1

Assessment of Plant Pest Risk for International Flower Developments Pty. Ltd. IFD524Ø1-4 and IFD-529Ø1-9 Rosa x hybrida (rose) varieties

International Flower Developments Pty. Ltd. (IFD) (Victoria, Australia) has petitioned 

APHIS for a determination that IFD-524Ø1-4 and IFD-529Ø1-9 roses (Rosa x hybrida) 

are unlikely to pose a plant pest risk and, therefore, are no longer regulated articles under 

regulations at 7 CFR part 340.

History of Development of IFD-524Ø1-4 and IFD-529Ø1-9 Rosa x hybrida varieties

Working with a rose grower in the U.S., Florigene and IFD have been researching and 

developing two lines of genetically engineered roses since 2004. Both of these rose lines 

include added genes for flavonoid 3’-5’ hydroxylase (from a black pansy, Viola tricolor) 

and anthocyanin 5-acyltransferase (from torenia, Torenia hybrida). These rose lines also 

contain the neomycin phosphotransferase gene (from the bacterium Escherichia coli)

which was used for selection in the laboratory. Both of these rose lines have been 

approved for commercial use, including environmental release, in Japan (petition pp. 51-

53). One line (IFD-524Ø1-4) has also been approved for commercial use/environmental 

release in Australia (petition p. 54) (Costs of the regulatory request for the other rose line 

and the small size of the Australian market led the company to only request approval for 

one line there). Addition of the Viola and Torenia genes alter the anthocyanin biosynthesis 

pathways and shunt some of these biochemicals toward production of the delphinidinbased anthocyanins, resulting in production of blue pigments in these rose lines. 

Production of these blue pigments alters the flower color of these rose lines (noted in 

Petition as the differences comparing Figure 14 (p. 39) with Figures 15 and 16 (pp. 40)). 

Description of added genes

Florigene’s IFD-524Ø1-4 and IFD-529Ø1-9 roses (Rosa x hybrida) were produced using 

disarmed Agrobacterium tumefaciens (Petition p. 24) and contain 3 transgene fragments 

(Petition Figure 5 and Table 6, pp. 25-26) from plasmid pSPB130 (Figure 5 and Table 6, 

pp. 25-26 of the Petition): 

(1) A nopaline synthase promoter/ neomycin phosphotransferase gene/ nopaline synthase 

terminator (NOS/NPT II/ NOS) fragment:

• The NOS promoter is from Agrobacterium tumefaciens and drives production of 

the npt II gene. 

• The npt II gene (from Escherichia coli) results in production of neomycin 

phosphotransferase and confers tolerance to the antibiotic kanamycin. Kanamycin 

is used to select for transgenic tissues under laboratory conditions. 

• The NOS terminator is from Agrobacterium tumefaciens. 

(2) A cauliflower mosaic virus 35S promoter/ flavonoid 3’, 5’- hydroxylase gene/ 

nopaline synthase fragment (CaMV35S/ F3’5’H/ NOS) fragment:
2

• The enhanced 35S promoter from cauliflower mosaic virus results in production of 

flavonoid 3’, 5’- hydroxylase.

• The flavonoid 3’, 5’-hydroxylase gene from Viola tricolor results in production of 

this biochemical. 

• The NOS terminator is from Agrobacterium tumefaciens. 

(3) A cauliflower mosaic virus 35S promoter/ anthocyanin 5-acyltransferase/ nopaline 

synthase fragment (CaMV35S/ 5AT/ NOS) fragment:

• The enhanced 35S promoter from cauliflower mosaic virus drives production of 

anthocyanin 5-acyltransferase.

• The anthocyanin 5-acyltransferase gene from Torenia hybrida results in 

production of this biochemical.

• The NOS terminator is from Agrobacterium tumefaciens. 

Production of F3’5’H and 5AT enzymes shunts production of anthocyanins in these rose 

lines away from the cyanidin based anthocyanins (pink) and toward production of 

delphinidin based anthocyanins (blue) (indicated in Figure 4, p. 22 of the Petition).

