Risk Assessment and Risk Management Plan - health.gov.au fileFSANZ Food Standards Australia New...

Risk Assessmentand

Risk Management Plan

Application for licence for dealings involving anintentional release into the environment

DIR 007/2001

Title: Improved alkaloid production in oilseed poppy(Papaver somniferum L.)

Applicant: Department of Agriculture, Western Australia

30 July 2002

Abbreviations

CaMv cauliflower mosaic virusCSIRO Commonwealth Scientific and Industrial Research OrganisationDEFRA The Department of Environment, Food and Rural Affairs, UKDIR dealing involving intentional releaseDNA deoxyribonucleic acidDPIWE Department of Primary Industries, Water and Environment TasmaniaFAO Food and Agriculture Organization of the United NationsFDA Food and Drug Administration (USA)FSANZ Food Standards Australia New Zealandg gramGM genetically modifiedGMAC Genetic Manipulation Advisory CommitteeGMO genetically modified organismGTTAC Gene Technology Technical Advisory CommitteeGUS β-glucuronidaseha hectareHPT Hygromycin phosphotransferase enzymehpt Hygromycin phosphotransferase geneIBC Institutional Biosafety CommitteeIOGTR Interim Office of the Gene Technology Regulatorm meterMAFF UK Ministry of Agriculture, Fisheries and FoodmRNA messenger ribonucleic acidNHMRC National Health and Medical Research CouncilNOS nopaline synthaseNPTII neomycin phosphotransferase II enzymenptII neomycin phosphotransferase II geneNRA National Registration Authority for Agricultural and Veterinary ChemicalsOECD Organisation for Economic Cooperation and DevelopmentOGTR Office of the Gene Technology Regulatorpara paragraphPC2 Physical containment 2PR planned releaseRARMP Risk assessment and risk management planRNA ribonucleic acidSPC standard plate countT0, T1, T2, T3 transformed generationsT-DNA transfer deoxyribonucleic acidTEV Tobacco etch virusWHO World Health Organisationµg micrograms

TABLE OF CONTENTS

PREFACE 1

THE REGULATION OF GENE TECHNOLOGY IN AUSTRALIA............................................................ 1SECTION 1 THE INITIAL CONSULTATION PROCESSES ............................................................. 1SECTION 2 THE EVALUATION PROCESS ................................................................................. 2SECTION 3 THE STRUCTURE OF THIS DOCUMENT .................................................................. 2

CHAPTER 1 INTRODUCTION 5

SECTION 1 THE LICENCE APPLICATION................................................................................. 5SECTION 2 SUBMISSIONS ON THE APPLICATION AND THE RARMP........................................... 5

CHAPTER 2 BACKGROUND ON THE APPLICATION AND PREVIOUS RELEASES 9

SECTION 1 THE APPLICATION COMPLIED WITH LEGISLATIVE REQUIREMENTS...................... 9SECTION 2 PREVIOUS LIMITED RELEASES OF THESE GMOS IN AUSTRALIA........................... 9SECTION 3 RESULTS FROM AUSTRALIAN RELEASES OF GM OILSEED POPPY....................... 11SECTION 3.1 POLLEN TRANSFER...............................................................................................11SECTION 3.2 CROSS-POLLINATION ...........................................................................................11SECTION 3.3 ALKALOID PRODUCTION ......................................................................................12SECTION 4 APPROVALS FOR GM OILSEED POPPY IN OTHER COUNTRIES ............................. 12

CHAPTER 3 INFORMATION ABOUT THE PARENT ORGANISM AND THE GMO 13

SECTION 1 THE PARENT ORGANISM.................................................................................... 13SECTION 2 INFORMATION ABOUT THE GMO....................................................................... 15SECTION 3 THE INTRODUCED GENES ................................................................................... 15SECTION 3.1 THE GENE FOR THE ALKALOID PATHWAY ENZYME.................................................15SECTION 3.2 THE HPT GENE .....................................................................................................16SECTION 4 POPPY ALKALOID PATHWAY.............................................................................. 16SECTION 5 METHOD OF GENE TRANSFER ............................................................................ 17SECTION 6 CHARACTERISATION OF THE INSERTED GENETIC MATERIAL AND STABILITY OF

THE GENETIC MODIFICATION ............................................................................ 18SECTION 7 EXPRESSION OF THE INTRODUCED PROTEINS .................................................... 18

CHAPTER 4 RISK ASSESSMENT 19

SECTION 1 THE RISK ANALYSIS FRAMEWORK .................................................................... 19SECTION 2 THE RISK ASSESSMENT PROCESS ....................................................................... 19SECTION 3 SUMMARY OF RISK ASSESSMENT CONCLUSIONS ................................................. 20SECTION 3.1 RISK ASSESSMENT ...............................................................................................20SECTION 3.2 IDENTIFICATION OF ISSUES TO BE ADDRESSED FOR FUTURE RELEASES.....................21SECTION 4 HAZARD IDENTIFICATION.................................................................................. 22

CHAPTER 5 TOXICITY OR ALLERGENICITY 23

CHAPTER 6 WEEDINESS 29

CHAPTER 7 TRANSFER OF INTRODUCED GENES TO OTHER ORGANISMS 33

SECTION 1 TRANSFER OF INTRODUCED GENES TO OTHER PLANTS ...................................... 33SECTION 2 TRANSFER OF INTRODUCED GENES TO OTHER ORGANISMS (MICROORGANISMS

AND ANIMALS )................................................................................................... 38

CHAPTER 8 RISK MANAGEMENT PLAN 43

SECTION 1 SUMMARY OF RISK ASSESSMENT CONCLUSIONS ................................................. 43SECTION 2 RISK MANAGEMENT PLAN.................................................................................. 43SECTION 2.1 RISK OF TOXICITY OR ALLERGENICITY ..................................................................43SECTION 2.2 RISKS OF WEEDINESS OR GENE TRANSFER..............................................................44SECTION 2.3 GENERAL LICENCE CONDITIONS............................................................................44SECTION 2.4 MONITORING AND ENFORCEMENT OF COMPLIANCE BY THE OGTR .........................45SECTION 3 SPECIFIC RISK MANAGEMENT LICENCE CONDITIONS ......................................... 45

REFERENCES 47

APPENDIX 1 - SUMMARY OF PUBLIC SUBMISSIONS 53

APPENDIX 2 - LICENCE CONDITIONS 55

APPENDIX 3 REASONS FOR SPECIFIC LICENCE CONDITIONS 67

DIR 007/2001 - RISK ASSESSMENT AND RISK MANAGEMENT PLAN

PREFACE 1

PREFACE

THE REGULATION OF GENE TECHNOLOGY IN AUSTRALIA

1. The Gene Technology Act 2000 (the Act), which underpins the new regulatory system,took effect on 21 June 2001. The new system is Australia’s first national regulatory systemfor gene technology and is designed to protect the health and safety of people, and theenvironment, by identifying risks posed by, or as a result of, gene technology, and to managethose risks by regulating certain dealings with genetically modified organisms (GMOs). Thenew regulatory system replaces the former voluntary system overseen by the GeneticManipulation Advisory Committee (GMAC).

2. The legislation also established a statutory officer, the Gene Technology Regulator (theRegulator) to administer the legislation and make decisions under the legislation.

3. The Regulator is supported by the Office of the Gene Technology Regulator (OGTR), aCommonwealth regulatory body located within the Health and Ageing portfolio.

4. The Act prohibits persons from dealing with GMOs unless the dealing is exempt, aNotifiable Low Risk Dealing, on the Register of GMOs, or licensed by the Regulator (seeSection 31 of the Act).

5. The requirements under the new legislation for consultation and for considering andassessing licence applications and preparing risk assessment and risk management plans(RARMP) are discussed in detail in Division 4, Part 5 of the Act and Chapter 4 andsummarised below.

6. Detailed information about the national regulatory system and the gene technologylegislation is also available from the OGTR website at: www.ogtr.gov.au

SECTION 1 THE INITIAL CONSULTATION PROCESSES

Details of Section 49 consideration

7. In accordance with Section 50 of the Act, the Regulator must seek advice in preparingthis risk assessment and risk management plan from:

Ø the States and Territories;

Ø the Gene Technology Technical Advisory Committee (GTTAC);

Ø prescribed Commonwealth agencies (Regulation 9 of the Gene TechnologyRegulations 2001 refers);

Ø the Environment Minister; and

Ø the local council(s) where the release is proposed.

8. As a measure over and above those required under the Act, in order to promote theopenness and transparency of the new regulatory system, the Regulator notified receipt of theapplication to the public by posting a notice of the application's receipt on the OGTR websiteand directly advising those on the OGTR mailing list.


PREFACE 2

9. As required under Sections 51 and 52 of the Act, the Regulator has taken into accountall issues raised in written submissions relating to the protection of human health and safetyand the environment in finalising the RARMP, and in making a decision on whether or not toissue a licence for the proposed release as required under Section 56 of the Act (seeChapters 1 and 9, respectively).

SECTION 2 THE EVALUATION PROCESS

10. An assessment of the potential hazards and likely risks associated with the proposedrelease was carried out in accordance with the Act, using the Risk Analysis Frameworkdeveloped by the Regulator (see Chapter 4). A risk assessment and risk management planwas then prepared to address these risks.

11. In preparing the RARMP, information presented by the applicant, the scientificliterature, advice from scientific experts, as well as submissions and advice from theEnvironment Minister, State and Territory Governments, GTTAC, Commonwealth agencies(see Regulation 9 of Gene Technology Regulations 2001), other organisations and membersof public was considered and assessed by the Regulator.

12. The legislation requires the Regulator to consider a number of specific issues inpreparing the RARMP (see Chapter 4, Section 2). These include: the properties of the parentorganism; the effect of the genetic modification; the potential for dissemination or persistenceof the GMO or its genetic material in the environment and any provisions for limiting this;the extent or scale of the proposed dealings; and any likely impacts of the proposed dealingson the health and safety of people.

13. The legislation also requires the Regulator to consider the potential of the GMO, in theshort and long term, to: be harmful to other organisms; adversely affect any ecosystems;transfer genetic material to other organisms; spread or persist in the environment; have aselective advantage in the environment; and be toxic, allergenic or pathogenic to otherorganisms.

14. Further details about the application can be found in Chapter 3 of this document.

SECTION 3 THE STRUCTURE OF THIS DOCUMENT

15. The document sets out the various matters that were considered by the Regulator inaccordance with Section 51 of the Act in preparing this RARMP. The Regulator has nowtaken into consideration written submissions on the RARMP as stated above.

16. The structure of the document reflects the matters which the Act and Regulationsrequire the Regulator to consider in preparing the RARMP. This document:

Ø provides an introduction to the licence application and summary of thesubmissions received on the application and the RARMP and how these havebeen taken into account. Chapter 1 and Appendix 1 refers.

Ø summarises how the licence application complied with legislative requirementsand provides background information relating to previous intentional releases ofthe GMOs and other related GMOs. Chapter 2 refers.


PREFACE 3

Ø provides detailed information about the parent organism(s), the GMO(s), and theintroduced genes. Chapter 3 refers.

Ø details the risk assessment undertaken to date in accordance with the RiskAnalysis Framework developed by the Regulator. Chapter 4 refers.

Ø details the risk assessment on the hazard of toxicity and allergenicity of GMoilseed poppy. Chapter 5 refers.

Ø details the risk assessment on the hazard of weediness of GM oilseed poppy.Chapter 6 refers.

Ø details the risk assessment on the hazard of transfer of introduced genes to otherorganisms. Chapter 7 refers.

Ø sets out the conclusions reached as a result of the risk assessment and presents arisk management plan to manage the identified risks and provides brief outline ofconditions that have been included in the licence to give effect to the riskmanagement plan. Chapter 8 refers.

Ø summaries of issues raised in public submissions on the RARMP and where theyare addressed in the RARMP. Appendix 1 refers.

Ø provides the licence conditions. Appendix 2 refers.

Ø provides the reasons for the specific licence conditions. Appendix 3 refers.



CHAPTER 1 INTRODUCTION

17. The Gene Technology Act 2000 (the Act) sets out requirements which the GeneTechnology Regulations 2001 (the Regulations) must follow when considering an applicationfor a licence to intentionally release a genetically modified organism (GMO) into theenvironment. Section 51 of the Act requires the Gene Technology Regulator (the Regulator)to prepare a RARMP for each licence application. Details of the process which the Regulatormust follow are set out in the Preface.

SECTION 1 THE LICENCE APPLICATION

18. This RARMP has been prepared in response to the licence application from theDepartment of Agriculture, Western Australia (WA) for the limited and controlled release ofgenetically modified (GM) oilseed poppy (DIR 007/2001).

19. The Department of Agriculture, WA is proposing to carry out a limited and controlledrelease on one site in the shire of Wyndham / East Kimberley, WA, comprising a total area of0.2 hectares in one season.

20. Oilseed poppy was genetically modified by introducing a gene (alkaloid pathwayenzyme gene) in the alkaloid production pathway to enhance alkaloid production. Inaddition, the GM oilseed poppy also contains an antibiotic resistance gene as a selectablemarker.

21. The purpose of this release is to conduct field evaluations to determine the effect of amodification to the alkaloid production pathway on alkaloid production and to produce seedfor assaying and possible future releases. Any future releases would be subject to separateapplications and assessments.

22. None of the GM oilseed poppy plants, or their by-products, will be used in human oranimal food or therapeutics or for any other commercial purposes.

23. More detailed information about the GMO, the parent organism, the geneticmodification process, the genes that have been introduced and the new proteins expressed inthe GMO is set out in Chapter 3.

SECTION 2 SUBMISSIONS ON THE APPLICATION AND THE RARMP

24. Comments received in written submissions on the application and the RARMP are veryimportant in shaping the final RARMP and in forming the Regulator’s final decision on theapplication.

25. Inputs from prescribed agencies, other government bodies and expert groups on theapplication and the RARMP and the seven submissions received in response to the publicconsultation undertaken as required by Section 50 of the Act (see Preface) were taken intoaccount in finalising the RARMP. These suggested that the RARMP should address thefollowing issues:

Ø clarify differences between oilseed poppy and ornamental poppy (see Chapter 3);



Ø pollen dispersal (see Chapter4; Chapter 7; Chapter 8);

Ø supporting data for efficacy of pollen trap to prevent insect-mediated cross-pollination (see Chapter 4);

Ø seed dormancy and seed dispersal (see Chapter 3; Chapter 4; Chapter 6;Chapter 8);

Ø potential for weediness of oilseed poppy in northern tropical region and relevantpreventative measures to prevent it becoming a weed (see Chapter 6, Chapter 8);

Ø potential of transfer of antibiotic resistance gene and alkaloid pathway enzymegene to other plants and other organisms and the subsequent impact (seeChapter 7; Chapter 8);

Ø impact of transfer of alkaloid pathway enzyme gene on the alkaloid productionpathway of other Papaver species (see Chapter 7);

Ø data on individual alkaloid (eg. morphine, codeine, thebaine, sanguinarine) levels(including overall alkaloid levels) in various tissues (eg. seeds, pollen, roots,exudates, leaves, etc) (see Chapter 4);

Ø persistence of alkaloids in the environment (see Chapter 4; Chapter 5);

Ø impact of increased alkaloids on the environment, honey, pesticide metabolism,herbivores, pathogens and on native species (see Chapter 4; Chapter 5;Chapter 8);

Ø sufficiency of the monitoring/exclusion period to eliminate all seed reservoirs inthe soil (Chapter 6, para 7 to 9; Chapter 8, para 5 to 7);

Ø potential for out-crossing of the GMO with non-GM poppy relatives (seeChapter 7; Chapter 8);

Ø the potential instability of inserted DNA sequences and any possible deleteriouseffects that may result from the movement of the insert within the genome (seeChapter 3);

Ø potential for horizontal gene transfer due to CaMV 35S promoter (see Chapter 7);

Ø determine location and extent of honeybees as it could affect outcrossing rates(see Chapter 5);

Ø specify conditions for use of isolation zone (see Appendix 2);

Ø data on potential for adverse effects on workers handling GM oilseed poppy (seeChapter 4).

26. Queries were also received by OGTR on:

Ø whether antibiotic hygromycin was used in clinical settings (see Chapter 5;Chapter 7);

Ø whether the hygromycin resistance selectable marker was sensitive to detectprogeny of out-crossings with non-GM plants (see Chapter 3);

Ø weed status of oilseed poppy and existence of related poppy species at the releasesite (see Chapter 3; Chapter 7).



