<p style="text-align: justify;"><b>Field Testing</b></p>

<p style="text-align: justify;">Maize event DAS-59122-7 was field tested in the United States in 2001, 2002 and at 18 separate sites in 2003. Agronomic characteristics of hybrids derived from DAS-59122-7 such as seed dormancy, vegetative vigour, early stand establishment, time to maturity, flowering period, susceptibilities to various pests and pathogens, and seed production were compared to those of unmodified counterparts. Nutritional components of DAS-59122-7, such as proximates, amino acids and fatty acids were compared with those of unmodified counterparts.</p>

<p style="text-align: justify;"><b>Outcrossing</b></p>

<p style="text-align: justify;">Pollen production and viability were unchanged by the genetic modification resulting in DAS-59122-7, therefore pollen dispersal by wind and outcrossing frequency should be no different than for other maize varieties. Gene exchange between DAS-59122-7 and other cultivated maize varieties will be similar to that which occurs naturally between cultivated maize varieties at the present time. In the United States and Canada, where there are no plant species closely-related to maize in the wild, the risk of gene flow to other species appears remote. Feral species in the United States related to corn cannot be pollinated due to differences in chromosome number, phenology (periodicity or timing of events within an organism&rsquo;s life cycle as related to climate, <i>e.g</i>., flowering time) and habitat.</p>

<p style="text-align: justify;">Maize (<i>Zea mays</i> ssp. <i>mays</i>) freely hybridizes with annual teosinte (<i>Zea mays</i> ssp. <i>mexicana</i>) when in close proximity. These wild maize relatives are native to Central America and are not present in the United States, except for special plantings. <i>Tripsacum</i>, another genus related to <i>Zea</i>, contains sixteen species, of which twelve are native to Mexico and Guatemala. Three species of <i>Tripsacum</i> have been reported in the continental United States: <i>T. dactyloides</i>, <i>T. floridanum</i> and <i>T. lanceolatum</i>. Of these, <i>T. dactyloides</i>, Eastern Gama Grass, is the only species of widespread occurrence and of any agricultural importance. It is commonly grown as a forage grass and has been the subject of some agronomic improvement (i.e., selection and classical breeding). <i>T. floridanum</i> is known from southern Florida and <i>T. lanceolatum</i> is present in the Mule Mountains of Arizona and possibly southern New Mexico. Even though some <i>Tripsacum</i> species occur in areas where maize is cultivated, gene introgression from maize under natural conditions is highly unlikely, if not impossible. Hybrids of <i>Tripsacum</i> species with <i>Zea mays</i> are difficult to obtain outside of the controlled conditions of laboratory and greenhouse. Seed obtained from such crosses are often sterile or progeny have greatly reduced fertility.</p>

<p style="text-align: justify;"><b>Weediness Potential</b></p>

<p style="text-align: justify;">The history and biology of maize indicates that non-transgenic plants of this species are not invasive in unmanaged habitats. Maize does not possess the potential to become weedy due to traits such as lack of seed dormancy, the non-shattering nature of corn cobs, and the poor competitive ability of seedlings. The data generated from field trials shows that DAS-59122-7 and derived hybrids are similar to their counterparts in this respect. Data submitted by the developer on the reproductive and survival biology of hybrids derived from DAS-59122-7 determined that early stand establishment, flowering period, vegetative vigor, time to maturity and seed production were within the normal range of expression of these traits currently displayed by commercial hybrids. No competitive advantage was conferred to DAS-59122-7, other than that conferred by resistance to rootworm and tolerance to glufosinate herbicide. These traits were demonstrated not to render maize weedy or invasive of natural habitats since none of the reproductive or growth characteristics were modified. The above considerations lead to the conclusion that DAS-59122-7 has no altered weed or invasive potential compared to currently commercialized corn.</p>

<p style="text-align: justify;"><b>Secondary and Non-Target Adverse Effects</b></p>

<p style="text-align: justify;">The agronomic characteristics of DAS-59122-7 hybrids were shown to be within the range of values displayed by currently commercialized maize hybrids, and indicate that the growing habit of maize will not be inadvertently altered by cultivation of DAS-59122-7. Field observations did not indicate modifications of disease and pest susceptibilities, other than to rootworm, which is not known to be a principal factor restricting the establishment or distribution of maize.</p>

<p style="text-align: justify;">Some of the genetic elements introduced into DAS-59122-7 were derived from known plant pathogens, but in all cases the genes responsible for the pathogenic qualities of the pathogen were not introduced. Therefore, the introduction of genetic material for <i>Diabotica spp.</i> resistance and glufosinate tolerance would not be expected to result in DAS-59122-7 expressing novel pathogenic characteristics.</p>

<p style="text-align: justify;">The history of use and literature suggest that the bacterial Cry34Ab1 and Cry35Ab1 toxins are not toxic to humans, other vertebrates, and non-coleopteran invertebrates. This protein is active only against specific coleopteran insects and, as the specificity of the Cry34Ab1 and Cry35Ab1 protein insecticidal activity is dependent upon their binding to specific receptors present in the insect mid-gut, these insecticidal proteins are not expected to adversely affect other invertebrates or vertebrate organisms, including non-target birds, mammals and humans. Laboratory and field studies on representative species supported these expectations.</p>

