<p style="text-align: justify;"><b>Field Testing:</b>&nbsp;&nbsp;LLCotton25 was field tested in the United States and Puerto Rico at 43 locations over three years (1999-2001). These trials were conducted to evaluate the agronomic performance, crop quality, and plant pest potential of LLCotton25. Seed and lint yield and quality data, and evaluations of stand counts, seedling vigour, plant height, height to node ratio, leaf morphology, and plant maturity supported the conclusion that LLCotton25 was as safe to grow as other cotton varieties and did not demonstrate the potential to become a plant health risk.</p>

<p style="text-align: justify;"><b>Outcrossing:</b>&nbsp;&nbsp;Cotton (<i>G. hirsutum</i>) is mainly a self-pollinating plant, but pollen is also routinely transferred by insects, particularly bumblebees and honey bees. The pollen is heavy and sticky and the range of natural crossing is limited. Outcrossing rates of up to 28% to other cotton cultivars grown directly adjacent to the pollen source have been observed under field conditions when sufficient insect pollinators have been present. The rate of outcrossing declines rapidly with increased distance from the pollen source. Results of outcrossing research trials in the United States have shown that pollen movement decreases rapidly at 12 metres from the pollen source. Nevertheless, certified seed cotton growers in the United States must maintain an isolation distance of 202 metres between fields. If LLCotton25 were to be grown in proximity with other cotton cultivars, and sufficient insect pollinators were present and active, the possibility would exists for introgression of the <i>bar</i> gene into these cultivars.</p>

<p style="text-align: justify;">In the United States, species genetically compatible with LLCotton25 include <i>G. hirsutum</i> (wild or under cultivation), <i>G. barbadense</i> (cultivated Pima cotton), and <i>G. tomentosum</i>. There have been reports of outcrossing from <i>G. hirsutum</i> to <i>G. barbadense</i> under suitable conditions, such as the presence of suitable pollinators. <i>G. thurberi</i> is another wild species found in the United States, but it is genetically incompatible with <i>G. hirsutum</i>. While <i>G. hirsutum</i> and <i>G. tomentosum</i> are genetically compatible (both possess the AADD genome), the possibility of gene transfer is unlikely due to the non-synchronous flowering periods (day for <i>G. hirstum</i> and night for <i>G. tomentosum</i>), and lack of common pollinators. Populations of wild <i>Gossypium</i> species occur in southern Florida (<i>G. hirsutum</i>), Hawaii (<i>G. tomentosum</i>), Puerto Rico and the U.S. Virgin Islands. In other cotton growing areas of the world, species that could intercross with <i>G. hirsutum</i> include <i>G. mustelinum</i> in Brazil, and <i>G. lanceolatum</i> in Mexico.</p>

<p style="text-align: justify;"><b>Weediness Potential:</b>&nbsp;&nbsp;Cultivated <i>Gossypium hirsutum</i> is not typically considered a weed species in the United States or other countries, but it is listed as a potential weed in southern Florida. Although the cotton plant is cultivated as an annual crop, it behaves as a perennial plant in undisturbed environments and suitable climatic conditions. Cotton does not tolerate cold conditions thereby limiting its overwintering potential to southern Florida, Hawaii, and Puerto Rico.<br />
No competitive advantage was conferred to LLCotton25 other than tolerance to glufosinate ammonium. Tolerance to glufosinate ammonium will not render LLCotton25 weedy or invasive of natural habitats since none of the reproductive or growth characteristics of this line have been modified. In the event of the formation of herbicide tolerant hybrids, there would no competitive advantage conferred on any hybrid progeny in the absence of sustained glufosinate ammonium use. The herbicide tolerant plants could be controlled by cultural, or mechanical means, or by using herbicides other than glufosinate ammonium. The same treatments would apply to cotton volunteers appearing in a following crop.</p>

<p style="text-align: justify;">LLCotton25 did not exhibit any increased tendency towards weediness, compared to its unmodified parental line, as demonstrated by the nonsignificant differences in parameters such as seed germination, seedling vigour, time to first bloom, and plant maturity.</p>

<p style="text-align: justify;"><b>Secondary and Non-target Adverse Effects:</b>&nbsp;&nbsp;Field trial observations of LLCotton25 revealed no negative effects on nontarget organisms; no alterations in the population levels of beneficial insects and birds were observed. LLCotton25 did not display any altered susceptibility to diseases such as <i>Rhizoctonia</i> sp. and <i>Phymototricum</i> sp. leading to the conclusion that its plant pest potential (<i>e.g</i>., increased potential to harbour a disease or become an alternate host) was not changed. A similar conclusion was reached with regard to susceptibility to insect pests such as budworm, boll weevil, thrips, cotton aphids, plant bugs, whiteflies, cutworms, stinkbug, and cotton bollworm. Data on germination, seedling vigour, plant growth and morphology, and agronomic performance indicated no significant differences among LLCotton25, its unmodified parental line, and other cotton cultivars, leading to the conclusion that LLCotton25 would not be expected to grow in new habitats and displace other species. The PAT enzyme is expressed at very low levels in the plant tissues, has a high substrate specificity to L-phosphinothricin, is rapidly degraded in simulated gastric and intestinal fluid conditions, and does not have any sequence similarities to known allergens or toxins. This enzyme is also present in the environment since it is produced by species of the soil bacterium <i>Streptomyces,</i> with no known adverse effects on humans or animals. This information led to the determination that LLCotton25 will not result in altered impacts on non-target organisms, compared to conventional cotton varieties.</p>

<p style="text-align: justify;"><b>Impact on Biodiversity:</b>&nbsp;&nbsp;The genetic modification that resulted in LLCotton25 did not confer any novel phenotypic characteristics that would increase its survival in either unmanaged habitats, or in areas outside the current geographic range of cotton production. Since the potential of gene transfer to wild relatives in the United States is quite minimal, the risk of transferring the herbicide tolerance trait to species in unmanaged habitats is insignificant.</p>
