ENVIRONMENTAL RISK MANAGEMENT AUTHORITY
DECISION
Application Code
GMF98011
Date
7 December 1999
Hearing Date
12 October 1999
Considered by
Special Committee of the Authority appointed under section 19(2)(b) of the
Hazardous Substances and New Organisms Act 1996.
Application Details
Application Code
Applicant
Purpose
Date Application Received
GMF98011
Carter Holt Harvey Ltd
To field test, in the Waikato, pre-reproductive Pinus radiata, in order to
study factors influencing gene expression and to assess the influence of
genetic modifications, involving the insertion of marker genes, on the
growth and morphology of trees.
18 June 1999
ERMA New Zealand Contact
Elizabeth Beale
Decision
The application is Approved with controls.
The organism approved is:
Pinus radiata D. Don (radiata pine)
Construct: Plasmid, pCW122, containing a gus (β-glucuronidase) gene, derived from Escherichia coli
(E. coli) under the control of a CaMV (Cauliflower Mosaic Virus) tandem 35S promoter, and an
nptII (neomycin phosphotransferase) gene, derived from E. coli, controlled by a CaMV 35S
promoter. Refer to Annex 1 for further details of the genetic construct, including a copy of the
plasmid map of pCW122.
Phenotype: The nptII gene confers resistance to the aminoglycoside group of antibiotics (including
neomycin, kanamycin, and geneticin). The gus gene is a reporter gene that upon testing will produce
a blue colour.
Application Process
The application was formally received on 18 June 1999, and verified on 20 July 1999, following
additional information requests.
The application was publicly notified on 24 July 1999 in The Dominion, The New Zealand Herald,
The Press and The Otago Daily Times.
Public submissions closed on 3 September 1999. Thirteen submissions were received.
The documents available for the evaluation and review of the application by ERMA New
Zealand included: the application (including supporting documentation and confidential
information provided), public submissions received, and submissions and comment from other
government agencies (including the Department of Conservation and the Ministry of Agriculture
and Forestry).
In accordance with section 19(2)(b) of the Hazardous Substances and New Organisms (HSNO)
Act 1996, the Authority appointed a Special Committee to determine the application. The
Committee comprised Authority members: Professor Barry Scott (Chairman), Helen Hughes,
and Dr Lindie Nelson.
A public hearing was held on 12 October 1999 in Wellington.
Hearing Review
The following parties made presentations to the Special Committee:
For the applicant:
1.
2.
Murray Parrish
Bryan Parkes
Carter Holt Harvey Forests (Manager- Business Communications)
Cater Holt Harvey Forests (Manager- Biotechnology Research)
For ERMA New Zealand:
1.
Elizabeth Beale
Project Leader
Submitters:
1.
Tikitu Tutua-Nathan
2.
3.
George Asher
Susie Lees
4.
5.
Don Murray
Graeme Tyree
Tuwharetoa Māori Trust Board (Project Manager, Environmental
Services Division).
Manager, Lake Taupo Forest Trust (Ngāti Tuwharetoa).
Also on behalf of Oraina Jones and Berylla Berylla (both
submitters).
Witness to Susie Lees.
On behalf of Nelson Greens and Nelson Bays Sustainable Living
Trust Inc. [and Nelson Peace Group (affiliated to Peace
Movement Aotearoa)]
Relevant Legislative Criteria
The application was lodged pursuant to section 40 of the HSNO Act 1996, and determined in
accordance with section 45 and the additional matters contained in sections 37 and 44.
Consideration of the application followed the relevant provisions of the Hazardous Substances
and New Organisms (Methodology) Order 1998 (the Methodology).
The Application
The application is for approval to field test Pinus radiata D. Don (radiata pine) modified with two
marker genes derived from the bacterium E. coli, β-glucuronidase (or gus) as a reporter for gene
expression, and neomycin-phosphotransferase or nptII for resistance against the aminoglycoside
Page 2 of 20
group of antibiotics including neomycin, kanamycin, and geneticin. The two genes are under the
control of a promoter sequence derived from the cauliflower mosaic virus (CaMV).
The purpose of the field test is to evaluate the expression of the marker genes in juvenile
genetically modified radiata pine trees, in order to determine the stability, level, and zones of
gene expression and what influence field conditions have on that expression. In addition, the
field trial is aimed at determining whether the transformation techniques used have any
detrimental effect on the growth and morphology of the genetically modified trees.
The development of the genetically modified radiata pine to be field tested in this trial, has been
approved by the Carter Holt Harvey Ltd (CHH) Institutional Biological Safety Committee
(IBSC) under delegated authority from the Environmental Risk Management Authority (the
Authority). The development was approved in March 1999 (ERMA Approval Code
GMD000034).
The field test will comprise up to one hundred and twenty genetically modified radiata pine trees
that will be grown for approximately two years and six months in the field. The trial will be
terminated and all above ground trees material removed from the trial site before the trees reach
reproductive maturity, ie before the trees reach 3 years 6 months of age.
The genetically modified radiata pine seedlings will be maintained in CHH Forest’s (CHHF)
greenhouse and subsequently in a shadehouse on the Forest Research campus in Rotorua for 11
months (plus or minus 5 months depending on growth rates) prior to planting in the field. The
field test site is located on CHH freehold land near Tokoroa (in the Waikato region).
The greenhouse, shadehouse, and the field test site are required to be registered by the Ministry
of Agriculture and Forestry (MAF) as containment facilities under the Biosecurity Act 1993 and
operated in accordance with the controls imposed in this decision.
Key Issues
The Committee’s consideration of the application encompassed those issues relevant to the
application, and included:
1.
The adequacy of the proposed containment regime, including:
i.
The ability of the organism (or any heritable material) to escape from containment,
including:

