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Risk Assessment andRisk Management Plan for

DIR 135

Limited and controlled release of sugarcane genetically modified for enhanced sugar content

Applicant: The University of Queensland

August 2015

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DIR 135 – Risk Assessment and Risk Management Plan (August 2015) Office of the Gene Technology Regulator

Summary of the Risk Assessment and Risk Management Plan

for

Licence Application No. DIR 135DecisionThe 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 135Applicant: The University of Queensland (UQ)Project Title: Limited and controlled release of sugarcane genetically modified for

enhanced sugar contentParent organism: Sugarcane (Saccharum spp. hybrid)

Introduced genes and modified traits:

Genes involved in sugar metabolism: Two genes from sugarcane1

Selectable marker genes from Escherichia coli: Neomycin phosphotransferase type II gene (nptII, also referred

to as aminoglycoside phosphotransferase gene, aphA) Beta-lactamase gene (bla)

Proposed release dates:

August 2015 – May 2020

Proposed locations: Two sites in the local government area of Burdekin (Queensland)Proposed release size:

Five hectares of field planting plus 1250 m2 for plant handling, analysis and waste storage

Primary purpose: To evaluate the field performance of GM sugarcane and to identify plants with enhanced sugar content

UQ proposes a field trial of sugarcane genetically modified for enhanced sugar content at Sugar Research Australia’s (SRA) Durre farm in Burdekin. A nursery at the SRA Burdekin Station and a waste holding area at Durre farm will also be used. This field trial would continue the characterisation of GM plants produced in glass houses to evaluate their suitability for commercial production.

1 The identities of the genes and gene constructs have been declared Confidential Commercial Information (CCI) under s185 of the Gene Technology Act 2000.

Summary I


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 unintended exposure to the GM plant material, increased potential for spread and persistence of the GM plants relative to the non-GM parent and transfer of the genetic modification to non-GM sugarcane or other sexually compatible 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 separate the GM sugarcane from other sugarcane; to prohibit the use of GM plant material in human food and animal feed; to destroy GMOs not required for further studies; to transport GMOs with appropriate care; and to conduct post-harvest monitoring at the trial site to ensure all GMOs are destroyed.

Summary II


Table of ContentsSummary of the Risk Assessment and Risk Management Plan....................................................I

Decision.....................................................................................................................................IThe application...........................................................................................................................IRisk assessment.........................................................................................................................IRisk management plan..............................................................................................................II

Table of Contents........................................................................................................................IIIAbbreviations ............................................................................................................................VChapter 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 (size, locations, duration and people)................23.2 The proposed controls to restrict the spread and persistence of the GMOs in the environment...........................................................................................................................3

Section 4 The parent organisms...........................................................................................4Section 5 The GMOs, nature and effect of the genetic modification..................................5

5.1 Introduction to the GMOs...........................................................................................55.2 Toxicity/allergenicity of the proteins associated with the introduced genes..............75.3 Characterisation of the GMOs....................................................................................7

Section 6 The receiving environment..................................................................................76.1 Relevant abiotic factors...............................................................................................96.2 Relevant biotic factors................................................................................................96.3 Relevant agricultural practices....................................................................................96.4 Presence of related plants in the receiving environment..........................................106.5 Presence of introduced gene sequences in the environment.....................................11

Section 7 Relevant Australian and international approvals...............................................117.1 Australian approvals.................................................................................................117.2 International approvals of GM sugarcane.................................................................11

Chapter 2 Risk assessment.................................................................................................12Section 1 Introduction.......................................................................................................12Section 2 Risk Identification.............................................................................................13

2.1 Risk source................................................................................................................132.2 Causal pathway.........................................................................................................142.3 Potential harm...........................................................................................................152.4 Postulated risk scenarios...........................................................................................15

Section 3 Uncertainty........................................................................................................24

Table of Contents


Section 4 Risk Evaluation..................................................................................................25Chapter 3 Risk management plan......................................................................................26

Section 1 Background........................................................................................................26Section 2 Risk treatment measures for substantive risks...................................................26Section 3 General risk management..................................................................................26

3.1 Licence conditions to limit and control the release..................................................263.2 Other risk management considerations.....................................................................29

Section 4 Issues to be addressed for future releases..........................................................31Section 5 Conclusions of the consultation RARMP..........................................................31

References ...........................................................................................................................32Appendix A Summary of submissions from prescribed experts, agencies and authorities...36

Table of Contents


AbbreviationsAPVMA Australian Pesticides and Veterinary Medicines AuthorityCCI Confidential Commercial Information as declared under section 185 of

the Gene Technology Act 2000DIR Dealings involving Intentional ReleaseFSANZ Food Standards Australia New ZealandGM Genetically modifiedGMO Genetically modified organismha HectareLGA Local government aream MetresNLRD Notifiable low risk dealingsOGTR Office of the Gene Technology RegulatorPC2 Physical Containment level 2RARMP Risk Assessment and Risk Management PlanRegulations Gene Technology Regulations 2001Regulator Gene Technology RegulatorUQ University of QueenslandSRA Sugar Research AustraliaTGA Therapeutic Goods Administrationthe Act The Gene Technology Act 2000

Abbreviations V

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 parameters 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 (Summary of parameters used to establish the risk assessment context).

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 which 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 Regulator was not required to consult with prescribed experts, agencies and authorities before

Chapter 1 – Risk assessment context 1


preparation of the Risk Assessment and Risk Management Plan (RARMP; see section 50 of the Act).

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 2013) 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), Australian Pesticides and Veterinary Medicines Authority (APVMA), Therapeutic Goods Administration (TGA), National Industrial Chemicals Notification and Assessment Scheme and Department of Agriculture, Biosecurity. 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. The University of Queensland (UQ) proposes to release up to 1000 lines of genetically modified (GM) sugarcane lines into the environment under limited and controlled conditions.

9. The purpose of the trial is to evaluate the GM sugarcane lines grown under field conditions for key changes to agronomic characteristics such as sugar yield.

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

conducting experiments with the GMOs

propagating the GMOs

growing or culturing the GMOs

transporting the GMOs

disposing of the GMOs and

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

11. These dealings are detailed further below.

3.1 The proposed limits of the dealings (size, locations, duration and people)12. The applicant proposes to conduct dealings with the GM sugarcane until May 2020.

13. The dealings with the GMOs are proposed to occur at SRA Burdekin Station and at SRA Durre farm in Burdekin (Table 1). The field trial site of 5 ha and a GM waste holding area (1000 m2) that would be used during the course of the licence are located on Durre farm. The nursery (200 m2; used for hardening tissue cultured seedlings and germination of one-eye setts) is located at the SRA Burdekin Station.


http://www.ogtr.gov.au/


Table 1 Proposed localities for GM sugarcane field trials

Trial sites Burdekin 1 Burdekin 2

LGA Burdekin Shire Council Burdekin Shire CouncilGeographical location SRA Burdekin Station,

Bruce Highway, BrandonSRA Durre farm, 35 Sayers Rd, Barratta

Total area 220 m2 5.1 haPlanting area N/A 5 ha

14. In addition, the applicant also proposes to use a juice laboratory (for juicing sugarcane) at SRA Burdekin Station and a shed adjacent to the waste holding area at SRA Durre farm for analysis of harvested sugarcane.

15. The applicant proposes to use the hot water treatment facility provided by Burdekin Productivity Services to treat sugarcane stalks before planting at the nursery as a prophylactic measure against disease development.