<p style="text-align: justify;">A series of diet bioassays was conducted with microbially-expressed Cry34Ab1 and Cry35Ab1 proteins to characterize the insecticidal specificity. Test species were: northern corn rootworm (<i>Diabrotica barberi</i>), western corn rootworm (<i>Diabrotica virgifera virgifera</i>), southern corn rootworm (<i>Diabrotica undecimpunctata howardi</i>), European corn borer (<i>Ostrinia nubilalis</i>), corn earworm (<i>Helicoverpa zea</i>), black cutworm (<i>Agrotis ipsilon</i>) and corn leaf aphid (<i>Rhopalosiphum maidis</i>). Only corn rootworm (<i>Diabrotica</i> spp.) experience high mortality when exposed to microbially-expressed Cry34 and Cry35 proteins.</p>

<p style="text-align: justify;">Acute dietary toxicity studies were conducted in laboratory tests on the effect of the Cry34 and Cry35Ab1 proteins on non-target invertebrates, including honeybee larvae, green lacewing larvae, ladybird beetles (<i>Hippodamia convergens</i> and <i>Coleomegilla maculate</i>), parasitic wasp, ground beetle, daphnia, collembola, and earthworm. Data were also submitted on non-target vertebrates including mice and the rainbow trout. Cry34 and Cry35Ab1 proteins were demonstrated to be safe to these indicator species when ingested alone or in combination, at doses exceeding the levels of direct or indirect exposure to Cry34 and Cry35Ab1 proteins from corn 59122 tissues. Although growth inhibition was reported when <i>Coleomegilla maculate</i> larvae were fed Cry34Ab1 protein at about 10 times the field pollen value, no delay in development or weigh reduction were observed when larvae were fed a diet composed of 50% corn line 59122 pollen, which represents a conservative estimate of the potential exposure in the field. Therefore, no sub-lethal effects on <i>C. maculate</i> ladybird beetle are expected from exposure to event DAS-59122-7.</p>

<p style="text-align: justify;">Field based observational studies were also performed on the stability and abundance of non-target arthropods communities and individual species where Cry34 and Cry35Ab1 expressing corn is grown. The species studied were representatives of various functional groups (predators, parasitoids, herbivores and detritivores) and were sampled using visual observations of corn plants, sticky traps, pitfall traps or litterbag traps. The studies did not reveal any negative impact on the abundance of these non-target organisms relative to the non-transgenic control hybrid. The Cry34 and Cry35Ab1 protein was shown to degrade readily in soil, indicating that it is unlikely to accumulate and persist in soil.</p>

<p style="text-align: justify;">The herbicide tolerance trait of event DAS-59122-7 is conferred by expression of the phosphinothricin N-acetyltransferase (PAT) gene. The PAT protein has been studied extensively and has been found to be safe for non-target organisms, including human beings. This gene has been used in a number of other approved transgenic maize events, such as T25, BT11, DLL25 and DAS-06275-8.</p>

<p style="text-align: justify;">Maize is not known for the production of significant levels of endogenous toxins and the transformation event that produced event DAS-59122-7 would not be expected to induce their synthesis. Maize is however, known to produce low levels of trypsin inhibitor and phytic acid and the levels of these compounds in DAS-59122-7 were found to be within the range of conventional corn lines. The genetic modification, therefore did not alter the expression of endogenous anti-nutrients.</p>

<p style="text-align: justify;"><b>Impact on Biodiversity</b></p>

<p style="text-align: justify;">DAS-59122-7 has no novel phenotypic characteristics which would extend its use beyond the current geographic range of maize production. Since maize does not out-cross to wild relatives in the united States or Canada, there will be no transfer of novel traits to unmanaged environments. In addition the novel traits were determined to pose minimal risks to non-target organisms.</p>

<p style="text-align: justify;">DAS-59122-7 provides an alternative method to existing methods of control of rootworms, an important agricultural pest of maize. The control of agricultural pest species is a common practice that is not restricted to the environmental release of transgenic plants. Therefore, the reduction in local pest species as a result of the cultivation of DAS-59122-7 does not present a significant change from existing agricultural practices. At present, the use of chemical insecticides to control rootworm is permitted, although crop rotation represents a major method of rootworm control in some countries.</p>

<p style="text-align: justify;">DAS-59122-7 also provides an alternative method of weed control in maize production. The use of broad spectrum herbicides has the intended effect of reducing local weed populations within agricultural fields and this may reduce local weed species biodiversity, and possibly other trophic levels which utilize these weed species. It must be noted, however, that reduction in weed biodiversity in agricultural fields is not unique to the use of transgenic plants, and is a common practice in virtually all modern agricultural systems. It was therefore concluded that DAS-59122-7 does not present a significantly altered impact on biodiversity in comparison to maize varieties currently being grown.</p>