failure of biological containment (as a result of failure to detect and remove any
reproductive structures);

breach of containment following deliberate or inadvertant action;

breach of containment during transportation or transplantation; and

transfer of DNA, including antibiotic resistance gene (nptII) and CaMV 35S
promoter, via horizontal gene transfer to other organisms, for example:
(a) transfer of DNA to soil microorganisms;
(b) transfer of DNA to gut bacteria of herbivores following consumption;
and
(c) uptake of DNA by mycorrhizal fungi.
ii.
The ability of the organism to establish a self-sustaining population.
Page 3 of 20
iii.
2.
The effects of the organism, including:
i.
ii.
3.
The ease of eradication of any population established.
Risks to the environment and human health and safety, including:

risks from the use of the cauliflower mosaic virus (CaMV) 35S promoter;

development of antibiotic resistance; and

long-term unanticipated environmental and health effects.
Risks to the relationship of Māori and their culture and traditions with their ancestral
lands, water, sites, waahi tapu, valued flora and fauna, and other taonga.
The benefits of the application.
Adequacy of the Proposed Containment Regime
Ability of the organism to escape from containment
In considering the ability of the organism to escape from containment, the Committee
considered, inter alia:
i.
failure of biological containment (as a result of failure to detect and remove any reproductive
structures);
ii.
breach of containment following deliberate or inadvertent action; and
iii. breach of containment during transportation or transplantation.
In addition, the Committee considered the following issues with respect to containing genetically
modified material:
iv. transfer of DNA, including antibiotic resistance gene (nptII) and CaMV 35S promoter, to
soil microorganisms;
v.
transfer of DNA to gut bacteria of herbivores following consumption; and
vi. uptake of DNA by mycorrhizal fungi.
i.
Failure of biological containment (as a result of failure to detect and remove any
reproductive structures)
The genetically modified radiata pine trees to be field tested in this trial are pre-reproductive and
therefore are not expected to produce pollen or seed. This is the basis of biological containment with
respect to the escape of genetically modified material from the trial site. The radiata pine plants
will be destroyed prior to reproductive maturity.
There is considerable variation in the duration of the juvenile phase, but Pinus radiata seedlings
are not expected to reach reproductive maturity until 4 to 6 years of age, and therefore are not
expected to flower during the period of this trial.
Reproductive buds can be differentiated from other vegetative structures prior to reproductive
competence. The time period from when an immature pollen catkin is able to be identified until
Page 4 of 20
pollen is released is approximately 3 months. The time taken for the development of female seed
cones is approximately 2 years.
The applicant has included in the trial plan, an inspection regime in order to detect and remove
any precocious reproductive structures that may form, and this includes specific checks to be
made of strobili development in April and May (the period in which structures are most readily
identifiable). However, in view of the uncertainty associated with the timing of reproductive
structure formation, an additional control condition has been included that requires an extension
of the monitoring time and frequency to cover any unexpected outcomes. This control requires
the applicant to inspect the radiata pine between February and July (at monthly intervals) for the
development of precocious reproductive structures, and to remove and record any found.
In addition, the controls require that the plants be destroyed by the time they reach 3 years 6
months of age. Phase change from juvenile to sexually mature plants is under genetic control and
can be influenced by unusual environmental conditions. Cessation of the trial at this stage
provides additional assurance that no reproductive structures will be able to form.
Taking into account the pre-reproductive nature of the radiata pine plants in this field test and
