Appendix 2: Sample evaluations of the potential for new plant products to be regulated under Part V of the Seeds Regulations

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In order to assist proponents in determining whether or not their products are PNTs and regulated under Part V of the Seeds Regulations, the following examples are provided in the following pages for reference purposes:

  • Appendix 2A: A variety of soybean (Glycine max Dowling) that possesses resistance to Asian soybean aphid (Aphis glycines Matsumura), conferred through an allele of the Rag1 gene and introduced through breeding Canadian germplasm with that of a foreign cultivar possessing the resistance allele.
  • Appendix 2B: A variety of canola (Brassica napus PM1 and PM2) that possesses tolerance to the herbicide imidazolinone, introduced through chemical mutagenesis of AHAS/ALS, the gene responsible for the tolerance trait.
  • Appendix 2C: A variety of sunflower (Helianthus annuus Clearfield™ Hybrid X81359) that possesses tolerance to the herbicide imidazolinone, introduced through breeding Canadian germplasm with a member of a wild population that possessed a mutated AHAS/ALS gene.
  • Appendix 2D: A variety of field pea (Pisum sativum CDC Mozart) that possesses resistance to powdery mildew (Erysiphe pisi).
  • Appendix 2E: A variety of spring wheat (Triticum aestivum Goodeve, previously known as BW841) that possesses resistance to orange wheat blossom midge (Sitodiplosis mosellana), conferred through an allele of the Sm1 gene and introduced through breeding with a cultivar possessing the resistance allele.
  • Appendix 2F: A hypothetical case involving a variety of asparagus (Asparagus officinalis L.) that possesses elevated levels of rutin, an antioxidant compound that has potential human health benefits.

Appendix 2A: Asian soybean aphid resistance in Dowling soybean

PNT Determination Work Sheet
A companion document to Regulatory Directive 2009-09:
Plants with Novel Traits Regulated under Part V of the Seeds Regulations

Species: Glycine max Dowling
Trait: Resistance to Asian soybean aphid (Aphis glycines Matsumura) conferred through breeding with foreign germplasm.

1. Was the trait bred into the plant from, or present in, germplasm (of the same species) cultivated* in Canada prior to 1996, or previously authorized by the PBO for use in a plant of the same species?

No – Soybean expressing resistance to Asian soybean aphid (Aphis glycines Matsumura) via the Rag1 allele in question has not been grown in the Canadian environment prior to 1996. It is, however, possible that the trait had been present in Canadian-adapted germplasm, but had not been noticed or selected for prior to the unintentional introduction of this aphid.


Diaz-Montano, J., J.C. Reese, J. Louis, L.R. Campbell, and W.T. Schapaugh. 2007. Feeding behaviour by the soybean aphid (Hemiptera: Aphididae) on resistant and susceptible soybean genotypes. J. Econ. Entomol. 100: 984-989

Fu, Y.-B., G.W. Peterson, and M.J. Morrison. 2007. Genetic diversity of Canadian soybean cultivars and exotic germplasm revealed by simple sequence repeat markers. Crop Sci. 47: 1947-1954

Gizlice, Z., Carter, T.E., Jr., and Burton, J.W. 1994. Genetic base for North American public soybean cultivars released between 1947 and 1998. Crop Sci. 34: 1143–1151.

USDA-GRIN. 2007. United States Department of Agriculture-Germplasm Resources Information Network. Available from: [Accessed October 18th, 2007].

* The Plant Biosafety Office interprets "cultivated" to mean "grown by an individual as a crop." This means that seed multiplication plots and breeding lines are considered "cultivated."

If the answer to question 1 is "yes," then STOP. The plant is not a PNT and is not subject to regulation under Part V of the Seeds Regulations. Otherwise, continue:

2. Does the plant have a potential to have a significant negative environmental impact, relative to an appropriate Canadian comparator line (or lines), in terms of:  
2a. Weediness potential: Is there an increased potential that the plant will become a weed of agriculture or be invasive in the Canadian environment?

No – In Canada, soybean is not considered to be a weed of agricultural ecosystems or invasive of natural habitats. The foreign cultivar parental line is used extensively in the development of germplasm cultivated in the United States (Gizlice et al. 1994), and none of these varieties have documented increases in weediness or invasiveness.