16. The applicant is proposing that only trained and authorised staff would be permitted to deal with the GM sugarcane.

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 sugarcane lines and the introduced genetic material in the environment. These include:

surrounding the field trial site by a 2 m guard row of non-GM sugarcane

keeping a barren isolation zone of 6 m around the guard row

locating the trial sites at least 50 m away from natural waterways

clearly identifying and separating all GM plants and plant material from non-GM plants and plant material

inspecting the field trial site for related species and flowering sugarcane once per month and increase the inspections to twice a week beginning in April

harvesting GM sugarcane at a time before flowering is likely to occur

removing any sporadic flowers manually, bagging and destroying them in the GM waste holding area

harvesting and processing (juicing) GM sugarcane separately from any other sugarcane

cleaning of equipment after use with GM sugarcane

surrounding the GM waste holding area with a 1 m high chicken wire and electric fence

destroying plant material not required for experimentation or propagation by herbicide treatment, burning or a combination of both

following cleaning of sites, monitoring for and destroying any GM sugarcane that may grow for at least 12 months and until the site is free of volunteers for a continuous 6-month period

permitting only people trained in the licence conditions to deal with GM plants or plant material



not allowing the GM plant material or products to be used in human food or animal feed.

18. The applicant proposes to transport and store the GM plant material in accordance with Regulator’s Guidelines for the Transport, Storage and Disposal of GMOs (2011). For transport of plant material within research stations the applicant proposes alternative procedures. It is proposed for trays of seedlings to be transported in covered trailers. For transport of sugarcane plant material such as stalks, the applicant proposes to use a covered flat bed of a truck ensuring no material is lost.

19. Figure 1 shows the proposed planting layout including some of these controls. The controls, and the limits outlined above, have been taken into account in establishing the risk assessment context (this Chapter), and their suitability for containing the proposed release is evaluated in Chapter 33.1.

Figure 1. Field trial layout including some of the controls proposed by the applicant (not drawn to scale)

Section 4 The parent organisms20. The parent organism is cultivated sugarcane, Saccharum spp., which is an interspecific hybrid of S. spontaneum and S. officinarum. Sugarcane is exotic to Australia and is commercially cultivated on the east coast of Australia from northern New South Wales (NSW) to far north Queensland (Qld).

21. Approximately 80% of the chromosomes in commercial sugarcane cultivars are derived from S. officinarum, 10% from S. spontaneum with the remainder being hybrid chromosomes from the two species (D'Hont et al. 1996; Piperidis et al. 2000). Repeated back-crossing of initial hybrids to a female S. officinarum parent produced the current commercial cultivars with higher sugar accumulation, cane yield and many desirable agronomic and disease-resistance traits.

22. Detailed information about the parent organism is contained in the reference document The Biology of the Saccharum spp. (Sugarcane) (OGTR 2011), which was produced to inform the risk assessment process for licence applications involving GM sugarcane plants. This document is available from the OGTR website or on request from the OGTR.


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


Section 5 The GMOs, nature and effect of the genetic modification5.1 Introduction to the GMOs23. The applicant proposes to release up to 1000 lines of GM sugarcane. The lines were produced using biolistics-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.

24. The insertion of genes into sugarcane using biolistics has been shown to be stable (Bower et al. 1996; Hansom et al. 1999). However, it is possible that expression of genetic changes may alter during successive cycles of vegetative propagation through various epigenetic effects such as change in methylation patterns and gene silencing phenomena. It is also possible that some GM sugarcane plants are chimeric and vegetative reproduction may result in loss of the genetic change.

25. The genetic modifications are expected to confer enhanced sugar content to the GM sugarcane.5.1.1 The introduced gene sequences and other introduced genetic elements

26. Up to four gene sequences have been introduced in the GM sugarcane lines. Two gene sequences are from sugarcane and two gene sequences are from Escherichia coli (Table 2). The identities of the introduced sugarcane gene sequences were declared Confidential Commercial Information (CCI) under section 185 of the Gene Technology Act 2000. The confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application. The GM sugarcane lines contain the antibiotic resistance selectable marker genes, nptII and bla, which were originally derived from the common gut bacterium Escherichia coli. The bla gene encodes β-lactamase, which confers resistance to ampicillin. This gene has a bacterial promoter that does not function in plants, so the gene is not expressed in the GM sugarcane plants. The gene was used to select for bacteria containing the desired genes, in the laboratory, prior to the production of the genetically modified plants. The nptII gene encodes the enzyme neomycin phosphotransferase and confers kanamycin or neomycin resistance on the GM plant. The nptII gene was used as a selective marker during early stages of development of the GM plants in the laboratory. Further information about marker genes can be found in the document Marker genes in GM plants , available from the Risk Assessment References page on the OGTR website.

27. Short regulatory elements (promoters, enhancers, introns and terminators) are used to control expression of the introduced genes. Regulatory elements used in GM sugarcane come from maize (Zea mays), E. coli, Agrobacterium tumefaciens, rice (Oryza sativa) and sugarcane (Table 2). The identity of some regulatory elements have been declared CCI by the Regulator. The confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

28. Promoters are DNA sequences that allow RNA polymerase to bind and initiate correct transcription. The promoters proposed to be used in the GM sugarcane lines are listed in Table 2. The Ubiquitin1 (Ubi1) promoter used for the nptII selectable marker gene was obtained from maize. It is a constitutive promoter which has been frequently used in plant genetic modification (Christensen et al. 1992).

29. Also required for gene expression in plants are messenger ribonucleic acid (mRNA) terminators, including a poly-adenylation signal. The mRNA terminators proposed to be used in the GM sugarcane lines were derived from the nopaline synthase (nos) gene from A. tumefaciens and other terminators from rice and sugarcane. The nos terminator has been used in a wide variety of constructs for plant genetic modification (Reiting et al. 2007).


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

http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/marker-genes-ref-1-htm



Although A. tumefaciens is a plant pathogen, the regulatory sequences comprise only a small part of its total genome, and are not capable of causing disease.



Table 2 Genetic elements introduced in the GM sugarcane. Red text denotes information that has been declared CCI by the Regulator. The confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

CategoryPromoter Intron Gene elements Terminator

Genetic element Origin Genetic

element Origin Function Genetic element Origin Function Genetic

element Origin

IW Maize

Intron Sugarcane Expression manipulation

Gene I

Sugarcane

Selected elements for manipulating

gene expression

Specific elements to terminate

gene expression

A. tumefaciens, rice or sugarcane

X Sugarcane

IIY Sugarcane

Gene 2Z Sugarcane

Selection marker

Ubiquitin (Ubi) Maize

Intron from shrunken-1

locusMaize

Enhanced nuclear export of transcripts

Aminoglycosi-dephospho-transferase

(aph A = npt II)

E. coliSelection of transformed

plant cells

Nopaline synthase

(nos)A. tumefaciens

Selection marker

Beta-lactamase

(bla)E. coli

Selection of transformed

bacterial cells



5.2 Toxicity/allergenicity of the proteins associated with the introduced genes30. The introduced gene sequences were obtained from sugarcane and E. coli, a common bacterium, and homologues of their encoded proteins occur naturally in a range of organisms, including plants and other food crops widely consumed by people and animals. On this basis, humans and other organisms have a long history of exposure to these gene sequences, or highly homologous variants, and their expressed proteins. Information about the identity of the introduced sugarcane gene sequences was declared CCI by the Regulator and was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

31. Analyses of the npt II and bla sequences were performed for previous licence applications (eg DIR 019/2002, DIR 070/2006) and information is available at the OGTR website. It was concluded that there is no evidence to indicate that the proteins encoded by the npt II and bla are toxic and they have a low likelihood to be allergenic (OGTR website).

32. No studies on the toxicity or allergenicity of the GM sugarcane lines and their products have been undertaken to date as the proposed trial is at an early stage. Such studies may have to be conducted if approval was sought for the GMOs or their products to be considered for human food and/or animal feed in Australia.

5.3 Characterisation of the GMOs5.3.1 Stability and molecular characterisation

33. As this is early stage research, no information about cross generation stability of the GM trait was provided.

34. Studies on other genes in sugarcane show that the introduced genes and traits remain stable across generations and transmit through progenies (Butterfield et al. 2002; Casu et al. 2005; Rae et al. 2005).5.3.2 Phenotypic characterisation

35. The applicant states that the growth of the GM lines is identical to non-GM parental lines. Following harvest and juicing, GM sugarcane plants and extracted juice would be subjected to compositional analysis.