controls in place to monitor for precocious development of reproductive structures, the
Committee concluded that the probability of pollen or seed derived from genetically modified
radiata pine plants escaping is very low.
ii. Breach of containment following deliberate or inadvertent action
The greenhouse and shadehouse where genetically modified radiata pine plants are to be
maintained prior to planting in the field are within the Forest Research Campus in Rotorua. The
site is off the road and subject to regular surveillance by personnel within the campus and those
specifically involved in the management of the trial. The probability of deliberate entry to the
campus and the potential for such an act to result in a breach of containment is considered to be
low.
Entry to the greenhouse and shadehouse is restricted to authorised personnel, thereby
minimising the possibility of inadvertant removal of genetically modified plants. In addition, the
controls imposed in this decision require a log of entry to the facilities to be maintained and be
available for inspection.
The relatively inaccessible location of the field test site within CHH freehold land near Kinleith,
the exact location information being provided to the Authority in confidence, and the fact that
this trial will be indistinguishable from other radiata pine field tests, reduces the possibility of
deliberate action to breach containment, to a low level.
As a part of the management of the field trial and controls imposed on this approval, the
applicant is required to inspect the field test site on a regular basis for the duration the plants are
in the field. In the course of this regular monitoring, should CHHF detect any disturbance it is
required to destroy any plant material recovered in order to minimise the possibility of any such
action resulting in the further loss of any genetically modified material.
In addition, as the trees are pre-reproductive and unlikely to survive physical disturbance,
adverse consequences resulting from removal of the plants, such as pollen release, are highly
unlikely.
Page 5 of 20
Under the monitoring regime and the controls imposed in this decision, the Committee concluded
that the probability of sabotage or inadvertent action resulting in the loss of plants or genetically
modified material from the greenhouse/shadehouse or the field test site is low.
iii. Breach of containment during transportation or transplantation
Genetically modified seedlings are to be transported from the Forest Research campus to the
field trial site in secure containment, comprising transportation in clearly identifiable potting
containers within closed boxes. Boxes containing juvenile seedlings are to be transported in an
enclosed vehicle.
In addition, a control on this approval requires an inventory of genetically modified radiata pine
plants to be undertaken prior to departing from the Forest Research campus and upon unloading
of the vehicle at the field test site. This inventory must be recorded in the logbook.
Under these conditions, the Committee concluded that the probability of a loss of genetically
modified material during transportation or transplantation from the greenhouse/shadehouse or the
field test site is low.
iv. Horizontal gene transfer to soil microorganisms
This issue is of particular relevance in the context of this field test as the applicant proposes,
following completion of the trial, removing above ground plant material and leaving the stumps
and roots to rot in the ground.
In order for horizontal gene transfer to occur from genetically modified radiata pine to soil
microorganisms, a sequence of individual events would have to occur (see below), that current
scientific evidence suggests, if they occur at all, would only occur at low frequencies.
In this case, in order for horizontal gene transfer to result in the transfer of a gene capable of
producing an active gene product (protein), the following sequence of events would have to
occur:

DNA (containing a modified gene) would have to survive in the soil intact;

the DNA would have to be transferred or be taken up by other organisms from the roots of
the plant, or from shed plant material into the surrounding soil;

a complete and active form of one of the modified genes would have to be taken up by the
microorganism;