27. Suggestions were also received regarding entry to the release site; fencing and birdnetting the release site to prevent access to animals and birds, disposal of waste includingfrom the cleaning site; monitoring of the sites; observation and destruction of volunteers;existence of waterways and other crops in the isolation zone, cultivation of pasture in therelease site and segregation, storage and transport of harvested GM seed.

28. Other issues addressed by the submissions referred to the information which would berequired for a large scale release and these matters were taken into account in preparing therisk assessment and risk management plan (see Chapter 4, para 9).

29. Some submissions also raised issues that related to matters that are the responsibility ofother regulatory authorities, in particular:

Ø labelling and safety of foods derived from GMOs; and

Ø inclusion of GM products in food without labelling.

30. These are issues that are dealt with by Food Standards Australia New Zealand(formerly called Australia New Zealand Food Authority). Further information about foodsafety assessments and food labelling is available from FSANZ:

Food Standards Australia New ZealandPO Box 7186

Canberra Mail Centre ACT 2610

Phone: (02) 6271 2222Fax: (02) 6271 2278

E-mail: [email protected]://www.foodstandards.gov.au

31. Some of the submissions also raised a number of broader issues that are outside thedirect scope of the gene technology legislation and the risk assessment process and so havenot been considered, in particular:

Ø moratorium on genetic engineering and GMOs until considered safe; and

Ø other countries should conduct trials on GM oilseed poppy before Australia.

32. The issues raised in the consultation process were considered carefully, and weighedagainst the body of current scientific information, in reaching the conclusions set out in thisdocument. A summary of seven public submissions and where issues they raised relating tothe protection of human health and safety and environment have been addressed in theRARMP is provided at Appendix 1.



CHAPTER 2 BACKGROUND ON THE APPLICATION ANDPREVIOUS RELEASES

33. This part of the document provides information about the background to the legislativerequirements of the application and information about previous releases into the environmentof relevant GMOs.

SECTION 1 THE APPLICATION COMPLIED WITH LEGISLATIVE REQUIREMENTS

34. The proposal was submitted in accordance with the requirements of Section 40 of theAct. As required by Schedule 4, Part 2 of the Regulations, the application includedinformation about:

Ø the parent organism;

Ø the GMO;

Ø the proposed dealing with the GMO;

Ø interaction between the GMO and the environment;

Ø risks the GMO may pose to the health and safety of people;

Ø risk management;

Ø previous assessments of approvals; and

Ø the suitability of the applicant.

35. The application also contained:

Ø additional information required for a GMO that is a plant; additional informationfor a GMO that is intended to be used as food for human or vertebrate animalconsumption (noting that material from this release will not be permitted to beused for human or animal food, therapeutics or for any other commercialpurpose); and

Ø supporting information from the Institutional Biosafety Committee. A copy ofthe application is available on request from the OGTR.

SECTION 2 PREVIOUS LIMITED RELEASES OF THESE GMOS IN AUSTRALIA

36. Five previous limited and controlled releases (field trials) of GM oilseed poppy(genetically modified for similar or other traits) have been carried out in the Australianenvironment. The first release of GM oilseed poppy was in 1998 in Tasmania. The releaseswere carried out under conditions to limit the spread or persistence of the GMO or its geneticmaterial in the environment.



37. The GM oilseed poppy has been grown in Tasmania and Western Australia.Organisations that have been involved in carrying out the releases include TasmanianAlkaloids Pty Ltd in collaboration with Commonwealth of Scientific and Industrial ResearchOrganisation (CSIRO), Glaxo Wellcome Australia Ltd1 in collaboration with the Departmentof Primary Industries and Fisheries, Tasmania 2 and with Agriculture Western Australia3. Inthe largest trial, the approved planting area was approximately 0.08 hectares.

38. These previous releases were assessed and conducted under the former voluntarysystem overseen by GMAC and in accordance with GMAC guidelines. These releases wereassessed by GMAC as not posing any significant risks. No adverse effects on human healthand safety or the environment have been reported in connection of any of these releases.

39. The releases assessed by GMAC included:

Ø PR-91, to determine the distance of pollen transfer from GM oilseed poppy plantsunder field conditions. PR-91 also sought to examine the potential for cross-pollination between the GM oilseed poppy plants and wild related species. TheGM oilseed poppy was modified to contain a reporter gene (GUS) from thebacterium Escherichia coli, encoding the enzyme β-glucoronidase, and aselectable marker gene (hpt) encoding resistance to antibiotic hygromycinallowing identification of the GM oilseed poppies from non-GM oilseed poppies.The applicants were Department of Primary Industries and Fisheries, Tasmaniaand Glaxo Wellcome Australia Ltd;

Ø PR-103, to assess the potential for gene flow from GM oilseed poppies to non-GM oilseed poppies and related species. GM oilseed poppy was modified tocontain a selectable marker gene encoding the enzyme phosphinothricin acetyltransferase from the bacterium Streptomyces viridochromogenes. This geneconfers resistance to phosphinothricin-based herbicides such as glufosinateammonium, thus allowing selection of the GM plants from non-GM plants. Theapplicants were CSIRO Plant Industry and Tasmanian Alkaloids Pty Ltd;

Ø PR-129: to assess in the field the effect of a modification to the alkaloidproduction pathway (oilseed poppy modified to contain a gene which modifiesthe alkaloid production pathway and a selectable marker gene conferringresistance to the antibiotic hygromycin) and to collect further data on genetransfer from the GM oilseed poppies to non-GM oilseed poppies and relatedplants (oilseed poppy modified to contain a reporter gene (GUS) from thebacterium Escherichia coli, encoding the enzyme b-glucoronidase). Theapplicants were Department of Primary Industries and Fisheries, Tasmania andGlaxoWellcome Australia Ltd;

Ø PR-129X, to assess in the field the effect of a genetic modification on the level ofalkaloid production. The GM plants contained an antisense (i.e. reverse order)copy of a gene for a poppy alkaloid pathway enzyme and a selectable markergene conferring resistance to the antibiotic hygromycin. The applicants wereDepartment of Primary Industries and Fisheries, Tasmania, and GlaxoWellcomeAustralia Ltd; and

1 now called GlaxoSmithKline Australia Pty Ltd2 now called Department of Primary Industries, Water and Environment, Tasmania (DPIWE)3 now called the Department of Agriculture, Western Australia



Ø PR-146, to assess the effect of modifications of genes involved in the alkaloidsynthesis pathway and a selectable marker gene that confers resistance to theantibiotic hygromycin, and measure the extent and distance of pollen movementfrom GM plants (oilseed poppy modified to contain a reporter gene (GUS) fromthe bacterium Escherichia coli, encoding the enzyme β-glucoronidase). Theapplicants were Agriculture Western Australia and GlaxoWellcomeAustralia Ltd.

SECTION 3 RESULTS FROM AUSTRALIAN RELEASES OF GM OILSEED POPPY

40. Factors assessed in the previous releases included measuring the distance of pollentransfer from GM oilseed poppy plants under field conditions, examining the potential forcross-pollination between the GM oilseed poppy plants and wild, related species andassessing the effect of modifications of genes involved in the alkaloid synthesis pathway onalkaloid production.

Section 3.1 Pollen transfer

41. Pollen transfer studies in PR-103 demonstrated that the likelihood of pollination beyond2.5 m is very low even when the plants are completely sexually compatible (See Chapter 7,Section 1B).

42. Pollen of P. somniferum is considered to be heavy and sticky and therefore highlyunlikely to be dispersed by wind and requires an insect vector for outcrossing (Gilbert 1977).Results from the PR-91 and PR-146 to determine distance for pollen transfer wereinconclusive.

Section 3.2 Cross-pollination

43. Cross-pollination studies in PR-103 demonstrated that the likelihood of pollination ofwild species (P. somniferum ssp setigerum, P. rhoeas, P. hybridum, P. argemone andP. dubium) is extremely low. The wild species were grown between rows of GM oilseedpoppy such that they were within 30cm of the GM oilseed poppy and yet no cross-pollinationwith wild species was detected. In the outcrossing study using the pat gene (conferringresistance to glufosinate ammonium), the amount of outcrossed seed in the buffer rowsdeclined very rapidly with distance from the transgenic plants. At 0.5 m the average amountof outcrossed seed was 2.35% that declined to 0.3% at 1.0 m and 0% at 2.5 m and 5 m (SeeChapter 7, Section 1B).

44. In PR-129, no cross-pollination was observed between P. somniferum and emasculatedbuds of P. rhoeas and P. dubium. Emasculation of buds of P. rhoeas and P. dubium wasdone to increase the chances of detecting hybridisation with P. somniferum by preventingself-fertilisation and encouraging cross-fertilisation in P. rhoeas and P. dubium (SeeChapter 7, Section 1B).

45. In PR-91, flower buds of non-GM oilseed poppy were emasculated to determine theextent of cross-pollination. Despite abundant pollen seed set was low demonstrating a lowrate (0.5%) of outcrossing under field conditions (See Chapter 7, Section 1B).



Section 3.3 Alkaloid production

46. As field research is in its early stages, there is lack of conclusive data on alkaloidproduction in plants in the field.

SECTION 4 APPROVALS FOR GM OILSEED POPPY IN OTHER COUNTRIES

47. No field trials of GM oilseed poppy have been reported in the literature for any othercountry.



CHAPTER 3 INFORMATION ABOUT THE PARENT ORGANISMAND THE GMO

48. In preparing the RARMP, the Regulator is required by Section 51 of the Act to considerthe matters set out in Section 49 (2) of the Act and Regulation 10 of the Gene TechnologyRegulations regarding the properties of the parent organism and the effects of the geneticmodification.

49. This part of the document addresses these matters and provides detailed informationabout the parent organism, the GMO(s) for intentional release, the genetic modificationprocess, the genes that have been introduced and the new proteins that are expressed in theGMO.

SECTION 1 THE PARENT ORGANISM

50. The parent organism is Papaver somniferum subsp somniferum (oilseed poppy). Thename oilseed poppy is derived from the high oil content in the seed. This organism is exoticto Australia and is believed to have originated in the eastern Mediterranean region but hasdispersed across most of the populated parts of the world.

51. Oilseed poppy is the sole commercial source of several important medicinal alkaloidsincluding morphine, codeine and thebaine.

52. Oilseed poppy is believed to have been introduced into Australia by Europeanimmigrants (Tetenyi 1995). However, cultivation is prohibited without a licence. Oilseedpoppy has been grown commercially in Tasmania since 1970 for production ofpharmaceutically used alkaloids such as codeine and morphine. It is grown within allrecognised cropping areas within Tasmania (http://www.justice.tas.gov.au/pacb/).

53. The Single Convention on Narcotic Drugs 1961 (as amended by the 1972 protocol)strictly regulates worldwide production, manufacture and sale of narcotic based drugs.Australia is a signatory to the Single Convention. The Single Convention requires stringentcontrol and supervision of the growing and production of opiates, with the aims of combatingdrug abuse and illicit traffic, preventing diversion of illicit opiates and ensuring adequatesupply of medical and scientific use.

54. In order to meet Australia's commitment to the 1961 Convention, the Poppy AdvisoryControl Board (the Board) was established. The Board, an agency of the Department ofJustice, Tasmania, is the State Government authority responsible for licensing of poppy cropsin Tasmania. The Board regulates poppy cultivation in Tasmania using the powers derivedfrom the Poisons Act 1971 to regulate the industry. Import, export and manufacture of thedrugs derived from the poppies are licensed by the Commonwealth under the Narcotic DrugsAct 1971.

55. The Board is also responsible for coordinating security and control of the industry.Because of the narcotic content of plants, strict controls are maintained over all aspects ofgrowing through to processing. Board field officers regularly patrol crops and liaise with theTasmanian Police Poppy Task Force, which operates each season.



56. Oilseed poppy is not considered as a weed in Australia. There is no indication thatoilseed poppy has ever become established as a weed in wilderness areas of Tasmania(CSIRO, Canberra application 2002). The invasiveness of oilseed poppy is limited due to itsrequirement for broken ground for establishment, good soil drainage, high soil pH, andfreedom from frosts during flowering.

57. Although there is a risk of seed dormancy with oilseed poppy, the seed of thecommercially grown oilseed poppy in Tasmania is enclosed in indehiscent (i.e. do not splitopen when ripe) capsules that limits the spread of the seed. Oilseed poppy seed can survivefor several years under ideal conditions, however, seed of commercial cultivars is large andrelatively fragile and germinates readily under field conditions if the seed is close to the soilsurface. Commercial oilseed poppy is always grown from seed and cannot propagatevegetatively.

58. Apart from possible seed dormancy, oilseed poppy grown in Tasmania lacks many ofthe characteristics of weeds (see above). However, these characteristics have not beenevaluated to the same extent in Western Australia, and may not be relevant, for instance,freedom from frost during flowering may not be an issue in northern Western Australia.

59. Commercial cultivation of oilseed poppy does not occur in Western Australia.Cultivation of oilseed poppy for experimental purposes is regulated under the PoisonsAct 1964 and is strictly licensed, controlled and monitored. Under State legislation it isillegal to grow oilseed poppy plants without a licence. In Western Australia oilseed poppycan be grown for experimental studies. However, a licence to grow oilseed poppy is requiredfrom the Western Australian Health Department. Since the proposed time of release for GMoilseed poppy is April to October 2003, Department of Agriculture will apply for the licencefrom the Western Australian Health Department in January 2003.

60. In assessing the potential risks for GM oilseed poppy, the risks of the non-GM parentorganism, i.e. Papaver somniferum subsp somniferum, were considered. It is considered thatthere are no significantly different risks (eg potential weediness, or the occurrence ofalkaloids) for GM oilseed poppy from non-GM oilseed poppy currently available inAustralia.

61. Further details relating to the potential of GM oilseed poppy to be a weed and thepotential for transfer of introduced genes to other organisms, including wild and relatedplants, are provided in Chapters 6 and 7 respectively.

62. Ornamental poppy, described as or P. paeoniaeflorum in seed catalogues and otherlisting is actually P. somniferum subsp somniferum. Although it is similar to oilseed poppy,it has two striking characteristics that make it look different from oilseed poppy:

Ø large, showy, brightly coloured petals; and

Ø dehiscent capsules (ripe capsules open up to release seed when the stem bearingthe capsules is shaken by the wind) (Laughlin and Morris, undated).

63. Although not confirmed, the reference to existence of P. somniferum subsp somniferumin the states and territories of Australia refers to ornamental poppy and not to oilseed poppy.



64. Small flowered opium poppy, known as P. setigerum is actually P. somniferum subsp.setigerum. It differs from oilseed poppy in having leaves which are more noticeably toothedat the margins and by the presence of conspicuous bristles on the back of the mid rib(Laughlin and Morris, undated).

65. Since cultivation of P. somniferum is prohibited without a licence, it follows thatcultivation of oilseed poppy, ornamental poppy and small flowered opium poppy isprohibited without a licence as all are subspecies of P. somniferum.

66. The parent cultivar V87 was derived from the GlaxoSmithKline Pty Ltd's (collaboratorof Department of Agriculture, WA) breeding program. It was cultivated for 3 seasons but isno longer in commercial use as it has been superseded by other material developed in theconventional breeding program (Department of Agriculture, Western Australiaapplication 2001).

67. The GM oilseed poppy to be released under this licence was developed by geneticmodification of V87, specifically by the introduction of an alkaloid pathway enzyme geneand a selectable marker gene (see Section 5 of this Chapter).

SECTION 2 INFORMATION ABOUT THE GMO

68. The organism to be released is GM oilseed poppy (Papaver somniferum subspsomniferum). The GM oilseed poppy contains an additional gene for an alkaloid pathwayenzyme that may lead to an increase in alkaloid production. Further details are provided inSections 3 and 4 of this Chapter.

69. The GM oilseed poppy also contains an antibiotic resistance gene. The antibioticresistance gene is the hygromycin phosphotransferase (hpt) gene, which is derived from thebacterium Streptomyces hygroscopicus and confers resistance to the antibiotic hygromycin.The antibiotic resistance gene is discussed in more detail in Section 3 of this Chapter.Potential risks relating to the transfer of this gene to other microorganisms are discussed inChapter 7, Section 2.

70. None of the regulatory or coding sequences introduced into the GM oilseed poppy areknown to code for pathogenic proteins (see Section 7).

71. The methods used to introduce the genes into oilseed poppy are discussed in Section 5of this Chapter.

SECTION 3 THE INTRODUCED GENES

Section 3.1 The gene for the alkaloid pathway enzyme

72. The gene of interest introduced to the GM oilseed poppy is a gene consisting of thecoding region of an alkaloid pathway enzyme gene.



