Barb’s Note: Notice that, as with the Alfalfa, the plant (tree in this instance) has a caveat. The planter is not supposed to allow the tree to flower. this is the same requirement for the planting of genetically engineered Alfalfa. Have you ever opened a bale of alfalfa? There is absolutely no way that you can control the flowering of each and every plant in an open field. I just bought Alfalfa from a local farmer, which was cut at the highest peak of protein development. This creates the highest yield for nutritional benefits. The more flowers, the less protein content in the hay. And guess what, it contains some flowers. http://www.forages.psu.edu/agfacts/agfact7.pdf
‘Confined’ field releases of Eucalyptus neither confined nor safe
‘Confidential Business Information’ makes a mockery of risk assessment
An application from ArborGen has taken a sinister turn in declaring most, if not all of the genetically modified (GM) constructs in a Eucalyptus hybrid clone to be tested in environmental release ‘Confidential Business information’, thereby precluding any meaningful independent safety and environmental assessment of the GM trees, or appropriate remedial action and identification in case of harm caused to the environment and innocent bystanders. The USDA/ APHIS (United States Department of Agriculture/Animal and Plant Health Inspection Service) has yet again made a highly flawed Environmental Assessment (EA) on the proposed release, dismissing every issue on safety largely on a priori assumptions and in the absence of real data .
ArborGen, LLC, based in Summerville, South Carolina, has applied for the ‘confined’ environmental release of the clone EH1 of a Eucalyptus grandis × Eucalyptus urophylla hybrid genetically modified with various constructs at six locations encompassing a total of 14.7 acres in the States of Alabama, Florida, Mississippi, and South Carolina . (As of September 2011, ArborGen is growing a total of approximately 67 acres of GM trees on 18 of the 32 permitted locations.) Five of the locations for the current release already have active APHIS permits for GM trees granted previously. The sixth site in South Carolina has been listed as a holding site for GM trees in previous APHIS permits and notifications, and is a new location for the release of GM Eucalyptus. ArborGen is requesting that trees be allowed to flower at four locations in Alabama, Florida and Mississippi. At two locations in South Carolina, ArborGen has requested to release trees in containers and have indicated they will not allow the trees to flower.
The stated purpose of the field release is to assess the effectiveness of different gene constructs intended to confer cold tolerance, to alter lignin biosynthesis, to alter growth rate, along with testing the efficacy of the barnase gene designed to alter fertility. In addition, the trees have been engineered with a selectable marker that confers resistance to the antibiotic kanamycin. With the exception of the C-Repeat Binding Factor (CBF) gene (see later), the barnase gene, and nptII gene, all genes are claimed as Confidential Business Information (CBI), even though they are different constructs from those in trees previously permitted for environmental releases by APHIS.
The designation of the majority of transgenic modifications CBI certainly prevents any rational, independent evaluation of the impact of those genes and the GM Eucalyptus on the environment, and on human and animal health. Unfortunately, USDA does not appear to have a mechanism for identifying and discarding frivolous CBI designations. The avalanche of CBI designations of transgenic crop and tree modifications suggests that the designation is being used to avoid doing proper risk assessment and also making it impossible for independent risk assessments that could otherwise be done; and in addition, prevent the detection of adverse side effects due to the modifications, which would also make it impossible to take appropriate remedial action. This is clearly unacceptable for protection of public health and the environment. The USDA should open the CBI designations to independent adjudication.
A number of risks were assessed in a perfunctory fashion and dismissed based to varying degrees on a priori assumptions in the absence of real data from dedicated investigations.
Alteration in susceptibility to diseases or insects
This was dismissed largely on basis of the statement in ArborGen’s application that “none of the genes being engineered into the Eucalyptus plants are expected to alter the susceptibility of the transgenic Eucalyptus plants to disease of insect damage.”
We have previously pointed out that reducing lignin in trees would make them more susceptible to attack by insects and pathogens (see  Low Lignin GM Trees and Forage Crops, SiS 23).
Risks from transgenes
Kanamycin resistance Risks from the kanamycin resistance gene is dismissed even though we have pointed out that kanamycin is still in clinical use and also kanamycin resistance cross-reacts with new antibiotics  (Kanamycin Still Used and Cross-Reacts with New Antibiotics, ISIS Report).