Gizlice, Z., Carter, T.E., Jr., and Burton, J.W. 1994. Genetic base for North American public soybean cultivars released between 1947 and 1998. Crop Sci. 34: 1143–1151.

2b. Gene flow: Are there negative consequences to environmental safety resulting from the production of hybrids between the plant and any domestic or wild sexually compatible relatives that are present in Canada?

No – Soybeans are self-pollinating, and in Canada there are no native wild relatives or naturalized populations of non-native wild relatives. The aphid resistance trait is not associated with altered floral morphology, which governs the self-pollination mechanism in soybean. This cultivar behaves as expected in controlled crosses (Hill et al. 2006; Li et al. 2007).


Hill, C.B., Li, Y., and Hartman, G.L. 2006. A single dominant gene for resistance to the soybean aphid in the soybean cultivar Dowling. Crop Sci. 46: 1601–1605.

Li, Y., Hill, C.B., Carlson, S.R., Diers, B.W., and Hartman, G.L. 2007. Soybean aphid resistance genes in the soybean cultivars Dowling and Jackson map to linkage group M. Mol. Breed. 19:25–34.

2c. Plant pest potential: Does the plant have increased potential to harbour and/or facilitate the spread of a pest or pathogen of the Canadian environment?

No – The Rag1 allele is not known to be linked to any unusual susceptibility to pathogens, and the foreign cultivar parental line is susceptible to a variety of soybean pests for which there are sources of resistance in soybean (Sij et al. 1999). There is some resistance to soybean rust and stem canker (Bowers et al. 1993); however, there is considerable variation among Canadian soybean varieties in terms of response to various pathogens (Ontario Oil and Protein Seed Crop Committee 2007).


Bowers, G.R., Ngeleka, K., and Smith, O.D. 1993. Inheritance of stem canker resistance in soybean cultivars Crockett and Dowling. Crop Sci. 33: 67–70.

Ontario Oil and Protein Seed Crop Committee. 2007. Homepage [on- line]. Available from [Cited October 23 2007].

Sij, J.W., Bowers, G.R., Way, M.O., and Boykin, D.L. 1999. Insect defoliation and population dynamics in blends of resistant and susceptible soybean cultivars. Agron. J. 91: 82–87.

2d. Potential negative impacts on non-target organisms: Could the plant have negative impacts on non-target organisms interacting directly or indirectly with it, including humans as workers or bystanders?

No – Direct effects: The exact mode of action for Rag1 is uncharacterized (Diaz-Montano et al. 2006), and no information is available on whether Rag1 affects other insects in a similar manner to how the Asian soybean aphid is affected. However, pests that consume soybean in North America are either generalists or introduced species, since soybean is an introduced species itself. Generalist pests of soybean can easily move to other non-aphid-resistant plant species. Soybean-specific pests that are affected by the uncharacterized mode of action will be adversely affected; however, it is unclear at this time if there are any other such pests in Canada.

Tritrophic effects: Several generalist predators consume Asian soybean aphids, but there are no known Asian soybean aphid-specific predators in Canada. The Asian multicoloured ladybeetle (Harmonia axyridis) is the most common aphid predator in Ontario soybean fields (Mason et al. 2006). There is no peer-reviewed literature on the subject of tritrophy in predators in Canada that have consumed Asian soybean aphids that have consumed the foreign cultivar parental line.

There is also at least one generalist parasitoid wasp (Aphelinus albipodus) of Asian soybean aphid (Ellingson and Hogg 2003). When wasps were reared on aphids that had fed on resistant soybean germplasm, there was no difference in their body size or percent parasitoid emergence compared to those reared on aphids that had fed on susceptible soybean germplasm (Crompton and Ode 2005). Although it is unclear whether this wasp is present in Canada, the results of the research can be extrapolated to the Canadian environment and associated non-target organisms because of similarities in soybean cultivation practices and locations.

No reports of adverse effects to humans have been reported due to the cultivation of the foreign cultivar that is the source of this trait, i.e., there is a history of safe cultivation in another country.