Southern blots were used to analyze gene insertion from plasmid pSPB130 into the 

rose genome and to examine the integrity and expression of the inserted DNA. APHIS

evaluated Florigene’s Southern blot data and came to several conclusions regarding 

insertion of the various genes into these 2 rose lines. Based on gel quantitation analysis of 

DNA, there are between 1 and 4 copies of each of the genes (npt II, F3’5’H, and 5AT) in 

both IFD-524Ø1-4 and IFD-529Ø1-9. Additionally, the Southern blots show the presence 

of multiple DNA bands (Figures 6-10, pp. 30-34 of the Petition) indicating that it is also 

likely that some gene rearrangements have occurred. The DNA banding patterns does

indicate that these genes were inserted differently in each of the two rose lines. 

Florigene also provided Northern analyses documenting production of RNA from the 

inserted genes (Figure 13, p 38 of the Petition). Each gene was expressed in the transgenic 

plants and there was no expression noted in the control non-transformed rose line. 

Further, Florigene presented data on production of delphinidin and cyanidin in the petals 

of these roses. Both transgenic lines showed accumulation of delphinidin (a blue 

anthocyanin) and much decreased production of cyanidin (a red anthocyanin) (Table 5, p. 

23 of the Petition). The control non-transformed rose did not produce delphinidin but did 

produce cyanidin. 

Plant Pest Risk Assessment

This plant pest risk assessment is to determine whether IFD-524Ø1-4 and IFD-529Ø1-9 

are likely to pose plant pest risks. Under the current Part 340 biotechnology regulations, if 

APHIS determines that a GE organism is unlikely to pose a plant pest risk, APHIS then 

has no regulatory authority under Part 340 or the plant pest provisions of the Plant 

Protection Act to continue to regulate that organism.
3

APHIS administers the regulations 7 CFR part 340 under the authority of the Plant 

Protection Act of 2000 (PPA). The PPA states that:

“The term ‘plant pest’ means any living stage of any of the following that can 

directly or indirectly injure, cause damage to, or cause disease in any plant or 

plant product:

(A) A protozoan.

(B) A nonhuman animal.

(C) A parasitic plant.

(D) A bacterium.

(E) A fungus.

(F) A virus or viroid.

(G) An infectious agent or other pathogen.

(H) Any article similar to or allied with any of the articles 

specified in the preceding subparagraphs.

The Animal and Plant Health Inspection Service of the United States Department of 

Agriculture (USDA-APHIS) has prepared a Plant Pest Risk Assessment in response to a 

petition (APHIS No. 08-315-01p) from International Flower Developments Pty Ltd. 

APHIS regulation 7 CFR 340.6(c) (4) stipulates the information needed to be considered 

in a petition for nonregulated status. APHIS uses information submitted by the applicant 

related to plant pest risk characteristics, disease and pest susceptibilities, expression of the 

gene product, new enzymes, or changes to plant metabolism, weediness of the regulated 

article, any impacts on the weediness of any other plant with which it can interbreed, and 

the transfer of genetic information to organisms with which it cannot interbreed for IFD524Ø1-4 and IFD-529Ø1-9 roses. Issues related to agricultural or cultivation practices and 

the effects of the regulated article on non-target organisms will be considered in the 

Environmental Assessment for these rose lines. 

Based on information on the biology of rose

(http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/riskassessments-1, accessed 

1/26/10), data presented in the Petition and scientific data relevant to a discussion of plant 

pest risk, APHIS has concluded the following regarding IFD-524Ø1-4 and IFD-529Ø1-9 

roses: 

Potential impacts of altered disease and pest susceptibilities 

USDA-APHIS assessed whether IFD-524Ø1-4 and IFD-529Ø1-9 roses are likely to have 

significantly altered disease and pest susceptibilities. The assessment encompasses a 

consideration of introduced traits and interactions with pests and diseases. 

Hybrid tea roses (Rosa x hybrida) are not considered plant pests in the United States. 