Section 6 The receiving environment36. The receiving environment includes: any relevant biotic/abiotic properties of the geographic regions where the release 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 2013).

37. The factors relevant to the growth, distribution and cultivation of commercial sugarcane can be found in The Biology of the Saccarum spp. (Sugarcane) (OGTR 2011).

38. The proposed dealings involve planting of GM sugarcane at the SRA Durre farm (near Ayr) in Burdekin (Figure 2). Hardening, growing (in pots) and juice extraction for analysis of GM plants is proposed to occur at the SRA Burdekin Station, Burdekin.





Figure 2 Geographic distribution of the Australian sugarcane industry. Red dot ( ) indicates the Burdekin region



39. The SRA sites are in agricultural areas on the outskirts of a rural town and can only be accessed via private roads. Commercial sugarcane is grown in the immediate vicinity of the SRA Durre farm and SRA Burdekin Station. Other GM and non-GM sugarcane trials are also conducted on these properties.

6.1 Relevant abiotic factors40. The abiotic factors relevant to the growth and distribution of commercial sugarcane in Australia are discussed in The Biology of the Saccharum spp. (sugarcane) (OGTR 2011). They include nutrient stress, temperature, salt tolerance, aluminium tolerance and water availability.

41. The release is proposed to take place in the Qld local government area of Burdekin. The region has a tropical climatic type as defined by the Koeppen Classification system used by the Australian Bureau of Meteorology.

42. The proposed field trial site is on flat arable land not subject to flooding. The GM sugarcane field trial site would be located at least 50 m from the nearest waterway. To date, no incidents of dispersal of GM sugarcane from field trials as a result of cyclones or storms have been reported to the OGTR.

6.2 Relevant biotic factors43. The biotic factors pertaining to the growth and distribution of commercial sugarcane are discussed in The Biology of the Saccharum spp. (sugarcane) (OGTR 2011). In addition, the following points are of particular relevance to this release:

The proposed release site is on SRA Durre station which is surrounded by commercial sugarcane growing fields.

Invertebrates, vertebrates and microorganisms are expected to be exposed to the GM sugarcane. Fauna, including rats and feral pigs may have access to the sites. To date there is no record of dispersal of sugarcane by these vertebrates.

Cut sugarcane stalks left in the field are degraded by microorganisms.

6.3 Relevant agricultural practices44. In Qld, commercial sugarcane is planted from autumn to spring and harvested after 12-18 months. The GM sugarcane is proposed to be grown in the field, and data collection and harvest would likely occur before flowering. GM sugarcane plants are either ratooned or killed.

Planting material

45. GM sugarcane plants proposed for release have been, or will be, produced in Physical Containment Level 2 (PC2) laboratories and raised and tested in glasshouses. No plants would be raised from seeds. Test plants would be transported to the nursery at the SRA Burdekin Station to be hardened on seedling benches for 8-10 weeks prior to planting and then transported to the trial site at SRA Durre farm.

46. Hot water treatment prior to planting of stem cuttings (setts) is a best management practice in commercial sugarcane cropping to prevent fungal infection. The applicant has proposed that the hot water treatment facility approved under licences DIR 096 and DIR 129 would also be used for DIR 135. Burdekin Productivity Services (BPS) will be contracted to provide this treatment prior to planting of setts in the field. GM sugarcane will be treated in tanks housed in secure access facilities. The procedure involves two steps, hot water treatment and cold water treatment. Whereas usually each treatment is conducted in a separate tank, one tank would be used for both treatments of the GM sugarcane. This is proposed to minimise the opportunity of accidental dispersal of GM sugarcane material.



47. Stem cuttings would be germinated in the nursery and transported to the field trial site. Field preparation and planting of GM plants will be similar to non-GM plants following standard techniques used to grow sugarcane.

Post-harvest procedures

48. The applicant proposes that all harvested plant material not used for experimental purposes or propagation would be stored in the GM waste holding area and destroyed as soon as practical. After each harvest, the GM trial site would be burnt. After the final harvest, the GM trial site would be burned, deep-ploughed and then inspected for sugarcane volunteers. Any volunteers would be destroyed by herbicide treatment, or ploughing, or both. All field sites would be inspected for at least 12 months and until volunteer-free for at least 6 months continuously.

6.4 Presence of related plants in the receiving environment49. The field trial is located within a commercial sugarcane growing region, and commercial sugarcane is grown adjacent to the proposed field trial site. The SRA Durre farm is used for breeding of commercial cultivars and for field trials of GM herbicide tolerant sugarcane authorised under DIR 096. This licence was issued to Sugar Research Australia and planting is permitted until November 2015. The proposed UQ GM sugarcane field trial site would be located at least 6 m from other all other sugarcane, both GM and non-GM.

50. Sugarcane is known to cross with other species within the Saccharum genus, however of these species, only S. spontaneum and S. officinarum are known to occur in Australia (OGTR 2011). Both species occur as discrete populations. Other members of the genus are kept in various Australian germplasm collections where S. officinarum, S. spontaneum and S. robustum plants are maintained. These species are all sexually compatible with cultivated sugarcane. Naturalised populations of S. spontaneum have been recorded at several locations in northern Qld, including in sugarcane growing areas along a significant part of the Mulgrave and Herbert rivers (reviewed by Bonnett et al. 2007; reviewed by Bonnett et al. 2010; Bonnett et al. 2008). Preliminary molecular analysis of the Mulgrave River population suggested the plants have reproduced vegetatively (Bonnett et al. 2008). The Herbert River population is genetically diverse, and seed from that population is the product of cross-pollination (Pierre et al. 2015).

51. Sugarcane has been reported to produce hybrids with a number of species of closely related genera in a group known as the Saccharum complex. However, these hybrids have usually occurred under controlled experimental conditions, often with Saccharum, in particular S. officinarum, as the female (Aitken et al. 2007; Nair 1999; Wang et al. 2009). Genera for which hybridisation has been established are Erianthus and Miscanthus, but these exotic species do not occur in Qld (reviewed by Bonnett et al. 2008). Possible hybridisations to the genera Narenga, Imperata, Schlerostachya and Miscanthidium have been reported, but these events have not been verified by molecular methods (reviewed by Bonnett et al. 2008). Blady grass (Imperata cylindrica) is common throughout Qld coastal areas, however it is not known if it is also present at the proposed field trial site.

52. Sugarcane has been reported to cross with a number of species not considered to be close relatives (reviewed in OECD 2013). A small number of hybrids between sugarcane and sorghum (Sorghum bicolor) have been generated from experiments in which large amounts of S. officinarum pollen were used to pollinate male sterile sorghum flowers (Nair 1999). Crosses of sorghum as the pollen donor with commercial Saccharum hybrids have been reported, but also under controlled conditions. The few hybrids produced by the above crosses lacked vigour and showed slow growth (Nair 1999). Hybrids between Saccharum and sorghum have not been observed under natural conditions (reviewed by Bonnett et al. 2008). Similarly, following hand



pollination of thousands of S. officinarum florets with maize pollen, only a single hybrid plant was produced (reviewed by Bonnett et al. 2008). Wild Sorghum species occur as widespread weeds of cultivated areas in Qld. According to the applicant, maize and sorghum are not cultivated in close proximity to the proposed trial sites although maize is grown commercially in the Burdekin region. Although hybridisation between Saccharum and Bambusa (bamboo) has been reported (Rao et al. 1967), subsequent analysis has suggested the hybridisation was not genuine (Grassl 1980).

6.5 Presence of introduced gene sequences in the environment53. The introduced gene sequences were isolated from the naturally occurring plant sugarcane, and from E.coli, a common gut bacterium. People routinely encounter these gene sequences and their gene products (or homologues) through everyday activities, including the consumption as food.

54. Short regulatory sequences are derived from plants (including maize, rice and sugarcane) and common bacteria (A. tumefaciens and E. coli). Some of the sequences are derived from a plant pathogen (A. tumefaciens) but comprise only a small part of the pathogen’s total genome.