Crompton, D.S., and Ode, P.J. 2005. Implications of the Rag1 soybean aphid resistance gene on biocontrol. Proceedings of the Entomological Society of Manitoba 61: 26.

Diaz-Montano, J., Reese, J.C., Louis, J., Schapaugh, W.T., and Campbell, L.R. 2006. Characterization of antibiosis and antixenosis to the soybean aphid (Hemiptera: Aphididae) in several soybean genotypes. J. Econ. Entomol. 99: 1884– 1889.

Ellingson, B., and Hogg, D.B. 2003. Potential of Aphelinus albipodus for biological control of soybean aphid. Proceedings of the 2003 Entomological Society of America Annual Meeting.

Mason, P., Baute, T., and Broadbent, B. 2006. Impact of predators on soybean aphid in Ontario. Société d'Entomologie du Québec–Entomological Society of Canada Joint Annual Meeting.

2e. Other potential negative impacts on biodiversity: Does the plant have any other potential negative impacts on biodiversity, including changes to environmentally sustainable crop management practices*

No – Dowling is not a variety that is adapted to cultivation in the Canadian environment (it requires a much longer growing season) (reviewed in Gizlice et al. 1994), and any Canadian breeding program using Dowling will require extensive backcrossing. The lack of adaptability of this germplasm in the Canadian environment will effectively minimize any potential unintended effects with respect to the five main environmental criteria (effects on weediness, gene flow and consequences, plant pest potential, non-target organisms, and biodiversity) that could be introgressed into Canadian-adapted germplasm through a conventional breeding program. Since soybean has no wild relatives with which it can outcross in Canada, there will be no transfer of the novel traits to other species in unmanaged environments. In addition, based on the available information, it is unlikely that the novel trait will have an adverse effect on non-target organisms.

Currently, the Ontario Ministry of Agriculture, Food, and Rural Affairs (OMAFRA) advises that natural enemies are a helpful way to control the Asian soybean aphid, and that foliar insecticides are only warranted in extreme infestations, as the insecticides kill the natural enemies and cause rebounds in the aphid population (OMAFRA 2002). The registered insecticides are highly toxic to non-target organisms, including beneficial insects and aquatic animals (OMAFRA 2007, Alberta Agriculture and Food 2007). Introduction of a genetic resistance to Asian soybean aphid in soybean may lead to a reduction in the application of these insecticides, which may in turn support biodiversity (Galvan et al. 2005). It may also supply an additional mode of action to control this insect pest, which may help delay the development of insecticide resistance. Therefore, the reduction in local pest species as a result of the aphid resistance trait does not present a significant change from existing agricultural practices.


Alberta Agriculture and Food. 2007. Agdex 606-1: Crop protection 2007. Alberta Agriculture and Food Information Packaging Centre, Edmonton, Canada.

Galvan, T.L., Koch, R.L., and Hutchison, W.D. 2005. Toxicity of commonly used insecticides in sweet corn and soybean to multicoloured Asian lady beetle (Coleoptera: Coccinellidae). J. Econ. Entomol. 98: 780–789.

OMAFRA. 2002. Publication 811: Agronomy Guide for Field Crops. Queen's Printer for Ontario, Toronto, Canada.

OMAFRA. 2007. Publication 812: Field crop protection guide 2007– 2008. Queen's Printer for Ontario, Toronto, Canada.

If the answer to any part of question 2 is "yes" or is unclear, then contact the Plant Biosafety Office: the plant may be a PNT and may be regulated under Part V of the Seeds Regulations.

* The Plant Biosafety Office interprets "environmentally sustainable crop management practices" to mean "crop management practices that promote long-term maintenance of ecosystem components and functions for future generations."

Please note: Depending on the product, data requirements for some criteria may be more extensive than others. Evidence, such as experimental data or peer-reviewed literature, should be available to support the rationale provided in this document.
The PBO reserves the right to request that more extensive data be supplied in support of a determination or to confirm the determination by the proponent.

Conclusion: This plant is not a PNT and is not regulated under Part V of the Seeds Regulations.

The trait of interest in this product does not appear to have the potential for significant negative environmental impact; the PBO does not have to be notified.

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