None of the gene sequences derived from the plant pests (Agrobacterium and cauliflower 

mosaic virus) that were incorporated into these rose lines result in the production of 

infectious agents or disease symptoms in plants, and so they are unlikely to pose a plant 

pest risk. The description of the genetic modifications, including genetic elements, 
4

expression of the gene products and their functions in these roses has been summarized 

above.

Cultivated hybrid tea roses are susceptible to numerous insect pests and diseases. 

Florigene has noted some of the major ones (Petition Addendum, p. 1). A variety of 

insecticides, miticides and other pesticides are commonly used on roses 

(http://urbanext.illinois.edu/roses/disease.cfm, accessed 1/25/10; 

http://www.ipm.ucdavis.edu/PMG/PESTNOTES/pn7466.html, accessed 1/25/10). 

Florigene has contracted for growing these rose lines in greenhouses in California for over

4 years. The grower (Jackson and Perkins Wholesale, Inc.) has noted that these rose lines

grow normally in all respects (Petition Addendum, p. 2). They further make note of the 

major pests of roses in California: two spotted spider mites, powdery mildew and downy 

mildew. They also indicate that all of these pests are controlled using common treatments

and conventional protocols (Petition Addendum, p. 2). These rose lines have also been 

grown extensively in Columbia under typical rose cultivation conditions there. The 

grower there indicates that both the transgenic and non-transgenic parental lines are 

susceptible to mildew but that there was no difference between the transgenic and nontransgenic plants (Petition Addendum, p. 2). 

Finally, this information submitted by Florigene indicates that there are no significant 

differences in terms of pest or disease susceptibilities between these rose lines and their 

non-transgenic counterparts.

Potential impacts from new gene products, changes to plant metabolism or 

composition

As a point of reference, Figure 4 in the Petition (p. 22) presents the basic biochemical 

pathways for production of anthocyanins in plants. 

New proteins produced in both IFD-524Ø1-4 and IFD-529Ø1-9 include the following:

1. Neomycin phosphotransferase (NPT II)

2. Flavonoid 3’, 5’-hydroxylase (F3’5’H)

3. Anthocyanin 5- acyltransferase (5AT)

NPT II is a common protein found in a number of genetically engineered plants that have 

been widely planted across the U.S. and in other parts of the world. In every case, no 

issues related to health or environmental safety have been noted (APHIS petitions 04-317-

01p, 04-264-01p, 01-137-01p, 01-206-02p, 01-206-01p, 95-352-01p, 96-051-01p, 95-045-

01p, 94-308-01p). NPT II confers tolerance to the antibiotic kanamycin and is used in a 

laboratory setting to select tissues transformed with the genes of interest. 

Flavonoid 3’, 5’-hydroxylase (F3’5’H) is an enzyme that is widely found in nature in 

plants producing anthocyanins, most often blue colors. This enzyme can be found in 

grapes (Bogs 2006), petunia (Toguri 1993), eggplant (Solanum melongena) (Chapple 

1998), gentian, torenia, campanula and many other plants (Tanaka 2006). F3’5’H is in the 
5

cytochrome P450 family of enzymes (designated in the CYP75A subfamily) (Nelson 

2009, http://drnelson.utmem.edu/CytochromeP450.html, accessed 1/26/10). Plant species 

lacking flavonoid 3’-5’-hydroxylase, such as non-engineered roses, do not make the blue 

delphinidin-based anthocyanins (Deng 2001).

Anthocyanin 5-acyltransferase (5AT) is also an enzyme that is widely found in nature in 

plants producing anthocyanins. In the anthocyanin biosynthesis pathways, this enzyme, as 

well as related anthocyanin acyltransferases, act to alter the biochemical structure of 

anthocyanin glucosides (AGS) (such as pelargonidin GS, cyanidin GS, and delphinidin 

GS) and make the resulting pigment (anthocyanin) more chemically stable in the plant cell 

(Nakayama et al 2003). 