this complete gene sequence would then have to be inserted behind an appropriate
promoter and other regulatory sequences, in the genome of the other species, in this case a
soil microorganism.
The Authority considers that the probability of this sequence of events occurring, based on
current scientific evidence, is low. No new information has arisen that would cause it to
reconsider its previously stated position on this issue, which is that while the scientific evidence
available is inconclusive, horizontal gene transfer from transgenic plants to soil microorganisms is unlikely (to
occur).
Page 6 of 20
v.
Horizontal gene transfer to gut bacteria
A similar sequence as described above would have to occur for genetic material to transfer from
genetically modified radiata pine to microorganisms present in the gut of herbivores, following
consumption of genetically modified radiata pine.
Following ingestion of plant material, the excised gene would have to survive the digestion
processes of the animals’ gut in order to be taken up intact by gut microflora/fauna. A complete
and active form of a modified gene would have to be taken up by the microorganism, and this
gene sequence would then have to be inserted behind an appropriate promoter and other
regulatory sequences, in the genome of the microorganism.
The Committee considers that the probability of this sequence of events occurring is low and
therefore considers the cumulative probability of the transfer of genes derived from these
genetically modified radiata pine trees to be low.
vi. Uptake of DNA by mycorrhizal fungi
Beneficial mycorrhizal fungi are found in association with the root systems of Pinus radiata. These
mycorrhizal fungi are able to penetrate the root cells of radiata pine plants so consequently are in
close contact with root cells. However, currently there is no known mechanism or evidence
available for the transfer of genetic material from plants to fungi or other microorganisms.
Given the small number of trees involved in this trial, and the short duration of the trial, the
Committee concluded that the probability of the transfer of genetic material via this route is very
low.
The Committee noted that currently there is very little empirical data on gene transfer from
plants to soil microorganisms. Such information would be particularly valuable in the event that
an application to release genetically modified radiata pine was made. The Committee noted
statements by the applicant that it would be feasible to undertake testing of soil microorganisms
in the context of this trial.
Ability of the organism to establish a self-sustaining population
Radiata pine is a monoecious species that relies on wind to disperse pollen. Seed is normally
contained in serotinous cones that open under hot dry conditions and shed seed. Radiata pine is
therefore capable of establishing a self-sustaining population via the spread of seed. In addition,
escape of pollen could fertilise non-modified trees, and some resultant seedlings could carry the
genetic modifications. However, in this field test, plants will be destroyed prior to reproductive
maturity, and therefore no pollen will be produced.
Since radiata pine in this trial will be destroyed prior to reproductive maturity, the probability of
the establishment of a self-sustaining population as a result of seed escape, or pollen fertilising
another population, from this trial, is low.
Ease of eradication of any population that established
Radiata pine plants are susceptible to a range of common herbicides and can be destroyed by
cutting off the plant at ground level. Coppicing of stumps or regeneration from roots has never
been observed in Pinus radiata.
Page 7 of 20
Effects of the Organism (Risks to the Environment and
Human Health and Safety)
Risk to the Environment and Public Health
For any effects on the environment to be realised as a result of this field test, the organism or
heritable material must first escape into the uncontrolled environment. The principal issue to be
considered therefore is whether the escape of such material is possible, and whether controls can
be imposed that would effectively prevent the organism or its heritable material from escaping
containment.
The Committee considered the following issues, with respect to potential risks to the
environment and public health:
i.
Risks from the use of the cauliflower mosaic virus (CaMV) 35S promoter
Submitters on this application have raised concerns that the use of the CaMV 35S promoter
could:

lead to the development of CaMV in Pinus radiata; and

recombine with infecting viruses to produce virulent new diseases, either in Pinus radiata, or
in other organisms (ie insects, herbivores, microorganisms) through transfer of the CaMV
35S promoter via horizontal gene transfer.
Evidence available suggests that Pinus radiata is not susceptible to viruses, and in particular is not
susceptible to infection by CaMV. Presence in the modified radiata pine of a promoter sequence
derived from the CaMV in itself cannot cause a CaMV infection.
Therefore the probability of the development of either CaMV in genetically modified radiata
pine or any other novel viral infection, as a result of the use of the 35S promoter region from
CaMV is small.
In its previous decisions involving organisms containing the CaMV promoter the Authority has
only once commented on the safety or otherwise of its use. In the decision on applications
GMF98005 and GMF98006 (in respect of field testing of genetically modified maize), the
Authority noted that to date there is no evidence that the CaMV promoter, which has been widely used for
many years, has any risk associated with its use.
ii. Development of antibiotic resistance
Submitters have raised the prospect that the transfer of antibiotic resistance (to soil bacteria) will
result in an amplification of antibiotic resistance genes in the biosphere, with the potential to lead
to adverse consequences for human health.
The Committee considered that the transfer of genetic material from radiata pine to soil
microorganisms in this trial is unlikely.
In addition, evidence suggests that antibiotic resistance (studies cited related to kanamycin
resistance1) is not a novel characteristic for a soil ecosystem, and therefore the effect of any
Nap, J-P., Bijvoet, J., Stiekema, W.J. 1992. Biosafety of kanamycin-resistant transgenic plants. Transgenic Research
1: 239-249.
1
Page 8 of 20
horizontal gene transfer of the nptII gene would be insignificant and would not alter or disturb
such a soil ecosystem.
The Committee concluded that the risk of the use of the nptII gene in genetically modified plants
contributing to increasing levels of antibiotic resistance is negligible, when compared to the
incidence of intrinsically resistant microorganisms, including human gut bacteria and soil
microorganisms.
iii. Long-term unanticipated environmental and health effects
As was the case in respect of applications GMF98005 and GMF98006, submissions were
received expressing concerns that the processes and consequences of genetic modification are
insufficiently established for the applicant to be able to provide assurance that there will be no
unanticipated long term adverse effects on either the environment or human health.
These submissions covered several grounds:

the uncertainty of genetic modification as a science obliged the Authority to take a
precautionary approach under the Act; and