Section 7 Relevant Australian and international approvals7.1 Australian approvals7.1.1 Approval by the Regulator

55. The GM sugarcanes proposed for release have not been released previously in Australia.

56. The Regulator has previously issued licences for the limited and controlled release (field trial) of GM sugarcane with different introduced traits. Information on previous DIR licences can be found on 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 Approval by other government agencies

57. The Regulator is responsible for assessing risks to the health and safety of people and the environment as a result of gene technology. Other government regulatory requirements may also have to be met in respect of release of GMOs.

58. Food Standards Australia New Zealand (FSANZ) is responsible for human food safety assessment and food labelling, including GM food. The applicant does not intend to use materials from the GM sugarcane lines in human food, and, accordingly no application has been submitted to FSANZ. FSANZ approval would need to be obtained before materials from these GM sugarcane lines could be sold as food or food ingredients.

7.2 International approvals of GM sugarcane59. None of the GM sugarcane lines have been released overseas.


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


Chapter 2 Risk assessmentSection 1 Introduction60. 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 3). 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 3 The risk assessment process

61. 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 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.

62. 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 2013). 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). In addition, relevant risk scenarios postulated in previous RARMPs prepared for licence applications of the same and similar GMOs are also considered.

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

Chapter 2 – Risk assessment 13


64. 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 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 Identification65. Postulated risk scenarios are comprised of three components:

the source of potential harm (risk source)

a plausible causal linkage to potential harm (causal pathway)

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

66. 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

characteristics of the parent organism(s).

67. Some information relevant to the risk assessment has been declared CCI by the Regulator and was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

2.1 Risk source68. 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.

69. As discussed in Chapter 1, the proposed GM sugarcane lines will be modified by the introduction of sugarcane gene sequences. These GM sugarcane lines with enhanced sugar content are considered further as potential sources of risk.

70. In addition, the GM sugarcane lines also contain selectable marker genes neomycin phosphotransferase type II gene (nptII, also referred to as aminoglycoside phosphotransferase gene, aphA) and beta-lactamase gene (bla). These genes and their products 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 marker genes can be found in the document Marker genes in GM plants available from the Risk Assessment References page on the OGTR website. As these genes have not been found to pose a substantive risk to either people or the environment, their potential effects will not be further considered for this application.

71. The introduced gene sequences are controlled by regulatory sequences, which are derived from maize (Zea mays), sugarcane (Saccharum. spp), rice (Oryza sativa) and A. tumefaciens. Such regulatory sequences are naturally present in all 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 one




bacterial sequence is derived from a plant pathogen, it only constitutes a very small fraction of the genome and cannot itself cause disease. However, regulatory sequences control gene expression and can thereby influence biochemical pathways in the GM plants. The effects of any potential changes, in particular potential changes to toxicity and allergenicity, will be discussed below.

2.2 Causal pathway72. 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 (eg climate, soil and rainfall patterns)

tolerance to biotic stressors (eg pest, pathogens and weeds)

tolerance to cultivation management practices

gene transfer to sexually compatible organism

gene transfer in a manner other than by sexual reproduction (horizontal gene transfer)

unauthorised activities.

73. 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.

74. The potential for unauthorised activities to lead to an adverse outcome has been considered in many previous RARMPs, most recently in 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 for this application.





2.3 Potential harm75. 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.

76. 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 scenarios77. Three risk scenarios were postulated and screened to identify substantive risk. These scenarios are summarised in Table 3 and more detail of these scenarios is provided later in this Section. Postulation of risk scenarios considers impacts of the GM sugarcane 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.

78. 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.

79.



Table 3 Summary of risk scenarios from dealings with sugarcane genetically modified for enhanced sugar content

Risk scenario

Risk source

Causal pathway Potential harm Substantive risk?

Reason

1 GM sugarcane with enhanced sugar content

Cultivation of GMOs at trial sites

Exposure of people who deal with the GMOs or other organisms that come into contact with the GMOs at the trial sites

Allergic reactions in people or toxicity in people and other desirable organisms

No The limited scale, short duration and other proposed limits and controls minimise exposure of people and other organisms to the GM plant material.

GM plant material would not be used in human food or animal feed.

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

2 GM sugarcane with enhanced sugar content

Persistence of the GMOs at the field trial site or dispersal of GM seed and plant material outside trial limits

Spread and persistence of populations of GM plants

Exposure of people or other organisms to GM plant material

Allergic reactions in people or toxicity in people and other desirable organismsReduced establishment and yield of desirable plantsReduced biodiversity

No The proposed limits and controls minimise the likelihood for dispersal of GMOs.

Environmental factors are expected to limit the spread and persistence of sugarcane in the area proposed for release.

3 Introduced genes for enhanced sugar content in the GM sugarcane

Dispersal of pollen outside the trial limits

Vertical transfer of introduced gene sequences to other sexually compatible plants


Allergic reactions in people or toxicity in people and other desirable organismsReduced establishment and yield of desirable plantsReduced biodiversity

No Sugarcane pollen viability is very low under natural conditions.

The GM sugarcane would be harvested before flowering.

Other proposed limits and controls would further minimise the likelihood of pollen flow to any sexually compatible plants.



2.4.1 Risk scenario 1

Risk source GM sugarcane with enhanced sugar content

Causal pathway

Cultivation of GMOs at trial sites

Exposure of people who deal with the GMOs or other organisms that come into contact with the GMOs at the

trial sites

Potential harm Allergic reactions in people or toxicity in people and other desirable organisms

Risk source

80. The source of potential harm for this postulated risk scenario is the GM sugarcane with enhanced sugar content.

Causal pathway

81. People involved in propagating, cultivating, harvesting, transporting and processing (juicing) the GM sugarcane or in experimenting with the GM sugarcane may be exposed to the GM plant material or products. As the applicant proposes that only authorised staff deal with the GM sugarcane, 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 GM plant material or its products, as the applicant proposes that GM plant material or its products would not be used in food. GM plant material that could be airborne and inhaled includes pollen. The applicant proposes to harvest the GM sugarcane before flowering is likely to occur and, therefore, the potential for exposure through inhalation of pollen is low. Workers could come into skin contact with GM plant material if they touch damaged plants where cell contents have been released. However, sugarcane plants possess leaves with sharp edges and, as such, workers typically wear protective clothing which would reduce dermal contact.

82. In addition to people, other organisms may be exposed directly to the GM plant material 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 GM plant material as it is not to be used as animal feed. The limited scale and duration of the proposed field trial would restrict the exposure of animals to the GM sugarcane.

83. After harvest, the applicant proposes to destroy all the GMOs not used for further research purposes or future planting.

84. The applicant has also proposed other measures, such as monitoring and inspecting the trial sites, cleaning equipment, and disposing of waste material, all of which would further reduce exposure of people and the environment to the GM sugarcane.

Potential harm

85. Expression of the introduced gene constructs in plant tissues modifies the expression of other genes. Changes in gene expression lead to changes in the level of sugars in the GM sugarcane. This could lead to changes in the GMOs that could increase allergenicity to humans and toxicity of the GM plants to humans and other desirable organisms.

86. 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).



87. The genetic modifications could lead to the production of substances in the GM sugarcane that are toxic or allergenic for people or toxic to other desirable organisms. All known food allergens are proteins, those derived from plants coming chiefly from peanut, tree nuts, wheat and soybean (Delaney et al. 2008; Herman & Ladics 2011). The introduced gene sequences are from sugarcane, which is widespread in the Australian environment (Chapter 1Section 4). The introduction of the gene constructs does not lead to expression of a novel protein which is from any class of proteins having known toxic or allergenic members (Radauer & Breiteneder 2007). Sugarcane pollen was the major allergen when field workers with respiratory disorders in India were tested for allergic responses (Chakraborty et al. 2001). However, there are no reports of any major allergic responses to commercial hybrid cultivars of sugarcane in Australia. In these circumstances, there is no reasonable expectation that the introduced constructs will lead to an increase in the level of any endogenous compound in the GM sugarcane that has toxic or allergenic properties.