Cold tolerance C-Repeat Binding Factor (CBF) genes are transcription factors belonging to the AP2/EREBP family of DNA binding proteins, which recognize a cold- and drought-responsive DNA regulatory sequence designated the C-repeat (CRT)/dehydration-responsive element (DRE), found in the promoter regions of many cold-inducible genes. When CBF genes are overpressed constitutively, as when placed under the control of the CaMV 35S promoter, it was associated with stunting, reduced flowering and lack of tuber production in potatoes. However, when the CBF gene was placed under the control of a cold-induced promoter, rd29A, it increased tolerance to freezing while restoring growth and tuber production to wild-type levels. The GM Eucalyptus trees tested have the CBF gene under the control of a cold inducible promoter, which causes the gene to be expressed only when cold, and hence “not expected to produce any toxic substance and is not expected to alter the characteristics of the engineered plants”.
This large assumption is far from justified as the Eucalyptus version, EguCBF1, when over expressed, not only results in cold tolerance, but also increased water retention, higher oil gland density and wax deposition, and over expression of anthocyanin pigments . The gene network influenced by CBF gene modification produces highly pleiotropic effects. But there does not appear to have been any investigation on the production of unintended metabolites, proteins, or nuclei acids in the modified trees, all of which could have health and environmental impacts.
Gene for altered fertility We have commented on the dangers of the barnase gene on numerous previous occasions, most recently in 2008  USDA FONSI for Transgenic Poplars Absurd & Dangerous, SiS 38). It is a well-known cytotoxic protein that breaks down RNA. Because they are from a soil bacterium, and unrelated to any mammalian RNAses, they are not susceptible to eukaryotic RNAse inhibitors. Consequently, they are highly toxic, and are actually being engineered currently as a means of killing cancer cells . But USDA has dismissed the dangers of this gene in the current EA as in previous EAs. This is unconscionable. Although the barnase is being used to prevent pollen formation, this is not 100 effective, and many beneficial insects and other wild-life could well be affected.
Apart from the above genes, the genes for altered lignin (3 of them), genes for altered growth (4), non-coding sequences (undisclosed number derived from plants and plant pathogens), were all not mentioned or explicitly not disclosed under CBI, and consequently, not risk assessed at all before stating that they are not expected to pose any risks.
Mode of transformation and hazards of horizontal gene transfer
One aspect that needs to be highlighted is the mode of transformation of the GM Eucalyptus involved Agrobacterium. This is a serious unresolved hazard in genetic modification that we have drawn attention to for years, most recently in 2011  Scientists Discover New Route forGM-gene ‘Escape’ (SiS 50). Research commissioned by the UK Department of the Environment, Food and Rural Affairs (DEFRA) in the 1990s had already revealed that it is very difficult, if not impossible to get rid of the Agrobacterium vector used in creating the transgenic plant. The bacterium is likely to remain dormant even after the transgenic plants are transplanted into the soil. Hence, it is expected to facilitate horizontal gene transfer, in the first instance, to wild-type Agrobacterium in the soil, and further afield, to other bacteria and fungi in the soil. It now transpires that Agrobacterium can enlarge their host range to infect other species and exchange genes with them through hormones produced at the site of plant wounds ( Scientists Discover New Route for GM-gene ‘Escape’, SiS 50).
The applicant has indicated that they are not aware of any commercial plantings of sexually compatible Eucalyptus species within 1 000 meters of the proposed test plot location at any of these sites. Therefore, based upon the limited distance that viable pollen is likely to occur outside a tree stand, it is deemed highly unlikely that gene flow would occur outside of the confined field test sites at these locations. An Australian study, however, found that remnant populations of Eucalyptus were connected by pollen dispersal to pollen sources up to 1.94 kilometers away .
It is by now obvious that transgenes can also spread horizontally to all species interacting with the trees and pollen, in the air, in the soil and in the water, as we have repeatedly pointed out to regulators . Needless to say, horizontal gene transfer was dismissed.
Deadly yeast in Eucalyptus
Cryptococcus neoformans gattii is a yeast pathogen hosted by a variety of Eucalyptus species as well as other tree species. It causes systemic fungal infections in humans, leading to fungal meningitis and death. C. neoformans gattii has been found on a number of Eucalyptus hosts, some being grown in commercial plantations and imported and exported for ornamental use. People have contracted and died from cryptococcosis in India, Africa, Taiwan, South America and California. C. neoformans gattii infections are found particularly in AIDS patients due to their weakened immune systems. Infections with this fungus are rare in those with fully functioning immune systems. For this reason, C. neoformans gattii is sometimes referred to as an opportunistic fungus. There was an outbreak of cryptococcal disease on the eastern part of Vancouver Island, British Columbia in 1999. The disease was previously only known to occur in tropical or semi-tropical climates. The risk that these field trials will result in a higher incidence of the fungus in the US and thereby pose a risk to human health is considered negligible for the following reasons. First, there is not a clear association between E. grandis or E. urophylla and C. gattii. Second, there is no reason to believe that the genetic modification of the hybrids will alter the association of the trees with C. gattii. Third, the scale of the field tests is miniscule compared to the vast expanses of native trees that could potentially harbour the pathogen .
