Table of Contents - OGTR | Office of the Gene … · Web viewVips are secreted during vegetative growth stages and sporulation, whereas the Cry proteins are expressed by Bt only during

Risk Assessment andRisk Management Plan for

DIR 133

Limited and controlled release of cotton genetically modified for insect resistance and herbicide tolerance

Applicant: Bayer CropScience Pty Ltd

May 2015

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DIR 133 – Risk Assessment and Risk Management Plan (May 2015) Office of the Gene Technology Regulator

Summary of the Risk Assessment and Risk Management Plan

for

Licence Application No. DIR 133DecisionThe Gene Technology Regulator (the Regulator) has decided to issue a licence for this application for a limited and controlled release of a genetically modified organism (GMO) into the environment. A Risk Assessment and Risk Management Plan (RARMP) for this application was prepared by the Regulator in accordance with the requirements of the Gene Technology Act 2000 (the Act) and corresponding state and territory legislation, and finalised following consultation with a wide range of experts, agencies and authorities, and the public. The RARMP concludes that this field trial poses negligible risks to human health and safety and the environment and that any risks posed by the dealings can be managed by imposing conditions on the release.

The applicationApplication number DIR 133

Applicant Bayer CropScience Pty Ltd

Project title Limited and controlled release of cotton genetically modified for insect resistance and herbicide tolerance

Parent organism Cotton (Gossypium hirsutum L.)

Introduced genes and modified traits

Three insect resistance genes Cry1Ab gene from Bacillus thuringiensis (Bt) Cry2Ae gene from Bacillus thuringiensis (Bt) Vip3Aa19 gene from Bacillus thuringiensis (Bt)

1. Two herbicide tolerance genes bar gene from Streptomyces hygroscopicus for glufosinate

ammonium tolerance 2mepsps gene from Zea mays (maize) for glyphosate toleranceOne selectable marker gene aph4 from Escherichia coli for resistance to antibiotic hygromycin B

Proposed locations Sites are to be selected from 56 possible local government areas in New South Wales, Queensland and Western Australia.

Proposed release size† Up to 12 sites per year for the first two years, at a maximum area of 10 hectares (ha) per site, and up to 20 sites per year for the remaining four years, at a maximum area of 30 ha per site.

Proposed release dates July 2015 – July 2021

Primary purpose To assess the agronomic characteristics of the genetically modified (GM) cotton grown under field conditions.To conduct trait development trials, breeding trials and seed production for future releases (subject to further approvals).

† Proposed release size was reduced after release of the consultation RARMP.

Summary III


Bayer proposes to conduct a field trial of GM cotton lines‡ containing introduced insect resistance and/or herbicide tolerance genes, and of GM cotton stacks produced by conventional breeding between the GM cotton lines. The proposed release is a continuation of the field trial authorised by licence DIR 113.

Risk assessmentThe risk assessment concludes that there are negligible risks to the health and safety of people, or the environment, from the proposed release.

The risk assessment process considered how the genetic modification and proposed activities conducted with the GMOs might lead to harm to people or the environment. Risks were characterised in relation to both the seriousness and likelihood of harm, taking into account current scientific/technical knowledge, information in the application (including proposed limits and controls), relevant previous approvals and advice received from a wide range of experts, agencies and authorities consulted on the RARMP. Both the short and long term impact were considered.

Credible pathways to potential harm that were considered included exposure to the GM plant material, dispersal of GM seed leading to spread and persistence of the GMOs, and transfer of the introduced genetic material to sexually compatible cotton plants. Potential harms associated with these pathways included toxicity or allergenicity to people, toxicity to other desirable organisms, and environmental harms due to weediness.

The principal reasons for the conclusion of negligible risks are that the proposed limits and controls effectively contain the GMOs and their genetic material and minimise exposure; the introduced genetic modifications are unlikely to cause harm to people or the environment; and the introduced genes are common in the environment.

Risk management planThe risk management plan describes measures to protect the health and safety of people and to protect the environment by controlling or mitigating risk. The risk management plan is given effect through licence conditions.

As the level of risk is considered negligible, specific risk treatment is not required. However, since this is a limited and controlled release, the licence includes limits on the size, locations and duration of the release, as well as controls to prohibit the use of GM plant material in human food or animal feed, to minimise dispersal of GM seed or GM pollen from trial sites, to transport GMOs in accordance with the Regulator’s guidelines, to destroy GMOs not required for testing or further planting, and to conduct post-harvest monitoring at trial sites to ensure all GMOs are destroyed.

‡ The term ‘line’ is used to denote plants derived from a single plant containing a specific genetic modification resulting from a single transformation event.

Summary IV


Table of ContentsSUMMARY OF THE RISK ASSESSMENT AND RISK MANAGEMENT PLAN..................................................III

DECISION ................................................................................................................................................................IIIRISK ASSESSMENT.......................................................................................................................................................IVRISK MANAGEMENT PLAN............................................................................................................................................IV

TABLE OF CONTENTS...................................................................................................................................................V

ABBREVIATIONS...........................................................................................................................................................VI

CHAPTER 1 RISK ASSESSMENT CONTEXT...........................................................................................................1

SECTION 1 BACKGROUND..........................................................................................................................................1SECTION 2 REGULATORY FRAMEWORK....................................................................................................................1SECTION 3 THE PROPOSED DEALINGS......................................................................................................................2

3.1 The proposed limits of the dealings (duration, size, locations and people)..................................33.2 The proposed controls to restrict the spread and persistence of the GMOs in the environment

..................................................................................................................................................................3SECTION 4 THE PARENT ORGANISM..........................................................................................................................4SECTION 5 THE GMOS, NATURE AND EFFECT OF THE GENETIC MODIFICATION....................................................4

5.1 Introduction to the GMOs......................................................................................................................45.2 The introduced genes, encoded proteins and their associated effects.........................................55.3 Toxicity/allergenicity of the proteins associated with the introduced genes.................................75.4 Characterisation of the GMOs.............................................................................................................8

SECTION 6 THE RECEIVING ENVIRONMENT...............................................................................................................86.1 Relevant abiotic factors.........................................................................................................................86.2 Relevant biotic factors...........................................................................................................................96.3 Relevant agricultural practices.............................................................................................................96.4 Presence of related plants in the receiving environment.................................................................96.5 Presence of similar genes and encoded proteins in the environment.........................................10

SECTION 7 RELEVANT AUSTRALIAN AND INTERNATIONAL APPROVALS...............................................................107.1 Australian approvals............................................................................................................................107.2 International approvals........................................................................................................................11

CHAPTER 2 RISK ASSESSMENT.............................................................................................................................12

SECTION 1 INTRODUCTION.......................................................................................................................................12SECTION 2 RISK IDENTIFICATION.............................................................................................................................13

2.1 Risk source............................................................................................................................ 132.2 Causal pathway......................................................................................................................142.3 Potential harm........................................................................................................................ 142.4 Postulated risk scenarios.......................................................................................................15

SECTION 3 UNCERTAINTY........................................................................................................................................22SECTION 4 RISK EVALUATION.................................................................................................................................22

CHAPTER 3 RISK MANAGEMENT PLAN................................................................................................................24

SECTION 1 BACKGROUND........................................................................................................................................24SECTION 2 RISK TREATMENT MEASURES FOR SUBSTANTIVE RISKS.....................................................................24SECTION 3 GENERAL RISK MANAGEMENT..............................................................................................................24

3.1 Licence conditions to limit and control the release.........................................................................243.2 Other risk management considerations...........................................................................................27

SECTION 4 ISSUES TO BE ADDRESSED FOR FUTURE RELEASES...........................................................................29SECTION 5 CONCLUSIONS OF THE RARMP...........................................................................................................29

REFERENCES.................................................................................................................................................................30

APPENDIX A.............SUMMARY OF SUBMISSIONS FROM PRESCRIBED EXPERTS, AGENCIES AND AUTHORITIES........................................................................................................................................34

Table of Contents


AbbreviationsAPVMA Australian Pesticides and Veterinary Medicines AuthorityBayer Bayer CropScience Pty LtdBt Bacillus thuringiensisCry CrystalDIR Dealings involving Intentional ReleaseDNA Deoxyribonucleic acidEPSPS 5-enolpyruvylshikimate-3-phosphate synthaseFSANZ Food Standards Australia New ZealandGM Genetically modifiedGMO Genetically modified organismha HectareHGT Horizontal gene transferHPT Hygromycin phosphotransferasekm Kilometresm MetresNLRD Notifiable Low Risk DealingNSW New South WalesOGTR Office of the Gene Technology RegulatorPAT Phosphinothricin acetyltransferasePC2 Physical Containment level 2RARMP Risk Assessment and Risk Management PlanRegulations Gene Technology Regulations 2001Regulator Gene Technology Regulatorthe Act The Gene Technology Act 2000Vip Vegetative insecticidal protein

Abbreviations VI

PROPOSED DEALINGSProposed activities involving the GMOProposed limits of the releaseProposed control measures

PARENT ORGANISMOrigin and taxonomyCultivation and useBiological characterisationEcology

PREVIOUS RELEASES

GMOIntroduced genes (genotype)Novel traits (phenotype)

RISK ASSESSMENT CONTEXT

LEGISLATIVE REQUIREMENTS(including Gene Technology Act and Regulations)

RISK ANALYSIS FRAMEWORK

OGTR OPERATIONAL POLICIES AND GUIDELINES

RECEIVING ENVIRONMENTEnvironmental conditionsAgronomic practicesPresence of related speciesPresence of similar genes


Chapter 1 Risk assessment contextSection 1 Background1. An application has been made under the Gene Technology Act 2000 (the Act) for Dealings involving the Intentional Release (DIR) of genetically modified organisms (GMOs) into the Australian environment.

2. The Act in conjunction with the Gene Technology Regulations 2001 (the Regulations), an inter-governmental agreement and corresponding legislation that is being enacted in each State and Territory, comprise Australia’s national regulatory system for gene technology. Its objective is to protect the health and safety of people, and to protect the environment, by identifying risks posed by or as a result of gene technology, and by managing those risks through regulating certain dealings with genetically modified organisms (GMOs).

3. This chapter describes the context within which potential risks to the health and safety of people or the environment posed by the proposed release are assessed. The risk assessment context is established within the regulatory framework and considers application-specific parameters (Figure 1).

Figure 1. Summary of parameters used to establish the risk assessment context

Section 2 Regulatory framework4. Sections 50, 50A and 51 of the Act outline the matters that the Gene Technology Regulator (the Regulator) must take into account, and the consultation required when preparing the Risk Assessment and Risk Management Plans (RARMPs) that inform the decisions on licence applications. In addition, the Regulations outline further matters the Regulator must consider when preparing a RARMP. In accordance with section 50A of the Act, this application is considered to be a limited and controlled release application, as its principal purpose is to enable the applicant to conduct experiments and the applicant has proposed limits on the size, locations and duration of the release, as well as controls to restrict the spread and persistence of the GMOs and their genetic material in the environment. Therefore, the

Chapter 1 – Risk assessment context 1


Regulator was not required to consult with prescribed experts, agencies and authorities before preparation of the Risk Assessment and Risk Management Plan.

5. Section 52 of the Act requires the Regulator to seek comment on the RARMP from the States and Territories, the Gene Technology Technical Advisory Committee, Commonwealth authorities or agencies prescribed in the Regulations, the Minister for the Environment, relevant local council(s), and the public. The advice from the prescribed experts, agencies and authorities and how it was taken into account is summarised in Appendix A. No public submissions were received.

6. The Risk Analysis Framework (OGTR 2013a) explains the Regulator’s approach to the preparation of RARMPs in accordance with the legislative requirements. Additionally, there are a number of operational policies and guidelines developed by the Office of the Gene Technology Regulator (OGTR) that are relevant to DIR licences. These documents are available from the OGTR website.

7. Any dealings conducted under a licence issued by the Regulator may also be subject to regulation by other Australian government agencies that regulate GMOs or GM products, including Food Standards Australia New Zealand (FSANZ), the Australian Pesticides and Veterinary Medicines Authority (APVMA), the Therapeutic Goods Administration, the National Industrial Chemicals Notification and Assessment Scheme and the Department of Agriculture. These dealings may also be subject to the operation of State legislation declaring areas to be GM, GM free, or both, for marketing purposes.

Section 3 The proposed dealings8. Bayer CropScience Pty Ltd proposes to release genetically modified (GM) cotton lines and crosses between these lines into the environment under limited and controlled conditions.

9. The purpose of the release is to assess the GM cotton grown under Australian field conditions. Trait development trials would generate data on agronomic performance, herbicide tolerance and insect resistance for product development and future regulatory submissions. Breeding trials would evaluate the yield and disease tolerance of different GMO genetic backgrounds for product development. Some sites would also be used for seed production in preparation for commercial release of the GMOs (subject to further approvals).