88. 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; Weber et al. 2012).

89. The introduction into sugarcane of any of the genes that are the subject of this application is unlikely to result in the production (directly or indirectly) of a novel toxin or allergen. However, there is some uncertainty as to what extent the genetic modification could alter the characteristics of the GMO and this uncertainty is addressed in Chapter 3. In summary, the range of possible unintended effects produced by genetic modification is not likely to be greater than that from accepted traditional breeding techniques (Bradford et al. 2005; Committee on Identifying and Assessing Unintended Effects of Genetically Engineered Foods on Human Health 2004). More detail on the potential for unintended effects as a result of the process of genetic modification can be found in the document Methods of plant genetic modification available from the Risk Assessment References page on the OGTR website.

90. Further information about the GMOs relevant to consideration of the risk scenario was declared CCI by the Regulator. This confidential information was made available to the prescribed experts and agencies that were consulted on the RARMP for this application.

Conclusion

91. Risk scenario 1 is not identified as a substantive risk because the introduced genetic elements are already widespread in the Australian environment, GM plant material would not be used in human food or animal feed, the biosynthetic pathway they affect is involved in the production of compounds that have not been associated with toxic or allergenic reactions, and the proposed limits and controls would minimise exposure of people and other organisms to the GM plant material. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.




2.4.2 Risk scenario 2

Risk source GM sugarcane with enhanced sugar content

Causal pathway

Persistence of the GMOs at the field trial site or dispersal of GM seed and plant material outside trial limits

Spread and persistence of populations of GM plants


Potential harm

Allergic reactions in people or toxicity in people and other desirable organismsor

Reduced establishment and yield of desirable and yield of desirable plantsor

Reduced biodiversity

Risk source

92. The source of potential harm for this postulated risk scenario is the GM sugarcane with enhanced sugar content.

Causal pathway

93. People and other organisms may be exposed to GM plant materials beyond the limits of the trial. This could occur due to persistence of viable GM seeds or stalks at the trial site after the proposed trial or through physical movement of GM seeds and stalks to areas outside the trial sites.

94. Baseline information on the characteristics of weeds in general, and the factors limiting the spread and persistence of non-GM sugarcane plants in particular, is given in The Biology of the Saccharum spp. (sugarcane) (OGTR 2011). In summary, the document concludes that modern cultivars of non-GM sugarcane (Saccharum spp.) are not invasive in natural undisturbed environments and do not appear to be a problem as volunteer weeds in Australia and occur almost exclusively as a managed agricultural crop.

95. The establishment, spread and persistence of sugarcane populations is likely to be limited by complex interactions involving weed competition, pest infestation, disease infection, moisture stress, soil fertility and other environmental conditions (OGTR 2011; Bakker 1999; Hogarth & Allsopp 2000; Pierre et al. 2014).

96. Persistence of GM sugarcane at the field trial site could occur if GM plant material, such as stem segments or seed, were to remain on site after harvest. The applicant proposes a number of control measures to prevent persistence of GM sugarcane at the field trial site. These include destroying GMOs that remain in the trial sites after harvest, post-harvest monitoring 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. These measures would minimise the potential for persistence of GM sugarcane after the trial and the potential for its dispersal. It is not expected that the introduced gene sequences would significantly increase dormancy levels or otherwise increase the ability of the GMOs to survive these standard control measures.

97. Dispersal of GM plant material outside the limits of any trial site could occur through the activity of people (including the use of agricultural equipment), the activity of animals such as rodents, feral pigs, herbivores and birds, or through extremes of weather such as flooding or high winds.

98. If reproductive plant material and seed was dispersed outside any of the trial sites, these could give rise to plants expressing the introduced genetic modification. These plants could spread and persist in the environment outside the trial limits, and people and other organisms may be exposed to GM plant materials.



99. As discussed in the RARMPs for DIR 096 and DIR 129, viable sugarcane stems could be unintentionally dispersed from the sites during transportation. Sugarcane volunteers have been found growing along roadsides or railways in sugarcane cultivation areas. These volunteers are believed to have originated from stem cuttings fallen from vehicles during transportation and generally consist of only a few stools (a group of stems growing from a single original plant base) and normally do not become self-perpetuating or result in further spread.

100. Transport of GM sugarcane plants and setts to and from SRA Burdekin station and SRA Durre farm is proposed to be conducted according to transport guidelines issued by the Regulator. The applicant has proposed different protocols for transport within the SRA Burdekin Station and SRA Durre farm.

101. For transport of sugarcane material such as stalks or waste, the applicant proposes to double contain, label and secure GM plant material in a truck or trailer. The truck will be followed to ensure that no material is lost during transport. This would reduce the risk of dispersal of the GMOs. In the unlikely event that viable GM plant material was dispersed it would be unlikely to germinate, as appropriate environmental conditions are necessary for survival and persistence of any dispersed setts. For example, soil-borne fungal infections are known to reduce sett germination, leading to the common practice of treating the setts with fungicide prior to planting (FAO 2004; FAO 2001).

102. In the nursery at SRA Burdekin station, the applicant proposes to clearly label and separate the GM sugarcane from any other sugarcane. Thus clear identification of GM plants is proposed to reduce the potential for accidental mixing through human error.

103. During harvesting of GM sugarcane, accidental mixing with non-GM sugarcane could occur through, for example, harvester driver error. This could lead to the GM sugarcane being inadvertently treated as non-GM sugarcane, thus allowing dispersal from the site as a component of a non-GM sugarcane harvest, or mixed with non-GM sugarcane that may be used for replanting at other locations. At the field site, the applicant proposes to separate GM sugarcane from any adjacent sugarcane plots by a 2 m guard row of non-GM sugarcane and a 6 m isolation zone. The isolation zone would clearly separate the GM sugarcane from neighbouring non-GM sugarcane. In addition, the applicant proposes to harvest the GM sugarcane before the start of harvest of commercial sugarcane in the area. These measures minimise the likelihood of accidental mixing at harvest.

104. On completion of the trial the sugarcane plant material would be destroyed by a combination of harvesting, deep ploughing and burning. Stalks of harvested GM sugarcane may be inadvertently left in the field. Small stem pieces are not expected to remain viable over long periods in the field, as they are degraded rapidly by soil micro-organisms under tropical conditions. Normally, stem cuttings are dipped in fungicide to increase their chances of survival (McMartin 1949).

105. The sexual reproduction of sugarcane has been little studied in the field, as seed production is not important for sugarcane cropping. To minimise any risk of dispersal by seeds, the applicant proposes to harvest the GM sugarcane plants before any flowering occurs.

106. Sugarcane seed is short-lived, losing 90% of viability within 80 days at 28ºC unless it is desiccated (Rao 1980). More recent data suggests that seed can remain viable for at least 2-3 months when stored at room temperature and that viability is somewhat cultivar dependent (Powell et al. 2008). Sugarcane seed needs appropriate environmental conditions for germination (eg. temperature and humidity) and these are seldom present in Australia without human intervention (Pierre et al. 2014). Furthermore, seedlings require particular favourable environmental conditions (eg. temperature and humidity) to survive for the first three to four weeks after germination (Breaux & Miller 1987) and normally human intervention is required



to successfully establish sugarcane seedlings. It is therefore highly likely that environmental factors, such as humidity and fluctuations in temperature, would affect seed viability and limit the survival and establishment of any dispersed seeds.

107. Observations over a number of decades indicate a lack of seed germination in regions south of the Burdekin region (the Burdekin region refers to the region around Ayr, south of Townsville) (Sugarcane Researchers 2008), and only low numbers of seedlings north of the Burdekin region (see Figure 2). For example, there are isolated reports of seeds germinating naturally in the Mulgrave, South Johnstone and Herbert River sugarcane growing regions, but not further south (Bonnett et al. 2010). It is unknown whether any field-germinated seedlings survive to maturity.