As described above, compositional assessment data supplied in the Petition regarding 

production of DNA, RNA and delphinidin supports the conclusion that IFD-524Ø1-4 and 

IFD-529Ø1-9 roses contain introduced f3’5’h and 5at genes from Florigene plasmid 

pSPB130. 

Florigene also provided an assessment of delphinidins found in common foods and 

common flowers (Tables 17 and 18, pp. 48-49 of the Petition). As noted in the tables, 

many foods, which are consumed widely, and ornamental flowers, which are grown 

widely, contain measurable quantities of delphinidin. None of the foods or ornamental 

plants noted are known to pose unique environmental risks because of the presence of 

delphinidin, its precursor biochemicals or catalytic enzymes (i.e., F3’5’H or 5AT) in the 

anthocyanin pathways. Specific data on toxicity and potential environmental effects of 

F3’5’H and 5AT is sparse but information on the chemistry of delphinidins and other 

anthocyanins is noted (Beheshti 2008; Vilanova 2009; Yu 2006). None of the documents 

identified or noted raise environmental concerns related to new gene products, changes in 

plant metabolism or plant composition. 

Based on all the noted considerations, APHIS concludes that IFD-524Ø1-4 and IFD529Ø1-9 roses pose no more of a plant pest risk from new gene products, changes to plant 

metabolism or composition than conventional hybrid tea roses.

Potential impacts from outcrossing of IFD-524Ø1-4 and IFD-529Ø1-9 roses 

to wild relatives 

Florigene has described the biology of roses, including hybrid tea roses, in its Petition (pp. 

10-20). They note that the first hybrid tea roses were produced in France in 1867 and that 

since then, hundreds of new cultivars of hybrid tea roses have been introduced. They also 

note that cultivated rose is the most widely produced cut flower crop in the world and that 

over 60 million rose plants are planted each year just to meet demand for cut flower 

production. The Australian Office of Gene Technology Regulator (OGTR) has also 

produced an extensive document on the biology of Rosa x hybrida

(http://www.health.gov.au/internet/ogtr/publishing.nsf/Content/rose3/$FILE/biologyrose09.pdf, accessed 1/25/10) and completed a risk assessment of one of 

these rose lines (IFD-524Ø1-4) in 2009 

(http://www.ogtr.gov.au/internet/ogtr/publishing.nsf/Content/dir090-
6

4/$FILE/dir090rarmp.doc, accessed 1/25/10). They describe the taxonomy of roses, rose 

species, centers of diversity and domestication, development and commercialization of 

modern rose cultivars, cultivation practices, breeding work (including use of “sports” as 

well as irradiation induced mutations), plant morphology, plant development and 

reproduction, biochemistry, biotic and abiotic interactions, disease and insect pests and 

weediness. Most of the information in those documents is directly relevant to this 

application and plant pest risk assessment and is incorporated here by reference. 

Florigene’s description and analysis of IFD-524Ø1-4 and IFD-529Ø1-9 roses indicates 

that they are both L1 periclinal chimeras1. The significance of this is that the f3’5’h and 

5at genes introduced are only found and expressed in the epidermal tissues and cannot be 

passed on to progeny through cross pollination (i.e., outcrossing). Florigene demonstrated 

this in the various pollination and in situ hybridization experiments they conducted during 

development of these lines (Petition pp. 35-37). Florigene analyzed over 100 seeds of 

hybrid pollinations with grandiflora and floribunda roses and found none that contained 

the IFD-524Ø1-4 and IFD-529Ø1-9 transgenes. They also analyzed 19 seeds from 

pollinations with wild rose (Rosa multiflora) and found none that contained the introduced 

genes. They also conducted specific RNA hybridization experiments using petal tissues of 

control and transgenic lines and identified stained cells only in the epidermis of the 

transgenic rose lines (Petition p. 37). 

In assessing the risk of gene introgression from IFD-524Ø1-4 and IFD-529Ø1-9 roses into 

its sexually compatible relatives, APHIS considers two primary issues: 1) the potential for 

gene flow and introgression and, 2) the potential impact of introgression. 