10. The dealings involved in the proposed intentional release include:

conducting experiments with the GMOs

breeding the GMOs

propagating the GMOs

using the GMOs in the course of manufacture of a thing that is not a GMO

growing the GMOs

importing the GMOs

transporting the GMOs

disposing of the GMOs

possession, supply or use of the GMOs for any of the purposes above.

11. These dealings are detailed further below.

12. The applicant also proposes that GMOs may be exported for experimentation. This will not be considered in the RARMP because the Gene Technology Regulator does not regulate



export of GMOs. Instead, the applicant would apply for an authorisation to import the GMOs from appropriate regulatory agencies in the destination country.

3.1 The proposed limits of the dealings (duration, size, locations and people)13. The applicant proposes to grow GM cotton plants over six years from July 2015 to July 2021.

14. In the first two years, the proposed trial would include up to 12 sites per year with a maximum area of 10 hectares (ha) per site. In the third to sixth years, the trial would include up to 20 sites per year with a maximum area of 30 ha per site. The maximum cumulative planting area would be 2640 ha over the period of the trial.

15. Sites for the proposed trial may be located in any of 56 local government areas (listed in Table 1) in current commercial cotton growing regions of NSW, Queensland and Western Australia.

Table 1 Local government areas proposed for field trials of the GMOsNew South Wales Queensland Western Australia

Balranald Jerilderie Balonne Wyndham-East KimberleyBerrigan Lachlan Banana BroomeBland Leeton Bundaberg RegionalBogan Liverpool Plains BurdekinBourke Moree Plains Central HighlandsBrewarrina Murray Goondiwindi RegionalCarrathool Murrumbidgee Isaac RegionalCentral Darling Narrabri Lockyer Valley RegionalCoolamon Narrandera Maranoa RegionalCoonamble Narromine ParooConargo Parkes Rockhampton RegionalDeniliquin Mildura South Burnett RegionalForbes Urana Southern Downs RegionalGilgandra Walgett Toowoomba RegionalGriffith Wagga Wagga Western Downs RegionalGunnedah Warren Whitsunday RegionalGwydir WarrumbungleHay WeddinInverell Young

16. Only authorised employees or contractors who are trained and/or experienced would be permitted to deal with the GM cotton.

3.2 The proposed controls to restrict the spread and persistence of the GMOs in the environment17. The applicant has proposed a number of controls to restrict the spread and persistence of the GM cotton and the introduced genetic material in the environment. These include:

locating trial sites at least 50 m away from natural waterways

separating trial sites from other cotton crops either by a 20 m pollen trap of non-GM or commercially released GM cotton, or by a 100 m monitoring zone and 3 km exclusion zone

treating any non-GM or commercially released GM cotton planted at the site or in the pollen trap as if it were the GM cotton being trialled

cleaning all equipment used with the GM cotton before using it for any other purpose

ginning the GM cotton separately from any other cotton crop

cultivating trial sites after harvest to encourage germination or decomposition of



remaining seed

post-harvest monitoring and destruction of any volunteer cotton at trial sites for at least 12 months and until sites have been free of volunteers for at least six months

destroying all GMOs from the trial that are not required for testing or future planting

transporting and storing the GMOs in accordance with the Regulator’s Guidelines for the Transport, Storage and Disposal of GMOs

not using GM plant material or products for human food or animal feed.

Section 4 The parent organism18. The parent organism is upland cotton (Gossypium hirsutum L.), which is the most commonly cultivated cotton species worldwide. Cotton is exotic to Australia and is grown as an agricultural crop in New South Wales and Queensland, with occasional trial or small-scale cultivation in Victoria, northern Western Australia and in the Northern Territory. In the 2013-14 growing season more than 99% of the Australian cotton crop was GM, and more than 95% of the national cotton crop contained stacked insect resistance and herbicide tolerance GM traits (Roth 2014).

19. Cotton is grown as a source of textile and industrial fibre, cottonseed oil for food use, and cottonseed meal for animal feed. Detailed information about the parent organism is contained in a reference document, The Biology of Gossypium hirsutum L. and Gossypium barbadense L. (cotton), which was produced to inform the risk assessment process for licence applications involving GM cotton plants (OGTR 2013b). The document is available from the OGTR website or on request from the OGTR.

Section 5 The GMOs, nature and effect of the genetic modification

5.1 Introduction to the GMOs20. The applicant proposes to release five cotton lines that are genetically modified for insect resistance and/or herbicide tolerance, along with stacks, which are crosses between any two or more of these lines. An overview of the genetic modification events is provided in Table 2, more detail on the introduced genes is provided in Table 3, and more detail on the introduced regulatory elements is provided in Table 4.

Table 2. Genetic elements of events proposed for release.

Event Promoter(s) 5’ leader (s) Transit peptide (s) Gene(s) Terminator(s)

COT102 PAct2PUbq3

intron1 Act2intron1 Ubq3

vip3Aa19aph4

3’nos3’nos

GHB119 PcsvmvXYZP35S2 5’cab22L TPssuAt

barcry2Ae

3’nos3’35S

GHB614 Ph4a748At intron1 h3At TPotp C 2mepsps 3’histonAtLL25 P35S3 bar 3’nos

T304-40 P35S3Ps7s7 5’e1

barcry1Ab

3’nos3’me1


http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/home-1

http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/home-1


Table 3. Genes introduced into the GM cotton lines proposed for release.

Gene Protein produced Source Intended functionaph4 Hygromycin-B phosphotransferase Escherichia coli Tolerance of antibiotic hygromycin Bbar phosphinothricin acetyltransferase

(PAT)Streptomyces hygroscopicus

Tolerance of herbicide glufosinate ammonium

cry1Ab crystal protein 1Ab Bacillus thuringiensis Insecticidal activitycry2Ae crystal protein 2Ae Bacillus thuringiensis Insecticidal activity2mepsps 5-enolpyruvylshikimate-3-phosphate

synthase (double mutant)Zea mays Tolerance of herbicide glyphosate

vip3Aa19 vegetative insecticidal protein 3A Bacillus thuringiensis Insecticidal activity

Table 4. Regulatory sequences introduced into the GM cotton lines proposed for release.

Element Description Function Source5’cab22L Leader sequence of chlorophyll a/b binding

protein genePromotes expression Petunia hybrida

5’e1 Leader sequence of tapetum specific E1 gene

Promotes expression Oryza sativa

intron1 Act2 First intron of actin 2 gene Promotes expression Arabidopsis thalianaintron1 h3At First intron of histone H3 gene Promotes expression Arabidopsis thalianaintron1 Ubq3 First intron of ubiquitin 3 gene Promotes expression Arabidopsis thalianaPAct2 Actin 2 promoter Constitutive promoter Arabidopsis thalianaPcsvmvXYZ Cassava vein mosaic virus promoter Constitutive promoter Cassava vein mosaic virusPh4a748At Histone H4 gene promoter Constitutive promoter Arabidopsis thalianaPs7s7 Genome segment 7 promoter Constitutive promoter Subterranean clover stunt virusP35S2 Cauliflower mosaic virus 35S promoter Constitutive promoter Cauliflower mosaic virusP35S3 Cauliflower mosaic virus 35S promoter Constitutive promoter Cauliflower mosaic virusPUbq3 Ubiquitin 3 promoter Constitutive promoter Arabidopsis thaliana3’histonAt Histone H4 gene terminator Stop signal Arabidopsis thaliana3’me1 NADP-malic enzyme terminator Stop signal Flavaria bidentis3’nos Nopaline synthase terminator Stop signal Agrobacterium tumefaciens3’35S Cauliflower mosaic virus 35S terminator Stop signal Cauliflower mosaic virusTPotp C Optimised transit peptide from RuBisCO

geneTargets protein to plastids

Zea mays, Helianthus annuus

TPssuAt Transit peptide from RuBisCO gene Targets protein to plastids

Arabidopsis thaliana

21. All GM cotton lines were produced by Agrobacterium tumefaciens-mediated plant transformation. Information about this transformation method can be found in the document Methods of plant genetic modification available from the Risk Assessment References page on the OGTR website. Stacks between the GM cotton lines were produced by conventional cross-breeding.

5.2 The introduced genes, encoded proteins and their associated effects5.2.1 The introduced genes for insect resistance and their associated proteins

22. The bacterium Bacillus thuringiensis (Bt) produces a range of insecticidal proteins, including the crystal (Cry) proteins (also known as delta-endotoxins) and vegetative insecticidal proteins (Vips). Vips are secreted during vegetative growth stages and sporulation, whereas the Cry proteins are expressed by Bt only during sporulation and form crystalline inclusions in spores (reviewed by Estruch et al. 1997). A survey of gene distribution in Bt strains found that 45% of the isolates contained a combination of cry1A, cry2 and vip3 genes (Hernandez-Rodriguez et al. 2009).

23. Both Cry proteins and Vips become active when ingested and cleaved by proteases in the insect midgut. In susceptible species, the activated toxins bind to specific receptors on the brush border membrane of the midgut epithelium, leading to formation of membrane pores, cell lysis, and eventual insect death (Bravo et al. 2007; Yu et al. 1997). Cry1A, Cry2A and



Vip3A protein classes all bind to different specific binding sites on the epithelial membrane surface (Gouffon et al. 2011; Sena et al. 2009).

24. The cry1Ab gene in the GM cotton was isolated from the Bt subspecies berliner (Höfte et al. 1986) and the gene sequence has been modified for expression in plants. The amino acid sequence is identical to the native protein except that it is truncated at the C-terminal end and an alanine has been inserted at the N-terminal end. The truncated Cry1Ab contains the region responsible for insecticidal activity.

25. The cry2Ae gene in the GM cotton was isolated from Bt subspecies dakota and the gene sequence has been modified for expression in plants.

26. The vip3Aa gene in the GM cotton was isolated from Bt strain AB88 (Estruch et al. 1996) and the gene sequence was modified for expression in plants. The amino acid sequence is identical to the native protein except that a glutamine residue has replaced a lysine residue at position 284.5.2.2 The introduced genes for herbicide tolerance and their associated proteins

27. Glufosinate-ammonium is the active ingredient in a number of proprietary broad-spectrum herbicides that have been registered for use in Australia (Australian Pesticides and Veterinary Medicine Authority).These herbicides function in plants by inhibiting the enzyme glutamine synthase. Inhibition of this enzyme both prevents the synthesis of the amino acid glutamate and causes toxic accumulation of its precursor, ammonia, in plant tissues (Evstigneeva et al. 2003).

28. Glufosinate-ammonium is a synthetic analogue of an antimicrobial secondary metabolite called bialaphos that is produced naturally by the soil bacterium Streptomyces hygroscopicus. To avoid the toxicity associated with biaphalos production, S. hygroscopicus expresses a biaphalos resistance gene known as bar. The bar gene encodes phosphinothricin acetyltransferase (PAT), an enzyme that acetylates the free amino groups of glufosinate-ammonium and renders it inactive (Thompson et al. 1987). Thus, plants containing the bar gene are expected to tolerate glufosinate-ammonium herbicide. The gene sequence of the bar gene in the GM cotton lines was modified for expression in plants.

29. Glyphosate is the active ingredient in a number of broad-spectrum systemic herbicides that have been approved for use in Australia (Australian Pesticides and Veterinary Medicine Authority). The mode of action of glyphosate is to specifically inhibit the function of the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Glyphosate binding to EPSPS in plants blocks biosynthesis of essential aromatic compounds, including the amino acids phenylalanine, tyrosine and tryptophan (Dill 2005).

30. The 2mepsps gene in the GM cotton was developed from a maize (Zea mays) gene that encodes an EPSPS enzyme. Site-directed mutagenesis of the wild-type maize epsps gene resulted in two amino acid changes in the encoded protein (substitution of threonine by isoleucine at position 102 and substitution of proline by serine at position 106). These changes greatly reduce the affinity of the maize EPSPS enzyme for glyphosate (Lebrun et al. 1987). Thus, GM cotton lines containing the 2mepsps gene are expected to tolerate glyphosate.5.2.3 The introduced gene for antibiotic resistance and its associated protein

31. The aph4 gene in the GM cotton was isolated from the common gut bacterium Escherichia coli. The aph4 gene encodes a hygromycin phosphotransferase (HPT) enzyme which inactivates the antibiotic hygromycin B. This antibiotic resistance trait was used as a selectable marker during plant transformation. The gene aph4 (also known as hph or hpt) is commonly used as a marker gene in GM plants. Further information about this gene can be found in the document Marker genes in GM plants available from the Risk Assessment References page on the OGTR website.


http://apvma.gov.au/





5.3 Toxicity/allergenicity of the proteins associated with the introduced genes5.3.1 The introduced proteins for insect resistance

32. The Cry1Ab, Cry2Ae and Vip3A proteins for insect resistance are derived from Bacillus thuringensis (Bt). Bt is naturally found worldwide in soil, on plant surfaces and in animals, and microbial preparations of Bt have been used as a commercial pesticide for over 60 years (OECD 2007). Thus, people and other organisms have a long history of safe exposure to Bt insecticidal proteins.