73. Expression of the introduced alkaloid pathway enzyme gene is driven by the highlyefficient 35S promoter from cauliflower mosaic virus (CaMV). A promoter is a small pieceof DNA that controls the level of expression of genes, acting like a switch. The expression ofthe alkaloid pathway enzyme gene is also effected by the 5' untranslated leader sequencefrom tobacco etch virus (TEV) which promotes high translation efficiency (Carrington andFreed 1990). The mRNA polyadenylation signals (poly (A) signals), which are required forgene expression in plants, are provided by the poly (A) signal from 35S gene of CaMV(Restrepo et al. 1990). None of these DNA sequences are known to encode pathogenicproteins. The properties of the alkaloid pathway enzyme protein are discussed in Section 4 ofthis Chapter and Chapter 5.

Section 3.2 The hpt gene

74. The selectable marker gene is the hpt antibiotic resistance gene. The hpt gene was firstisolated from the bacterium Streptomyces hygroscopicus (Leboul and Davies 1982). Itencodes the enzyme hygromycin phosphotransferase (HPT) that confers resistance to theantibiotic hygromycin.

75. The HPT enzyme catalyses the phosphorylation of the 4-hydroxy group on thehyosamine moiety, thereby inactivating hygromycin (Rao et al. 1983) and preventing it fromkilling cells producing HPT. The hpt gene was used as selectable marker gene in the earlylaboratory stages of development of the plants to enable selection of plant cells containing thedesired genetic modification. It would also be used to identify transgenics by enzymaticassay, and for selection of putative transgenic seeds.

76. Expression of the hpt gene is driven by the nopaline synthase (nos) promoter. The3’ polyadenylation signal is derived from Agrobacterium tumefaciens gene 7 (Dhaeseet al. 1983; Velten and Schell 1985).

SECTION 4 POPPY ALKALOID PATHWAY

77. Alkaloid biosynthesis is not a random process. It is highly ordered with respect to plantdevelopment, controlling the expression of pathways within organs or within organellesinside cells (De Luca and Pierre 2000). Over 40 alkaloids have been identified inP. somniferum (Bentley 1971; Santavy 1970).

78. Biogenesis of alkaloids begins with the conversion of two molecules of tyrosine totyramine and then to dopamine and4-hydroxyphenylacetaldehyde (Loeffler et al. 1987; Stadler et al. 1988, Kutchan 1998).Through a series of steps, the stereospecific condensation of 4-hydroxyphenylacetaldehydeand dopamine produces (S)-reticuline, a key intermediate involved in the synthesis of severalalkaloids including protoberberine (eg. berberine), benzophenanthridine (eg. sanguinarine)and morphinandienone (eg. morphine) alkaloids (Stadler et al., 1987). (S)-reticuline isconverted to (R)-reticuline and forms morphine through a multistep pathway involvingsalutaridine, thebaine and codeine. (S)-reticuline is converted to berberine through amultistep pathway involving (S)-scoulerine and columbamine. (S)-reticuline is converted tosanguinarine through a multistep pathway involving (S)-scoulerine, (S)-stylopine anddihydrosanguinarine.



79. The primary aim of genetically modifying oilseed poppy is to alter the alkaloid contentof the crop. The alkaloid biosynthetic pathway is regulated by both environmental anddevelopmental influences (Facchini and Bird 1998). It is theoretically possible to increasethe production of particular alkaloids in the pathway given a detailed knowledge of the poppyisoquinoline biosynthetic pathway (alkaloid production pathway) by modifying production ofthe enzymes that control the various steps in the pathway.

SECTION 5 METHOD OF GENE TRANSFER

80. The GM oilseed poppy contains the genes for a modified alkaloid pathway enzyme andhygromycin phosphotransferase, originally inserted into P. somniferum via a binary vectorpBIG in Agrobacterium tumefaciens.

81. The Agrobacterium-mediated DNA transformation system is well understood and usedextensively in genetic transformation of plants (Zambryski 1992).

82. Agrobacterium tumefaciens is a common gram-negative soil bacterium that causescrown gall disease in a wide variety of plants. The molecular biology of crown gall diseaseshows that plants can be genetically transformed by the transfer of DNA (T-DNA, locatedbetween specific border sequences) from A. tumefaciens through the mediation of the genes(vir region) of Ti plasmids. Disarmed Agrobacterium strains have been constructedspecifically for plant transformation. The disarmed strains do not contain the genes for theoverproduction of auxin and cytokinin, which are required for tumour induction and rapidcallus growth and so cannot cause crown gall (Klee and Rogers 1989).

83. A useful feature of the Ti plasmid is the flexibility of the vir (virulence) region to act ineither cis or trans configurations to the T-DNA. This has allowed the development of twotypes of transformation systems and both provide functionally equivalent transformationsystems:

Ø co-integration vectors that join the T-DNA that is to be inserted into the plant andthe vir region in a single plasmid (Stachel and Nester 1986);

Ø binary vectors that have the T-DNA and vir regions segregated on two plasmids(Bevan 1984).

84. Transformation was accomplished using the A. tumefaciens strain EHA105 that isderived from A281 containing the pTiBO542 plasmid (Hood et al. 1986). The EHA105strain was transformed with the hygromycin resistance binary vector pBIG (Becker 1990)into which the gene construct for the modified alkaloid pathway enzyme was cloned.

85. Experiments were conducted by GlaxoSmithKline Pty Ltd (collaborator of Departmentof Agriculture, WA) to test for the presence of A. tumefaciens in the T0, T1, T2 and T3(transformed generations) seeds. T0, T1 and T2 seeds were grown in antibiotic-free media toensure that growth of A. tumefaciens was not inhibited. No growth of A. tumefaciens wasobserved thus confirming its absence in transformed generations. There is no reason toconsider that growth of A. tumefaciens would occur in further generations.



SECTION 6 CHARACTERISATION OF THE INSERTED GENETIC MATERIAL ANDSTABILITY OF THE GENETIC MODIFICATION

86. GM oilseed poppies have been grown through three seed generations and continue toexpress the modified alkaloid pathway enzyme and hygromycin resistance. The presence ofthe transgene was determined by polymerase chain reaction, northern gel blot analysis andsouthern gel blot analysis. Mendelian inheritance indicates that a single copy of thetransgene construct was randomly introduced into the poppy genome. The plants to bereleased and assayed would be both homozygous and hemizygous (Department ofAgriculture, Western Australia application 2001).

SECTION 7 EXPRESSION OF THE INTRODUCED PROTEINS

87. The mRNA from the introduced alkaloid pathway enzyme gene would beindistinguishable from the endogenous transcript except for the presence of the TEV leaderreplacing the normal 5' untranslated leader. It will be expressed at higher levels than theendogenous gene and constitutively, where as the endogenous gene is expressed in a tissue-specific manner. The alkaloid pathway intermediates are expected to increase depending onthe flux at each step of the alkaloid production pathway. The proponent states that theincrease in alkaloid production in various parts of the GMO should be proportional to theoriginal levels in the parent organism resulting in a similar distribution to the parentorganism. However, this should be confirmed by testing of the various plant parts. Thisinformation would be requested before any large scale release could be approved.

88. Three seed generations have shown expression of the hygromycin resistance, indicatingtranscription of the hpt gene and translation of mRNA transcripts. The nos promoter whichdrives expression of the antibiotic resistance gene, is believed to be constitutively active(Lichtenstein and Fuller 1987, An et al. 1990).



CHAPTER 4 RISK ASSESSMENT

89. This part of the document discusses the risk analysis framework (Section 1), explainsthe risk assessment process (Section 2), summarises the conclusions of the risk assessment(Section 3), outlines the potential hazards that have been identified (Section 4) and provides adetailed discussion of the risks posed by these hazards (Chapters 5, 6 and 7).

SECTION 1 THE RISK ANALYSIS FRAMEWORK

90. The risk assessment was carried out in accordance with the Gene Technology Act 2000(the Act) and Gene Technology Regulations 2001, using the Risk Analysis Framework (theFramework) developed by the Regulator (available on the OGTR website). It also takes intoaccount the guidelines and risk assessment strategies used by related agencies both inAustralia and overseas. The Framework was developed in consultation with the States andTerritories, Commonwealth government agencies, GTTAC and the public. Its purpose is toprovide general guidance to applicants and evaluators and other stakeholders in identifyingand assessing the risks posed by GMOs and in determining the measures necessary to manageany such risks.

SECTION 2 THE RISK ASSESSMENT PROCESS

91. In undertaking the risk assessment, the following were considered and analysed:

Ø the data presented in the proponent’s application, including additionalinformation supplied by Department of Agriculture, WA (see Chapter 2,Section 1);

Ø data provided previously to GMAC or the IOGTR in respect of previousapplications for release of GM oilseed poppy (see Chapter 2, Sections 2 to 3);

Ø advice from prescribed agencies, other Government agencies and theEnvironment Minister (see Chapter 1, Section 2);

Ø advice from GTTAC (see Chapter 1, Section 2);

Ø submissions from the public (see Chapter 1, Appendix 1);

Ø information from other national regulatory agencies; and

Ø current scientific knowledge and the scientific literature.

92. In considering this information and preparing the RARMP, the following specificmatters were taken into account, as required by section 51 of the Act and set out insection 49:

Ø the risks posed to human health and safety or risks to the environment;

Ø the properties of the organism to which the dealings relate before it became, orwill become, a GMO (see Chapter 3, Section 1);

Ø the effect, or the expected effect, of genetic modification that has occurred, orwill occur, on the properties of the organism (see Chapter 3, Section 3, 7);



Ø provisions for limiting the dissemination or persistence of the GMO or its geneticmaterial in the environment (see Chapter 8, Section 2);

Ø the potential for spread or persistence of the GMO or its genetic material in theenvironment (see Chapters 6 and 7);

Ø the extent or scale of the proposed dealings (see Chapter 1, Section 1); and

Ø any likely impacts of the proposed dealings on the health and safety of people(see Chapter 5 and 7).

93. In accordance with Regulation 10 of the Regulations, the following were also taken intoaccount:

Ø any previous assessment, in Australia or overseas, in relation to allowing orapproving dealings with the GMO (see Chapter 2, Sections 2, 4);

Ø the potential of the GMO concerned to:

• be harmful to other organisms (see Chapter 5);

• adversely affect any ecosystems (see Chapters 6 and 7);

• transfer genetic material to another organism (see Chapter 7);

• spread, or persist, in the environment (see Chapter 6);

• have, in comparison to related organisms, a selective advantage in theenvironment (see Chapter 6); and

• be toxic, allergenic or pathogenic to other organisms (see Chapter 5).

94. Regulation 10 also requires the Regulator to consider both the short and long termwhen taking these factors into account.

95. Through the risk assessment process, a number of potential hazards were identified.The risks posed by these hazards were evaluated by considering:

Ø the likelihood of the hazard occurring;

Ø the likely consequences if the hazard were to be realised; and

Ø the availability of mechanisms for effectively managing identified risks.

SECTION 3 SUMMARY OF RISK ASSESSMENT CONCLUSIONS

Section 3.1 Risk assessment

96. In summary, a number of potential hazards were identified and the risks which mayoccur should these hazards be realised were considered. It is concluded that no substantiveadditional risks would be posed to public health and safety or to the environment by theproposed release as a result of the genetic modification of GM oilseed poppy, compared tothose posed by non-GM oilseed poppy because:



Ø the GM oilseed poppy is not likely to prove more allergenic or toxic to humans orother organisms than non-GM oilseed poppy. Although possible increase inalkaloid content may cause an increase in toxicity, it is unlikely to be a risk asexposure to toxins will be minimal. The alkaloid content in non-GM oilseedpoppy has been increased 3-fold over the last several years through conventionalbreeding and has been managed such that the risk of exposure to the toxins hasnot increased.

Ø the risk of the GM oilseed poppy establishing as a weed is low and furtherrestricted due to the tight controls imposed on the release ; and

Ø the likelihood of transfer of the introduced genes to other organisms is low, buteven if such transfer occurred would be unlikely to pose any hazard to humanhealth and safety or the environment.

Section 3.2 Identification of issues to be addressed for future releases

97. During the evaluation process, a range of data and information requirements wereidentified which would be required before any future commercial release could becontemplated. In part these data deficiencies relate to the need for more detailed informationon the parent organism. These include further information and data on:

Ø confirmation of distance of pollen flow;

Ø efficacy of pollen trap and isolation zone to prevent pollen flow;

Ø weediness potential of the GM and non-GM oilseed poppy in the proposed areasof commercial release;

Ø viability of pollen and seed in different environments;

Ø the expression levels and distribution of the introduced proteins, the individualand overall alkaloid content in various plant tissues;

Ø the potential for toxic / allergenic exposure for humans and animals, includingpathogens and soil biota; and

Ø confirmed absence of Agrobacterium tumefaciens from future generations of GMoilseed poppy.

98. The general issue of the use of antibiotic resistance marker genes may also need to beconsidered in the longer term. This issue has been addressed recently by international foodstandard setting bodies, including the FAO/WHO Expert Consultation on Foods Derivedfrom Biotechnology (29 May-2 June 2000, Geneva Switzerland) and the Codex Ad HocIntergovernmental Taskforce on Foods Derived from Biotechnology (November 2000,Tokyo) and the OECD.



99. These international bodies accept that there is no evidence of human health and safetyproblems resulting from the use of antibiotic resistance marker genes in GM foods (e.g. thehpt gene). However, they have also stated that alternative transformation technologies thatdo not result in antibiotic resistance marker genes in foods are to be encouraged in the futuredevelopment of recombinant DNA plants, where such technologies are available anddemonstrated to be safe. However, this issue is not directly relevant to the current applicationas neither the GM oilseed poppy nor any plant parts or by-products will be used for human oranimal food or therapeutics.

SECTION 4 HAZARD IDENTIFICATION

100. This part of the risk analysis presents a summary of the possible hazards that wereidentified and assessed, and the conclusions that were drawn.

101. A number of potential hazards arising from the genetic modification of oilseed poppywere identified through: assessment of the application; review of the scientific literature; andreview of data from other regulatory bodies and overseas bodies as referenced in regulation 9of the Regulations and Section 3. The potential hazards identified were that:

Ø the GM oilseed poppy might be harmful to organisms, because it may be moretoxic or allergenic than non-GM oilseed poppy as a result of the novel geneproducts and possible enhanced expression of alkaloids in the plants or indifferent parts of the plant or because of unforeseen or unintended effects (seeChapter 5);

Ø the GM oilseed poppy might be harmful to the environment because of inherentweediness or increased potential for weediness (see Chapter 6); and

Ø the new genes introduced into the GM oilseed poppy might transfer to non-GMoilseed poppy crops and naturalised Papaver species or to other organisms, withadverse consequences (see Chapter 7).



CHAPTER 5 TOXICITY OR ALLERGENICITY

102. Each potential hazard identified in Section 4 of Chapter 4 is addressed in detail below,in three parts:

Ø Part A explains the nature of each potential hazard and any adverse impacts thesemight cause;

Ø Part B examines the likelihood of the potential hazard occurring; and

Ø Part C draws conclusions about the risks and their possible impacts.

A: Nature of the potential toxicity or allergenicity hazard

103. The GM oilseed poppy might be more toxic or allergenic to organisms than non-GMoilseed poppy. This could occur because of the novel gene products, possible enhancedexpression in the plant of the introduced gene or differential expression in various plant partsof the introduced genes, or because of unforeseen, unintended effects.

TOXICITY OR ALLERGENICITY FOR HUMANS

104. If the GM oilseed poppy is toxic or allergenic, there could be impacts relating to:

Ø health of people due to ingestion of alkaloid enhanced poppy;

Ø occupational health and safety of people (for example, for farm workers, orresearch workers involved with GM oilseed poppy);

Ø environmental exposure (for example, people breathing poppy pollen); and

Ø eating animals that have consumed GM oilseed poppy or honey from beesforaging on the GM oilseed poppy.

TOXICITY FOR OTHER ORGANISMS

105. If GM oilseed poppy is toxic for other organisms, there could be potential impactsrelating to:

Ø toxicity for insects or soil biota, with potential to harm the natural environment(for example, adverse impacts on native biodiversity); and

Ø toxicity for grazing animals, including native animals.

