wait plants. The vaiidity of the initiai design of the monitoring protocol should be the above considerations, prior to the

37、nnal definition of the monitoring protocol before execution. 6.4 Execution of the monitoring protocol 6.4.1 General The monitoring protmol should be executed in three based on a review of following stages: a) sampling (see 6.4.2); b) obtaining data (see 6.4.3); c) valid analysis of data (see 6.4.4).

38、 6.4.2 Sampling It is essential that samphg regimes for collection of relevant material andor observations for monitoring purposes ase valid and realistic. Sampling should be carried out in accordance with EN 12305. 6.4.3 Obtaining data Monitoring daa should be obtained by the method(s) determined i

39、n the monitoring protocol, over the appropriate timescale and relevant area. The frequency and extent of monitoring during the release should be adequate to ensure that the objectives of the monitoring strategy can be met, and should take into account the growth rate, stage of maturity and flowering

40、 season of relevant plants. Monitoring should be carried out over the relevant area, in accordance with the objectives of the experimental design; in general monitoring should concentmte on the release site and the likely dispersal area. Post-release monitoring can be carried out, if required for th

41、e monitoring simtem, after the release has been completed and the plants harvested After harvest, the genetically modified plant andor the inserted gene may or may not be present in the release site as residual ungerminated seeds, shed seed (from the released genetically modified plants, or from pol

42、len compatible species), or other plant material or organ capable of vegetative multiplication The monitoring can vary with the species andor variety of plant used in the experiment, the novel trait(s) expressed and the monitoring objectives. 6.4.4 Valid analysis data The data ardysis, regardless of

43、 the method(s) applied, should be regarded as valid if: a) the experiment has been performed according to the written protocol or any deviations from the protocol have been indicated; b) samples and appropriate contsols are analysed shultaneousiy using the same methods, c) all controls give expected

44、 resulk, d) sensitivity and reproducibility of the applied method(s) are appropm, e) where required, a Mcaily relevant number of samples has been analysed; f) relevant external fadors which can afect the analysis andor interpretation of the results have been taken into account and g) results are rec

45、orded and appropriately expressed and realistically assessed with regard to the information value of the applied method. CP BSI 1998 STD-ES1 ES EN 12468-ENGL 1998 lb24bb9 Ob93017 b94 Page 7 EN 12468:1997 6.6 Record keeping A record should be kept of the monitoring strategy. This should include: a) d

46、escription of any sampling protocol employed (see EN 12305); b) description of the monitoring protocol - identification of the experiment; - identity of the persons performing the monitoring; - identification and description of the release site; - method(s) chosen for monitoring; - identification of

47、 plant species; - origin of samples; - controls used; - number of samples analyse6 - deviations from the strategy; - registration of unexpected incidents, c) description of the analysis and results of the execution of the monitoring of the genetically modified plant; d) any other specific detaiis wh

48、ich can be relevant to the specic monitoring strategy and subsequent interpretation of results obtained. NOTE This should not be considered as an exhaustive list Annex A (informative) Relationship between a sampling and monitoring valid strategy When genetically modified plants are subject to experi

49、mental release into the environment, the sampling strategy is dependent upon the objectives of the monitoring sirate. Therefore, the monitoring strategy encompasses the sampling strategy both of which are required to meet the objectives of the monitoring. The relationship between the sampling and the monitoring strategy for deliberate releases of genetically modified plants is given in Figure Al. * * rn O BSI 1998 STD-BSI BS EN 12468-ENGL 1998 = 1624bb9 Ob93038 520 H Page 8 EN 124681997 Figure A.l - Development of a valid monitoring and sampling st