33. Cry and Vip proteins are toxic to susceptible insects through a mechanism of binding to specific receptors found on the midgut epithelium (Bravo et al. 2007; Lee et al. 2006). The proteins are not expected to be toxic to any organism lacking these specific receptors. The Cry2Ae and Vip3A proteins are only known to be toxic to lepidopteran insects. The Cry1Ab protein has confirmed toxicity to a range of lepidopteran insects and a hemipteran species. There are reports in the scientific literature that Cry1Ab is toxic to species in other insect orders, but the results were equivocal and/or not reproducible (van Frankenhuyzen 2013).

34. Acute oral toxicity studies of the purified Cry1Ab and Cry2Ae proteins reported no adverse effects in mice. Both proteins showed minimal oral toxicity to five representative non-target arthropods. Both proteins were degraded in simulated gastric or intestinal fluids. Neither protein had amino acid sequence similarity to known allergens (Bushey et al. 2008).

35. Mice and ten other representative non-target organisms showed no adverse effects following oral administration of high levels of purified Vip3A protein (Raybould & Vlachos 2011). The Vip3A protein had no sequence similarity to known protein allergens, and was degraded in simulated gastric fluid (Hill et al. 2003).

36. The effects of GM cotton expressing the Vip3A protein on arthropods were studied in Australian field trials. No major differences in species richness or diversity of beneficial and non-target arthropods were found in comparison to non-GM cotton (Whitehouse et al. 2007). Field measurements of insect abundance in GM corn crops expressing stacked Vip3A and Cry1Ab proteins showed no significant difference in overall biodiversity compared to non-GM crops without insecticide treatment. There were changes in density of some non-target taxa, for example due to reduced lepidopteran prey abundance, but these did not carry over to the subsequent growing season (Dively 2005).

37. Food derived from cotton varieties containing the Cry1Ab, Cry2Ae and Vip3A proteins have been approved by FSANZ as safe for human consumption (under applications A509, A615, A1028 and A1040, FSANZ website).5.3.2 The introduced proteins for herbicide tolerance

38. Purified PAT protein was not toxic to mice when administered intravenously at high doses. No sequence homology was found between PAT and any known toxic or allergenic proteins. PAT is rapidly degraded in simulated gastric or intestinal fluid (Herouet et al. 2005). FSANZ has approved food derived from GM cotton varieties expressing PAT protein as safe for human consumption (under applications A533, A1028 and A1040, FSANZ website). The Regulator has also previously approved the commercial release of GM cotton lines expressing the PAT protein (DIR 062/2005 and DIR 091).

39. The 2mepsps gene is a variant of the native maize epsps gene, and the modified 2mEPSPS protein is 99.5% identical to the native maize protein. Maize has been safely consumed by humans and other animals for centuries. The 2mEPSPS protein has no sequence similarity to known toxins or allergens, is rapidly degraded in simulated gastric or intestinal fluids and had no detrimental effect on mice when purified protein was administered orally or intravenously (Herouet-Guicheney et al. 2009). FSANZ has approved food derived from GM


http://www.foodstandards.gov.au/code/applications/pages/default.aspx



cotton varieties expressing 2mEPSPS as safe for human consumption (under application A614, FSANZ website). 5.3.3 The introduced protein for antibiotic resistance

40. HPT protein purified from E. coli had no adverse effects in an acute oral toxicity study on mice (Zhuo et al. 2009). The protein has no sequence similarity to known allergens and is rapidly digested by simulated gastric fluids (Lu et al. 2007). FSANZ has approved food derived from a GM cotton variety expressing the HPT protein as safe for human consumption (under application A509, FSANZ website).

5.4 Characterisation of the GMOs5.4.1 Stability and molecular characterisation

41. Southern blot analyses were used to determine the copy number in each parental GM cotton line. For events COT102, GHB119, GHB614 and LL25, the GM cotton lines contain one intact copy of the intended introduced DNA. For event T304-40, the GM cotton line contains one nearly intact copy of the intended introduced DNA (the 3’nos and 3’me1 terminators are both incomplete) and one adjacent partial copy containing a partial Ps7s7 promoter, the 5’e1 leader sequence, the cry1Ab gene and the 3’me1 terminator.

42. In all parental GM cotton lines, the insertions were found to be stably located within the genome over multiple generations. Standard Mendelian inheritance of the introduced genes was observed (information supplied by applicant).5.4.2 Phenotypic characterisation

43. Some of the introduced genes in the GM cotton lines provide resistance against herbivory by target insect species. These GM cotton plants therefore sustain less pest damage than non-GM plants that have not been treated with insecticide. Some of the introduced genes in the GM cotton lines provide post-emergence tolerance to glyphosate or glufosinate-ammonium herbicides. GM cotton plants that were grown in the greenhouse, or in preliminary field trials under licence DIR 113, had no other reported changes to normal cotton phenotype. Limited phenotypic data has been collected in field trials to date, but further data would be collected during the proposed release.

Section 6 The receiving environment44. The receiving environment includes: any relevant biotic/abiotic properties of the geographic regions where the field trial would occur; intended agricultural practices, including those that may be altered in relation to normal practices; other relevant GMOs already released; and any particularly vulnerable or susceptible entities that may be specifically affected by the proposed release (OGTR 2013a).

6.1 Relevant abiotic factors45. The abiotic factors relevant to the growth and distribution of commercial cotton in Australia are discussed in The Biology of Gossypium hirsutum L. and Gossypium barbadense L. (cotton) (OGTR 2013b). To summarise, factors restricting where cotton can be grown in Australia are water availability (through rainfall or irrigation), soil suitability and, most importantly, temperature. Cotton seedlings may be killed by frost, growth and development of cotton plants below 12°C is minimal, and a long, hot growing season is crucial for achieving good yields.

46. The local government areas where the release is proposed to take place (see Table 1, Section 3.1) have a broad range of climate types, as defined by the Köppen Classification system used by the Australian Bureau of Meteorology. Proposed planting areas include


http://www.bom.gov.au/




subtropical, grassland and temperate climate types, with areas of tropical climate in northern Queensland and northern Western Australia. The proposed cotton growing areas in Western Australia and coastal Queensland may experience the effects of tropical cyclones between November and April. There is a possibility of extreme weather events and flooding moving into inland Queensland and northern NSW (Bureau of Meteorology). The applicant has stated that they will select trial sites that are not prone to flooding.

6.2 Relevant biotic factors47. The biotic factors relevant to the cultivation of cotton in Australia are discussed in The Biology of Gossypium hirsutum L. and Gossypium barbadense L. (cotton) (OGTR 2013b). To summarise, cotton is very susceptible to competition from weeds; the most important pests of cotton in southern Australia are the caterpillars of Helicoverpa armigera and Helicoverpa punctigera and the spider mite Tetranychus urticae; and a number of fungal pathogens are significant diseases of cotton in Australia. These issues are managed by various strategies in commercial cotton cultivation.

6.3 Relevant agricultural practices48. GM cotton is proposed to be grown during the standard summer cotton season, and would also be grown during the winter in Western Australian trial sites. Both GM cotton lines and non-GM comparator cotton lines would be grown at the field trial sites. Small trial sites would be planted using specialised small plot seeders and harvested by hand. Large trial sites would be planted and harvested with commercial equipment. Most trial sites would be irrigated, using similar irrigation practices to commercial cotton crops.

49. GM cotton lines containing bar and/or 2mepsps genes would be treated with glyphosate and/or glufosinate ammonium herbicides to determine their levels of herbicide tolerance. GM cotton would also be treated with other herbicides for volunteer control.

50. GM cotton lines containing cry1Ab, cry2Ae and/or vip3A genes would be exposed to larvae of target pests to determine their levels of insect resistance. Introduced infestations of Helicoverpa armigera (cotton bollworm) and/or Helicoverpa punctigera (native budworm) would be contained inside mini-cages and destroyed prior to harvest.

51. The applicant proposes to sell cotton lint from the GMOs grown at some trial sites.

52. Proposed controls to restrict the spread and persistence of the GMOs and their genetic material are listed in Section 3.2 of this chapter.

6.4 Presence of related plants in the receiving environment53. Cotton is grown as a large-scale commercial crop in the majority of the Queensland and NSW local government areas where the release is proposed to take place (Cotton Australia). Cotton is grown as an experimental or small-scale crop in proposed Western Australia and northern Queensland growing areas. Commercial cotton grown in Australia is either Gossypium hirsutum or Gossypium barbadense, with 99% of cotton planted in 2006 being G. hirsutum (OGTR 2013b). The GM G. hirsutum proposed for release is capable of crossing with both species of commercially grown cotton.

54. Ephemeral populations of cotton volunteers can be found on cotton farms, by roadsides where cotton seed is transported, or in areas where cotton seed is used as livestock feed (Addison et al. 2007; Eastick & Hearnden 2006). Small populations of naturalised cotton exist in northern Australia (OGTR 2013b). Most reports of naturalised G. hirsutum populations are from tropical areas of the Northern Territory, while reports of naturalised G. barbadense are mainly from coastal Queensland (data from Australian Virtual Herbarium).


http://www.anbg.gov.au/avh/

http://cottonaustralia.com.au/australian-cotton/basics/where-is-it-grown


55. There are 17 native species of Gossypium in Australia, and the distribution of some of these species overlaps the areas where the GM cotton release is proposed to take place. However, well established genetic incompatibility prevents crossing of native cotton species with cultivated cotton in the natural environment (discussed in OGTR 2013b).

6.5 Presence of similar genes and encoded proteins in the environment56. The cry1Ab, cry2Ae and vip3A genes were isolated from Bacillus thuringiensis (Bt). Bt is a natural soil bacterium in Australia. Also, microbial preparations of Bt are used as insecticide sprays in Australia, particularly in organic agriculture and domestic gardening (Australian Pesticides and Veterinary Medicine Authority). Therefore, these genes and their encoded proteins are widespread in the Australian environment.

57. The bar gene was isolated from Streptomyces hygroscopicus, which is a natural soil bacterium in Australia. The bar gene is also present in Liberty Link® GM cotton, which has been commercially planted in Australia since 2008, as up to 2% of Australia’s cotton crop (information reported to the OGTR).

58. The 2mepsps gene was isolated from Zea mays (maize or corn), which is widely grown as a food crop in Australia. The DNA sequence of the maize EPSPS-encoding gene was modified to generate the 2mepsps gene, however, the encoded protein has only two amino acids that differ from the native maize protein.

59. The aph4 gene was isolated from the common bacterium E. coli, which is part of the normal flora of human and animal guts. Therefore, these genes and their encoded proteins are widespread in the Australian environment.

Section 7 Relevant Australian and international approvals

7.1 Australian approvals7.1.1 Approvals by the Regulator

60. All GM cotton lines included in this application have been previously approved by the Regulator for release in Australia. The relevant authorisations are shown in Table 5.

Table 5. Previous releases of the GM cotton lines in Australia.Event Gene/s Field trial licences Commercial licences

COT102 vip3A, aph4DIR 017/2002, DIR 025/2002, DIR 034/2003, DIR 036/2003, DIR 058/2005, DIR 065/2006, DIR 073/2007, DIR 101, DIR 113, DIR 120

DIR 124(not yet commercially planted)

GHB119 bar, cry2Ae DIR 087, DIR 113GHB614 2mepsps DIR 113

LL25 bar DIR 015/2002, DIR 036/2003, DIR 038/2003, DIR 056/2004, DIR 113

DIR 062/2005(commercially planted from 2008)

T304-40 bar, cry1Ab DIR 087, DIR 113

61. All stacks between GM cotton lines included in this application have been previously approved by the Regulator for field trials under licence DIR 113.

62. Bollgard® II GM cotton, which contains two cry insect resistance genes similar to the cry genes in events GHB119 and T304-40, and Roundup Ready Flex® GM cotton, which contains a glyphosate tolerance epsps gene similar to the epsps gene in event GHB614, are approved for commercial release under licence DIR 066/2006. In 2013-14, over 96% of the Australian cotton crop contained Bollgard® II technology and 99% of the Australian cotton crop contained Roundup Ready Flex® technology (Roth 2014).





63. Information on these licences is available from the GMO Record on the OGTR website. There have been no reports of adverse effects on human health or the environment resulting from any of these releases.7.1.2 Approvals by other government agencies

64. FSANZ is responsible for human food safety assessment and food labelling, including GM food. FSANZ has approved the use of food derived from each of the GM cotton lines included in this application. Assessments of events COT102 (application A509), GHB119 (application A1040), GHB614 (application A614), LL25 (application A533) and T304-40 (application A1028) are available from the FSANZ website. However, the applicant does not intend to use GM plant material generated in the proposed release in human food.

65. APVMA has regulatory responsibility for agricultural chemicals, including herbicides and insecticidal products, in Australia. Some of the GM cotton lines and stacks proposed for release meet the definition of an agricultural chemical product under the Agricultural and Veterinary Chemicals Code Act 1994, due to their production of insecticidal substances. The applicant holds a permit from APVMA (PER13358) to plant these GM cotton varieties in field trials.