B: Likelihood of the toxicity or allergenicity hazard occurring

106. In assessing the likelihood of adverse impacts due to toxicity or allergenicity of GMoilseed poppy, a number of factors were considered including:

Ø the toxicity of GM oilseed poppy for humans and other animals and soil biota;and



Ø information about the likely levels and routes of exposure to GM oilseed poppyand the introduced proteins, for example in food or feed, in non-food products, inresidues generated in manufacturing processes, or through direct contact with thecrop or contact with soil in which the crop is grown, and through the food chain,pollen or honey.

TOXICITY OR ALLERGENICITY OF THE INTRODUCED PROTEINS AND ALKALOID LEVELS

107. The GM oilseed poppy differs from non-GM oilseed poppy in the possible enhancedexpression of the alkaloid pathway enzyme from the modified alkaloid production pathwayand an additional new protein HPT. These have all been considered for their potentialtoxicity and allergenicity.

Alkaloids

108. The purpose of this release is to obtain data on alkaloid production as a result ofmodification to the alkaloid production pathway under field conditions. The geneticmodification is aimed to increase alkaloid production of the GM oilseed poppy. The possibleenhanced alkaloid production in GM oilseed poppy is from a modification that does notexpress a new protein, rather it possibly increases expression of an endogenous enzyme andtherefore possibly increase the level of naturally occurring alkaloids. The coding region ofthe introduced gene is identical to that of the endogenous gene in oilseed poppy. Therefore,the alkaloid pathway enzyme expressed from the introduced gene will be indistinguishablefrom that produced by the endogenous gene (Department of Agriculture, WAapplication 2001).

Toxicity and allergenicity of alkaloids for animals including humans

109. Oilseed poppy, if ingested, is known to be moderately toxic to animals and humans dueto its morphinan alkaloid content. The alkaloid content of commercial non-GM oilseedpoppy has been increased 3-fold over the past 30 years through conventional plant breedingmethods.

110. There have been no reports of additional adverse effects on animals or insects despitewide cultivation over a number of years of non-GM oilseed poppies with increased alkaloidlevels derived through conventional breeding.

111. No adverse effects are expected on humans from potential enhanced production ofalkaloids in GM oilseed poppy due to strict regulatory controls (See Chapter 3, Section 2).No GM oilseed poppies or by-products from this release will be used for human food oranimal feed or therapeutics.

Toxicity of alkaloids for soil biota

112. The toxicity for soil biota from alkaloids in GM oilseed poppy is unlikely to besubstantially changed from that due to non-GM oilseed poppy. Alkaloids are readilymetabolised in the environment.



113. GlaxoSmithKline had conducted a preliminary experiment to study the effect ofalkaloids on soil microflora (microbial populations in soil) where poppy plants werecultivated. Samples were analysed for standard plate count (SPC), yeast and mould. Thedata obtained was limited and the results were inconclusive (GlaxoSmithKline 2001).

114. The Regulator would require further data on the potential for toxic/allergenic exposurefor humans and animals including soil biota, herbivores and pathogens before large scalerelease can be considered.

Alkaloid Pathway Enzyme

115. The GM oilseed poppy contains an additional gene for an alkaloid pathway enzyme thatmay lead to an increase in alkaloid production.

Toxicity and allergenicity of alkaloid pathway enzyme for animals including humans

116. The alkaloid pathway enzyme which may be overexpressed is identical to a geneproduct in the parent and therefore should not present any new issues of toxicity orallergenicity. The alkaloid pathway enzyme is not a known toxin or allergen.

HPT Protein

117. Resistance to the antibiotic hygromycin is conferred by HPT, first isolated fromStreptomyces hygroscopicus (Leboul and Davies, 1982). Hygromycin is an aminoglycosideantibiotic that is active against both prokaryotic and eukaryotic cells (Gonzales et al. 1978).It has been reported to interfere with translocation step of elongation (Cabanas et al. 1978;Gonzales et al. 1978) and to cause misreading of mRNA (Eustice and Wilhelm 1984a,1984b).

Toxicity and allergenicity of HPT protein for animals including humans

118. The HPT protein is not known to be toxic to humans or animals as the current range ofmarker genes including hpt do not code for any protein with known toxicity and are thereforeconsidered safe (Malik and Saroha 1999). However, there is no detailed safety evaluationavailable for hygromycin resistance genes (Karenlampi 1996).

119. Hygromycin is not in current clinical use, therefore the potential for inactivation of anorally dosed hygromycin in humans by consuming plant material containing the marker geneis not an issue. Hygromycin is used in veterinary medicine (Karenlampi, 1996). Given thesmall scale of the release, the likelihood of any animal consuming the GM plant material isextremely low.

POTENTIAL FOR EXPOSURE TO GM OILSEED POPPY AND THE INTRODUCED PROTEINS

120. The level of exposure to the GMO and novel proteins in the genetically modified cropis not likely to be significant. The potential for exposure of humans, other animals and soilbiota is discussed in detail below. Please note that, as stated above, no GM oilseed poppiesor by-products from this release will be used for human or animal food or therapeutics.



Occupational and environmental exposure for humans

Alkaloids

121. The morphinan alkaloids are expressed in laticifer cells that occur predominantly inaerial organs of oilseed poppy plants such as the capsule, stem and leaf tissue (Nessler andMahlberg 1977, 1978). As noted previously, conventional oilseed poppies have toxicproperties. In the GM oilseed poppy, increases in alkaloids in other plant parts (wherealkaloids are not produced at high levels) would be likely to be similar to those in non-GMoilseed poppy because only a single alkaloid biosynthetic enzyme will be expressedconstitutively and not the whole alkaloid production pathway that has tissue-specificexpression. As such it is unlikely that there will be increased levels of alkaloids in tissues ofGM oilseed poppy other than in tissues where alkaloids are already expressed in higherlevels. However, before any large-scale release can be considered this hypothesis should betested.

122. Occupational asthma has been reported in 6 out of 28 workers in a pharmaceuticalfactory extracting morphine and other alkaloids from shells of non-GM oilseed poppy(Moneo et al. 1993).

Flower and Pollen

123. Oilseed poppy is predominantly self-pollinating. The pollen is heavy and sticky andnot easily dispersed by wind. Poppy pollen is therefore not likely to be present in theatmosphere, limiting any possible human exposure to poppy pollen as a potential aeroallergenor toxin.

124. Allergic contact urticaria has also been reported from non-GM Papaver rhoea flowers(Gamboa et al. 1997). The use of protective overalls and face masks when handling GMoilseed poppy is therefore advisable. The Regulator has imposed licence conditions to ensurethat if the workers are required to handle the GMO during flowering, they wear masks,gloves and disposable overalls. The overall level of exposure to transgenic proteins forworkers handling the GM oilseed poppy is therefore likely to be negligible.

125. The limited scale of the proposed release (0.2ha) will further limit direct occupationalor environmental exposure to transgenic proteins and alkaloids in GM plants for farmworkers, factory workers, or farming communities.

Dietary exposure for humans

Poppy Seeds

126. There are reports of poppy seed allergy caused by ingestion of poppy seed (Crivellaroet al. 1999, Gloor et al. 1995). There will be no dietary exposure to animals or humans fromthis release since the oilseed poppy and its by-products will not be used for animal or humanfood or therapeutics. As such any incidents of toxicity / allergenicity through consumablesubstances will not occur.

127. Oilseed poppy is a restricted crop in Australia. Possession of poppy or poppy plantswithout appropriate authority is a criminal offence. Since access by members of the public topoppy and poppy products is restricted, incidents of ingestion by humans and subsequenttoxicity / allergenicity are unlikely to occur.



Pollen

128. It is possible that limited amounts of GM oilseed poppy pollen might be incorporated inhoney, since honey may contain small amounts of pollen. As a standard practice, commercialhives are generally not placed near crops because of the potential for exposure and loss offoraging bees or contamination of honey with the pesticides normally applied to crops.

129. The Department of Environment, Food and Rural Affairs (DEFRA), UK has carried outa study using honey containing pollen from genetically modified canola that expressed thenptII gene. They estimated that a 500 g pot of honey would contain up to 0.005 µg of NPTIIprotein (DEFRA 1997). To put this in perspective, it has been estimated that amounts as lowas 100 µg of peanut protein may cause mild reactions in a limited number of hypersensitiveindividuals, (amounts of 2000 to 5000 µg are required to produce significant reactions in thesame patients) (cited in MAFF, 1997). Compared to the minimum amount of peanut protein(100 µg) required to cause mild reaction, the NPTII protein found in honey is in significantlylower amount (0.005 µg) and is even less likely to cause adverse reaction in individuals.

130. There is no evidence to suggest that the proteins and alkaloids expressed in the pollenof the GM oilseed poppy would be more allergenic than from non-GM oilseed poppy.Alkaloid content in non-GM oilseed poppy has increased 3-fold by conventional breedingand there have been no reports of adverse effects from eating honey produced by beesforaging on non-GM oilseed poppy. Although the introduced protein is driven by theconstitutive promoter, there is no evidence that the alkaloid content of nectar or pollen wouldbe significantly increased from its present levels.

131. In addition, very small quantity of honey is produced in Wyndham. Beekeepers in thearea are usually pollination providers for cucurbit growers and so the beehives are alongsidecucurbit crops. Optimal foraging theory used in the ecological study of a wide range ofanimal species predicts that bees will forage on the best available food sources as near to thehives as possible (Delaplane and Mayer 2000). The closest beehives are about 1 kilometreaway from the Frankwise Research Institute where the proposed release site is located. It istherefore unlikely that the bees will forage on GM oilseed poppy.

Exposure of livestock and wildlife, including animals, fish and birds

132. None of the products of the GM oilseed poppy will be released for use as animal feed.As part of risk management, the proponent will also be required to use a rabbit-proof fence toenclose the release site and cover the fence and the release site with a bird net to further limitthe possibility of ingestion by preventing the exposure of livestock and wild life to GMplants.

133. The pollen and hand harvested GM seed (seeds are produced in indehiscent capsuleswhich do not split open) is not expected to enter aquatic habitats in any significant quantity,and therefore aquatic species will not be exposed. The release site is not prone to floodingand there are no natural waterways within 50 m of the release site. Any exposure will befurther limited because of the controlled and limited scale of the release.



134. Invertebrates may be exposed directly, through feeding on the GM oilseed poppyplants, or indirectly through eating other organisms, that feed on the plants. Exposure is notexpected to be significant, particularly to pollen and nectar. Exposure will be further limitedby the controlled and limited scale of the release.

Exposure of soil biota

135. The hygromycin resistance gene has been sourced from Streptomyces hygroscopicus, acommon soil bacterium. As such soil biota are likely to be exposed to the HPT proteinnaturally present in the soil and increased exposure from GM oilseed poppy is not expectedto be significant.

136. The alkaloids are readily biodegradable. In addition, exposure to transgenic proteinswould be further limited because of the limited scale of the release. Tissues with highestconcentration of alkaloids (ie. capsules) will be harvested and the remaining plant materialwill be destroyed.

C: Conclusions regarding toxicity and allergenicity

137. Oilseed poppy, if ingested, is known to be moderately toxic to animals and humans dueto its morphinan alkaloid content. However, it is considered that the likelihood of adversetoxic or allergenic impacts on humans or other species, as a result of possible enhancedtoxicity or allergenicity of GM oilseed poppy in the proposed release is very low.

138. Due to the various factors outlined above, exposure to the GM oilseed poppy and theintroduced proteins would be minimal. The Regulator has imposed conditions on the licenceto restrict use of material from the release such that no GM plants from the release or theirby-products would be used for animal feed or human food, or therapeutics or for any othercommercial use (see Chapter 9, Section 3). The scale of the proposed release is relativelysmall (0.2 ha) on an agricultural scale, and any environmental impacts due to toxicity arelikely to be localised to the specific release site and will, therefore, be manageable.

139. The level of alkaloids has been increased over 3-fold in non-GM oilseed poppy overlast 30 years through conventional breeding. This increase in alkaloid and resultant toxicityhas been managed without any adverse consequences. Oilseed poppy is grown under tightlycontrolled and monitored conditions (Poisons Act 1964 WA; Poisons Act 1971, Tasmania).In the release proposed by the Department of Agriculture WA, there may be an increase inalkaloids but it is considered that if there are any risks associated with this increase to humanhealth and safety and to the environment, these would be able to be managed.

140. It should be noted that oilseed poppy has been grown for centuries for use inpharmaceutical and medical applications. There have been no reports of adverse impactsfrom the previous releases of GM oilseed poppy. There is a risk that the GM oilseed poppymay prove toxic or allergenic to a very small number of individuals but given the small scaleand isolated location of the release, the likelihood of exposure is very limited. In addition, onthe basis of the risk assessment above, the risks are no greater than those created byconventional breeding and could be managed.

DIR - 007/2001 - RISK ASSESSMENT AND RISK MANAGEMENT PLAN


CHAPTER 6 WEEDINESS

A: Nature of the weediness hazard

141. The possibility was considered that GM oilseed poppy might have the potential to beharmful to the environment because of inherent weediness or increased potential forweediness.

142. There is also the possibility that the genetic modification has, either directly or as aresult of pleiotropic (i.e. a single gene responsible for a number of distinct and seeminglyunrelated effects) effects, increased the weediness of the oilseed poppy plants. This couldresult from changes such as increased fitness or increased fecundity.

B: Likelihood of the weediness hazard occurring

WEEDINESS OF NON-GM OILSEED POPPY

143. Oilseed poppy has been grown for centuries throughout the world without any reportsthat it is a serious weed pest. Oilseed poppy is not generally found growing in the wild, beingmostly domesticated (Kapoor 1995).

144. Oilseed poppy is not listed as a weed in Australia. Tasmania is the only State in whichoilseed poppy has been grown commercially. There is no indication that oilseed poppy hasever become established as a weed in wilderness areas of Tasmania (CSIRO Plant Industryapplication 2002). Oilseed poppy is not considered to have invasive weedy characteristics asan annual plant grown in Australia. It occurs exclusively as a managed cultigen in Australia.Volunteer oilseed poppy plants can be easily controlled in an agricultural environmentthrough the application of broad spectrum and selective herbicides to which these plants aresensitive (Experts Group on Gene Technology 2001). The invasiveness of oilseed poppy islimited due to its requirement for broken ground for establishment, good soil drainage, highsoil pH, and freedom from frosts during flowering.

145. Although there is a risk of seed dormancy with oilseed poppy, the seed of thecommercially grown oilseed poppy in Tasmania is enclosed in indehiscent (i.e. do not splitopen when ripe) capsules that limits the spread of the seed. Oilseed poppy seed can survivefor several years under ideal conditions, however, seed of commercial cultivars is large andrelatively fragile and germinates readily under field conditions if the seed is close to the soilsurface. Commercial oilseed poppy is always grown from seed and cannot propagatevegetatively.

146. Apart from possible seed dormancy, oilseed poppy grown in Tasmania does not exhibitmany of the characteristics of weeds (see above). These characteristics have not beenevaluated to the same extent in Western Australia. However, they may not be relevant, forinstance, susceptibility to frost during flowering may not be an issue in northern WesternAustralia.



147. Commercial cultivation of oilseed poppy does not occur in Western Australia.Cultivation of oilseed poppy for experimental purposes is regulated under the PoisonsAct 1964 and is strictly licensed, controlled and monitored. Under State legislation it isillegal to grow oilseed poppy plants without a licence. In Western Australia oilseed poppycan be grown for experimental studies and a licence to grow oilseed poppy is required fromthe Western Australian Health Department. Since the proposed time of release for GMoilseed poppy is April to October 2003, Department of Agriculture will apply for the licencefrom the Western Australian Health Department in January 2003.

148. Ornamental poppy, described as or P. paeoniaeflorum in seed catalogues and otherlistings is actually P. somniferum subsp somniferum. Although it is similar to oilseed poppy,it has two striking characteristics that make it look different from oilseed poppy:

Ø large, showy, brightly coloured petals; and

Ø dehiscent capsules (ripe capsules open up to release seed when the stem bearingthe capsules is shaken by the wind) (Laughlin and Morris, undated).

149. Although not confirmed, references to the occurrence of P. somniferum subspsomniferum in the states and territories of Australia refers to ornamental poppy and not tooilseed poppy.

150. Small flowered opium poppy, known as P. setigerum is actually P. somniferum subsp.setigerum. It differs from oilseed poppy in having leaves which are more noticeably toothedat the margins and by the presence of conspicuous bristles on the back of the mid rib(Laughlin and Morris, undated).

151. Since cultivation of P. somniferum is prohibited without a licence, it follows thatcultivation of oilseed poppy, ornamental poppy and small flowered opium poppy isprohibited without a licence as all are subspecies of P. somniferum.