108. The spread and persistence of the GM sugarcane plants would also be limited by a number of environmental factors that limit the spread and persistence of sugarcane in Australia such as humidity, temperature, , nutrients and pests and diseases, in addition to sugarcane’s low intrinsic competitive ability, low fertility and seed viability, poor ability of seedlings to establish and compete without human intervention (OGTR 2011; Bakker 1999; Hogarth & Allsopp 2000; Pierre et al. 2014).

109. The impact of the genetic modifications on survival of the GM sugarcane plants is uncharacterised under field conditions; however some predictions can be made based on the genetic modification. These predictions do not indicate that the GM sugarcane have any selective advantage compared to the non-GM parent as the genetic modification is not expected to directly influence sugarcane characteristics involved in environmental stress tolerance. However, this is an area of uncertainty. Water stress and prolonged periods of waterlogging can result in a reduction in sugar production, while high soil salinity can affect the sucrose content of the stalk (FAO 2004; Rozeff 1995). It is not known if enhanced sugar content will in turn influence the ability of sugarcane plants to tolerate drought and salinity. It is also possible that sugar content may influence degradation of sugarcane stem segments by microorganisms.

110. In addition, a reduction in plant vigour is routinely observed in sugarcane plants which have undergone tissue culture. Data provided by the applicant from field trials under licence DIR 070/2006 show that plant height, stalk number, stem diameter and cane yield were reduced in GM sugarcane expressing nptII compared to untransformed sugarcane. These effects are expected to generally decrease the competitiveness of GM sugarcane.

111. Severe weather conditions such as flooding may cause dispersal of GM plant material. However, control measures have been proposed by the applicant to minimise dispersal by flooding. These include locating the proposed release field site at least 50 m from natural waterways on land that is not known to flood. Sugarcane stalks generally do not break upon flooding, unless significant water flow occurs.

Potential Harm

112. If the GM sugarcane plants were to establish or persist better in the environment than non-GM sugarcane, then this could lead to one or more harms. The potential for increased allergenicity in people or toxicity in people and other desirable organisms as a result of contact with GM plant materials at the trial sites has been considered in Error: Reference source not found and was not considered to be a risk that would be greater than negligible.

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



114. Potential toxicity of the GM sugarcane could reduce biodiversity. However, as discussed in Risk Scenario 1, the introduced gene sequences are already present in non-GM sugarcane and are not expected to result in the production (directly or indirectly) of a novel toxin or allergen. Still, there is some uncertainty about unintended effects due to the genetic modification that could lead to an increase in potentially toxic metabolites.

Conclusion

Risk scenario 2 is not identified as a substantive risk because the biochemical pathway the introduced genetic elements affect is not associated with toxic or allergenic compounds, the introduced gene sequences are already present in the environment and the modified trait is not associated with weediness. Further, the proposed limits and controls are designed to minimise exposure of people and other organisms to the GM plant material and restrict plant dispersal. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.2.4.3 Risk scenario 3

Risk source Introduced genes for enhanced sugar content in the GM sugarcane

Causal pathway

Dispersal of GM pollen outside trial limits

Vertical transfer of introduced genes to other sexually compatible plants


Potential harm

Allergic reactions in people or toxicity in people and other desirable organismsor

Reduced establishment and yield of desirable plantsor

Reduced biodiversity

Risk source

115. The source of potential harm for this postulated risk scenario is the introduced genes for enhanced sugar content in the GM sugarcane.

Causal pathway

116. The first step of this pathway is that pollen from the GM sugarcane could be dispersed outside the limits of the trial. Pathways include:

(a) Dispersal of pollen from GM sugarcane to sexually compatible plants during the trial, or

(b) Dispersal of pollen outside the trial limits to sexually compatible plants leading to the persistence of the genetic modification in the environment.

117. Sexually compatible plants include commercial non-GM sugarcane as well as GM sugarcane trialled under other DIR licences. Other plant species are also somewhat compatible under controlled conditions (Chapter1 Section Chapter 16.4). Any hybrids resulting from fertilisation with GM sugarcane pollen could pass the genetic modification on to their offspring.

118. People and other organisms could be exposed to the GM sugarcane with enhanced sugar content through contact with or consumption of GM plant material.

119. Vertical gene flow is the transfer of genetic information from an individual organism to its progeny by conventional heredity mechanisms, both asexual and sexual. In flowering plants, pollen dispersal is the main mode of gene flow (Waines & Hegde 2003). For GM crops, vertical gene flow could therefore occur via successful cross-pollination between the crop and



neighbouring crops, related weeds or native plants (Glover 2002). Alternatively, if seed was dispersed outside the trial site, GM plants may grow and subsequently disperse pollen. Hybrid plants possessing the introduced genes may form the basis for the spread of these genes in other varieties of sugarcane, or other sexually compatible plant species.

120. It should be noted that vertical gene flow per se is not considered an adverse outcome, but may be a link in a chain of events that may lead to an adverse outcome.

121. Baseline information on vertical gene transfer associated with non-GM sugarcane plants can be found in The Biology of the Saccharum spp. (sugarcane) (OGTR 2011). In summary, sugarcane pollen viability is low in the areas proposed for release, commercial sugarcane varieties show low fertility, and crossing to plants outside of the Saccharum genus has rarely been observed. Thus, it is highly unlikely that crossing with sexually compatible plants would occur.

122. Sugarcane is cultivated within close proximity to the proposed trial site. GM sugarcane authorised under a different licence, non-GM and commercial sugarcane are cultivated within the SRA Durre farm at a minimum distance of 8 m from the proposed trial site. Within the SRA Burdekin Station, GM plants would be grown in pots and prepared for field planting; the GM sugarcane would not reach flowering at this location.

123. As described in The Biology of the Saccharum spp. (sugarcane) (OGTR 2011), sugarcane does not flower with great uniformity, and cultivars differ in their propensity to flower (Bonnett et al. 2007). Shortening day length appears to control the initiation of flowering (Moore & Nuss 1987) and environmental factors impact on the extent of flowering. The frequency of flowering in the Burdekin region and further south is generally lower than north of the Burdekin region; however, this has not been systematically recorded.

124. Sugarcane is principally a wind pollinated out-crosser with a low frequency of self-pollination. Environmental conditions including temperature, relative humidity and wind intensity have a great influence on pollen viability and pollen movement (discussed below). There is little information available on outcrossing rates and distances for sugarcane.

125. Sugarcane pollen desiccates rapidly and is not viable beyond 35 minutes at 26.5ºC and 65% relative humidity (Moore 1976; Venkatraman 1922). Sugarcane pollen viability is variable and is strongly influenced by temperature, being greatly reduced under night-time temperatures below 21°C (Berding 1981). In Qld, this leads to a general reduction in levels of pollen viability the further south sugarcane is cultivated. Pollen viability on inflorescences sampled from commercial crops in the Mulgrave region (north of the Burdekin region) was significantly higher than in samples from the Burdekin region, which showed very low pollen viability (Bonnett et al. 2007; Bonnett et al. 2010). These reports indicate that the frequency of flowering and production of viable pollen is very low in most parts of Qld except in northern parts of the state.

126. The applicant proposed to harvest all GM sugarcane before flowering, thus the proposed limits and controls of the trial (Chapter 1, Section Chapter 13.1 and Section Chapter 13.2) would restrict the potential for pollen development and pollen flow and gene transfer to sexually compatible plants. The applicant also proposes to perform post-harvest monitoring of the site for at least twelve months and until no volunteers are observed for a continuous six month period and to destroy any volunteer plants found at the site. This would ensure that any remaining GM sugarcane seeds or plants that were potentially the product of gene flow in these areas would be destroyed.

Potential harm



127. If the vertical transfer of the introduced genes from the GM plants causes the recipient species to spread and persist in the environment to a degree greater than normally found amongst these species, they may produce one or more harms. People who are exposed to the GM plant material through contact or consumption of GM plant material may show toxic or allergenic reactions, while other desirable organisms may show toxic reactions from consumption of GM plant material. The GM plants may act to reduce the establishment or yield of desired plants, and subsequently reduce biodiversity.