7.2 International approvals66. All of the GM cotton lines proposed for release in this application have been approved for commercial cultivation in other countries, as shown in Table 6.

Table 6. International approvals for commercial cultivation of the GM cotton linesEvent Gene/s Country Year

COT102 vip3A, aph4 United States 2011

GHB119 x T304-40 cry1Ab, cry2Ae, bar BrazilUnited States

20112012

GHB614 2mepsps United StatesBrazil

20092010

LL25 bar United StatesBrazil

20032008


http://www.foodstandards.gov.au/


Chapter 2 Risk assessmentSection 1 Introduction

67. The risk assessment identifies and characterises risks to the health and safety of people or to the environment from dealings with GMOs, posed by or as the result of gene technology (Figure 2). Risks are identified within the context established for the risk assessment (see chapter 1), taking into account current scientific and technical knowledge. A consideration of uncertainty, in particular knowledge gaps, occurs throughout the risk assessment process.

Figure 2 The risk assessment process

68. Initially, risk identification considers a wide range of circumstances whereby the GMO, or the introduced genetic material, could come into contact with people or the environment. Consideration of these circumstances leads to postulating plausible causal or exposure pathways that may give rise to harm for people or the environment from dealings with a GMO in the short and long term. These are called risk scenarios.

69. A number of risk identification techniques are used by the Regulator and staff of the OGTR, including checklists, brainstorming, reported international experience and consultation (OGTR 2013a). A weed risk assessment approach is used to identify traits that may contribute to risks from GM plants. In particular, novel traits that may increase the potential of the GMO to spread and persist in the environment or increase the level of potential harm compared with the parental plant(s) are used to postulate risk scenarios (Keese et al. 2014). Risk scenarios postulated in previous RARMPs prepared for licence applications of the same or similar GMOs are also considered.

70. Postulated risk scenarios are screened to identify those that are considered to have some reasonable chance of causing harm. Pathways that do not lead to harm, or could not plausibly occur, do not advance in the risk assessment process.

71. Substantive risks (i.e. those identified for further assessment) are characterised in terms of the potential seriousness of harm (Consequence assessment) and the likelihood of harm (Likelihood assessment). Risk evaluation then combines the Consequence and Likelihood assessments to

Chapter 2 – Risk assessment 12


estimate the level of risk and determine whether risk treatment measures are required. The potential for interactions between risks is also considered.

Section 2 Risk Identification72. Postulated risk scenarios are comprised of three components

i. The source of potential harm (risk source).

ii. A plausible causal linkage to potential harm (causal pathway).

iii. Potential harm to an object of value, people or the environment.

73. The risk context, including the following factors, is taken into account when postulating relevant risk scenarios:

the proposed dealings, which may be to conduct experiments, develop, produce, breed, propagate, grow, import, transport or dispose of the GMOs, use the GMOs in the course of manufacture of a thing that is not the GMO, and the possession, supply and use of the GMOs in the course of any of these dealings

the proposed limits including the extent and scale of the proposed dealings

the proposed controls to limit the spread and persistence of the GMO and

characteristics of the parent organism(s).

2.1 Risk source74. The sources of potential harms can be intended novel GM traits associated with one or more introduced genetic elements, or unintended effects/traits arising from the use of gene technology.

75. As discussed in Chapter 1, the GM cotton lines and stacks have been modified by the introduction of up to three insect resistance genes and up to two herbicide tolerance genes. These introduced genes are considered further as potential sources of risk.

76. In addition, some of the GM cotton lines and stacks contain the aph4 antibiotic resistance selectable marker gene (also known as hph or hpt). This gene and its product have already been extensively characterised and assessed as posing negligible risk to human or animal health or to the environment by the Regulator as well as by other regulatory agencies in Australia and overseas. Further information about this gene can be found in the document Marker genes in GM plants available from the Risk Assessment References page on the OGTR website. As this gene has not been found to pose a substantive risk to either people or the environment, its potential effects will not be further considered for this application.

77. The introduced genes are controlled by introduced regulatory sequences. The regulatory sequences are derived from plants, plant viruses and the bacterium Agrobacterium tumefaciens (see Table 4). Regulatory sequences are naturally present in plants, and the introduced elements are expected to operate in similar ways to endogenous elements. There is no evidence that regulatory sequences themselves have toxic or allergenic effects (EPA 1996). Although the viral and bacterial sequences are derived from plant pathogens, they only constitute small fractions of the genomes and cannot themselves cause disease. Hence, potential harms from the regulatory elements will not be considered further. However, the introduced regulatory sequences control gene expression and hence the distribution and concentration of the introduced proteins in the GM plants. The effects of protein levels, especially in relation to toxicity and allergenicity, will be considered below.

78. The genetic modifications have the potential to cause unintended effects in several ways including altered expression of endogenous genes by random insertion of introduced DNA in the genome, increased metabolic burden due to expression of the introduced proteins, novel traits arising out of interactions with non-target proteins and secondary effects arising from altered



substrate or product levels in biochemical pathways. However, these types of effects also occur spontaneously and in plants generated by conventional breeding. Accepted conventional breeding techniques such as hybridisation, mutagenesis and somaclonal variation can have a much larger impact on the plant genome than genetic engineering (Schnell et al. 2015). Plants generated by conventional breeding have a long history of safe use, and there are no documented cases where conventional breeding has resulted in the production of a novel toxin or allergen in a crop (Steiner et al. 2013). Therefore, unintended effects resulting from the process of genetic modification will not be considered further.

2.2 Causal pathway79. The following factors are taken into account when postulating plausible causal pathways to potential harm:

routes of exposure to the GMOs, the introduced gene(s) and gene product(s) potential effects of the introduced gene(s) and gene product(s) on the properties of the

organism potential exposure to the introduced gene(s) and gene product(s) from other sources in the

environment the environment at the site(s) of release agronomic management practices for the GMOs spread and persistence (invasiveness) of the GM plant, including establishment and

reproduction dispersal by natural means and by people tolerance to abiotic conditions (e.g. climate, soil and rainfall patterns) tolerance to biotic stressors (e.g. pest, pathogens and weeds) tolerance to cultivation management practices gene transfer to sexually compatible organisms gene transfer by horizontal gene transfer (HGT) unauthorised activities.

80. The potential for horizontal gene transfer (HGT) from GMOs to other organisms, and any possible adverse outcomes, have been reviewed in the literature (Keese 2008) and assessed in many previous RARMPs. HGT was most recently considered in the RARMP for DIR 108 (available from the GMO Record on the OGTR website). In previous assessments of HGT no substantive risk was identified, due to the rarity of these events and because the wild-type gene sequences are already present in the environment and available for transfer via demonstrated natural mechanisms. Therefore, HGT will not be further considered for this application.

81. The potential for unauthorised activities to lead to an adverse outcome has been considered in many previous RARMPs, most recently in the RARMP for DIR 117 (available from the GMO Record on the OGTR website). In previous assessments of unauthorised activities, no substantive risk was identified. The Act provides for substantial penalties for unauthorised dealings with GMOs or non-compliance with licence conditions, and also requires the Regulator to have regard to the suitability of an applicant to hold a licence prior to the issuing of the licence. These legislative provisions are considered sufficient to minimise risks from unauthorised activities. Therefore, unauthorised activities will not be considered further.

2.3 Potential harm82. Potential harms from GM plants include:

harm to the health of people or desirable organisms, including toxicity/allergenicity



reduced establishment or yield of desirable plants reduced products or services from the land use restricted movement of people, animals, vehicles, machinery and/or water reduced quality of the biotic environment (e.g. providing food or shelter for pests or

pathogens) or abiotic environment (e.g. negative effects on fire regimes, nutrient levels, soil salinity, soil stability or soil water table)

reduced biodiversity for nature conservation.

83. These harms are based on those used to assess risk from weeds (Keese et al. 2014; Standards Australia New Zealand & CRC for Australian Weed Management 2006). Judgements of what is considered harm depend on the management objectives of the land where the GM plant may spread and persist. A plant species may have different weed risk potential in different land uses such as dryland cropping or nature conservation.

2.4 Postulated risk scenarios84. Three risk scenarios were postulated and screened to identify substantive risk. These scenarios are summarised in Table 7 and more detail of these scenarios is provided later in this Section. Postulation of risk scenarios considers impacts of the GM cotton or its products on people undertaking the dealings, as well as impacts on people and the environment if the GM plants or genetic material were to spread and/or persist.

85. In the context of the activities proposed by the applicant and considering both the short and long term, none of the three risk scenarios gave rise to any substantive risks.Table 7. Summary of risk scenarios from dealings with GM cotton genetically modified for insect resistance and herbicide toleranceRisk scenario

Risk source Causal pathway Potential harm/s Substantive risk?

Reasons

1 GM cotton expressing introduced insect resistance or herbicide tolerance genes

Cultivation of GMOs at trial sites

Exposure of people who deal with the GMOs or of animals at the trial site to introduced proteins

Toxicity or allergenicity in people or toxicity to desirable organisms

No GM plant material will not be used in human food or animal feed.

The introduced proteins are already present and widespread in the Australian environment.

The introduced insect resistance proteins are only toxic to a limited range of insect species.

The insect resistant GM cotton has similar insecticidal activity to most existing commercial cotton in Australia.


Dispersal of GM seed outside trial limits

Spread and persistence of populations of GM plants containing the introduced proteins


Reduced establishment or yield of desirable plants

Reduced biodiversity

No The proposed controls minimise dispersal of GM cotton seeds.

Cotton has limited ability to survive outside agricultural settings and the introduced proteins are not expected to increase its weediness.

The GM cotton is susceptible to weed control measures used on commercial cotton volunteers.



Risk scenario

Risk source Causal pathway Potential harm/s Substantive risk?

Reasons


Pollen from GM plants fertilising other sexually compatible plants

Exposure to or dispersal of hybrid seeds containing the introduced proteins


Reduced establishment or yield of desirable plants


No Cotton has limited ability to outcross.

The proposed controls would minimise pollen flow to cotton plants outside the trial sites.

Hybrids between the GMOs and commercial GM cotton lines would have similar properties to the GMOs.

2.4.1 Risk scenario 1Table 8. Steps in Risk Scenario 1.

Risk source GM cotton expressing introduced insect resistance or herbicide tolerance genes

Causal pathway

Cultivation of GMOs at trial sites

Exposure of people who deal with the GMOs or of animals at the trial site to introduced proteins

Potential harm Toxicity or allergenicity in people or toxicity to desirable organisms

Risk source

86. The source of potential harm for this postulated risk scenario is GM cotton expressing introduced insect resistance or herbicide tolerance genes.

Causal pathway

87. Workers who cultivate, harvest, gin, transport, experiment or conduct other dealings with the GM cotton grown at trial sites would be exposed to cotton plant material. As the applicant proposes that only authorised staff deal with the GM cotton, other people are not expected to be exposed to the GM plants. Potential pathways of exposure to the introduced proteins are ingestion, inhalation or dermal contact. There is little potential for human ingestion of the introduced proteins, as the applicant proposes that no GM plant material would be used as food. GM plant material that could potentially be airborne and inhaled includes pollen or cotton dust produced during the harvesting or ginning processes. However, cotton pollen is heavy, sticky and not easily dispersed by wind (OGTR 2013b), and people who enter commercial cotton gins typically wear protective face masks (International Fibre Centre website). Workers could come into skin contact with the introduced proteins if they touch damaged plants where cell contents have been released.

88. The applicant proposes to sell lint (long fibres) from GM cotton. Processed cotton lint contains over 99% cellulose and does not contain detectable protein (OGTR 2013b). Therefore, people wearing cotton clothing or using other products made from GM cotton would not be exposed to the introduced proteins. Cotton lint is used for textile manufacture and does not enter human food. The applicant does not propose to sell cotton linters (very short fibres), which are sometimes used as additives in processed foods (OGTR 2013b).

89. Non-human organisms may be exposed directly to the introduced proteins through ingesting the GM plants, or exposed indirectly through the food chain, or exposed through contact with dead plant material (soil organisms). Livestock would not be expected to ingest the introduced proteins as the GM plant material is not to be used as animal feed. Wild mammals and birds generally avoid feeding on cotton plants, in particular finding the seed unpalatable because of its high gossypol content (OGTR 2013b). A range of invertebrates would be expected to ingest GM cotton plant material. The limited scale and duration of the proposed field trial would restrict the total number of invertebrates exposed to the introduced proteins.

Potential harm


http://www.ifc.net.au/


90. Toxicity is the adverse effect(s) of exposure to a dose of a substance as a result of direct cellular or tissue injury, or through the inhibition of normal physiological processes (Felsot 2000). Allergenicity is the potential of a substance to elicit an immunological reaction following its ingestion, dermal contact or inhalation, which may lead to tissue inflammation and organ dysfunction (Arts et al. 2006).