the possibility of long term adverse effects materialising well into the future has to be taken
into account in considering the well-being of future generations.
In terms of the Act the Authority can consider these kinds of issue in relation to, either:

the requirement under section 5(b) to recognise and provide for the maintenance and
enhancement of the capacity of people and communities to provide for their own economic,
social and cultural well-being and for the reasonably foreseeable needs of future generations;
or

the requirement under section 7 of the Act to take account of the need for caution in
managing adverse effects where there is scientific and technical uncertainty about those
effects.
The Committee does not dismiss any of the concerns expressed. However, concerns regarding
scientific uncertainty, and potential long term adverse impacts on future generations are more
relevant to release applications than to an application for a small-scale contained field test. There
may be some scientific uncertainty regarding the potential consequences of the genetic
modifications proposed in the present applications, but this will not result in adverse
consequences for the environment, human health, or future generations while the field test is
undertaken in containment. The caution required of the Authority relates to the adequacy of the
containment conditions and management regime. In this regard the Committee considers the
risks to be negligible for current and future generations alike.
The Committee considers however, that the applicant should take note of the concerns
expressed, and be prepared to address them in the event that an application is made to release
the genetically modified radiata pine. This decision relates only to the proposed contained field
test.
Risk to the Relationship of Māori and their Culture and Traditions
with Taonga
Ngā Kaihautū Tikanga Taiao, an advisory group set up to advise the Authority specifically of
issues raised in respect of section 6(d) and 8 of the Act did not consider it necessary to make a
Page 9 of 20
formal report on this application. However, in advice provided to ERMA New Zealand they
noted in respect of this application that it is the belief of many Maori that it is inappropriate for the
whakapapa of one species to be mixed with that of another species.
Ngā Kaihautū Tikanga Taiao also noted that Carter Holt Harvey representatives have consulted with
appropriate tangata whenua early to the satisfaction of tangata whenua. The Committee noted how willing
Carter Holt Harvey has been to engage in effective consultation with local and other nearby
Māori.
The applicant consulted with the two iwi who are tangata whenua in the area in which the field
test is to be undertaken, Te Arawa and Ngāti Raukawa. The Raukawa Trust Board noted that it
did not oppose the application, however reserved the right to oppose it once they had
considered the views of Te Arawa and other Māori that made submissions. Prior to submission
of the application Ngāti Raukawa had access to the views of Te Arawa, and were, following the
hearing, provided with copies of the presentations made by representatives from Ngāti
Tuwharetoa at the hearing. Ngāti Raukawa indicated that they had no further comments to make
on this material.
Te Arawa gave written support to the application dependent on certain conditions that Carter
Holt Harvey has met, or has undertaken to meet. This included Carter Holt Harvey’s
undertakings to maintain a regular on-going dialogue with Te Arawa and Ngāti Raukawa as the
trial proceeds and to provide them with copies of any results or summaries of research findings.
The Committee noted that these consultations should also be extended to include Ngāti
Tuwharetoa.
Representatives from the Tuwharetoa Māori Trust Board, a neighbouring iwi, spoke to their
submission at the hearing, noting their opposition to the application on the grounds that the
transfer of genetic material between species represents a significant compromise of whakapapa
(genealogy or lineage) and interference to the principles of whanaungatanga (kindred
relationships). In addition, Ngāti Tuwharetoa noted their forestry interests in the area and raised
the possibility of Forest Stewardship Council (FSC) certification of their forestry products being
revoked should genetically modified Pinus radiata be released in the area.
In essence, Ngāti Tuwharetoa has indicated that this application is against their tikanga.
However, in their submissions they also noted that at this stage they do not have all the
information that pertains to the potential cultural and spiritual implications that may occur.
The Committee accepts the views of Ngāti Tuwharetoa in respect of their statements that this
application is against their tikanga, however notes that the interpretation and significance of
traditional spiritual beliefs may vary amongst Māori, as may the consequences flowing from any
affront to those beliefs.
The Committee is of the view that for a containment application, information on the risks to the
relationship of Māori and their culture and traditions with taonga, is required primarily in respect
of local Māori (those who are tangata whenua), although there are close genealogical links
between Ngāti Tuwharetoa, Te Arawa and Ngāti Raukawa.
Benefits
The benefits outlined in the application relate to enhancing CHHF’s technological capability.
There are no direct financial or commercial benefits either to CHHF or to New Zealand from