128. In the rare event of the vertical transfer of the introduced genetic material from the GM plants to other GM or non-GM sugarcane plants or sexually compatible species, it is expected that this material will have the same or very similar properties in the recipient as in the GM sugarcane. As discussed in risk scenario 1, the introduced genetic elements are not expected to result in GM plants that are allergenic to humans or toxic to humans or other desirable organisms. Risk scenario 2 summarises the rationale as to why the genetic modification is unlikely to make the GM sugarcane weedier. These reasons are applicable to any plants to which the genes are transferred via hybridisation. In the case of GM sugarcane with herbicide tolerance planted under DIR 096, there is no reasonable expectation that a combination of the genetic modifications would result in combinational effects as the genetic modifications affect totally unrelated biochemical pathways.

Conclusion

129. Risk scenario 3 is not identified as a substantive risk due to the expected lack of toxicity, allergenicity or increased weediness in any offspring of the GM plants and other plants, either commercial sugarcane crops or other sexually compatible plants. Further, the proposed limits and controls are designed to minimise pollen dispersal and sugarcane pollen viability is very low in the areas proposed for release. Therefore, this risk could not be greater than negligible and does not warrant further detailed assessment.

Section 3 Uncertainty130. Uncertainty is an intrinsic property of risk and is present in all aspects of risk analysis2. 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 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.

131. 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.

132. For DIR 135, uncertainty is noted particularly in relation to:

potential for increased toxicity or allergenicity due to unintended effects

potential for increased weediness of the GMOs due to unintended phenotypic changes.

133. 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.

2 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


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

Section 4 Risk Evaluation135. 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.

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

risk criteria

level of risk

uncertainty associated with risk characterisation

interactions between substantive risks.

137. 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 3 and include:

limits on the size, locations and duration of the release proposed by UQ

controls proposed by UQ to restrict the spread and persistence of the GMO

widespread presence of the same and similar genes in the environment and lack of evidence of harm from them

limited ability of sugarcane to establish populations outside cultivation

limited ability and opportunity for the GM sugarcane plants to transfer the introduced genetic material to commercial sugarcane crops or sexually related species

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

138. Therefore, risks to health and safety of people, or the environment, from the proposed release of the GM sugarcane plants into the environment are considered to be negligible. The Risk Analysis Framework (OGTR 2013), which guides the risk assessment and risk management process, defines negligible as risks of no discernible concern with no present need to invoke actions for mitigation. Therefore, no controls in addition to those proposed in the application 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

139. 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.

140. 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.

141. 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.

142. 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 risks143. The risk assessment of the risk scenarios listed in Chapter 2 concluded that there are negligible risks to people and the environment from the proposed field trial of GM sugarcane. These risk scenarios were considered in the context of the scale of the proposed release and the proposed control measures (Chapter 1, Chapter 1Section 3), and the receiving environment (Chapter 1 Chapter 1Section 6), and considering both the short and the long term. The risk evaluation concluded that no controls in addition to those proposed by the applicant are required to treat these negligible risks.

Section 3 General risk management144. 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 the licence.

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

145. Sections 3.1 and Chapter 13.2 of Chapter 1 provide details of the limits and controls proposed by UQ in their application. These are taken into account in three risk scenarios postulated for the proposed release in Chapter 2. Many of these 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.

Chapter 3 – Risk management 27


146. The duration of the field trial would be limited to five years. The trial would be limited to two locations, with field planting occurring only at one location with a maximum area of 5 ha. The small size and relatively short duration of the trial would limit exposure to the GMOs (Error: Reference source not found).

147. 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).

148. GM sugarcane in the hot water treatment facility will be labelled, contained inside a shade cloth bag and placed in the water tank. Hot water treatment of the GM sugarcane will be conducted separately from any other sugarcane. The treatment water will be filtered before release and the tank will be cleaned after use. These measures reduce the likelihood of dispersal of any GMO (Risk scenario 2).

149. The trial sites would be located more than 50 m from the nearest natural waterways on land which has minimal risk of flooding, which would reduce the likelihood of GMOs being washed away from the sites (Risk scenario 2).

150. At the field site, the applicant proposes to surround the GM sugarcane with a 2 m guard row of non-GM sugarcane and a 6 m isolation zone, separating it from any other sugarcane (except guard row) by at least 8 m. This physical separation would help to prevent inadvertent mixing of cane in adjacent plots, for example, as a result of harvester driver error, at the time of harvest. From a risk perspective, the 2 m guard row is not required as the 6 m isolation zone is considered sufficient to separate GM sugarcane from any other sugarcane. The 6 m isolation zone as a method for physical separation was therefore included in the licence (Risk Scenario 2).

151. At the SRA Burdekin Station, where hardening and raising of GM sugarcane is to occur, the applicant has proposed to cultivate GM sugarcane plants on seedling benches prior to planting in the field, using areas set aside for this purpose. Non-GM sugarcane and other GM sugarcane plants would also be cultivated in these facilities. The applicant proposes to keep this GM sugarcane on separate benches from any other sugarcane, separated by at least 1 m and labelled as GM material, including the implementation of a barcoding system to allow for identification of the GM sugarcane plants during the various stages of assessment. In order to further reduce the potential of unintended dispersal of potted GM sugarcane by, for example, workers unaware of restrictions on their movement, the licence conditions include a requirement that the GM plants be clearly identifiable, and to have signs indicating which benches contain GM material.

152. The applicant has proposed that GM sugarcane will be harvested before flowering and if any sporadic flowering occurs, the flowers will be removed manually. The applicant has also proposed to inspect the field trial site once per month before April and twice per week from April onwards. As discussed in risk scenario 3, sugarcane shows variable flowering in the Burdekin area, has low pollen viability and low seed set. However, little information is available regarding the time from flower onset to opening of flowers. One study from Egypt (Rizk et al. 2007) showed that opening of flowers occurred between 32 and 47 days after initiation of flowering. Therefore, the licence specifies that the field trial site is inspected once per month before April and once every two weeks from April onwards until harvest (Risk Scenario 3).

153. The applicant has stated that any plant material taken off-site for experimental analysis would be transported according to the Regulator’s Guidelines for the transport, storage and disposal of GMOs. These are standard protocols for the handling of GMOs to minimise



exposure of people and other organisms to the GMOs (Risk scenario 1), dispersal into the environment and gene flow/transfer (Risk scenario 2 and Risk scenario 3). As discussed in Risk scenario 2, transport within the SRA Burdekin station and SRA Durre farm is proposed to be conducted differently. For transport of sugarcane material such as stalks or waste, the applicant proposes to contain the material with two layers of tarpaulin, label and secure it to a truck or trailer. The truck will be followed to ensure that no material is lost during transport. Sugarcane stems are considered propagative plant material, and can potentially give rise to new sugarcane plants. This proposed method of transportation is effectively a lower level of containment than specified by the Regulator’s Transport Guidelines. However, for the purpose of transporting GM sugarcane within the SRA station or field trial site this level of containment is considered appropriate and is included in the licence. Transport outside the field trial site and the SRA station is required to occur according to the Regulator’s Guidelines and this is included in the licence.

154. The applicant has proposed to destroy any viable GM plant material remaining at the field site after final harvest by deep ploughing, burning and/or by herbicide treatment (Risk scenario2). Similar disposal and/or destruction methods have been used previously for GM sugarcane under the licences for DIR 019/2002, DIR 051/2004, DIR 070/2006, DIR 078/2007, DIR 095 and DIR 096. These are also used in the industry as standard destruction methods (OGTR 2011). These methods of destruction would minimise exposure to the GM plant material (Error: Reference source not found) and limit the likelihood of spread and persistence (Risk scenario 2). Therefore, deep ploughing, herbicide treatment and burning have been specified in the licence as methods of destruction of GM sugarcane.