91. Non-GM cotton produces natural toxins for defence against herbivory including gossypol and cycloprenoid fatty acids (OGTR 2013b). The introduced proteins in the GM cotton have well understood modes of action that would not alter cotton metabolic pathways. Therefore, GM cotton is not expected to have increased levels of natural toxins.

92. The introduced herbicide tolerance genes were isolated from organisms that are widespread and prevalent in the Australian environment (Chapter 1, Section 6.5), so people and animals are regularly exposed to the proteins encoded by these genes. Available information does not suggest that the PAT or 2mEPSPS proteins are toxic or allergenic to people or toxic to other organisms (Chapter 1, Section 5.3.2).

93. The introduced insect resistance genes were isolated from Bt, which is widespread in the Australian environment (Chapter 1, Section 6.5). Available information does not suggest that the introduced Bt proteins are toxic or allergenic to people (Chapter 1, Section 5.3.1). Oral toxicity studies of the Cry1Ab, Cry2Ae and Vip3A proteins were conducted to test their effect on a range of non-target organisms. None of the proteins were toxic to representative vertebrates or to desirable invertebrates, including honeybees, earthworms, and predatory lacewing and ladybird insects, at doses well above the maximum expected environmental concentration (Bushey et al. 2008; CERA 2011; Raybould & Vlachos 2011).

94. Cry1Ab, Cry2Ae and Vip3A proteins are toxic to overlapping subsets of lepidopteran insect species, and Cry1Ab is also toxic to at least one hemipteran species (Chapter 1, Section 5.3.1). GM cotton expressing introduced insecticidal proteins would be expected to be toxic to the target lepidopteran pests, but could also be toxic to related non-target organisms, such as native Australian moth species, if these insects feed on cotton. In stacks containing two or three of the insect resistance proteins, it is possible that additive or synergistic effects could occur, potentially increasing the range of sensitive insects. Some uncertainty exists in this area due to data gaps. However, it is unlikely that the GM cotton stacks could be toxic to a wider range of insects than microbial Bt insecticidal products, which contain a similar or larger array of insect resistance proteins. The World Health Organisation’s International Programme on Chemical Safety evaluated the environmental safety of microbial Bt insecticides, and concluded that, because of the specificity of the mode of action of Bt toxins, Bt products are unlikely to pose any hazard to humans, other vertebrates, or the great majority of non-target invertebrates (International Programme on Chemical Safety 1999).

95. The commercial GM cotton Bollgard® II comprised over 96% of the Australian cotton crop in 2013-14 (Roth 2014). Bollgard® II contains the insecticidal Bt proteins Cry1Ac and Cry2Ab, which are homologous to the Cry1Ab and Cry2Ae proteins of this application. The Bollgard® II proteins are toxic to a range of lepidopteran insects, along with a few hemipteran and dipteran species (van Frankenhuyzen 2013). Therefore, the insect resistant GM cotton in this application has a similar range of insecticidal activity to most current commercial cotton, and is not expected to cause more harm to insects than existing cotton crops.

96. Conclusion: Risk scenario 1 is not identified as a substantive risk because GM plant material will not be used in human food or animal feed, the introduced proteins are already present and widespread in the Australian environment, the introduced insect resistance proteins are only toxic to a limited range of insect species, and the insect resistant GM cotton has similar insecticidal activity to most existing commercial cotton in Australia. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.



2.4.2 Risk Scenario 2Table 9. Steps in Risk Scenario 2.


Causal pathway

Dispersal of GM seed outside trial limits

Spread and persistence of populations of GM plants containing the introduced proteins

Potential harms

Toxicity or allergenicity in people or toxicity to desirable organismsor

Reduced establishment or yield of desirable plantsor


Risk source


Causal Pathway

98. The first step in the causal pathway for this risk scenario is dispersal of GM seed outside the trial limits. This could occur due to persistence of viable GM seeds at the trial sites after the intended duration of the trial, or through physical movement of GM seeds to areas outside the trial sites.

99. The applicant proposes a number of control measures to prevent persistence of GM seeds in the seed bank at the trial sites. These include destroying GMOs that remain in the trial sites after harvest, cultivating trial sites after harvest to encourage decomposition or germination of remaining seed, post-harvest monitoring of each trial site for at least twelve months and until the site has been clear of volunteers for six months, and destroying any volunteers found prior to flowering. It is not expected that expression of the introduced genes for insect resistance or herbicide tolerance would raise dormancy levels of cotton seeds or otherwise increase the ability of the GMOs to survive these standard control measures.

100. Cotton seeds are enclosed in large, heavy bolls that remain attached to the plant (OGTR 2013b), so they are not normally physically transported by wind or by runoff after rainfall or irrigation. The applicant proposes to select trial sites that are at least 50 m away from natural waterways and are not prone to flooding. Nonetheless, an extreme weather event such as a cyclone could physically disperse cotton seeds. In order to disperse viable seeds, the extreme weather event would need to occur in the period between seed sowing and germination (approximately one week) or in the period between maturation of the first bolls and harvest (approximately one month). As only a small proportion of proposed trial site locations are in northern Australian areas that could potentially be affected by cyclones, and there are narrow time windows when a cyclone could disperse viable seed, it is unlikely that GM cotton seeds would be dispersed by a cyclone.

101. Wild mammals and birds generally avoid feeding on cotton plants, in particular finding the seed unpalatable because of its high gossypol content (OGTR 2013b). Therefore, wild animals are unlikely to disperse GM cotton seeds from trial sites. GM cotton seeds would not be used as stock feed, so would not be dispersed by stock.

102. Dispersal of cotton seeds by authorised people entering the trial sites would be minimised by cleaning all equipment used with the GM cotton before using it for any other purpose. GM cotton would be ginned separately from any other cotton crop to avoid accidentally mixing GM cotton seed with other cotton seed, then dispersing the mixed cotton seed. The applicant proposes to contain GM seeds during transportation and storage in accordance with the Regulator’s Guidelines for the Transport, Storage and Disposal of GMOs, and in the unlikely event of a spill occurring, it would be managed by cleaning and monitoring the spill site.



103. The second step in the causal pathway for this risk scenario is establishment, spread and persistence of populations of GM cotton plants. However, the spread and persistence of non-GM cotton plants are limited by a number of biotic and abiotic factors, especially cold stress in southern Australia and water stress in non-irrigated environments throughout almost all of Australia. Feral cotton populations are sparse and ephemeral in all current cotton growing regions of Australia (OGTR 2013b). A study found that even when cotton was sown in cleared northern Australian sites with high water availability, the cotton plants did not establish stable naturalised populations (Eastick & Hearnden 2006). A few small populations of naturalised cotton are reported in northern Australia, but these are not derived from modern cultivars (OGTR 2013b), and may have a greater ability to survive outside agricultural settings than modern cotton cultivars.

104. It is not expected that the expression of the introduced herbicide tolerance genes would allow cotton to overcome the biotic and abiotic factors, such as cold and water stress, that limit spread and persistence. A herbicide tolerance gene does not provide any advantage to a plant except the ability to survive application of a specific herbicide in a managed environment. Expression of the introduced insect resistance genes could reduce herbivory of cotton plants by insects susceptible to the insecticidal proteins, primarily lepidopteran species. However, previous RARMPs (most recently DIR 124, available from the GMO Record on the OGTR website) have assessed whether lepidopteran insect herbivory is a significant factor limiting spread and persistence of cotton, and have concluded that it is not. Therefore, expression of the introduced insect resistance genes is not expected to increase the ability of GM cotton to spread or persist.

Potential harms

105. The potential harms from this risk scenario are toxicity or allergenicity in people or toxicity to desirable organisms, reduced establishment or yield of desirable plants, or reduced biodiversity.

106. As discussed in Risk Scenario 1, the introduced proteins in the GM cotton are not expected to be toxic or allergenic to people, or toxic to vertebrates, or toxic to desirable invertebrates.

107. The GM cotton could reduce the establishment or yield of desirable plants in agricultural settings if GM cotton volunteers grew in other crops. The introduced herbicide tolerance proteins could potentially reduce the effectiveness of weed management measures to control the cotton volunteers. However, glyphosate herbicide is not typically used to control cotton volunteers, as most commercial Australian cotton is genetically modified for glyphosate tolerance, and glufosinate-ammonium herbicide is not one of the three herbicides recommended as most effective for control of seedling cotton (Charles et al. 2013). Also, cultivation is an alternative method for cotton volunteer control. Therefore, GM cotton volunteers could be controlled by similar weed management measures as volunteers from commercial cotton.

108. The GM cotton could reduce the establishment or yield of desirable plants in the natural environment if the GM cotton spread and persisted as a weed in nature reserves, displacing native vegetation. However, as discussed above, cotton has limited potential to survive outside agricultural settings, and the introduced proteins are not expected to increase its ability to spread and persist.

109. The GM cotton could potentially reduce biodiversity through direct toxicity of the introduced insect resistance proteins to susceptible insects, and indirect effects on predators and parasites that depend on susceptible insects. However, studies of arthropod communities in Australian cotton crops expressing Cry1A, Cry2A and Vip3A proteins found only small differences in species richness or diversity between fields of GM crops or conventional crops (Whitehouse et al. 2014). The variance in composition of the invertebrate community was not statistically significant in half of the sites studied, and in the other half of the sites ranged from 4-20%, mostly due to reduced lepidopteran abundance. The effects of GM cotton on arthropod biodiversity would be of even less concern in cases where the GM cotton was part of a mixed plant community rather than cultivated as a monoculture.



110. Conclusion: Risk scenario 2 is not identified as a substantive risk because the proposed controls minimise dispersal of GM cotton seeds, cotton has limited ability to survive outside agricultural settings and the introduced proteins are not expected to increase its weediness, and the GM cotton is susceptible to weed control measures used on commercial cotton volunteers. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.2.4.3 Risk Scenario 3Table 10. Steps in Risk Scenario 3.


Causal pathway

Pollen from GM plants fertilising other sexually compatible plants

Exposure to or dispersal of hybrid seeds containing the introduced proteins

Potential harms

Toxicity or allergenicity in people or toxicity to desirable organismsor

Reduced establishment or yield of desirable plantsor


Risk source


Causal pathway

112. The first step in the causal pathway for this risk scenario is pollen from GM plants fertilising other sexually compatible plants. Cotton is predominantly self-pollinating, with pollen that is large, sticky and heavy and generally not dispersed by wind. Pollen can be transported by insect pollinators, chiefly honeybees, but gene flow studies have shown that outcrossing occurs at low levels and decreases rapidly with distance (OGTR 2013b). It is not expected that the introduced insect resistance or herbicide tolerance genes would alter the pollen dispersal characteristics of the GM cotton.

113. The applicant has proposed to restrict pollen flow by surrounding active trial sites either with a 20 m pollen trap of commercial cotton or with a 100 m monitoring zone (where any cotton plants are destroyed) and a 3 km exclusion zone (where no cotton crops are planted). In addition, any post-harvest cotton volunteers on the trial sites would be destroyed before flowering. These controls would minimise the potential for pollinators to transfer pollen from GMOs to related plants outside the trial sites. Some outcrossing is expected to occur between the GMOs and related plants grown at the trial sites, i.e. non-GM comparator cotton plants and cotton plants from pollen traps.

114. The second step in the causal pathway for this risk scenario is exposure to or dispersal of hybrid seeds containing the introduced proteins. As non-GM cotton plants grown in trial sites and cotton plants from pollen traps are expected to produce a small proportion of hybrid seeds, the applicant has proposed that non-GM cotton or commercially authorised GM cotton planted at the location and/or in the pollen trap will be treated as if it were the GMOs. The limits and controls proposed for the GMOs would minimise exposure to hybrid seeds (Risk Scenario 1), and minimise dispersal of hybrid seeds (Risk Scenario 2).

115. In the unlikely event that the GMOs overcame barriers to pollen transfer outside trial sites, outcrossing could occur only with Gossypium hirsutum or G. barbadense plants, as native Gossypium species are not sexually compatible with cotton (Chapter 1, Section 6.4). If the GMOs pollinated volunteer or feral cotton, hybrid seeds would be unlikely to establish a stable population, as discussed in Risk Scenario 2. If the GMOs pollinated a commodity cotton crop, a small amount of hybrid seed containing the introduced proteins could enter human food and animal feed. If the



GMOs pollinated a cotton crop intended for seed production, hybrid seed could be purposefully dispersed and grown, leading to wider exposure to the introduced proteins.

Potential harms

116. The potential harms from this risk scenario are toxicity or allergenicity in people or toxicity to desirable organisms, reduced establishment or yield of desirable plants, or reduced biodiversity.

117. As discussed in Risk Scenario 1, the introduced proteins in the GM cotton are not expected to be toxic or allergenic to people, or toxic to vertebrates, or toxic to desirable invertebrates. They are only toxic to a limited range of insect species. This is also expected to be the case if the introduced proteins are expressed in hybrids with non-GM cotton.