conducting this trial.
Page 10 of 20
The applicant has indicated that there may be potential commercial benefits (to Carter Holt
Harvey and New Zealand) from subsequent research and development for which further
applications would be required. Any future work in the area of tree improvement through
biotechnology is reliant on the company perfecting the techniques via the work in this
application.
Submitters questioned the benefits associated with this application given that a similar field trial
of genetically modified Pinus radiata is currently being undertaken by Forest Research, involving
the same marker gene constructs as are the subject of this application. Carter Holt Harvey has
indicated that the genetically modified radiata pine to be field tested under this application were
produced using different transformation and propagation protocols and include a number of
different genotypes.
Conclusion
In terms of clause 26 of the Methodology the Authority may, taking into account the measures
available for risk management, approve an application where an organism poses negligible risks
to the environment and human health and safety if it is evident that the benefits of the
application outweigh the costs.
The Committee formed the view that the risks associated with this field test are negligible. In this
case, the fact that the application has been made indicates that the benefits to the applicant
outweigh the costs. In the absence of any costs to other parties, the Committee concludes that
benefits outweigh costs.
Pursuant to section 45(1)(a)(i) of the Act, the Committee was satisfied that this application was for
one of the purposes specified in section 39(1) of the Act, being section 39(1)(b): Field testing any new
organism.
The Committee is satisfied that the proposed containment regime and the control conditions
imposed in this decision will adequately contain the organism.
Page 11 of 20
Controls
In order to provide for the matters detailed in Part I of the Third Schedule to the Act, Containment
Controls for Development and Field Testing of Genetically Modified Organisms, this application is approved
subject to the following controls:
1.
To limit the likelihood of any accidental release of any organism or any
viable genetic material2:
Controls relating to the Greenhouse/Shadehouse
1.1
The greenhouse and shade house used to store genetically modified radiata pine trees to
be used as a part of this approval shall be registered by the Ministry of Agriculture and
Forestry (MAF) as containment facilities under the Biosecurity Act 1993.
1.2
The operation, management and construction of the containment facilities (greenhouse and
shadehouse) shall be in accordance with the Australian New Zealand Standard AS/NZS
2243.3:1995 Safety in Laboratories: Part 3: (Microbiology), at Plant House Physical
Containment Level 1 (PC1) and the controls imposed in this decision.
The requirements of AS/NZS 2243.3:1995 Plant House (PC1) and the controls imposed
in this decision constitute the standard applicable for the registration of these places as
containment facilities under section 39 of the Biosecurity Act 1993.
1.3
The greenhouse and shadehouse shall be approved as containment facilities for a
specified time, being the duration of the field test approved under this decision.
1.4
ERMA New Zealand and the facility Supervisor3 shall be advised of the date of the
transfer of radiata pine seedlings from the laboratory to the greenhouse and shadehouse.
1.5
All genetically modified radiata pine material shall be properly and adequately identified
at all times.
1.6
No genetically modified radiata pine plants shall be removed from the
greenhouse/shadehouse (unless for transportation to the field test site) except with
approval from the Supervisor.
1.7
A register of plants held in the greenhouse and shadehouse containment facilities shall be
maintained. The following records shall be made for each plant:
i.
ii.
iii.
iv.
v.
the identity of plant(s) and details of genetic modification;
unique identification number allocated to the accession or modification;
date of accession into the facility;
date of development; and
date and method of final disposal of plants.
Viable Genetic Material is biological material that can be resuscitated to grow into tissues or organisms. It can be
defined to mean biological material capable of growth even though resuscitation procedures may be required, eg
when organisms or parts thereof are sublethally damaged by being frozen, dried, heated, or affected by chemical.
2
3
An inspector appointed under the Biosecurity Act.
Page 12 of 20
1.8
ERMA New Zealand and the facility Supervisor shall be informed in writing prior to
removal of genetically modified radiata pine from the containment facilities for planting
in the field test site.
1.9
An inventory of radiata pine material removed from the greenhouse/shadehouse shall be
taken and recorded prior to removal of seedlings for transport to the field test site. An
inventory of seedlings planted at the field test site shall be taken and recorded, and be
available for inspection by the Supervisor.
1.10
Genetically modified radiata pine seedlings shall be transported in secure containment
from the greenhouse/shadehouse to the field test site to ensure that there is no spillage
of modified material outside the trial site. Radiata pine seedlings shall be transported in
clearly identifiable potting containers within closed boxes, in an enclosed vehicle.