WEEDINESS OF GM OILSEED POPPY

152. The agronomic characteristics (e.g. germination, seed survival, vigour, yields, diseasesusceptibility) of the GM oilseed poppy have been evaluated in glasshouse and previous fieldreleases, and found to be within the range for current commercial non-GM varieties(Department of Agriculture, WA application 2001).

153. Two releases of GM poppies in Tasmania received 'deemed' licences under thetransitional provisions of the Gene Technology Act 2000 (PR-129 and PR-129X). These havebeen regularly monitored by OGTR following harvest of the GM crop and no GM oilseedpoppy volunteers have been detected.

154. One previous release of GM oilseed poppy (PR-146) has occurred at the same site asthe proposed release. The first planting of this trial failed and the second planting alsoexperienced some problems with germination of the plants and their general vigour. OGTRmonitoring visits have confirmed that the site was compliant with its licence conditions.

155. In addition, the seed of GM oilseed poppy is enclosed in indehiscent capsules. Thecapsules do not split open when ripe and will be hand harvested thus limiting the spread ofthe GM seed.



156. There is also no indication that the potential weediness of the GM plant has increasedas a result of the genetic modifications. The modification to the alkaloid production pathwayis not expected to impact on any of the factors which contribute to the weediness of a plant.The alkaloid pathway enzyme gene could lead to an increase in alkaloids and thus increasethe toxicity. Increased toxicity could only confer a selective or competitive advantage to theGMO if it decreased consumption by herbivores where herbivores controlled the plant orinhibited the growth of pathogens where pathogens controlled the plant. There is no evidencethat this could occur as oilseed poppies only occur as a managed cultigen in Australia andwhere they occur as volunteers in the agricultural environment they are controlled byherbicides. It should be noted that alkaloid content has been increased in non-GM oilseedpoppies over 3-fold through conventional breeding methods in the last 30 years (Departmentof Agriculture, WA application 2001) but oilseed poppy has not emerged as a weed problem.

157. The only differences that would be expected between the GM and non-GM oilseedpoppy are the expression of the hygromycin transferase and alkaloid pathway enzyme andpossible over-expression of endogenous alkaloids. There is no evidence, nor any reason tobelieve that these changes in the GM oilseed poppy would alter any of the characteristic weedattributes listed above.

158. The HPT protein, encoding resistance to hygromycin, would not confer a selectiveadvantage on the oilseed poppy, since antibiotics are not used on commercial oilseed poppycrops or in any environment in which the poppy may establish.

159. In addition, before a commercial release could be approved, the RARMP has identifieda range of data that would be required, including more data on the weediness potential of theGM oilseed poppy in the proposed areas of commercial release.

160. These points, together with tight controls on the proposed release, argue against anyweed problems arising due to GM oilseed poppy.

C: Conclusions regarding weediness

161. Any volunteer poppies that may grow would be able to be detected and removed easilyand quickly. In addition the risks would be further minimised by the licence conditionsimposed by the Regulator to ensure that the GMO would not spread from the release site orpersist in the environment or at the site after harvest, thus reducing the potential for the GMOto establish as a weed outside the release site (see Chapter 8, Section 3).

162. Oilseed poppy grown in Tasmania does not exhibit weedy characteristics (apart frompossible seed dormancy). Although these characteristics have not been as extensivelyevaluated in Western Australia, the risk of oilseed poppy becoming a weed in WesternAustralia is negligible due to the strict regulatory controls imposed by the State legislation,the small and limited scale of the release, and the licence conditions imposed by theRegulator.



163. It is concluded that the risk of the GM oilseed poppy spreading into the environmentand causing harm to the environment is very low and not likely to be greater than for non-GM oilseed poppy. In summary, the reasons for these conclusions are that the introducedgenes in oilseed poppy are not likely to increase the weedy potential of the plants. Oilseedpoppy is not a weed and is grown as a restricted crop in Australia. It is, therefore, highlyunlikely that the GM oilseed poppy will become a weed problem.



CHAPTER 7 TRANSFER OF INTRODUCED GENES TO OTHERORGANISMS

164. In general terms, the types of hazards that might result from transfer of the genesintroduced into GM oilseed poppy to other organisms could include the alteration ofnaturalised Papaver species to produce enhanced alkaloid levels and antibiotic-resistantpathogens with potential to harm human or animal health.

165. The potential hazards are addressed in the following sections, with respect specificallyto:

Ø other plants (Section 1 of this Chapter); and

Ø other organisms (Section 2 of this Chapter).

SECTION 1 TRANSFER OF INTRODUCED GENES TO OTHER PLANTS

A: Nature of the gene transfer hazard

TO OTHER OILSEED POPPY PLANTS

166. Transfer of the introduced genes to other non-GM oilseed poppy plants via crossfertilisation would present the same hazards and have the same potential impacts as thepresence of the genes in GM oilseed poppy (see Chapter 3, Sections 1 and 3). If transferoccurred, this would increase the possibility that the genes could spread in the environment.

TO OTHER PLANT SPECIES

167. Transfer of the introduced genes into other plant species, in particular to native flora,might have adverse effects on biodiversity if the genes conferred a selective advantage andallowing the recipient to outcompete other plants or by being toxic to other species. Otherpotential hazards specific to the transferred gene sequences are as follows:

Ø Alkaloid pathway enzyme gene:Plants could become toxic due to an increased alkaloid content.

Ø Antibiotic resistance marker gene:Plants could become resistant to the antibiotic.

Ø CaMV 35S promoter and other regulatory sequences:Some of these sequences are derived from plant pathogens (cauliflower mosaicvirus and Agrobacterium tumefaciens) and therefore the possibility should beconsidered that they might have pathogenic properties. If gene transfer did occur,there could be unintended or unexpected effects if the introduced genes andregulatory sequences altered the expression of endogenous plant genes. If suchperturbation of normal plant gene expression did occur, the impact would dependon the resultant phenotype.



B: Likelihood of the gene transfer hazard occurring

TRANSFER OF GENES TO OTHER OILSEED POPPY PLANTS

Outcrossing rates for oilseed poppy

168. The transfer of genes from GM oilseed poppy to non-GM oilseed poppy requires thetransfer of GM pollen to non-GM oilseed poppy. Oilseed poppy pollen is considered to beheavy and sticky and requires an insect vector for outcrossing. Wind dispersal is negligible(Gilbert 1977).

169. Oilseed poppy is considered to be predominantly self-pollinating (Iljina 1962). Whengrown in large fields there is about 30% insect-mediated cross-pollination (Loof 1966)although cross-pollination studies conducted in Australia under field conditions with GMoilseed poppy indicate a much lower crosspollination rate of 0.5% to 2.35%(GlaxoWellcome 1999; Tait and Fist 2002).

170. Insect prevalence, especially of honeybees, strongly influences outcrossing rates foroilseed poppy (Gilbert, 1977; Department of Agriculture, WA application, 2001 and CSIROPlant Industry application 2002).

171. Cross-pollination studies for oilseed poppy were carried out under field conditions in aprevious release of GM oilseed poppies (PR-91). Flower buds of non-GM oilseed poppyplants were emasculated just prior to flowering at a range of locations and distances from theGM oilseed poppy plants. The emasculated non-GM oilseed poppy plants were surroundedin all cases by non-GM oilseed poppy plants that had not been emasculated. Seed set resultsindicated that on average 30.6 seeds were produced per emasculated capsule as compared to6600 seeds per non-emasculated capsule. This indicates that the level of outcrossingobserved under field conditions in Australian study of GM oilseed poppy crop is extremelylow, in the order of 0.5%, even though abundant pollen was available from surroundingplants (GlaxoWellcome 1999).

172. In another outcrossing study (under PR-103) using the pat gene (conferring resistanceto glufosinate ammonium), seed was collected from open pollinated non-GM oilseed poppyplants grown in various locations in the field trial of GM oilseed poppy. This seed was usedto grow plants in trays in the greenhouse. Outcrossing was estimated by scoring survival ofthe seedlings when sprayed with glufosinate ammonium solution. Seedlings containing thepat gene survived the treatment with glufosinate ammonium solution, whilst the seedlingslacking the pat gene developed chlorosis and died. The results suggest that the amount ofoutcrossed seed in the buffer rows declined very rapidly with distance from the transgenicplants. At 0.5 m the average amount of outcrossed seed was 2.35% that declined to 0.3% at1.0 m and 0% at 2.5 m and 5 m. A single plant grown from seed collected at 10 m scored astransgenic from southern side of the trial. Unfortunately it was not confirmed whether thisplant was a contaminant (GM seed mixed in the non-GM seed control) or a true transgenic(Tait and Fist 2002).



Pollen dispersal distances for oilseed poppy

173. The results from the above study indicate that no confirmed transgenics were obtainedbeyond 2.5 m (apart from the single unconfirmed transgenic plant obtained at 10 m) eventhough the bees were active during the trial. As honeybee foraging behaviour is complex andis influenced by factors like quality of the food source, the attractiveness of flowers, theavailability of other food sources, etc (Delaplane & Mayer 2000), the failure to detectpollination beyond 2.5 m suggests either one or all of the above factors or decline in pollenviability or unimportance of bees in pollination leading to predominantly self-pollination mayinfluence pollen dispersal in oilseed poppy.

174. The absence of any separation distance for certified commercial seed production ofoilseed poppy reflects the industry consensus that pollen dispersal occurs over extremelyshort distances as observed in Australian studies.

Isolation from non-GM oilseed poppy crops

175. Oilseed poppy is grown commercially only in Tasmania as a managed cultigen. Poppygrowers must comply with general conditions of licence set by the Board to grow oilseedpoppy in Tasmania. These conditions include clauses intended to limit the occurrence ofoilseed poppy as a weed. Additional measures are undertaken by the companies to ensurethat the persistence and spread of oilseed poppy is minimised to avoid the possibility of itestablishing as a weed.

176. No naturalised populations of oilseed poppy exist in Australia including WesternAustralia because oilseed poppy is a restricted crop. In Western Australia oilseed poppy canbe grown for experimental studies. However, a licence to grow oilseed poppy is requiredfrom the Western Australian Health Department.

177. Physical isolation and pollen traps of non-GM oilseed poppy could be used to providefurther containment of pollen. The Department of Agriculture WA will be required tosurround the GM crop by a 10 m pollen trap (buffer) of non-GM oilseed poppy. The pollentrap and its seed would be destroyed at the end of the trial in the same way as GM crop. Inaddition, there are no commercial poppy crops within several thousand kilometres of therelease site, so the likelihood of transfer of the introduced gene to non-GM oilseed poppy iseffectively zero.

TRANSFER OF GENES TO OTHER PLANT SPECIES

178. The most likely possibility of gene transfer to other plant species would be transfer torelated Papaveraceae and this is discussed below. Transfer to unrelated plant species can beconsidered highly improbable, and there is no evidence for any horizontal gene transfermechanism by which this could occur.



Distribution of Papaveraceae species in Western Australia

179. The Western Australian Florabase contains the following naturalised members ofPapaveraceae: Argemone ochroleuca (Mexican poppy), Argemone ochroleuca subspochroleuca, Eschscholzia californica (Californian poppy), Glaucium corniculatum, Papaverhybridum (rough poppy), P. rhoeas (field poppy), P. somniferum subsp. setigerum (smallflowered opium poppy), P. somniferum subsp. somniferum (ornamental poppy), Romneyacoulteri (California tree poppy) and Romneya trichocalyx (Western Australian Herbarium(1998). FloraBase - Information on the Western Australian Florabase, Department ofConservation and Land Management, http://www.calm.wa.gov.au/science/florabase.html).

180. The Western Australian Florabase also lists P. somniferum (opium poppy) as existing inWestern Australia. This reference is to the species that contains two subspeciesP. somniferum subsp. setigerum (small flowered opium poppy) and P. somniferum subsp.somniferum (ornamental poppy). P. somniferum subsp. somniferum (oilseed poppy) is notwild, naturalised or grown as ornamental in Western Australia although it is grown forexperimental work which is regulated under the Poisons Act 1964.

Potential for gene transfer to naturalised Papaveraceae in Western Australia

181. As with non-GM cultivated oilseed poppy, most risk accrues from the deposition ofpollen from GM oilseed poppy on the stigmas of naturalised Papaveraceae. The reciprocalevent has significant risk implications only if hybrids between naturalised species andP. somniferum backcross to the naturalised parent.

182. Although attempts have been made to do so, there are no known reports of successfulenforced cross-pollination (ie. requiring human intervention to obtain the cross) betweenP. somniferum and any of the wild related species that occur in Australia except forP. somniferum ssp. setigerum. Viable hybrids were obtained in crosses with P. somniferumssp. setigerum where P. somniferum was used as both female and male parent. Hybridisationstudies indicate that P. somniferum ssp. setigerum can cross with P. somniferum to producetriploid hybrids with reduced fertility (Department of Agriculture, WA application 2001,CSIRO Plant Industry application 2002).

183. An apparent incidence of viable hybrids from a cross between P. dubium (femaleparent) x P. somniferum (male parent) is currently being investigated. Controlled crossesbetween P. somniferum and P. bracteatum have produced F1 hybrids that grow very slowlycompared to parents and set very few seeds, and with F2 generation being even less viablethan F1 (CSIRO Plant Industry application 2002). Controlled crosses between P. somniferumand P. orientale produced F1 hybrids with poor pollen production, which were progressed toproduce fertile F4 plants (Jonsson and Loof 1973). Controlled crosses between P.somniferum and P. pseudo-orientale produced F1 hybrids with high pollen sterility (Rousiand Ojala 1984).



184. P. dubium, P. bracteatum, P. orientale and P. pseudo-orientale are not found in WA.P. somniferum ssp setigerum does not occur within several thousands of kilometres of therelease site effectively excluding the possibility of gene transfer through pollen flow. In theextremely unlikely event that cross-pollination of P. somniferum (2n=22) with P. somniferumssp setigerum (2n=44) occurred, it would result in a triploid plant that would be expected tohave low fertility and be a poor competitor. In the extremely unlikely event that cross-pollination of naturalised Papaveraceae occurred, it is not expected that such an event wouldimpart any environmental benefit or disadvantage to the wild species in terms of vigour(Department of Agriculture, WA application 2001). The likelihood of fertile hybridsoccurring, surviving to reproductive maturity and backcrossing to the parental native is,therefore, extremely low. Any consequences in the event of such transfer on the alkaloidproduction pathway of alkaloid producing Papaver species would be further considered forlarge scale or commercial releases.

C: Conclusions regarding gene transfer to other plants

185. The likelihood of gene transfer from the GMO proposed to be released into other plants(including other oilseed poppy crops, and related naturalised species, with associatedpotential adverse impacts on biodiversity) is negligible.

186. The Regulator has applied various measures, including pollen traps and isolation zone,to limit any possible outcrossing to other oilseed poppy crops and to further limit genetransfer. Furthermore, the Regulator has imposed additional licence conditions to ensureappropriate control measures are in place (see Chapter 9, Sections 2 and 3).

187. The conclusions with respect to transfer of the specific gene sequences are as follows:

Ø Alkaloid pathway enzyme gene:If this gene was transferred to other oilseed poppy crops or related naturalisedspecies, the plants would not have any survival advantage. They may haveincreased toxicity (Chapter 5, section B) but that would not provide any selectiveor competitive advantage. This is unlikely to occur, as expression of the gene inplants other than oilseed poppy would only represent one step of the multiplesteps in the alkaloid production pathway (see Chapter 7, Section 2A). Thusproduction of alkaloids or enhanced production of alkaloids is unlikely to beconferred.

Ø Antibiotic resistance genes:There would be no adverse consequences even if outcrossing occurred. The HPTprotein is derived from a soil bacterium Streptomyces hygroscopicus and isexpected to be prevalent in the environment. Any plant that did receive theantibiotic resistance marker would not have a selective advantage as antibioticsare not generally used on plants outside of the laboratory.



Ø CaMV 35S promoter and other regulatory sequences:Although some of the regulatory sequences transferred to the plants are derivedfrom plant pathogens, they only represent a very small proportion of the pathogengenome. The probability of a hazard arising due to outcrossing of thesesequences to other plants is remote, given the low likelihood of gene transfer byoutcrossing. Plants are already exposed in nature to the bacteria and viruses fromwhich these sequences are derived. In addition, none of the regulatory sequencesfrom these organisms introduced into GM oilseed poppy code for any knownpathogenic functions. It should be noted that CaMV is already ubiquitous in theenvironment and in the human diet (Hodgson 2000a).