118. If the GMOs included in this application outcrossed with commercially released GM cotton lines, the hybrid offspring could contain stacks between the introduced genes of this application and introduced genes in commercial GM cotton. The GM cotton lines approved for commercial release (licences DIR 062/2005, DIR 066/2006, DIR 091, DIR 118 and DIR 124) have the traits of insect resistance and/or herbicide tolerance. Their introduced insect resistance and herbicide tolerance genes are either identical or homologous to the genes in this application (Bt cry1 genes and a cry2A gene, an epsps gene). In hybrids containing stacked Bt insect resistance proteins from both this application and commercial GM cotton, it is possible that additive or synergistic effects could occur, potentially increasing the range of sensitive insects. Some uncertainty exists in this area due to data gaps. However, it is unlikely that the hybrids could be toxic to a wider range of insects than microbial Bt insecticidal products, which are considered unlikely to pose any risk to humans, other vertebrates, or the great majority of non-target invertebrates (International Programme on Chemical Safety 1999). In addition, FSANZ has approved food derived from all GM cotton lines in this application and all commercially released GM cotton lines as safe for human consumption. No separate approval is necessary for food derived from breeding between approved GM parent lines, so any hybrids would also be considered safe for human consumption (FSANZ website).

119. The potential for the GMOs to reduce establishment or yield of desirable plants was discussed in Risk Scenario 2. Cotton plants expressing the introduced proteins are unlikely to spread and persist in nature reserves or to survive standard weed management practices for cotton volunteers in agricultural settings. The herbicide tolerant GMOs in this application and commercial herbicide tolerant GM cotton lines tolerate the same herbicides (glyphosate and glufosinate ammonium), so there is no potential for stacked hybrids to have greater resistance to weed management.

120. Risk Scenario 2 discussed the potential of cotton containing the introduced proteins to reduce arthropod biodiversity through direct toxicity and indirect effects, and found that there was unlikely to be significant reduction of biodiversity. As any insect resistant hybrids between the GMOs and commercial GM cotton would have a similar range of insect toxicity (discussed above), the stacks would not be expected to have greater impact on biodiversity.

121. Conclusion: Risk scenario 3 is not identified as a substantive risk because cotton has limited ability to outcross, the proposed controls would minimise pollen flow to cotton plants outside the trial sites, and hybrids between the GMOs and commercial GM cotton lines would have similar properties to the GMOs. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.

Section 3 Uncertainty122. Uncertainty is an intrinsic property of risk and is present in all aspects of risk analysis§. Uncertainty in risk assessments arises from sources such as incomplete knowledge and inherent biological variability. Uncertainty is addressed by approaches including balance of evidence, conservative assumptions, and applying risk management measures that reduce the potential for risk § A more detailed discussion of uncertainty is contained in the Regulator’s Risk Analysis Framework available from the OGTR website or via Free call 1800 181 030.


http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/risk-analysis-framework



scenarios involving uncertainty to lead to harm. If there is residual uncertainty that is important to estimating the level of risk, the Regulator will take this uncertainty into account in making decisions.

123. As field trials of GMOs are designed to gather data, there are generally data gaps when assessing the risks of a field trial application. However, field trial applications are required to be limited and controlled. Even if there is uncertainty about the characteristics of a GMO, limits and controls restrict exposure to the GMO, and thus decrease the likelihood of harm.

124. For DIR 133, uncertainty is noted particularly in relation to:

Potential toxicity to an increased range of insects due to combination of several introduced insect resistance proteins in stacked GM cotton lines.

Potential toxicity to an increased range of insects due to combination of several introduced insect resistance proteins in hybrids between the GMOs and commercially released GM cotton lines.

125. Additional data, including information to address these uncertainties, may be required to assess possible future applications with reduced limits and controls, such as a larger scale trial or the commercial release of these GMOs.

126. Chapter 3, Section 4, discusses information that may be required for future release.

Section 4 Risk Evaluation127. Risk is evaluated against the objective of protecting the health and safety of people and the environment to determine the level of concern and, subsequently, the need for controls to mitigate or reduce risk. Risk evaluation may also aid consideration of whether the proposed dealings should be authorised, need further assessment, or require collection of additional information.

128. Factors used to determine which risks need treatment may include:

risk criteria level of risk uncertainty associated with risk characterisation interactions between substantive risks.

129. Three risk scenarios were postulated whereby the proposed dealings might give rise to harm to people or the environment. In the context of the control measures proposed by the applicant, and considering both the short and long term, none of these scenarios were identified as substantive risks. The principal reasons for these conclusions are summarised in Table 7 and include:

none of the GM plant material or products will enter human food or animal feed supply chains

widespread presence of the introduced genes and their encoded proteins in the environment the proteins encoded by the introduced insect resistance genes are only expected to be toxic

to a limited range of insect species limited ability of the GM cotton plants to establish populations outside cultivation limited ability of the GM cotton plants to transfer the introduced genetic material to other

cotton plants suitability of controls proposed by Bayer to restrict the spread and persistence of the GM

cotton plants and their genetic material.

130. Therefore, risks to the health and safety of people, or the environment, from the proposed release of the GM cotton plants into the environment are considered to be negligible. The Risk Analysis Framework (OGTR 2013a), which guides the risk assessment and risk management



process, defines negligible risks as risks of no discernible concern with no present need to invoke actions for mitigation. Therefore, no additional controls are required to treat these negligible risks. Hence, the Regulator considers that the dealings involved in this proposed release do not pose a significant risk to either people or the environment.



Chapter 3 Risk management planSection 1 Background

131. Risk management is used to protect the health and safety of people and to protect the environment by controlling or mitigating risk. The risk management plan addresses risks evaluated as requiring treatment and considers limits and controls proposed by the applicant, as well as general risk management measures. The risk management plan informs the Regulator’s decision-making process and is given effect through licence conditions.

132. Under section 56 of the Act, the Regulator must not issue a licence unless satisfied that any risks posed by the dealings proposed to be authorised by the licence are able to be managed in a way that protects the health and safety of people and the environment.

133. All licences are subject to three conditions prescribed in the Act. Section 63 of the Act requires that each licence holder inform relevant people of their obligations under the licence. The other statutory conditions allow the Regulator to maintain oversight of licensed dealings: section 64 requires the licence holder to provide access to premises to OGTR inspectors and section 65 requires the licence holder to report any information about risks or unintended effects of the dealing to the Regulator on becoming aware of them. Matters related to the ongoing suitability of the licence holder are also required to be reported to the Regulator.

134. The licence is also subject to any conditions imposed by the Regulator. Examples of the matters to which conditions may relate are listed in section 62 of the Act. Licence conditions can be imposed to limit and control the scope of the dealings. In addition, the Regulator has extensive powers to monitor compliance with licence conditions under section 152 of the Act.

Section 2 Risk treatment measures for substantive risks135. The risk assessment of risk scenarios listed in Chapter 2 concluded that there are negligible risks to people and the environment from the proposed field trial of GM cotton. These risk scenarios were considered in the context of the scale of the proposed release (Chapter 1, Section 3.1), the proposed containment measures (Chapter 1, Section 3.2), and the receiving environment (Chapter 1, Section 6), and considering both the short and the long term. The risk evaluation concluded that no additional controls are required to treat these negligible risks.

Section 3 General risk management136. The limits and controls proposed in the application were important in establishing the context for the risk assessment and in reaching the conclusion that the risks posed to people and the environment are negligible. Therefore, to maintain the risk context, licence conditions have been imposed to limit the release to the proposed size, locations and duration, and to restrict the spread and persistence of the GMOs and their genetic material in the environment. The conditions are discussed and summarised in this Chapter and listed in detail in the licence.

3.1 Licence conditions to limit and control the release3.1.1 Consideration of limits and controls proposed by Bayer

137. Sections 3.1 and 3.2 of Chapter 1 provide details of the limits and controls proposed by Bayer in their application. These are taken into account in the three risk scenarios postulated for the proposed release in Chapter 2. Many of the proposed control measures are considered standard for GM crop trials and have been imposed by the Regulator in previous DIR licences. The appropriateness of these controls is considered further below.

138. The duration of the field trial would be limited to six years. In the first two years there would be up to 12 trial sites per year of up to 10 ha each, and during subsequent years there would be up to

Chapter 3 – Risk management 24


20 trial sites per year of up to 30 ha each. The size and duration of the trial would limit exposure to the GMOs in comparison to a commercial release (Risk Scenario 1).

139. The applicant proposes that only authorised and trained and/or experienced personnel would be permitted to deal with the GMOs. A standard licence condition requires that all persons dealing with the GMOs must be informed of any applicable licence conditions. These measures would limit the potential exposure of humans to the GMOs (Risk Scenario 1).

140. The applicant does not propose using any of the GM plant material for human or animal consumption. Therefore, a licence condition prohibits material from the trial from being used for human food or animal feed. This control will restrict exposure of humans and other organisms to the GMOs (Risk Scenario 1) and the potential for GM cotton seed to be dispersed outside the trial limits (Risk Scenario 2).

141. The applicant proposes to sell cotton lint (long fibres) from GMOs grown in the trial. As discussed in Risk Scenario 1, cotton lint is free of protein so is not a means of exposure to GM material. Therefore, licence conditions permit transport and sale of lint from GM cotton.

142. The applicant would clean all equipment used with the GMOs before using the equipment for other purposes. Equipment used on the trial sites would be cleaned on site. The GM cotton would be ginned separately from other cotton crops and the gin would be cleaned after use to prevent GM cotton seed mixing with other seed. These measures are appropriate to restrict potential dispersal of GM cotton seed outside the trial sites (Risk Scenario 2).

143. After the trial sites have been harvested, all GMOs will be destroyed except for plant material required for testing and cotton seed required for further authorised planting. Cotton seeds have low dormancy levels and do not generally form a viable seed bank, however, dormancy can be induced in cotton seeds by low soil temperature and/or soil moisture (OGTR 2013b). The applicant proposes post-harvest cultivation of the trial sites to promote cotton seed germination or decomposition. A licence condition has been imposed requiring tillage, followed by an irrigation event, in the spring or summer following the harvest, so that soil temperature and moisture will be suitable for cotton seed germination. These measures would restrict the persistence of a GM cotton seed bank after the duration of the trial (Risk Scenario 2).

144. The applicant proposes that each trial site will be monitored post-harvest at least every two months for a minimum of twelve months and until the site has been clear of volunteers for at least six months. During this period any cotton volunteers will be destroyed before flowering. However, the proposed frequency of monitoring may not be sufficient. The OGTR’s experience from previous GM cotton trials indicates that if there is vigorous growth of a post-harvest crop, cotton volunteers in the 2-4 leaf stage may be missed during inspections, and also that in particularly favourable weather conditions the period between the 4 leaf stage and the beginning of flowering can be less than 40 days. Therefore, a licence condition requires post-harvest monitoring at least every 35 days, with destruction of any cotton volunteers. These measures would restrict the persistence of GMOs after the duration of the trial (Risk Scenario 2).

145. The applicant proposes a number of methods to dispose of GMOs. Destruction by hand weeding, stalk pulling, ploughing, treatment with herbicide, burning/incineration, autoclaving or burial under at least one metre of soil are all methods that have been included in previous DIR licences for GM cotton. The proposed method of destruction by root cutting and mulching or slashing is expected to be effective in killing the GM cotton plants and preventing regrowth of ratoon cotton from the root stock. The proposed destructive analysis by grinding seeds is expected to be effective in making cotton seeds non-viable. The proposed method of destroying cotton linters by acid delinting is considered unnecessary. Non-viable plant parts are not a potential pathway for spread or persistence of GMOs. Therefore, under the licence, the applicant would only be required to destroy GMOs (ie live plants and plant parts that could grow into live plants, such as seeds and



root stock), by one of the proposed methods. Once linters have been separated from cotton seeds, the linters are not GMOs and there is no requirement to destroy them.

146. The applicant proposes that GMOs will be transported and stored according to the Regulator’s current Guidelines for the Transport, Storage and Disposal of GMOs (OGTR website). These protocols would restrict the potential for dispersal of GM seeds outside the trial sites (Risk Scenario 2). A licence condition also permits alternative methods for transport of harvested cotton modules or bales, using double wrapping or an enclosed chain bed truck designed for cotton transport. These methods have been authorised in previous cotton licences and are considered to provide secure containment of GM cotton during transport.