Personnel involved in transportation shall be familiar with the genetically modified
nature of the plants and the controls imposed by this decision.
1.11
No radiata pine plants shall be maintained in the greenhouse/shadehouse beyond the age
of 3 years 6 months. All radiata pine material contained in the greenhouse/shadehouse
and not planted in the field test site shall be destroyed by incineration.
Controls relating to the forest field test site
1.12
The trial site for the field testing of genetically modified radiata pine trees shall be
registered by the Ministry of Agriculture and Forestry (MAF) as a containment facility under
the Biosecurity Act 1993.
The controls imposed in this decision constitute the standard applicable for registration
of that place as a containment facility under section 39 of the Biosecurity Act 1993.
1.13
The field test site shall be approved as a containment facility for a specified time, being
the duration of the field test approved under this decision.
1.14
The exact location of the trial site shall be provided as confidential information to the
Authority and the facility Supervisor (MAF). The trial site shall be used solely for the
field testing of genetically modified radiata pine plants approved in this decision and nonmodified radiata pine control plants.
1.15
The field trial site shall be delineated by marker posts.
1.16
The number of genetically modified radiata pine plants in the field test site shall not
exceed 120 at any one time.
1.17
All radiata pine material to be used in this field test shall be properly and adequately
identified at all times.
1.18
A register of plants grown in the field trial shall be maintained. The following records
shall be made for each plant:
i. the identity of plant(s) and details of genetic modification;
ii. date of planting; and
iii. date and method of final disposal of plants.
1.19
All genetically modified radiata pine plants no longer required for this application or if
the experiment ceases shall be destroyed on-site by incineration.
Page 13 of 20
1.20
The field test shall be terminated on/or prior to genetically modified radiata pine trees
reaching 3 years 6 months of age.
1.21
The plants in the field test shall be inspected for strobili development in accordance with
the schedule in Annex 4 of this decision. This includes inspections at monthly intervals
from February to July inclusive.
1.22
To prevent unintended/accidental release of genetically modified radiata pine from the
field trial site, any reproductive structures identified on the plants shall be immediately
removed and transported in secure containment and destroyed (by incineration or
autoclaving).
1.23
Following completion of the field test all radiata pine plants shall be cut off immediately
above ground and incinerated on-site.
1.24
At the conclusion of the field test, remaining stumps of genetically modified radiata pine
shall be treated with herbicide.
1.25
Any genetically modified radiata pine plant material removed from the field test site for
the purposes of testing in an approved containment laboratory shall be transported in
secure containment, including storage in closed containers in a closed vehicle.
2.
To exclude unauthorised people from the facility:
2.1
At all times only persons authorised by the Operator/Manager (of the containment
facilities) shall have access to the greenhouse/shadehouse containment facilities.
2.2
A log of all persons accessing the greenhouse/shadehouse/field test containment
facilities shall be maintained and be available for inspection by the facility Supervisor.
3.
To exclude other organisms from the facility and to control undesirable
and unwanted organisms within the facility:
4.
To prevent unintended release of the organism by experimenters working
with the organism:
4.1
The applicant shall comply with the requirements contained in the standard listed in
control 1.2 relating to the prevention of unintended release of genetically modified
radiata pine by experimenters working with the radiata pine within the greenhouse and
shadehouse.
4.2
Handling of radiata pine plants used in this field test shall ensure that there is no spillage
of plant material outside the field trial site.
5.
To control the effects of any accidental release or escape of an organism:
5.1
In case of unintended or accidental release or escape of genetically modified radiata pine
plants involved in the field trial, the applicant shall use best endeavours to recover the
plants and if they cannot be replanted in the trial site, shall destroy them by incineration.
5.2
If for any reason a breach of containment occurs the applicant shall notify the facility
Supervisor, and ERMA New Zealand immediately the event is noticed (and at least
within 24 hours of the breach being detected).
Page 14 of 20
6.
Inspection and monitoring requirements for containment facilities:
6.1
Any incidence of interference with the field test site shall be reported to the facility
Supervisor and ERMA New Zealand immediately it is detected (and at least within 24
hours).
6.2
The Authority or its authorised agent or properly authorised enforcement officers, may
inspect the field trial site at any reasonable time.
A schedule of inspections to be undertaken by the Supervisor is included as Annex 2 to
this decision.
6.3
ERMA New Zealand and the facility Supervisor shall be advised of any changes to the
timetable for the field trial of genetically modified radiata pine if different than indicated