SECTION 2 TRANSFER OF INTRODUCED GENES TO OTHER ORGANISMS(MICROORGANISMS AND ANIMALS)

A: Nature of the gene transfer hazard

188. Potential hazards, with respect to the specific gene sequences, are as follows:

Ø Alkaloid pathway enzyme gene:Transfer of alkaloid pathway enzyme may make an organism more toxic orprovide a selective advantage.

Ø Antibiotic resistance gene:Transfer of the gene to animals (including humans) other than helminths ormicroorganisms other than bacteria (such as viruses) would not present a hazard.However, bacteria or helminths that acquired the antibiotic resistance gene couldbecome resistant to that antibiotic or a related antibiotic if they also acquired afunctional, appropriately aligned prokaryotic promoter (see chapter 5, section 16).The consequences of this would depend on:

• the pathogenicity of the microorganism or helminth;

• the use and significance of the antibiotic in clinical and/or veterinarypractice;

• whether resistance to the antibiotic is already widespread in themicrobial or helminth population.

Ø CaMV 35S promoter and other regulatory sequences:

Ø If gene transfer occurred, there could be unintended or unexpected effects if theintroduced regulatory sequences alter the expression of endogenous genes. Ifsuch perturbation of normal gene expression occurred, the impact would dependon the resultant phenotype.

Ø Some of these sequences are derived from plant pathogens (cauliflower mosaicvirus and Agrobacterium tumefaciens). The possibility should be considered thatthey might have pathogenic properties.

Ø The possibility that the regulatory sequences could recombine with the genome ofother microorganisms infecting the plants to create novel recombinantmicroorganisms should also be considered.



B: Likelihood of the gene transfer hazard occurring

TRANSFER OF GENES TO HUMANS OR OTHER ANIMALS INCLUDING HELMINTHS

189. No evidence has been identified for any mechanism by which the genes could betransferred from the GM oilseed poppy plants to humans or animals including helminths, norany evidence that this has occurred during evolutionary history, despite the fact that animalsand humans eat large quantities of plant DNA (Jonas et al. 2001).

TRANSFER OF GENES TO BACTERIA

190. Transfer of the introduced genes from GM oilseed poppy to bacteria is extremelyunlikely. Horizontal gene transfer from plants to bacteria has not been demonstratedexperimentally under natural conditions (Syvanen 1999, Nielsen et al. 1997, Nielsen et al.1998) and deliberate attempts to induce such transfers have so far failed (e.g. Schlüter et al.1995, Coghlan 2000). Transfer of plant DNA to bacteria has been demonstrated only underhighly artificial laboratory conditions, between homologous sequences under conditions ofselective pressure (Mercer et al. 1999, Gebhard and Smalla 1998, Nielsen et al. 1998), andeven then only at a very low frequency. Phylogenetic comparison of the sequences of plantand bacterial genes suggests that horizontal gene transfer from plants to bacteria duringevolutionary history has been extremely rare, if occurring at all (Doolittle 1999;Nielsen et al. 1998).

191. The hpt gene is prevalent in naturally occurring bacteria found in soil. Transfer of thegenes from these naturally occurring bacteria, through well documented mechanisms forhorizontal gene transfer between bacteria (Nielsen et al. 1998; Doblhoff-Dier et al. 2000), isfar more likely than transfer of the same genes from GM oilseed poppy.

192. The transfer of a gene from a genetically modified plant to bacteria in the human gutwould require a series of steps, each of which has a very low probability (Pittard 1997; USFDA Draft Guidance Document on Use of Antibiotic Resistance Marker Genes in TransgenicPlants 1998, Jonas et al. 2001). Even if transfer of an intact gene did occur, other factorsmitigate against the hpt gene being expressed or retained by the microorganism. An intactcopy of the gene would need to:

Ø survive degradation during processing of food in the gut, and by acid andnucleases in the stomach and intestines;

Ø excision of DNA fragments containing the gene and its promoter (would need tobe active in bacteria);

Ø be taken up by a bacterium in the digestive tract;

Ø survive efficient bacterial defence mechanisms for degrading foreign DNA;

Ø become stably integrated into the bacterial genome or on a plasmid, in precisealignment with a bacterial promoter (if this were not co-transferred, intact, fromthe plant); and

Ø maintenance and expression of the gene by the bacteria.



193. In addition, the hpt gene is driven by a promoter that would need to be active inbacteria. The hpt gene, even if stably integrated, would therefore be unlikely to be expressedunless it acquired a functional bacterial promoter in appropriate orientation, which is unlikelyto occur. Further, if gene transfer did occur, the hpt gene would be unlikely to be maintainedin the microorganisms since there would be no selective advantage to ensure that themicroorganisms retained the gene and persisted and multiplied in the gut or the environmentwith the introduced gene. The overall possibility of all of these above steps occurring isremote.

194. The transfer and subsequent expression of the hpt gene in microorganisms in the humandigestive tract is therefore highly unlikely because of the number and complexity of the stepsthat would need to take place consecutively. For similar reasons, transfer of the hpt gene inmicroorganisms in the animal digestive tract is also highly unlikely.

195. The aminoglycoside inactivating genes that confer resistance to antibiotics likehygromycin are prevalent in multiple ecosystems (Ainsa et al 1997, Kelch and Lee 1978).Introduction of aminoglycoside-resistant plants is therefore not likely to alter the distributionof these genes in nature, including soil organisms and intestinal microflora (Malik andSaroha 1999).

196. In any event the potential impact on human health and safety or the environment in theunlikely event of successful transfer would be negligible.

TRANSFER OF GENES TO VIRUSES

197. There is a theoretical possibility of recombination between sequences that have beenintroduced into the genome of genetically modified oilseed poppy and the genome of virusesthat might infect the oilseed poppy. Recombination between viral sequences and planttransgenes has only been observed in the laboratory at very low levels, and only betweenhomologous sequences under conditions of selective pressure, e.g. regeneration of infectiousvirus by complementation of a defective virus, containing a deletion mutation in its coatprotein, by sequences transcribed from viral coat gene introduced into a transgenic plantgenome (Greene and Allison 1994, Teycheney and Tepfer 1999).

C: Conclusions regarding gene transfer to other organisms

198. Horizontal gene transfer from plants to animals (including humans) or microorganismsis extremely unlikely. The conclusions, with respect to the specific gene sequences are asfollows:

Ø Alkaloid pathway enzyme gene:This would not present a hazard to human health or the environment. Thealkaloid biosynthesis pathway is a multiple step pathway and the modified generepresents only one of many genes required to produce alkaloids. The alkaloidbiosynthesis pathway and the related genes are not found in animals andmicroorganisms. Transfer of a single enzyme in the absence of the wholepathway would not be expected to have any metabolic impacts, and certainly notresult in the production of alkaloids.



Ø Benzylisoquinoline alkaloids are a group of heterocyclic nitrogenous compoundsfound only in plants due to the stereochemical complexity of the molecules(Facchini 2001). As such, it is unlikely that the gene could be integrated orpersist in the bacterial genome.

Ø Antibiotic resistance gene:Transfer of this gene to organisms other than bacteria or helminths would notpresent a hazard, since the antibiotic in question is only used to treat or preventbacterial or helminthic infections. Horizontal transfer to bacteria is alsoextremely unlikely and is considered to pose negligible risks to human health orthe environment for the following reasons:

Ø Hygromycin is not used in clinical medicine but is used in veterinary practice,and alternative antibiotics are readily available.

Ø The hpt gene is prevalent in naturally occurring bacteria found in soil. Transferof the genes from these naturally occurring bacteria, through well documentedmechanisms for horizontal gene transfer between bacteria (Nielsen et al. 1998;Doblhoff-Dier et al. 2000), is far more likely than transfer of the same genes fromGM oilseed poppy.

Ø The antibiotic resistance genes in these bacteria are often located on transmissiblegenetic elements that are readily transferable between bacterial species andtherefore widespread in the environment. So, in the unlikely event that the hptgenes were transferred from GM oilseed poppy to a bacterium, this would beunlikely to have any detectable impact on the existing level of resistance inmicrobial populations.

Ø CaMV 35S promoter and other regulatory sequences:As discussed above, horizontal gene transfer from plants to bacteria or animalsincluding humans, is extremely unlikely. The probability of recombination of theCaMV 35S or other regulatory sequences in the modified oilseed poppy withother microorganisms can be considered to be negligible because recombinationbetween microbial sequences and plant genes has only been observed at very lowlevels between homologous sequences under conditions of selective pressure.

Ø While Ho et al (2000) have postulated that there are risks posed throughrecombination of the CaMV 35S promoter with the genomes of other virusesinfecting the plants to create new viruses, or of integration of the CaMV 35Spromoter into other species causing mutations, cancer or reactivation of dormantviruses, these claims have been challenged in the scientific literature (eg Hodgson2000 a,b; Hull et al 2000; Morel and Tepfer 2000). CaMV is already ubiquitousin the environment and in the human diet and the CaMV 35S promoter isexpressed at far higher levels in naturally infected plants than in transgenic plants.Hull et al (2000) found that there are compelling arguments to support the viewthat there would be no more risks arising from potential recombination than thereare from existing non-transgenic crops.



CHAPTER 8 RISK MANAGEMENT PLAN

199. This part of the document recaps the main conclusions from the risk assessment relatingto risks to human health and safety or the environment, and details the risk management plandeveloped by the Regulator to manage these risks.

SECTION 1 SUMMARY OF RISK ASSESSMENT CONCLUSIONS

200. It has been concluded that the proposed release of the GM oilseed poppy in WesternAustralia would not pose any risks to human health and safety or to the environment as aresult of the genetic modification of the oilseed poppy that could not be managed. The mainconclusions from the risk assessment are that:

Ø The GM oilseed poppy is not likely to prove more allergenic or result in moretoxic exposure to humans or other organisms, than non-GM oilseed poppy asneither any plant part nor plant product is for human or animal consumption andbecause of the tight controls on the release;

Ø the risk of the GM oilseed poppy establishing as a weed is very low and not likelyto be greater than that of non-GM oilseed poppy;

Ø the potential for transfer of the introduced genes to non-GM oilseed poppy cropsis negligible;

Ø the potential for transfer of the introduced genes to naturalised members of familyPapaveraceae is extremely low because of the geographical isolation of therelease site and genetic incompatibility between oilseed poppy and the naturalisedspecies; and

Ø the likelihood of transfer of the introduced genes to other organisms is extremelylow, but even if such transfer occurred would be unlikely to pose any hazard tohuman health and safety or the environment.

SECTION 2 RISK MANAGEMENT PLAN

Section 2.1 Risk of toxicity or allergenicity

201. On the basis of the risk assessment, the risks of toxicity or allergenicity of the GMoilseed poppy are considered extremely low because of the limited scale of the release andthe highly regulated nature of the crop. None of the plant parts or by-products of GM oilseedpoppy will be used for human food or animal feed or therapeutics or for any othercommercial purpose. The following management strategies detailed below in paragraph 4 areconsidered adequate to manage risks of toxicity and allergenicity.

202. The licence conditions imposed by the Regulator require that products from this releasewould not be used in human food or animal feed or therapeutics or for commercial purposesand to restrict use of GM material from the release. As a further precaution the Regulator hasissued a licence that includes the requirement for workers to use facemasks, disposablegloves and overalls when handling the GMO. The proponent is also required to enclose therelease site with a rabbit-proof fence and bird net to prevent access to birds and animals.



Section 2.2 Risks of weediness or gene transfer

203. It has been concluded that the risks relating to weediness or gene transfer are extremelylow. Nevertheless, it was considered that the risks could be further reduced by implementingvarious strategies to minimise the spread and persistence of the GM oilseed poppy, or themodified genetic material, in the environment.

204. The licence includes a number of specific conditions relating to management ofpotential risks of the GM oilseed poppy becoming a weed or transferring the genetic materialto other organisms. The specific licence conditions include the requirement to:

Ø isolate the GM oilseed poppy crop from other non-GM oilseed poppy crops;

Ø have an isolation zone of 500 m around the release site;

Ø surround the release site with a 10 m pollen trap comprising of non-GM oilseedpoppy to limit the possibility of outcrossing to other oilseed poppy crops;

Ø fence and bird net the release site to prevent access to animals and birds;

Ø handpick the seed capsules (these are indehiscent, i.e. do not split and disperseseed when ripe);

Ø destroy by burning any viable material not required for subsequent releases(subject to further approvals) after the harvest;

Ø lightly cultivate and irrigate the release site after harvest of the GMO toencourage germination of volunteers;

Ø test for transfer of hygromycin resistance gene in any Papaver species that is notoilseed poppy detected after harvest in the isolation zone;

Ø destroy volunteers after harvest; and

Ø all the places required to be monitored must be monitored at least once everymonth for at least 2 years, after harvest and thereafter, until such time as theplaces to be monitored shows no presence of the volunteers for a period of12 months and the Regulator has agreed in writing that further monitoring is notrequired.

205. The release site is not prone to flooding and the absence of natural waterways in 50 mof the release site will prevent the spread of seed by water.

206. The specific licence conditions, and the reasons behind them, are set out in detail inSection 3 of this Chapter, Appendix 2 and Appendix 3.

Section 2.3 General licence conditions

207. In addition to management of potential risks by imposition of specific licenceconditions as given in Section 3 of this Chapter, the licence issued by the Regulator alsocontains a number of general conditions including statutory conditions relating torequirements under Sections 61 to 65 of the Act. These conditions apply to all licencesissued by the Regulator, for example, there are conditions that will require the licence holderto:



Ø identify the persons or classes of person covered by the licence;

Ø only undertake the specified dealings covered by the licence;

Ø require the applicant to inform people covered by the licence of their obligationsunder the licence:

Ø ensure that the Regulator, or persons authorised by the Regulator for the purposesof monitoring or auditing have access to the release site and any buffer orisolation zone sites;

Ø inform the Regulator if the applicant becomes aware of any additionalinformation about risks to human health or safety or to the environment, anyunintended effects of the release, or any contraventions of the licence conditions;and

Ø ensure appropriate training for persons covered by the licence.

Section 2.4 Monitoring and enforcement of compliance by the OGTR

208. It should be noted that, as well as imposing licence conditions, the Regulator hasadditional options for risk management. The Regulator has legislative capacity to enforcecompliance with licence conditions, and indeed, to direct a licence holder to take any stepsthe Regulator deems necessary to protect the health and safety of people or the environment.The OGTR also independently monitors release sites to determine whether the licence holderis complying with the licence conditions, and to assess whether there are any unforseenproblems.

SECTION 3 SPECIFIC RISK MANAGEMENT LICENCE CONDITIONS

209. The licence conditions set out in Appendix 2 are intended to manage the identifiedrisks, largely through preventing dissemination of the GMOs or their genetic material outsidethe release site. The licence conditions also include contingency provisions to cover anyunintended release of the GMOs outside the release site. Detailed reasons for the individuallicence conditions are set out in Appendix 3.

210. Qualified Department of Agriculture, Western Australia staff would be directlysupervising the proposed release if the Regulator issues a licence for the release. TheDepartment of Agriculture, Western Australia is required, under licence conditions, to beproactive in reviewing and assessing any new information that comes to light about the risksand the efficacy of the proposed management strategies during the course of the release. Thelicence issued to The Department of Agriculture, Western Australia will be able to be variedat any time to add new conditions, for instance to manage any new risks that are identified, orto improve the existing management strategies.

211. The Regulator will also be proactive in reviewing any new information about the risksof the proposed release and may amend licence conditions on the basis of this.