147. The applicant proposes that each trial site will be surrounded either by a 20 m wide pollen trap or by a 100 m wide monitoring zone and a 3 km exclusion zone, to control pollen flow from the GMOs to cotton plants outside the trial sites. As discussed in Risk Scenario 3, cotton is predominantly self-pollinating and outcrossing rates decrease rapidly with distance. A 20 m pollen trap around GM cotton was generally found to be an effective buffer under Australian conditions (Llewellyn et al. 2007). Cotton gene flow can be enhanced in the presence of unusually high honeybee populations, resulting in detectable levels of cross-pollination at distances greater than 20 m (Llewellyn et al. 2007; Van Deynze et al. 2005). Very high honeybee abundance might occur if GM cotton was planted adjacent to a bee-pollinated crop such as orchards or melons with introduced bee hives, or adjacent to bushland housing feral honeybee hives. In either of these cases, there would be reduced likelihood of the GM cotton also being adjacent to a commercial cotton field that could be cross-pollinated by the GMOs. Therefore, using a 20 m pollen trap would minimise gene transfer to cotton plants outside the trial sites (Risk Scenario 3). A licence condition specifies that if a pollen trap is used, the pollen trap must be at least 20 m wide, the plants within the pollen trap must be either non-GM cotton or GM cotton approved for commercial release, and they must be maintained to flower at the same time as the GMOs.

148. If monitoring and exclusion zones are used, the applicant proposes that the 100 m wide monitoring zones would be inspected every 30 days while GMOs are being grown and any cotton plants destroyed prior to flowering. However, this proposed period over which inspections would occur is longer than necessary, as GMOs cannot transfer pollen before they flower. A licence condition has been imposed requiring monitoring zones to be inspected from 14 days prior to the expected flowering of the GMOs, every 35 days until harvest of the GMOs. Monitoring zones of 100 m are considered appropriate to restrict pollen flow from GMOs to volunteer or feral cotton plants outside the trial sites (Risk Scenario 3).

149. If monitoring and exclusion zones are used, the applicant proposes that the exclusion zones would be 3 km wide and inspected every 30 days to ensure that no cotton crop has been planted in an exclusion zone while GMOs are growing. This proposed period over which inspections would occur is considered longer than necessary, and a licence condition requires exclusion zones to be inspected from 14 days prior to the expected flowering of the GMOs, every 35 days until all GMOs have finished flowering. In addition, the proposed exclusion zone width is considered larger than necessary. A study measured pollen-mediated gene flow between herbicide tolerant cotton fields and conventional cotton fields in California in the absence of a pollen trap, and found that average gene flow was less than 0.1% at 400 m and 0.04% at 1625 m (Van Deynze et al. 2005). Another study in Arizona found that GM cotton fields at distances of more than 750 m from the edge of monitored non-GM fields did not appear to contribute to outcrossing (Heuberger et al. 2010). Cotton outcrossing distances in Australia are not expected to be longer than in the United States, and may be significantly shorter because Australia lacks bumblebees, which are very effective pollinators of cotton (OGTR 2013b). Taking this literature into account, a licence condition specifies that exclusion zones must be at least 1.5 km wide. This measure would minimise pollen flow from GMOs to cotton crops outside the trial sites (Risk Scenario 3).


http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/transport-guide-1


150. The applicant intends to plant both GM and non-GM cotton in the trial sites, and may plant non-GM or commercially approved GM cotton plants in pollen traps. Under the licence, the applicant must treat all non-GM or commercially approved GM cotton from the trial sites or pollen trap as if it were the GM cotton in this application. This measure would minimise exposure to or dispersal of hybrid seed resulting from outcrossing between the GMOs and other cotton (Risk Scenario 3).

151. The applicant proposes that GMOs may be transported, stored or subjected to experiments in certified physical containment facilities under a Notifiable Low Risk Dealing (NLRD) authorisation in accordance with applicable requirements of the Gene Technology Regulations 2001. This activity is not included in the licence as it would be permitted by a separate valid authorisation. 3.1.2 Summary of licence conditions to be implemented to limit and control the release

152. A number of licence conditions have been imposed to limit and control the proposed release, based on the above considerations. These include requirements to:

limit the duration of the field trial to between July 2015 and July 2021

limit the field trial to a maximum area of 10 ha/site at up to 12 sites per year for the first two years and 30 ha/site at up to 20 sites per year in the third to sixth years.

locate trial sites at least 50 m away from natural waterways

restrict pollen flow using one of the following measures:

o surround the planting area with a 20 m pollen trap of non-GM cotton or GM cotton approved for commercial release, or

o surround the planting area with a 100 m monitoring zone and a 1.5 km exclusion zone

ensure that pollen trap plants flower for the same period of time as the GMOs

destroy any cotton plants growing in the monitoring zone prior to flowering

ensure that no cotton crops are grown in the exclusion zone

treat any non-GM or commercially approved GM cotton planted in the planting area or pollen trap as if it were the GMOs

clean all equipment used with the GMOs before using it for any other purpose

gin the GMOs separately from any other cotton crop

use tillage and irrigation to promote germination of any cotton seeds remaining in the trial sites after harvest

monitor the trial sites for at least 12 months after harvest and destroy any cotton volunteers until no volunteers are detected for a continuous 6 month period

destroy all GMOs from the trial that are not required for testing or future planting

transport and store the GMOs in accordance with the Regulator’s Guidelines for the Transport, Storage and Disposal of GMOs or other specified transport conditions

not allow GM plant material to be used for human food or animal feed.

3.2 Other risk management considerations153. All DIR licences issued by the Regulator contain a number of conditions that relate to general risk management. These include conditions relating to:

applicant suitability



contingency plans

identification of the persons or classes of persons covered by the licence

reporting requirements

access for the purpose of monitoring for compliance.3.2.1 Applicant suitability

154. In making a decision whether or not to issue a licence, the Regulator must have regard to the suitability of the applicant to hold a licence. Under section 58 of the Act, matters that the Regulator must take into account, for either an individual applicant or a body corporate, include:

any relevant convictions of the applicant

any revocation or suspension of a relevant licence or permit held by the applicant under a law of the Commonwealth, a State or a foreign country

the capacity of the applicant to meet the conditions of the licence.

155. On the basis of information submitted by the applicant and records held by the OGTR, the Regulator considers Bayer suitable to hold a licence. The licence includes a requirement for the licence holder to inform the Regulator of any information that would affect their suitability.

156. In addition, any applicant organisation must have access to a properly constituted Institutional Biosafety Committee and be an accredited organisation under the Act.3.2.2 Contingency plan

157. Bayer is required to submit a contingency plan to the Regulator before planting the GMOs. This plan will detail measures to be undertaken in the event of any unintended presence of the GM cotton outside permitted areas.

158. Bayer is also required to provide the Regulator with a method to reliably detect the GMOs or the presence of the genetic modifications in a recipient organism. This methodology is required before planting the GMOs.3.2.3 Identification of the persons or classes of persons covered by the licence

159. The persons covered by the licence are the licence holder and employees, agents or contractors of the licence holder and other persons who are, or have been, engaged or otherwise authorised by the licence holder to undertake any activity in connection with the dealings authorised by the licence. Prior to growing the GMOs, Bayer is required to provide a list of people and organisations that will be covered by the licence, or the function or position where names are not known at the time.3.2.4 Reporting requirements

160. The licence requires the licence holder to immediately report any of the following to the Regulator:

any additional information regarding risks to the health and safety of people or the environment associated with the trial

any contraventions of the licence by persons covered by the licence

any unintended effects of the trial.

161. A number of written notices are also required under the licence to assist the Regulator in designing and implementing a monitoring program for all licensed dealings. The notices include:

expected and actual dates of planting

details of areas planted to the GMOs



expected dates of flowering

expected and actual dates of harvest and cleaning after harvest

details of inspection activities.3.2.5 Monitoring for compliance

162. The Act stipulates, as a condition of every licence, that a person who is authorised by the licence to deal with a GMO, and who is required to comply with a condition of the licence, must allow inspectors and other persons authorised by the Regulator to enter premises where a dealing is being undertaken for the purpose of monitoring or auditing the dealing. Post-release monitoring continues until the Regulator is satisfied that all the GMOs resulting from the authorised dealings have been removed from the release site.

163. If monitoring activities identify changes in the risks associated with the authorised dealings, the Regulator may also vary licence conditions, or if necessary, suspend or cancel the licence.

164. In cases of non-compliance with licence conditions, the Regulator may instigate an investigation to determine the nature and extent of non-compliance. The Act provides for criminal sanctions of large fines and/or imprisonment for failing to abide by the legislation, conditions of the licence or directions from the Regulator, especially where significant damage to health and safety of people or the environment could result.

Section 4 Issues to be addressed for future releases165. Additional information has been identified that may be required to assess an application for a commercial release of these GM cotton lines, or to justify a reduction in limits and controls. This includes:

additional data on the potential toxicity of combinations of the insecticidal proteins included in this application

additional data on the potential toxicity of combinations of the insecticidal proteins included in this application with insecticidal proteins present in GM cotton lines that have been commercially released in Australia.

Section 5 Conclusions of the RARMP166. The RARMP concludes that the proposed limited and controlled release of GM cotton poses negligible risks to the health and safety of people or the environment as a result of gene technology, and that these negligible risks do not require specific risk treatment measures.

167. However, conditions have been imposed to limit the release to the proposed size, locations and duration, and to restrict the spread and persistence of the GMOs and their genetic material in the environment, as these were important considerations in establishing the context for assessing the risks.



ReferencesAddison, S.J., Farrell, T., Roberts, G.N., Rogers, D.J. (2007). Roadside surveys support predictions of negligible naturalisation potential for cotton (Gossypium hirsutum) in north-east Australia. Weed Research 47: 192-201

Arts, J.H.E., Mommers, C., de Heer, C. (2006). Dose-response relationships and threshold levels in skin and respiratory allergy. Critical Reviews in Toxicology 36: 219-251

Bravo, A., Gill, S.S., Soberon, M. (2007). Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon 49: 423-435

Bushey, D., Freyssinet, M., Poe, M., and Rinehardt, M. (2008). Petition for Determination of Nonregulated Status for Insect-Resistant and Glufosinate Ammonium-Tolerant cotton: TwinLink™ cotton. Bayer CropScience

CERA (2011). A review of the environmental safety of the Cry1Ab protein. Center for Environmental Risk Assessment

Charles, G., Roberts, G., Kerlin, S., and Hickman, M. (2013). WEEDpak: Controlling volunteer cotton. Cotton Research and Development Corporation

Dill, G.M. (2005). Glyphosate-resistant crops: history, status and future. Pest Management Science 61: 219-224

Dively, G.P. (2005). Impact of transgenic VIP3A x Cry1Ab Lepidopteran-resistant field corn on the nontarget arthropod community. Environmental Entomology 34: 1267-1291

Eastick, R., Hearnden, M. (2006). Potential for Weediness of Bt Cotton (Gossypium hirsutum) in Northern Australia. Weed Science 54: 1142-1151

EPA (1996). Plant pesticide inert ingredient CP4 Enolpyruvylshikimate-3-D and the genetic material necessary for its production in all plants. Report No. 61, US Environmental Protection Agency

Estruch, J.J., Carozzi, N.B., Desai, N., Duck, N.B., Warren, G.W., Koziel, M.G. (1997). Transgenic plants: an emerging approach to pest control. Nature Biotechnology 15: 137-141

Estruch, J.J., Warren, G.W., Mullins, M.A., Nye, G.J., Craig, J.A., Koziel, M.G. (1996). Vip3A, a novel Bacillus thuringiensis vegetative insecticidal protein with a wide spectrum of activities against lepidopteran insects. Proceedings of the National Academy of Sciences of the United States of America 93: 5389-5394

Evstigneeva, Z.G., Solov'eva, N.A., Sidel'nikova, L.I. (2003). Methionine sulfoximine and phosphinothricin: A review of their herbicidal activity and effects on glutamine synthetase. Applied Biochemistry and Microbiology 39: 539-543

Felsot, A.S. (2000). Insecticidal genes part 2: Human health hoopla. Agrichemical & Environmental News 168: 1-7

Gouffon, C., Van Viet, J., Van Rie, S., Jansens, S., Jurat-Fuentes, J.L. (2011). Binding sites for Bacillus thuringiensis Cry2Ae toxin on heliothine brush border membrane vesicles are not shared with Cry1A, Cry1F, or Vip3A toxin. Applied and Environmental Microbiology 77: 3182-3188

Appendix A 30


Hernandez-Rodriguez, C.S., Boets, A., Van Rie, J., Ferre, J. (2009). Screening and identification of vip genes in Bacillus thuringiensis strains. Journal of Applied Microbiology 107: 219-225

Herouet-Guicheney, C., Rouquié, D., Freyssinet, M., Currier, T., Martone, A., Zhou, J., Bates, E.E.M., Ferullo, J., Hendrickx, K., Rouan, D. (2009). Safety evaluation of the double mutant 5-enol pyruyvlshikimate-3-phosphate synthase (2mEPSPS) from maize that confers tolerance to glyphosate herbicide in transgenic plants. Regulatory Toxicology and Pharmacology 54: 143-153

Heuberger, S., Ellers-Kirk, C., Tabashnik, B.E., Carriere, Y. (2010). Pollen- and Seed-Mediated Transgene Flow in Commercial Cotton Seed Production Fields. PLoS ONE 5: e14128

Hill, K., Jiang, X., Lee, M., Mascarenhas, V., Mullins, M., Privalle, L., Rabe, S., Schriver, T., Stein, J., Vlachos, D., Walters, F., Ward, K., and Zawodny, J. (2003). Petition for the determination of non-regulated status: Lepidopteran insect protected VIP3A cotton transformation event COT102. Syngenta Seeds Incorporated North Carolina.