155. The applicant has proposed to monitor the field release site for 12 months post-harvest and destroy any volunteer sugarcane, until no volunteers are observed for a continuous six month period. Sugarcane seed has little dormancy (Simpson 1990), and stem pieces capable of giving rise to new plants are not expected to remain viable over long periods in the field, as they are degraded rapidly by soil micro-organisms under tropical conditions. Normally, stem cuttings are dipped in fungicide to increase their chances of survival (McMartin 1949). The proposed time period is therefore considered appropriate for minimising the persistence of the GMOs in the environment, potential for dispersal (Risk scenario 2) and gene flow (Risk scenario 3). Therefore, the requirement to monitor the field release site monthly for at least 12 months and until no volunteers are observed for a continuous six month period, is included as a licence condition.

156. The applicant does not propose using any of the plant material for human or animal consumption. FSANZ conducts mandatory premarket assessments of GM products in human foods. As the GM sugarcane has not been assessed by FSANZ, a condition in the licence prohibits material from the trial from being used in human food or animal feed.3.1.2 Summary of licence conditions to be implemented to limit and control the release

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

limit the field planting to a maximum area of 5 ha at one site

limit the duration of dealings with GM sugarcane to between August 2015 and May 2020

locate the field trial sites at least 50 m away from natural waterways

surround the field trial location by an isolation zone of at least 6 m which must be kept free of sugarcane and related species



separate GM sugarcane material from other sugarcane when propagating seedlings or setts on seedling benches, and clearly identifying the GM sugarcane

separate GM from other sugarcane in the nursery by at least 1 m

inspect field trial location once per month from planting, and once every two weeks from April onwards, until harvest

harvest and process the GM sugarcane separately from any other sugarcane and before expected flowering

remove any inflorescences manually and as soon as practical if flowering occurs early

establish a secure holding area for storage and destruction of GM sugarcane

maintain containment of the GM sugarcane stalks during hot water treatment

carry out analysis of plant materials at the SRA Burdekin Station, SRA Durre farm or in PC2 laboratories

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

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

monitor the trial site and destroy any GM sugarcane that may grow for at least 12 months after final harvest and until no volunteers are observed for a continuous six month period

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

not allow the GM plant material or products to be used as human food or animal feed.

3.2 Other risk management considerations158. 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

159. 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.

160. On the basis of information submitted by the applicant and records held by OGTR, the Regulator considers UQ 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.



161. 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

162. UQ 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 sugarcane outside of the permitted areas.

163. UQ 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

164. 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, UQ 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

165. 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.

166. 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

167. 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.

168. 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.



169. 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 releases170. Additional information has been identified that may be required to assess an application for a commercial release of these GM sugarcane lines, or to justify a reduction in limits and controls. This includes:

additional molecular and biochemical characterisation of the GM sugarcane lines, particularly with respect to production of potential toxins or allergens

additional phenotypic characterisation of the GM sugarcane lines, particularly with respect to traits that may contribute to weediness.

Section 5 Conclusions of the consultation RARMP171. The RARMP concludes that the proposed limited and controlled release of GM sugarcane 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.

172. 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.



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Appendix A Summary of submissions from prescribed experts, agencies and authorities3

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 Comment

1 Noted agreement with the overall conclusions of the RARMP.

Noted.

The Regulator should consider a provision that allows removal of developing flower heads that occur early on specific plants.

This provision was included in the consultation version of the licence, and is retained in the issued licence.

The Regulator should consider clarifying in the RARMP:

a. outcomes and likelihood of crosses with GM plants from other field trials

The Regulator has considered likelihood of crosses throughout the RARMP. GM sugarcane from this trial is unlikely to cross with any other sugarcane because it will be harvested before flowering.

b. the total area to be planted The total planting area is limited to 5 ha. The wording in the RARMP and licence has been amended to further clarify this.

c. the description regarding restrictions on the use of the GMO in food or feed

No GMOs from this trial will be used for human food or animal feed. The wording in the RARMP and licence has been amended to ensure clarity.

2 Notes that the licence will prohibit the use of material from the trials for human or animal consumption.

Noted.

3 Considers the risks to the environment are minimal given the biology and ecology of sugarcane and the limits and control measures proposed.

Noted.

Notes that the engineered product of the plants is not regarded as a toxic substance and none of the GM plants or plant material would be used for food or animal feed. However, the statement in paragraph 88 – ‘the introduction of the gene constructs does not lead to expression of a novel protein which could be toxic or allergenic’ – should be tightly linked to supporting information. Such evidence should include that the plant proteins are not from any class having known toxic (or allergenic) members (Radauer and Breitender 2007, Delaney, Astwood et al. 2008).

The RARMP notes that plant food allergens are proteins mainly derived from peanut, tree nuts, wheat and soybean (Delaney et al. 2008; Herman & Ladics 2011). Sugarcane falls outside of this group of plants. The RARMP has been updated with additional text to further support this statement and also include a reference to Radauer and Breitender 2007.

The genetic modification or trait is unlikely to increase the ability of the plants to spread and persist, confer a selective advantage or increase weediness in sugarcane.

Noted. The potential for spread and persistence of the GM sugarcane has been addressed in Chapter 2, Risk Scenario 2. Licence conditions have also been imposed to prevent the spread and persistence of the GMO.

3 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 37


Sub.No: Summary of issues raised Comment

The risk of gene transfer to non-GM sugarcane or related species is minimised due to the proposed limits and controls. However, the RARMP would benefit from an explanation of why an isolation zone of 6 m was decided on. Skinner (1959) recorded highly variable pollen dispersal and outcrossing rates, with viable pollen up to 20 m away, leading to recommended isolation distances of 100 m in open forest and 300 m in open ground.

The 6 m isolation zone is not intended to address pollen flow. As stated in Chapter 1 of the RARMP, and discussed in Chapters 2 and 3, the applicant proposed the 6 m of physical separation to prevent inadvertent mixing of cane in adjacent plots at the time of harvest. The Regulator considers that, from a risk perspective, the 6 m isolation is sufficient to avoid mixing of cane at harvest.The risk of gene transfer to non-GM sugarcane was considered in risk scenario 3. GM sugarcane from this trial is unlikely to cross with any other sugarcane because it will be harvested before expected flowering. In the unlikely event that the GM sugarcane did flower, it is unlikely that crossing with sexually compatible plants would occur because:1) sugarcane pollen viability is low in the area proposed for release;2) commercial sugarcane varieties show low fertility;3) crossing to plants outside of the Saccharum genus has rarely

been observed.The licence requires that any developing inflorescences be removed before flowering to further restrict the potential for pollen flow and gene transfer to sexually compatible plants.

Notes that there is always uncertainty regarding potential unintended effects of genetic modifications; generally agrees with further data requirements proposed in the RARMP for any future large scale or commercial release.Notes the statement that, prior to commercialisation, additional phenotypic characterisation may be required to address uncertainty around potential changes in weediness. Suggests that this may include data collection on pollen viability, seed dormancy, plant height, growth rate, outcrossing rates and tolerance to abiotic or biotic stresses.

The likelihood of the introduced trait leading to a selective advantage for the GM sugarcane compared to non-GM sugarcane is discussed in the RARMP (risk scenario 2).As field trials of GMOs are designed to gather data, there are generally data gaps. However, conditions have been imposed to prevent the spread and persistence of the GMO.Chapter 3 lists additional information that may be required to assess future applications for larger scale trials, reduced containment measures or commercial release. Applicants/licence holders liaise with the OGTR about collection of appropriate data, and the DIR application forms provide guidance on the information required.

Notes that paragraph 109 refers to uncertainty that sugar levels in the plants may affect degradation by microorganisms or drought and salinity tolerance. Reasons and supportive evidence for this statement should be provided

Additional text has been added to the RARMP to explain uncertainty in relation to the impact of altered sugar levels on stress tolerance.

Identified other issues regarding clarity and presentation of the RARMP.

Additional text has been added to the RARMP to improve clarity.

4 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 135.

Noted.

5 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.

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

Noted.

Appendix A 38

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