Given the demonstrated L-I chimeric nature of these rose lines, the potential for gene flow 

and introgression into sexually compatible relatives of rose is essentially zero. Since gene 

flow cannot occur, any potential impact of introgression, therefore, is also zero. 

Potential impacts based on the relative weediness of IFD-524Ø1-4 and IFD529Ø1-9 rose.

APHIS assessed whether Florigene IFD-524Ø1-4 and IFD-529Ø1-9 roses are any more 

likely to become weeds than the non-transgenic recipient rose line, or other rose currently 

cultivated. The assessment encompasses a thorough consideration of the basic biology of 

rose and an evaluation of unique characteristics of these rose lines.

The biology and cultivation practices of hybrid tea rose have been well described 

(http://www.health.gov.au/internet/ogtr/publishing.nsf/Content/rose3/$FILE/biologyrose09.pdf) and Florigene has also provided information on Rosa x 

hybrida cultivation, taxonomy, pollination, weediness and potential modes of gene flow 

(Petition pp. 10-20). Cultivated hybrid tea roses are all complex hybrids (Petition p. 11) 

derived from breeding work done over centuries from more than 5 different rose species 

 1 An L-I periclinal chimera is a type of plant chimera in which the L-I cell layer has a different genetic 

make-up than the L-II and L-III cell layers. The L-I cell layer gives rise to epidermal tissues of a plant while 

the L-II and L-III layers give rise to reproductive tissues, vascular tissues and other internal plant tissues. 
7

and numerous rounds of selection and cross hybridization at each step. Cut flower rose 

varieties are the most widely cultivated cut flower crop worldwide with sales of over 6 

billion stems per year (Petition p. 12). Florigene has noted that USDA lists over 100

species of Rosa in its Plants database (http://plants.usda.gov/). Only 3 of these Rosa

species are common weeds in the U.S. (Rosa arkansana, Rosa multiflora, and Rosa 

rugosa) (http://plants.usda.gov/java/noxiousDriver#state) and Rosa arkansana is native to 

the U.S. None of the Rosa species are listed as Federal noxious weeds. 

In the U.S., Rosa x hybrida is not listed as a weed in several major weed references 

(Crockett 1977; Holm et al. 1979; Muenscher 1980) nor is it listed as a noxious weed 

species by the U.S. Federal Government 

(http://www.aphis.usda.gov/plant_health/permits/organism/federal_noxious_weeds.shtml,

accessed 12/22/09). As noted by Florigene, these roses also do not readily propagate by 

vegetative means (Petition, p. 46). Because of the L-I chimeric nature of these rose lines, 

they also will not disperse the genes for f3’5’h and 5at by pollen. As is typical of hybrid 

tea roses grown for cut flowers, the likelihood of moving genes around by seed is very 

small as these roses are cut and harvested long before hips or seeds would mature. 

Compared to the non-transgenic progenitor roses, these attributes are no different. 

Weediness for the purposes of this part of the plant pest risk assessment is an attribute, 

which causes a crop to act as a weed due to the addition of genes, in comparison to the 

non-transgenic comparator. If the fitness of these rose lines improves in natural or 

agricultural ecosystems due to the inserted DNA, the potential for weediness could 

increase. The following analysis of the inserted DNA is intended to document that IFD524Ø1-4 and IFD-529Ø1-9 roses have a negligible likelihood of increased weediness. As 

described previously, these rose lines differ only in the expression of the F3’5’H and 5AT 

enzymes which ultimately result in production of blue delphinidin-based anthocyanins. 

Florigene collected data on numerous phenotypical features of these rose lines. Data 

collected related to plant height, flower stem length, flower height, flower diameter, petal 

length and width, number of pistils, number of stamens, and others (Petition p. 42). While 

some of these characteristics showed statistical differences between the non-transgenic 

and transgenic lines, none of these characteristics stand out as ones that would provide a 

fitness advantage to these lines in an unmanaged situation. Florigene also collected data 

on pollen viability, pollen grain germination, and pollen diameter (Petition pp. 43-45). 