REFERENCES 47

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APPENDIX 1 SUMMARY OF PUBLIC SUBMISSIONS 53

APPENDIX 1 - SUMMARY OF PUBLIC SUBMISSIONS

a Submission from: A: agricultural organisation; I: individual; E: environmentalorganisation; F: food interest organisation; C: consumer/public interest organisation;b Refers to Chapter (Ch.) and section (s.) numbering within risk assessment and managementplan; OSA: outside scope of the assessment; NR: No specific response; NA: Not applicable

SUB.NO:

TYPEA Summary of issues raised CONSIDERATIONOF ISSUEB

1 C • concerns that antibiotic resistance marker gene is beingused although WHO has advised that use of antibioticmarker genes should be phased out

• what is the need to genetically modify oilseed poppywhen it has been shown that by conventional breedingalkaloid production has already been increased threetimes in oilseed poppy

• whether Australian native animals as opposed to non-indigenous grazing animals such as horses, sheep andcattle have been taken into account for no reports ofadditional adverse effects on animals or insects despitewide cultivation over a number of years of non-GMoilseed poppy

• type of fence be stipulated to keep animals [includingnative animals like kangaroos, wombats] out

• CaMV 35S promoter should not be used until it has beendemonstrated beyond any doubt that there is no danger[from using it]

Chs 4, 7

Ch 3

Chs 4, 5

Appendix 2

Ch 7

2 I • moratorium on genetic engineering and GMOs untilspiritual, moral, health and legal issues are resolved

• everything GE should be comprehensively labelled

OSA. For healthissues see Ch 1;

Appendix 2

OSA

3 I • proposed release should be monitored within theguidelines of the legislative provisions and not solelyrely on voluntary system investigations

• any non-intentional release of modified alkaloid [poppy]with antibiotic resistance to be promptly monitored andlimited by appropriate response

Ch 8, s 2.5;Appendix 2

Ch 8, s 2.5;Appendix 2


APPENDIX 1 SUMMARY OF PUBLIC SUBMISSIONS 54

4 I • other countries should conduct trials on GM poppybefore Australia

• although trial will be well controlled to prevent [GMoilseed poppy] being used in food but other [GM]commodities have been included in [food] with orwithout labelling

• reason for trial

• birds, bees and butterflies could be killed due to toxicityof oilseed poppy.

OSA

OSA

Ch 3

Ch 5

5 A • nil tolerance to release of anything genetically modifieduntil further laboratory testing is conducted.

OSA

6 A • no comments NA

7 A • likelihood of horizontal spread of GM constructs tounrelated species

• risk of cross pollination and spreading should not exist[negligible in RARMP].

• possibility of vector CaMV 35S promoter to recombinewith other pathogens and create new and potentiallydangerous viruses

Ch 7

Ch 7

Ch 7


APPENDIX 2 LICENCE CONDITIONS 55

APPENDIX 2 - LICENCE CONDITIONSPART 1

This instrument, including its attachments, is a licence authorising dealings involving theintentional release of GMOs into the environment. It is issued by the Gene TechnologyRegulator pursuant to the Gene Technology Act 2000 (Cth).

Holder of Licence

1.1 The holder of this licence (‘the licence holder’) is the Department of Agriculture,Western Australia.

Project Supervisor

2.1 The Project Supervisor in respect of this licence is the person identified at ‘ProjectSupervisor’ at Attachment A.

Persons covered by licence

3.1 The persons covered by this licence are the licence holder and employees, agents orcontractors of the licence holder and other persons who are, or have been, engaged toundertake any activity in relation to a GMO grown in a Location.

(Explanatory Note: Each person covered by this licence is a ‘person covered by a GMOlicence’ for the purposes of the Gene Technology Act 2000 (Cth)).

Description of GMO covered

4.1 The GMO covered by this licence (‘the GMO’) is identified and described at ‘GMODescription’ at Attachment B.

Dealings authorised by licence

5.1 This licence authorises the licence holder and persons covered by the licence to dealwith the GMO as expressly authorised by this licence and subject to the limitations ondealing with the GMO that are contained in the conditions in this licence.

Period covered by licence

6.1 This licence remains in force until it is cancelled or surrendered. No dealings with theGMO are authorised during any period of suspension.



PART 2

Interpretation and Definitions

Words and phrases used in this licence have the same meanings as they do in the GeneTechnology Act 2000 (Cth) and the Gene Technology Regulations 2001.

Words importing a gender include any other gender.

Words in the singular include the plural and words in the plural include the singular.

Words importing persons include a partnership and a body whether corporate or otherwise.

References to any statute or other legislation (whether primary or subordinate) is to a statuteor other legislation of the Commonwealth of Australia as amended or replaced from time totime unless the contrary intention appears.

Where any word or phrase is given a defined meaning, any other part of speech or othergrammatical form in respect of that word or phrase has a corresponding meaning.

In this licence:

‘Clean’ (or ‘Cleaned’), as the case requires, means:(a) in relation to a Location or an area, the destruction of the GMO, Material from

the GMO, Pollen Trap plants or Material from Pollen Trap plants in thatLocation or area, to the reasonable satisfaction of the Regulator; or

(b) in relation to Equipment, the removal and destruction of the GMO andMaterial from the GMO, Pollen Trap plants or Material from Pollen Trapplants from the Equipment, to the reasonable satisfaction of the Regulator.

‘Destroy’, (or ‘Destroyed’ or ‘Destruction’), as the case requires, means killed by one of thefollowing methods:

(a) stalk pulling; or(b) uprooting by ploughing to a depth of not more than 50mm; or(c) burning; or(d) treatment with herbicide; or(e) slashing; or(f) mowing; or(g) hand weeding; or(h) a combination of the above.

Note: This definition is intended to require the killing of whatever is intended to bedestroyed. Depending on the circumstances, one or more of the techniques described abovemay not be appropriate. For example, in the case of killing the remains of harvest of theGMO, treatment of post-harvest remains by herbicide would not be a sufficient mechanism.

‘Equipment’ includes storage equipment, transport equipment (eg, trucks and tarpaulins),clothing and tools.

‘GM’ means genetically modified.

‘Isolation Zone’ means an area of land, extending outwards 500 metres in all directions fromthe outer edge of a Pollen Trap in respect of the Location.

‘Location’ means the area of land where the GMO is grown.

‘Material from Pollen Trap plants’ means seed, stubble, pollen or any other material(including parts of a plant) that is derived from or produced by a Pollen Trap plant.



‘Material from the GMO’ means GM seed, stubble, pollen or any other geneticallymodified material (including part of a GMO) that is derived from or produced by the GMO.

‘Monitoring Zone’ means the area of land extending outwards by 100m in all directionsfrom the outer edge of the Pollen Trap.

‘Natural Waterways’ means waterways other than irrigation channels, holding dams orstorage ponds used to collect water runoff from irrigated areas.

‘OGTR’ means the Office of the Gene Technology Regulator.

‘Pollen Trap’ means an area of land, extending 10 metres in all directions from the outsideedge of the Location, containing non-genetically modified Papaver somniferum somniferum(oilseed poppy) that is grown in such a way as to reasonably promote a dense and vigourousgrowth and flowering of the non-genetically modified Papaver somniferum somniferum(oilseed poppy) at the same time as the GMO.

‘Pollen Trap plant’ means Papaver somniferum somniferum (oilseed poppy) from a PollenTrap.

‘Related species’ means the following:

Eschscholzia californica (Californian poppy),Papaver dubium (long head poppy),Papaver hybridum (rough poppy),Papaver rhoeas (field poppy),Papaver argemone (pale poppy),Papaver somniferum subsp. setigerum (small flowered opium poppy),Papaver somniferum subsp. somniferum (ornamental poppy),Papaver bracteatum,Papaver orientale, andPapaver pseudo-orientale.

‘Volunteer plants’ means progeny of the GMO or a Pollen Trap plant, growing at theLocation, the Pollen Trap or the Monitoring Zone during post-harvest monitoring.



PART 3

Conditions of licence

The licence holder and persons covered by this licence must comply with the conditions ofthis licence.

Section 1: General Conditions

Informing people of their obligations

1.1 The licence holder must inform each person covered by this licence of the obligationsimposed on them as a result of the conditions in this licence.

1.2 The licence holder must provide the Regulator, on the Regulator’s written request, asigned statement from each person covered by this licence that the licence holder hasinformed the person of the conditions of this licence that apply to that person.

Material Changes in circumstances

2.1 The licence holder must immediately, by notice in writing, inform the Regulator of:(a) any relevant conviction of the licence holder occurring after the

commencement of this licence;(b) any revocation or suspension of a licence or permit held by the licence holder

under a law of the Commonwealth, a State or a foreign country, being a lawrelating to the health and safety of people or the environment;

(c) any event or circumstances occurring after the commencement of this licencethat would affect the capacity of the holder of his licence to meet theconditions in it.

Remaining an accredited organisation

3.1 The licence holder must, at all times, remain an accredited organisation and complywith any conditions of accreditation set out in the Guidelines for Accreditation ofOrganisations.

Changes to details

4.1 The licence holder must immediately notify the Regulator in writing if any of thecontact details of the Project Supervisor change.



Section 2: Specific Conditions

Locations and size of trial

1.1 The GMO may be grown.

1.2 The GMO must not be grown outside the Shire of Wyndham/East Kimberley inWestern Australia.

1.3 The GMO may be grown at a single Location within the Shire of Wyndham/EastKimberley in Western Australia. The Location must be within the FrankwiseResearch Institute, Durack Drive, Kununurra, WA. The total area of land comprisingthe Location at which the GMO may be grown must not exceed 0.2 hectares,cumulatively over the life of the licence. Prior to commencing to grow the GMO atthe Location, the Location’s GPS coordinates and either a street address, or otherdirections to the Location, must be provided to the Regulator by notice in writing.

1.4 Planting the GMO at the Location must be commenced between 1 March 2003 and30 June 2003.

1.5 The licence holder must be able to access and control the Location to the extentnecessary to comply with this licence, for the duration of the life of the licence.

Pollen Traps and Isolation Zones

2.1 A Pollen Trap and an Isolation Zone must completely surround the outer edge of theLocation.

2.2 The Pollen Trap must be planted at the same time as the planting of the GMO at theLocation. Once planted, Pollen Trap plants must be handled and controlled as if theyare GMOs under this licence.

2.3 The edge of the Pollen Trap farthest from the GMO (the ‘outer edge of the PollenTrap’) must not be within 50 metres of a Natural Waterway. Natural Waterways mustnot fall within the Location or the Pollen Trap.

2.4 The Pollen Trap and Isolation Zone must be able to be accessed and controlled by thelicence holder to an extent that is commensurate with the licence holder’s rights toaccess and control the Location.

2.5 No related species may be grown in the Isolation Zone while the GMO is beinggrown. Any related species occurring in the Isolation Zone while the GMO is beinggrown must be destroyed prior to it flowering.

2.6 The outer edge of the Pollen Trap must be surrounded by a rabbit-proof fence.

2.7 The Location and the Pollen Trap must be enclosed within a bird net that excludesbirds from the Pollen Trap and Location.

Protective clothing and control of access to Locations and Pollen Traps

3.1 Access to the Location and Pollen Trap must be restricted to those persons covered bythis licence.



3.2 Subject to 3.1 above, persons entering the Location or Pollen Trap during any periodof flowering of the GMO must wear facemasks, disposable gloves, rubber shoes andoveralls.

Research on Gene Flow and environment impacts

4.1 The licence holder must, in consultation with the OGTR, develop and implement aprogram to collect information on the effectiveness of the Pollen Trap and IsolationZone in preventing gene flow from the GMO to non-genetically modified Papaverspecies. This program must collect, among other things, information on transfer ofthe hygromycin resistance to Papaver species planted in the Isolation Zone.

Harvest of GMO and seed extraction

5.1 Oilseed poppy capsules, and only capsules, may be harvested from the Location andthe Pollen Trap.

5.2 Any oilseed poppy capsules harvested from the Location must be harvested by hand,separately from any other non-genetically modified Papaver somniferum subsp.somniferum (oilseed poppy).

5.3 If oilseed poppy capsules are harvested from the Pollen Trap, they must be harvestedwith and at the same time as any oilseed poppy capsules harvested from the Location.

5.4 Following harvest, all harvested oilseed poppy capsules (from either the Location orthe Pollen Trap) must, at all times, be kept and stored separately to maintain theiridentity.

5.5 Subject to 5.6, immediately following harvest, all harvested oilseed poppy capsules(from either the Location or the Pollen Trap) must be:

(a) Stored in a secure (locked) room at the Frankwise Research Institute, DurackDrive, Kununurra, WA, that is marked to indicate that genetically modifiedoilseed poppy capsules are stored in the room; or

(b) transported to the GlaxoSmithKline field service store at 14 Henry Street, LaTrobe, Tasmania (‘the field service store’), and stored in a secure (locked)room that is marked to indicate that genetically modified oilseed poppycapsules are stored in the room; or

(c) destroyed.

5.6 If harvested oilseed poppy capsules are transported to the field service store:

(a) seeds may be extracted from those capsules at the field service store. The seedmay be stored in a sealed container, within a secure (locked) room that ismarked to indicate that genetically modified oilseed poppy seed is stored in theroom. GM seed may be stored for the purposes of future experimentalresearch at the field service store.

(b) any de-seeded GM oilseed poppy capsules may be transported to the QAlaboratory, GlaxoSmithKline factory, Princes Highway, Port Fairy, Victoriaand subjected to analysis at the factory.



5.7 Following any processes described in 5.5 or 5.6 above, any remaining GMO, Materialfrom the GMO or Pollen Trap plants or Material from Pollen Trap plants, includingany waste materials, for example waste materials arising during the process ofextracting seed, must be destroyed.

Cleaning – post harvest and generally – GM plants and Pollen Trap plants

6.1 Where Equipment or a Location and Pollen Trap is used pursuant to this licence inrespect of GMOs, Material from GMOs, Pollen Trap plants or Material from PollenTrap plants, the Equipment, Location and Pollen Trap must be Cleaned.

6.2 Subject to 6.4, then either within 14 days of harvest of the GMO or by30 November 2003, whichever occurs first, the Location and Pollen Trap must beCleaned.

6.3 If Equipment is Cleaned, the area in which the Equipment is Cleaned must also beCleaned. (For the sake of clarity, it is not necessary for Equipment to be Cleaned onlyat a Location.).

6.4 Cleaning must occur immediately or as soon as practicable after the use and before itis used for any other purpose.

6.5 On the request of the Regulator, the Regulator must be provided with writtendocumentation of the procedures in place to ensure continuing compliance with thecleaning conditions in this licence.

Promotion of germination of the GMO after Cleaning

7.1 Following Cleaning of the GMO and any Material from the GMO, at the Location andPollen Trap, the Location and Pollen Trap must be lightly tilled and irrigated on 3separate occasions in such a way as to encourage the growth of any GMO or Materialfrom the GMO that is still at the Location or Pollen Trap.

7.2 For the purposes of this condition, ‘lightly tilled’ means ploughed to a depth of notmore than 50mm.

7.3 Subject to 7.4 below, light tilling must take place between:

(a) Harvest and 1 November 2003; and/or

(b) 1 April 2004 and 31 October 2004; and/or

(c) 1 April 2005 and 31 October 2005.

7.4 The 3 occasions of light tilling must be carried out at least 3 weeks apart.

7.5 A report on light tilling activities undertaken must be reported to the Regulator inwriting. Results must be reported to the Regulator within 35 days of any day onwhich light tilling occurs. Results of reporting must include:

(a) details of the Location (and Pollen Trap, if any) tilled; and

(b) details of the tillage methods used.

Note: The condition above recognises that the obligation to harvest and Clean Locations willnot ensure the complete eradication of the GMO from a Location, particularly any remainingseed bank.



It is possible that following the Cleaning of Locations, some seed or other viable GMO mightremain, for example in a seed bank. The purpose of this condition is to oblige licence holdersto encourage the germination of any remaining viable GMO after harvest and Cleaning,particularly so as to reduce and manage any seed bank remaining in the soil.

Light tilling may take place 3 times between April and October 2004, so long as a gap of atleast 3 weeks is left between each tilling, to allow any regeneration of the GMO to occur.

Monitoring – post harvest and generally

8.1 Following Cleaning of the GMO, Material from the GMO, Pollen Trap plants orMaterial from Pollen Trap plants, at the Location and Pollen Trap, the followingplaces must be monitored for the existence of Volunteer plants:

(a) the Location;

(b) the Pollen Trap;

(c) the Monitoring Zone; and

(d) any areas used to Clean Equipment used in connection with the GMO or todestroy the GMO, Material from the GMO, Pollen Trap plants or Materialfrom Pollen Trap plants.

8.2 Monitoring must be performed by a person who is able to recognise Volunteer plants.

8.3 All the places required to be monitored must be monitored at least once every monthfor at least 2 years, commencing on the last day of Cleaning of the Location, andthereafter, until such time as the place to be monitored shows no presence of theVolunteer plants for a period of 12 months and the Regulator has agreed in writingthat further monitoring is not required.

8.4 The results of monitoring activities must be reported to the Regulator in writing.Results must be reported to the Regulator within 14 days of any day on whichmonitoring occurs. Results of reporting must include:

(a) the names of the person or persons who undertook the monitoring and detailsof the experience, training or qualification that enabled them to recogniseVolunteer plants;

(b) the number of Volunteer plants observed, if any;

(c) details of the development stages reached by the Volunteer plants, if any; and

(d) details of methods used to destroy Volunteer plants, if any.