But there is already evidence among forest or urban trees that Eucalyptus species are homes for the deadly yeast. Furthermore, there is no vast expanse of native species in the US that are homes for the toxic yeast, according to the peer reviewed scientific publications [11-13]. The deadly yeast should have been studied in the transgenic trees rather than being groundlessly and a priori dismissed by USDA.
Issues raised in previous submissions on transgenic Eucalyptus still unresolved
The Institute of Science in Society previously submitted several briefs objecting to environmental releases of GM Eucalyptus, and dealing with other issues in more detail, such as the alteration in susceptibility to disease or insects , the potential of the Eucalyptus to harbour plant pests, the kanamycin resistance selectable marker gene, the barnase gene, genes for altered lignin, and the deadly yeast C. gattii [14-15] Field Testing Genetically Engineered Eucalyptus: Environment Assessment Still Inadequate, SiS 46, GM Eucalyptus Environmental Assessment Irregular, SiS 35]. None of the issues we raised have been properly addressed, let alone resolved.
We can only repeat our call  for a Moratorium on all GM Trees and Ban on GM Forest Trees (SiS 35).
- DEPARTMENT OF AGRICULTURE Animal and Plant Health Inspection Service [Docket No. APHIS–2011–0130] ArborGen, LLC; Availability of an Environmental Assessment for Controlled Release of a Genetically Engineered Eucalyptus Hybrid Federal Register Vol. 77, No. 28 Friday, February 10, 2012 Notices 7123.
- Eck C. Permit application 11-052-101rm received from ArborGen Field testing of genetically engineered Eucalyptus grandis X Eucalyptus urophylla Draft Environmental Assessment December 6, 2011 http://www.aphis.usda.gov/brs/aphisdocs/11_052101rm_pea.pdf
- Cummins J. Low lignin GM trees and forage crops. Science in Society 23, 38-39, 2004.
- Cummins J. Kanamycin still used and cross-reacts with new antibiotics. ISIS report, 27 May 2001, http://www.i-sis.org.uk/kanomycin.php
- Navarro M, Ayax C, Martinez Y, Laur J, El Kayal W, Marque C, Teulières C. Two EguCBF1 genes overexpressed in Eucalyptus display a different impact on stress tolerance and plant development. Plant Biotechnol J 2011, 9(1), 50-63.
- Cummins J and Ho MW. USDA FONSI for transgenic poplars absurd & dangerous. Science in Society 38, 40-41, 2008.
- Ulyanova V, Vershinina V and Ilinskaya O. Barnase and binase: twins with distinct fates. The FEBS Journal 2011, 3633-43.
- Ho MW. Scientists discover new route for GM-gene ‘escape’. Science in Society 50, 14-16, 2011.
- Knight CJ, Bailey AM, Foster GD. Investigating Agrobacterium-mediated transformation of Verticillium albo-atrum on plant surfaces. PLOS ONE 2010, 5(10): e13684. Doi:10.1371/journal.pone.0013684
- Sampson JF, Byrne M. Outcrossing between an agroforestry plantation and remnant native populations of Eucalyptus loxophleba. Mol Ecol 2008, 17(11), 2769-81.
- Chen M, Liao WQ, Wu SX, Yao ZR, Pan WH, Liao Y. Taxonomic analysis of cryptococcus species complex strain S8012 revealed Cryptococcus gattii with high heterogeneity on the genetics. Chin Med J (Engl). 2011, 124(13), 2051-6.
- Chowdhary A, Randhawa HS, Boekhout T, Hagen F, Klaassen CH, Meis JF. Temperate climate niche for Cryptococcus gattii in Northern Europe. Emerg Infect Dis. 2012, 18(1), 172-4. doi: 10.3201/eid1801.111190
- Crous PW, Groenewald JZ, Shivas RG, Edwards J, Seifert KA, Alfenas AC, Alfenas RF, Burgess TI, Carnegie AJ, Hardy GE, Hiscock N, Hüberli D, Jung T, Louis-Seize G, Okada G, Pereira OL, Stukely MJ, Wang W, White GP, Young AJ, McTaggart AR, Pascoe IG, Porter IJ, Quaedvlieg W. Persoonia 2011, 26, 108-56.
- Cummins J. Field testing genetically Engineered Eucalyptus: rnvironment assessment still inadequate. Science in Society 46, 36, 2010.
- Cummins J and Ho MW. GM eucalyptus environmental assessment irregular. Science in Society 35, 50, 2007.
- Cummins J and Ho MW. Moratorium on all GM trees and ban on forest trees. Science in Society 35, 32-34, 2007.