None of the data on pollen characteristics showed any statistical significance between the 

non-transgenic and transgenic lines. 

None of the characteristics that APHIS considers in its assessment of weediness point to 

the potential for these rose lines to become more weedy or invasive than the nontransgenic comparator lines. APHIS concludes that introduction of these rose lines do no 

result in increased plant pest risk related to weediness. 

Potential Impacts on Target and Non-target Organisms, Including Beneficial 

Organisms
8

Based on the data provided by the applicant and existing literature, APHIS evaluated the 

potential for deleterious effects or significant impacts of these rose lines on non-target or

beneficial organisms.

The genes introduced into these rose lines result directly in production of the F3’5’H and 

5AT proteins and indirectly in production of delphinidin, a blue pigment. Florigene notes 

that delphinidin and delphinidin derivatives are contained in many common foods in 

relatively large amounts (Petition, pp. 48-50). Anyone or anything consuming these foods, 

therefore, consumes delphinidin as well as the F3’5’H and 5AT proteins required for its 

production. The 5AT protein is also found in foods containing other related anthocyanin 

pigments (Petition, p. 50). As noted by Florigene, the amount of delphinidin found in 

these rose lines is approximately 100 times less than that found in fresh blueberries 

(Petition, p. 49). Florigene also notes that, as a group, anthocyanins have a very low 

toxicity (Petition, p. 48; http://www.inchem.org/, accessed 1/25/10). 

In addition to this lack of toxicity associated with these rose lines, Florigene notes that 

these roses will be grown in a limited number of locations under highly controlled 

conditions by experienced rose growers (Petition, p. 46). As such, exposure to organisms 

outside these conditions will be extremely limited. 

This data submitted by the applicant indicates that the interactions between these rose 

lines and other roses, including the control lines, are similar. Considering all this, APHIS 

concludes that Florigene rose lines IFD-524Ø1-4 and IFD-529Ø1-9 are unlikely to pose 

safety risks to non-target or beneficial organisms. 

Potential impacts from transferring genetic information from IFD-524Ø1-4 

and IFD-529Ø1-9 rose to organisms with which it cannot interbreed.

APHIS examined the potential for the new genetic material inserted into these rose lines 

to be horizontally transferred to other organisms without sexual reproduction and whether 

such an event could lead directly or indirectly to disease, damage, injury or harm to plants, 

including the creation of more virulent pathogens. First, many genomes (or parts thereof) 

have been sequenced from bacteria that are closely associated with plants including 

Agrobacterium and Rhizobium (Kaneko et al. 2000; Wood et al. 2001; Kaneko et al. 

2002). There is no evidence that these organisms contain genes derived from plants. 

Second, in cases where review of sequence data implied that horizontal gene transfer 

occurred, these events are believed to occur on an evolutionary time scale on the order of 

millions of years (Koonin et al. 2001; Brown 2003). Third, transgene DNA promoters and 

coding sequences are optimized for plant expression, not prokaryotic bacterial expression. 

Thus even if horizontal gene transfer occurred, proteins corresponding to the transgenes 

are not likely to be produced. Fourth, the FDA has evaluated horizontal gene transfer from 

the use of antibiotic resistance marker genes, and concluded that the likelihood of transfer 

of antibiotic resistance genes from plant genomes to microorganisms in the 

gastrointestinal tract of humans or animals, or in the environment, is remote (Council for 

Biotechnology Information, 2001; http://vm.cfsan.fda.gov/~dms/opa-armg.html, accessed 

1/26/10). Finally, a recent review of issues related to horizontal gene transfer concluded 

that this type of gene transfer is unlikely to occur and poses negligible risks to human 
9

health or the environment (Keese 2008). Therefore APHIS concludes that horizontal gene 

transfer is unlikely to occur and thus poses no significant environmental or plant pest risk. 