in the application
6.4
The applicant shall prepare and use a containment manual to implement the controls
imposed by this approval. The manual shall specify the containment system within the
greenhouse/shadehouse and the field test site. It shall also include the trial monitoring
regime, as required by these controls and defined in the application.
The containment manual shall also describe contingency plans in place to take account
of:
 the accidental release of plants outside the facilities; and
 fire or any other emergency.
6.5
The applicant shall monitor the field test site at the intervals identified in Annex 4 of this
decision. Such monitoring checks shall be recorded in a log book and be available for
inspection.
6.6
The applicant shall provide a comprehensive report to ERMA New Zealand in each July
on the progress of the field trial of genetically modified radiata pine, including an
inventory of plants involved in the field test. This report shall include information on
adherence with the containment controls outlined in this decision (including any
incidence of the development of precocious reproductive structures) and any incidents
that may be of interest to the public or ERMA New Zealand or MAF (such as a breach
of containment or any interference, other than by authorised personnel, of either the
greenhouse/shadehouse or the field test site).
6.7
A final report shall be provided to ERMA New Zealand by 15 July 2003 by when the
exploratory work outlined in the application is expected to be completed under this
approval. The report shall focus on adherence with the containment controls outlined in
this decision (including any incidence of the development of precocious reproductive
structures) and any incidents that may be of interest to the public or ERMA New
Zealand or MAF (such as a breach of containment or any interference, other than by
authorised personnel, of either the greenhouse/shadehouse or the field test site).
Page 15 of 20
7.
7.1
Qualifications required of the persons responsible for implementing
those controls:
The applicant shall inform all personnel involved in the field testing of genetically
modified radiata pine of the controls imposed in this decision.
_____________________
Chair
__________________
Date
_____________________
Name
Page 16 of 20
Annex 1
35S
npt II
ocs 3’
2 x 35 S
gus
Eco R V
Sph I
Bgl II
Xho I
Eco R I
Sma I
Bham H I
Xba I
Eco RV
Nco I
Sma I
Hind III
Sal I
BamH I
KpnI
Sma I
Pst I
Sst I
Kpn I
Nco I
Plasmid Map of pCW122
CaMV Poly A
pUC backbone
The structure of Construct pCW122.
Plasmid size : pCW122
Constructed by: Christian Walter
Construction data: 14/04/93
Comments: nptII gne from pANDY 3 cloned (KpnI) into pJI 166.
Source and function of the DNA used to produce pCW122
DNA
Source
Reference
Function
-glucuronidase
gene (gus-gene)
gus locus from
Escherichia coli
Jefferson et al., 1987
reporter (blue colour)
neomycinphosphotransferase
(npt II)
Escherichia coli
Herrera-Estrella et al.,
1983
resistance against the
aminoglycoside group of
antibiotics (including
neomycin, kanamycin, and
geneticin).
CaMV 35S promoter
Cauliflower Mosaic Virus
Chenault and Melcher,
1993
Promoter
Page 17 of 20
Annex 2
Schedule of Inspections
To be undertaken by the facility Supervisor (Ministry of Agriculture and
Forestry)
1.
One inspection to be undertaken whilst plants are in the greenhouse, ie approximately 6
months after the date of notification of the commencement of this trial.
2.
One inspection to be undertaken whilst plants are in the shadehouse, ie approximately 12
months after the date of notification of the commencement of this trial.
3.
Inspections shall be undertaken at the following points during the management of the field
trial:
i.
Immediately following planting of the trees in the field;
ii.
Once during the period whilst the trees are in the field, ie one year after planting; and
iii.
At the completion of the field trial, once plant material has been removed and
incinerated.
Page 18 of 20
Annex 3
Forest Field Test Site
Confidential
Page 19 of 20
Annex 4
Schedule of field test site monitoring
To be undertaken by the applicant
Activity
Placement into shade-house
Placement into Field Tests
Health Check
Assess growth / Sample tissue
Inspect for strobili development
Health Check
Inspect for strobili development
Inspect for strobili development
Inspect for strobili development
Inspect for strobili development
Assess growth / Sample tissue
Inspect for strobili development
Health Check
Assess growth / Sample tissue
Inspect for strobili development
Health Check
Inspect for strobili development
Inspect for strobili development
Inspect for strobili development
Inspect for strobili development
Assess growth / Sample tissue
Inspect for strobili development
Health Check
Termination age
Post-termination check
Timing4
Tree Age5
Mid February ‘01
Mid March ‘01
2.8 months
10 months
1 year 1 month
1 year 4 months
1 year 5 months
1 year 6 months
Mid April ‘01
Mid May ‘01
Mid June ‘01
Mid July ‘01
1 year 7 months
1 year 8 months
1 year 9 months
1 year 10 months
Mid February ‘02
Mid March ‘02
2 years 1 months
2 years 4 months
2 years 5 months
2 years 6 months
Mid April ‘02
Mid May ‘02
Mid June ‘02
Mid July ‘02
2 year 7 months
2 year 8 months
2 years 9 months
2 years 10 months
3 years 1 months
3 years 6 months
terminated
4
Approximate timing, dependent on the date at which the seedlings are planted in the field test site.
5
Approximate tree age depending on growth rates.
Page 20 of 20