8.5 Any Volunteer plant identified must be Cleaned prior to the plant flowering.

8.6 The destruction of any Volunteer plant must be performed in such a way as tofacilitate the licence holder to meet their obligations under 4.1 above.

Use of Locations post-harvest

9.1 If the GMO is grown at the Location, no other Papaver somniferum subsp somniferumof any kind may be grown at the Location, or the Pollen Trap, or the MonitoringZone, for a period of 3 years commencing from the date of conclusion of Cleaning ofthe GMO at the Location. [This condition ceases to operate if the Regulator agrees inwriting, pursuant to 8.3 above, that further monitoring is not required.]



9.2 If the GMO is grown at the Location, no plants may be planted at the Location or thePollen Trap or the Monitoring Zone for 3 years from the date of conclusion ofCleaning of the GMO at the Location unless:

(a) prior to planting the plants, the Regulator has received written notice of theproposed date of planting and the plant proposed to be planted; and

(b) the plants are grasses (grass pastures), or a plant agreed to in writing by theRegulator; and

(c) the Regulator is satisfied that monitoring and destruction of Volunteer plantswill not be adversely affected by the planting.

Transportation of the GMO, Material from GMO, Pollen Trap plants and Material fromPollen Trap plants

10.1 The GMO, Material from the GMO, Pollen Trap plants and Material from Pollen Trapplants (including oilseed poppy capsules, de-seeded GM oilseed poppy capsules andoileed poppy seed) may only be transported outside the Location and Pollen Trap ifcontained within a primary, sealed container that is packed in a secondary,unbreakable container.

10.2 All transportation of the GMO, Material from the GMO, Pollen Trap plants andMaterial from Pollen Trap plants (including oilseed poppy capsules and oilseed poppyseed), must be undertaken in accordance with the Guidelines for the Transport ofGMOs issued by the Regulator.

10.3 Every container used to transport GMO, Material from the GMO, Pollen Trap plantsand Material from Pollen Trap plants (including oilseed poppy capsules and oilseedpoppy seed) must be labelled:

(a) to indicate that it contains genetically modified oilseed poppy; and

(b) with telephone contact numbers for the licence holder and instructions tocontact the licence holder in the event that the container is broken ormisdirected.

10.4 The licence holder must have in place accounting procedures to verify whether thesame quantity of GMO, Material from the GMO, Pollen Trap plant or Material fromPollen Trap plants sent is delivered and must document methods and procedures usedfor transportation.

Contingency Plans

11.1 Within 30 days of the date of the commencement of this licence, a writtenContingency Plan must be submitted to the Regulator detailing measures to be takenin the event of the unintended presence of the GMO, Material from the GMO, PollenTrap plants and Material from Pollen Trap plants outside a Location that must bemonitored.

11.2 The Contingency Plan must include details of procedures to:

(a) ensure the Regulator is notified immediately if the licence holder becomesaware of the event;



(b) to destroy any of the GMOs, Material from the GMOs, Pollen Trap plants orMaterial from Pollen Trap plants;

(c) monitor and destroy any Volunteer plants that may exist as a result of theevent.

11.3 The Contingency Plan must be implemented in the event that the unintended presenceof the GMO, Material from the GMO, Pollen Trap plants or Material from PollenTrap plants is discovered outside an area that must be monitored.

Compliance Management Plan

12.1 Prior to growing the GMO, a written Compliance Management Plan must be providedto the Regulator. The Compliance Management Plan must describe in detail how thelicence holder intends to ensure compliance with these conditions and document thatcompliance.

Reporting

13.1 The licence holder must provide the Regulator with a written report within 90 days ofeach anniversary of this licence, in accordance with any Guidelines issued by theRegulator in relation to annual reporting.

Testing methodology

14.1 The licence holder must provide a written instrument to the Regulator describing anexperimental method that is capable of reliably detecting the presence of the GMOand any transferred genetically modified material that might be present in a recipientorganism. The instrument must be provided by 30 March 2003.

Statements from persons covered by the licence

15.1 The licence holder must inform each person covered by this licence of the obligationsimposed on them by the conditions of the licence. The licence holder must provide tothe Regulator, on the Regulator’s written request, a signed statement from each personcovered by this licence that the licence holder has informed the person of theconditions of this licence that apply to that person.

Notification of planting of the GMO

16.1 The licence holder must provide notices in writing to the Regulator in respect of eachof the following:

(a) the long term forecasted date or dates of commencement of planting of theGMO and Pollen Trap ('the long term forecast planting date notice'). Thisnotice must be provided at least 30 days before the forecasted date or dates ofcommencement of planting;

(b) for each Location, the short term forecasted date or dates of commencement ofplanting of the GMO and Pollen Trap ('the short term forecast planting datenotice'). This notice must be provided at least 7 days, and not more than 20days, prior to the dates notified to the Regulator in the long term forecastplanting dates notice; and



(c) for each Location, the actual date or dates of commencement of planting of theGMO and Pollen Trap ('the actual planting date notice'). This notice must beprovided within 7 days of commencement of planting of the GMO and PollenTrap.

Notification of commencement of flowering of the GMO

17.1 The licence holder must provide notices in writing to the Regulator in respect of eachof the following:

(a) the long term forecasted date or dates of commencement of flowering of theGMO and Pollen Trap ('the long term forecast flowering date notice'). Thisnotice must be provided at least 30 days before the forecasted date or dates ofcommencement of planting;

(b) the short term forecasted date or dates of commencement of flowering of theGMO and Pollen Trap ('the short term forecast flowering date notice'). Thisnotice must be provided at least 7 days, and not more than 20 days, prior to thedates notified to the Regulator in the long term forecast flowering date notice;and

(c) the actual date or dates of commencement of flowering of the GMO and PollenTrap ('the actual flowering date notice'). This notice must be provided within7 days of commencement of flowering of the GMO and Pollen Trap.

Notification of commencement of harvest of GMO

18.1 The licence holder must provide notices in writing to the Regulator in respect of eachof the following:(a) the long term forecasted date or dates of commencement of harvest of the

GMO ('the long term forecast harvest date notice'). This notice must beprovided at least 30 days before the forecasted date or dates of commencementof planting;

(b) the short term forecasted date or dates of commencement of harvest of theGMO ('the short term forecast harvest dates notice'). This notice must beprovided at least 7 days, and not more than 20 days, prior to the dates notifiedto the Regulator in the long term forecast harvest date notice; and

(c) the actual date or dates of commencement of harvest of the GMO ('the actualharvest date notice'). This notice must be provided within 7 days ofcommencement of harvest of the GMO.

GMO and GM products not to be consumed

19.1 Neither the GMO nor any Material from the GMO or Pollen Trap may be used forhuman food or animal feed.

19.2 Neither the GMO nor any Material from the GMO or Pollen Trap may be used for anytherapeutic or commercial purpose.



APPENDIX 3 REASONS FOR SPECIFIC LICENCE CONDITIONS

Specific conditions (Part 3, Section 2 of the licence):

A description of the basic reasons for inclusion of the specific licence conditions set out inPart 3, section 2 of the licence follow.

These reasons should be read in conjunction with the risk assessment and risk managementplan, which sets out details of the risks and the identified means by which those risks are ableto be managed.

The conditions that have been implemented in the licence are primarily intended to:

• Directly limit the potential for spread and persistence of the GM oilseed poppy in theenvironment outside the release site or the isolation zone, as well as any potential foradverse allergenic or toxicity related incidents, since this would increase the potential forrisks to human health and safety or the environment;

• Encourage and promote compliance with licence conditions (and thereby indirectly limitthe potential for spread and persistence of the GM oilseed poppy in the environmentoutside the release site or the isolation zone) by implementing efficient and effectivereporting of the licence holder’s conduct under the licence and monitoring of that conductby the OGTR.

Compliance with licence conditions is considered to be a fundamental component of theoverall management strategy for the proposed release. Accordingly, conditions throughoutthe licence require the provision of information to the Regulator so that compliance withlicence conditions can be monitored, and where licence conditions are not observed, othermeasures can be taken to manage any resulting risks. Where information is required to beprovided by a licence condition it is considered to be necessary and appropriate within thecontext of ensuring the successful monitoring of the licence by the OGTR.

Condition 1 limits the size, location and timing of the release.

The condition limits the dealings that may be undertaken by the licence holder to those thathave been sought by the licence holder and considered and assessed by the Regulator. TheRegulator has not considered risks associated with larger scale field trials, in differentlocations and at different times. The Regulator has not considered risks associated withdealings other than those dealings expressly authorised in the licence conditions. As theserisks have not been assessed, such other dealings are prohibited.

At the time of granting the licence, the licence holder had not determined the exact locationsproposed to be used for the trials. While the Regulator has assessed the risks associated withthe conduct of the proposed release in the Shire indicated in the licence and determined thatthey are able to be managed, the Regulator requires detailed information about site locationprior to commencement of any release in order to maintain the GMO Record, as well as to beable to effectively plan and implement OGTR monitoring of the release site. Accordingly,before any dealings authorised by the licence can take place, the licence holder must provideadditional, specific information about where the dealings will take place.



Historically, some difficulties have been encountered in managing the monitoring of GMOfield trials post-harvest, when agreements for the use of release sites (such as farming leases)have expired prior to all agreed monitoring taking place. For this reason, the Regulatorconsiders it important to require licence holders to obtain continuing access to field releasesites (and, where necessary, surrounding areas) to the extent necessary to meet anycontinuing licence conditions. In the absence of this condition, the Regulator considers thatrisks associated with licence holders failing to comply with other conditions in this licence,particularly post-harvest monitoring conditions, may not be able to be properly managed.

Condition 2 is intended to minimise gene flow outside the release site and to prevent thedissemination of the GMO by birds and animals. This is to limit the risks to human healthand safety and the environment, posed by the potential toxicity or allergenicity or weedinessof oilseed poppy, or by gene transfer from oilseed poppy to other organisms.

The imposition of a pollen trap, an isolation zone, a rabbit proof fence and a bird net isconsidered to be a satisfactory and appropriate method of managing these risks.

In the absence of these conditions, the Regulator considers that risks to human health and theenvironment posed by the potential toxicity or allergenicity or weediness of oilseed poppy, orby gene transfer from oilseed poppy to other organisms in relation to this release, may not beable to be managed.

The requirement to handle pollen trap plants as if they are GMOs is considered to be areasonable means by which to minimise the risk of gene flow from the GMO to plants in apollen trap and then on into sexually compatible plants in the broader environment.

The release site is not permitted to be situated near natural waterways because the Regulatorconsiders this necessary to manage risks that might otherwise arise as a result of entry ofpollen or other genetically modified material into a waterway.

The requirement for the licence holder to be able to access and control pollen traps andisolation zones is imposed for the same reasons that the licence holder is required to be ableto access and control the location. These reasons are discussed in more detail in thediscussion of condition 1 above.

Condition 3 seeks to manage risks associated with human-related dissemination of the GMO,as well as potential adverse impacts related to toxicity and allergenicity to GM pollen. In theabsence of the condition, it is considered that these risks can not be effectively managed.

Condition 4 requires the licence holder to develop and implement research on gene flow andenvironment impacts in consultation with the Regulator so that the Regulator can assesswhether the study is sufficiently rigorous to provide meaningful data, and request anyamendments in a timely fashion. The provision of this information to the Regulator isconsidered necessary, in the context of the Regulator’s oversight of all licenced dealings,because it facilitates the review of the effectiveness of licence conditions over time.

Condition 5 places restrictions on harvesting, storing, transporting, extracting seed, testingand destroying the GMO and pollen trap plants, to prevent the spread or persistence of theGMOs or pollen trap plants (some of which may be modified) in the environment. These



containment requirements are considered necessary to manage risks associated with thespread or persistence of the GMO, including, for example, by mixing with non-GM plants orseed, loss of identity as GM or dissemination in the course of transportation.

Conditions requiring the GMO from releases to be able to be traced are considered necessaryin the context of the Regulator’s overall monitoring of the licence.

The requirement to destroy remaining plants and material following experiments isconsidered necessary to limit the possibility of persistence of the GMO in the environment.

Condition 6 describes requirements to destroy GMOs at the release site and pollen trap, aswell as any GM plants or material that might be attached to equipment. This is considered tobe necessary to reasonably protect against the spread or persistence of the GMO outside therelease site, including at times well in the future.

The destruction of the GMO at the release site and the cleaning of equipment used inconnection with the release are considered to be one of the more fundamentally importantrisk management conditions.

The requirement to provide the Regulator with written documentation about the procedures inplace to clean sites and locations is considered necessary in the context of the Regulator’soverall monitoring of the licence.

Condition 7 describes procedures that must be followed to promote germination of the GMOafter harvest and cleaning. The purpose of these procedures is to reduce the seed bank of theGMO by measures that will promote the germination of oilseed poppy seed and reduce theproportion of seed that will enter secondary dormancy. Burial of seed increases thelikelihood of secondary dormancy, and requiring shallow tillage will reduce the risk of thisoccurring. These conditions are considered necessary to manage risks associated withpersistence of the GMO in the environment, including at times well in the future. Therequirement that the licence holder provide a written report on tiling activities to theRegulator is considered necessary in the context of the Regulator’s overall monitoring of thelicence.

Condition 8 describes the procedures for post-harvest monitoring. These conditions areconsidered necessary to limit the spread and persistence of the GMO in the environment afterthe release.

It is considered that post-trial monitoring and the destruction of volunteer GMOs post-harvestis one of the primary, fundamental risk management practices required in the licence.

The places required to be monitored are those where it is considered reasonably possible thatthe GMO might persist. The period of time over which post-trial monitoring must take placeis considered to be reasonable to manage the identified risk and takes account of advice fromthe GTTAC. It is also consistent with current scientific literature about the way in whichoilseed poppy continues to persist in the soil post-harvest.

The requirement that monitoring activities be reported to the Regulator is included for thesame reasons that other evidential documents are required to be produced in the course ofconducting dealings pursuant to the licence. The requirement to provide a report, as opposed



to merely requiring procedures to be documented, is considered to be reasonable havingregard to the high importance of this condition within the context of the overall riskmanagement strategy.

Condition 9 places restrictions on the use of the release site after harvest of the GMO. Theserestrictions are considered to be necessarily incidental to the requirements to perform post-trial monitoring, in order to reasonably ensure that post-harvest monitoring and destruction ofvolunteers is carried out effectively and effectively, thus preventing spread and persistence ofthe GMO outside the release site and the isolation zone. For example, planting of Papaversomniferum subsp somniferum would effectively frustrate any post-harvest monitoring anddestruction of volunteers. Accordingly, it is prohibited.

Condition 10 describes conditions for transport of the GMOs or GM material from theGMOs. These conditions are considered necessary to reasonably protect against any escapeand the spread or persistence of the GMO outside the release site or isolation zone that mightarise in the course of transportation of the GMO.

Conditions 11 recognises that notwithstanding that a rigourous risk assessment and a riskmanagement plan has been prepared, unforseen circumstances, as well as other situations,such as failures to comply with licence conditions, may result in the persistence of the GMOoutside the release site. It is considered necessary in these circumstances, in order to managerisks associated with the possible dissemination of the GMO, for written contingency plans tobe in place and ready to be implemented.

Conditions 12 and 13 require the licence holder to provide a compliance management planand annual reports on operations to the Regulator. This is so that the Regulator is aware ofthe procedures that the licence holder has in place to ensure and document compliance withthe licence conditions, as well as the actual performance of the licence holder. Theserequirements are considered necessary in the context of the Regulator’s overall monitoring ofthe licence.

Condition 14 requires the licence holder to provide the Regulator with a method that willenable the presence of the GM oilseed poppy or its genetic material to be detected withaccuracy. These requirements are considered necessary in the context of the Regulator’soverall monitoring of the licence. It is also a necessary for the licence holder to be able tocomply with other conditions in the licence that the licence holder be able to detect thepresence of the GMO with accuracy.

Conditions 16 to 18 require notification of the probable and actual dates of planting,flowering and harvesting. The Regulator requires these forecasts and actual dates in order toefficiently and effectively plan monitoring activities in connection with these times, whichare the times when dissemination, outcrossing and persistence of the GMO in theenvironment are most likely to occur. These requirements are considered necessary in thecontext of the Regulator’s overall monitoring of the licence.

Condition 19 requires that neither the GMO nor any GM products must be used for humanfood or animal feed or therapeutics or for commercial purpose to minimise the risk of toxicityand allergenicity. This is considered necessary in order to manage potential risks associatedwith possible toxicity and allergenicity in connection with the GMO.

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