Höfte, H., De Greve, H., Seurinck, J., Jansens, S., Mahillon, J., Ampe, C., Vandekerckhove, J., Vanderbruggen, H., Van Montagu, M., Zabeau, M., Vaeck, M. (1986). Structural and functional analysis of a cloned delta endotoxin of Bacillus thuringiensis berliner 1715. European Journal of Biochemistry 161: 273-280

International Programme on Chemical Safety (1999). Environmental Health Criteria 217: Bacilus thuringiensis. Report No. E, United Nations Environment Programme; International Labour Organisation; World Health Organization

Keese, P. (2008). Risks from GMOs due to horizontal gene transfer. Environmental Biosafety Research 7: 123-149

Keese, P.K., Robold, V., Myers, R.C., Weisman, S., Smith, J. (2014). Applying a weed risk assessment approach to GM crops. Transgenic Research 23: 957-969

Lebrun, M., Waksman, G., Freyssinet, G. (1987). Nucleotide sequence of a gene encoding corn ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcs). Nucleic Acids Research 15: 4360

Lee, M.K., Miles, P., Chen, J. (2006). Brush border membrane binding properties of Bacillus thuringiensis Vip3A toxin to Heliothis virescens and Helicoverpa zea midguts. Biochemical and Biophysical Research Communications 339: 1043-1047

Llewellyn, D.J., Tyson, C., Constable, G.A., Duggan, B., Beale, S., Steel, P. (2007). Containment of regulated genetically modified cotton in the field. Agriculture, Ecosystems & Environment 121: 419-429

Lu, Y., Xu, W., Kang, A., Luo, Y., Guo, F., Yang, R., Zhang, J., Huang, K. (2007). Prokaryotic expression and allergenicity assessment of hygromycin B phosphotransferase protein derived from genetically modified plants. Journal of Food Science 72: M228-M232

OECD (2007). Consensus document on safety information on transgenic plants expressing Bacillus thuringiensis - derived insect control protein. Report No. 42, Organisation for Economic Co-operation and Development Paris, France.

Appendix A 31


OGTR (2013a). Risk Analysis Framework. The Office of the Gene Technology Regulator, Canberra, Australia.

OGTR (2013b). The biology of Gossypium hirsutum L. and Gossypium barbadense L. (cotton) v2.1. The Office of the Gene Technology Regulator, Canberra, Australia.

Raybould, A., Vlachos, D. (2011). Non-target organism effects tests on Vip3A and their application to the ecological risk assessment for cultivation of MIR162 maize. Transgenic Research 20: 599-611

Roth, G. (2014). Australian grown cotton sustainability report 2014. Cotton Research and Development Corporation and Cotton Australia

Schnell, J., Steele, M., Bean, J., Neuspiel, M., Girard, C., Dormann, N., Pearson, C., Savoie, A., Bourbonniere, L., Macdonald, P. (2015). A comparative analysis of insertional effects in genetically engineered plants: considerations for pre-market assessments. Transgenic Research 24: 1-17

Sena, J.A., Hernandez-Rodriguez, C.S., Ferre, J. (2009). Interaction of Bacillus thuringiensis Cry1 and Vip3A proteins with Spodoptera frugiperda midgut binding sites. Applied and Environmental Microbiology 75: 2236-2237

Standards Australia New Zealand, CRC for Australian Weed Management (2006). National Post-Border Weed Risk Management Protocol.

Steiner, H.Y., Halpin, C., Jez, J.M., Kough, J., Parrott, W., Underhill, L., Weber, N., Hannah, L.C. (2013). Evaluating the potential for adverse interactions within genetically engineered breeding stacks. Plant Physiology 161: 1587-1594

Then, C. (2010). Risk assessment of toxins derived from Bacillus thuringiensis-synergism, efficacy, and selectivity. Environ Sci Pollut Res Int 17: 791-797

Thompson, C.J., Movva, N.R., Tizard, R., Crameri, R., Davies, J., Lauwereys, M., Botterman, J. (1987). Characterization of the herbicide-resistance gene bar from Streptomyces hygroscopicus. EMBO Journal 6: 2519-2523

Van Deynze, A.E., Sundstrom, F.J., Bradford, K.J. (2005). Pollen-mediated gene flow in California cotton depends on pollinator activity. Crop Science 45: 1565-1570

van Frankenhuyzen, K. (2013). Cross-order and cross-phylum activity of Bacillus thuringiensis pesticidal proteins. Journal of Invertebrate Pathology 114: 76-85

Whitehouse, M.E.A., Wilson, L.J., Constable, G.A. (2007). Target and non-target effects on the invertebrate community of Vip cotton, a new insecticidal transgenic. Australian Journal of Agricultural Research 58: 273-285

Whitehouse, M.E.A., Wilson, L.J., Davies, A.P., Cross, D., Goldsmith, P., Thompson, A., Harden, S., Baker, G. (2014). Target and nontarget effects of novel triple-stacked Bt-transgenic cotton 1: canopy arthropod communities. Environmental Entomology 43: 218-241

Yu, C.G., Mullins, M.A., Warren, G.W., Koziel, M.G., Estruch, J.J. (1997). The Bacillus thuringiensis vegetative insecticidal protein Vip3A lyses midgut epithelium cells of susceptible insects. Applied Environmental Microbiology 63: 532-536

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Zhuo, Q., Piao, J.Q., Tian, Y., Xu, J., Yang, X.G. (2009). Large-scale purification and acute toxicity of hygromycin B phosphotransferase. Biomedical and Environmental Sciences 22: 22-27

Appendix A 33


Appendix A Summary of submissions from prescribed experts, agencies and authorities**

Advice received by the Regulator from prescribed experts, agencies and authorities on the consultation RARMP is summarised below. All issues raised in submissions that related to risks to the health and safety of people and the environment were considered in the context of the currently available scientific evidence and were used in finalising the RARMP that formed the basis of the Regulator’s decision to issue the licence.

Sub. No: Summary of issues raised Comment1 Considers that the environmental risks of

the trial are likely to be low and manageable.

Noted.

Notes that the RARMP states that synergistic or additive effects between stacked insecticidal genes could pose risks to a greater range of non-target insects. Suggests that examples of stacking events with synergistic or additive effects would be beneficial to the RARMP.

Additive or synergistic effects between stacked Bt genes are reported in the scientific literature. For instance, synergism of Bt genes was reviewed by Then (Then 2010). However, no data was found regarding additive or synergistic effects on the selectivity of the specific Bt genes combined in the current application. Therefore, this was flagged in the RARMP as an area of uncertainty due to data gaps.

Suggests that RARMP could be improved by more detailed discussion of potential effects of stacked insecticidal genes on non-target organisms, particularly those native to Australia.

Text has been added to the RARMP to clarify that GM cotton could be toxic to certain non-target organisms, such as native Australian moth species, if these insects feed on cotton. The RARMP already notes that the limited scale and duration of the proposed field trial would restrict the number of invertebrates exposed to the introduced proteins.

Notes that the RARMP states the applicant may be required to collect further data about whether stacked insecticidal genes are toxic to an increased range of insects, prior to submitting a future application for commercial release. Suggests that if this issue is important, the data collection should be required.

The section of the RARMP listing issues to be addressed for future releases is intended to provide guidance to the applicant, rather than to impose requirements. In the event of a future application for commercial release, at that time the Regulator will evaluate whether the data provided in the application and/or published in the scientific literature is sufficient for risk assessment.

Suggests that RARMP could discuss the potential for insects to develop cross-resistance to the insecticidal genes.

The issue of development of resistance to the introduced insecticidal genes is not discussed in the RARMP because:a) the proposed release is a field trial, and it is expected that that

the limited scale and duration of the trial will minimise the potential for insects to develop resistance, and

b) the efficacy of insecticidal products and appropriate resistance management strategies are regulated by APVMA.

Suggests that reasons be provided for statements in the RARMP that the introduced proteins are not expected to increase the ability of cotton to spread and persist.

The section of the RARMP discussing the ability of GM cotton to spread and persist has been clarified and expanded.

Suggests that since the proposed trial is very large, and the applicant intends to use the trial for seed production and for sale of cotton lint, the Regulator should consider whether the trial should be characterised as a limited and controlled release.

After considering information supplied in the application, and additional information requested from the applicant, the Regulator was satisfied that the principal purpose of the application is to conduct experiments. Therefore, the application was accepted for assessment as a limited and controlled release.The applicant was made aware of the concerns regarding the scale of the trial, and reduced the proposed planting area by 16%.

** Prescribed agencies include the Gene Technology Technical Advisory Committee, State and Territory Governments, relevant local governments, Australian Government agencies and the Minister for the Environment.

Appendix A 34


Sub. No: Summary of issues raised CommentAdvises that although a 20 m pollen trap has been found to effectively prevent outcrossing in many parts of Australia, in one site in northern Australia outcrossing rates of 0.76-0.79% were observed at 50 m, possibly due to high local populations of pollinators (Llewellyn et al. 2007). Similarly, an experiment on outcrossing rates in California (Van Deynze et al. 2005) found low rates in regions with low pollinator populations (<0.1% at 30 m) and higher rates in regions with high pollinator activity (0.31% at 30 m). Suggests that for trials north of the Tropic of Capricorn a monitoring and exclusion zone be used instead of a 20 m pollen trap. Suggests that further information on outcrossing be collected during the trial to address this area of uncertainty.

Text has been added to the RARMP to clarify that the effectiveness of a 20 m pollen trap is reduced in the presence of unusually high honeybee populations. These high honeybee populations might occur if GM cotton was planted adjacent to a bee-pollinated crop with introduced bee hives, or adjacent to bushland housing feral honeybee hives. However, in either of these cases, there would be reduced likelihood of the GM cotton also being adjacent to a commercial cotton field that could be cross-pollinated by the GMOs.The location of trials in tropical versus non-tropical regions is not the key factor influencing honeybee abundance. Therefore, it is not considered appropriate to impose different isolation conditions in tropical regions.The purpose of collecting information during this trial to address uncertainty would be to aid risk assessment during a future application. However, it is anticipated that if there is a future application regarding these GMOs, it will be for a commercial release. Typically, no isolation conditions are imposed on a commercial release, so information on the efficacy of isolation measures would not be useful for risk assessment.

Considers that the RARMP would benefit from an examination of experience with conventionally bred crops that have herbicide tolerance or insect resistance traits.

Crops with the general traits of herbicide tolerance or insect resistance have many different properties, conferred by a wide range of unrelated mechanisms. For the purposes of accurate risk assessment of a specific GMO, it is preferable to use data that relates to the GMO itself, or to a close homologue. No conventionally bred crops were closely related to the GMOs in this application, so data from conventionally bred crops was not considered.

2 Opposed to trial being conducted within its Shire. Concerned that GM material will disperse into land surrounding the trial sites. Refers to a recent news story (Marsh vs Baxter case) where GM canola contaminated an adjacent organic farm.

Note that the Marsh vs Baxter case, in which an organic farmer claimed damages from a neighbouring GM grower for loss of organic certification, related to commercially approved GM canola, not to GMOs from a field trial.The current GM cotton application is for a field trial, which may only be conducted under limited and controlled conditions. Strict licence conditions have been imposed to restrict spread of the GMOs outside the trial sites. There has been no documented loss of containment from any field trial authorised by the Regulator.

3 Noted that the RARMP was circulated for comment and no adverse comments were received. Indicated no objection to the issue of a licence for DIR 133.

Noted

4 Notes that the licence will prohibit the use of material from the trials for human or animal consumption. No further comments on the licence application at this stage.

Noted

5 Agrees with the overall conclusions of the RARMP.

Noted

Suggests that the RARMP clarify potential for spread and dispersal in northern Australia due to cyclones.

The part of the RARMP discussing the potential for dispersal of GM seeds by a cyclone has been expanded.

Suggests that the licence define a natural waterway.

A definition of a natural waterway has been added to the licence.

Advises that workers that deal with cotton often do not wear protective clothing due to heat.

The RARMP has been altered in accordance with this advice.

6 Supported the conclusions of the RARMP that the proposed dealings pose negligible risk of harm to human health and the environment.

Noted

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Sub. No: Summary of issues raised Comment7 Supportive of the application as the

consultation RARMP indicates that the proposed release poses negligible risks to people or the environment. Understands that licence conditions will be imposed to limit and control the release.

Noted.

Appendix A 36

Documents

Table of Contents - OGTR | Office of the Gene … · Web viewVips are secreted during vegetative growth stages and sporulation, whereas the Cry proteins are expressed by Bt only during