Conclusion 

APHIS has reviewed and conducted a plant pest risk assessment on Florigene’s IFD524Ø1-4 and IFD-529Ø1-9 roses. Due to the lack of plant pest risk from the inserted 

genetic material, the lack of weediness characteristics of these rose lines, the lack of 

atypical responses to disease or plant pests, the lack of deleterious effects on non-targets 

or beneficial organisms in the agro-ecosystem, and the lack of horizontal gene transfer, 

APHIS concludes that IFD-524Ø1-4 and IFD-529Ø1-9 rose is unlikely to pose a plant 

pest risk.

Reference:

Bogs, J., A. Ebadi, D. McDavid, and S.P. Robinson. 2006. Identification of the Flavonoid 

Hydroxylases from Grapevine and Their Regulation during Fruit Development. 

Plant Phys. 140: 279-291. 

Beheshiti, E.H. 2008. Characterization of antioxidant activities from fruits rich in 

delphinidin or malvidin anthocyanins. M.S. Thesis in Food Science, The 

University of British Columbia.

Brown, J. R. 2003. Ancient horizontal gene transfer. Genetics. 4:121-132.

Chapple, C. 1998. Molecular-genetic Analysis of Plant Cytochrome P450-dependent 

Monooxygenases. Annu. Rev. Plant Physiol. Plant Mol. Biol. 49: 311-343. 

Council for Biotechnology Information. 2001. The Use of Antibiotic Resistance Markers 

to Develop Biotech Crops. 

(http://www.foodsafety.ksu.edu/articles/12/ant_res_mark_council4biotech.pdf, 

accessed 1/26/10). 

Crockett, L. (1977) Wildly Successful Plants: North American Weeds. University of 

Hawaii Press, Honolulu, Hawaii. 609 pp.

Deng, C and T.M. Davis. 2001. Molecular identification of the yellow fruit color (c) locus

in diploid strawberry: a candidate gene approach. Theor. Appl. Genet. 103: 316-

322.

Gebhard, F., Smalla, K. 1998 Transformation of Acinetobacter sp. strain BD413 by 

transgenic sugar beet DNA. Apl. Environ. Microbiol. 64, 1550-1554.

Holm, L., Pancho, J. V., Herbarger, J. P., and Plucknett, D. L. (1979) A Geographical 

Atlas of World Weeds. John Wiley and Sons, New York. 391 pp.
10

Johnson, R., Narvaez, J., AN, G., Ryan, C.A. (1989) Expression of proteinase inhibitors I 

and II in transgenic tobacco plants: Effects on natural defense against Manduca 

sexta larvae. Proc. Natl. Acad. Sci. USA 86:9871-75

Kaneko, T., Y. Nakamura, S. Sato, E. Asamizu, T. Kato, and S. Sasamoto. 2000. 

Complete genome structure of the nitrogen-fixing xymbiotic bacterium 

Mesorhizobium loti. DNA Research. 7:331-338. 

Kaneko, T., Y. Nakamura, S. Sato, K. Minamisawa, T. Uchiumi, S. Sasamoto, A. 

Watanabe, K. Idesawa, M. Iriguchi, K. Kawashima, M. Kohara, M. Matsumoto, S. 

Shimpo, H. Tsuruoka, T. Wada, M. Yamada, and S. Tabata. 2002. Complete 

genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium 

japonicum. USDA 110. DNA Research. 

Keese, P. 2008. Risks from GMOs due to Horizontal Gene Transfer. Environ. Biosafety 

Res. 7: 123-149. 

Koonin, E. V., K. S. Makarova, and L. Aravind. 2001. Horizontal gene transfer in 

prokaryotes: quantification and classification. Annual Review of Microbiology. 

55:709-742.

Muenscher, W. C. (1980) Weeds. Second Edition. Cornell University Press, New York 

and London. 586 pp.

Nakayama, T., H. Suzuki, and T. Nishino. 2003. Anthocyanin acyltransferases: 

specificities, mechanism, phylogenetics, and applications. J. Molec. Catalysis B: 

Enzymatic. 23: 117-132. 

Nelson, D.R. 2009. The Cytochrome P450 Homepage. Human Genomics 4: 59-65. 

(http://drnelson.utmem.edu/CytochromeP450.html)

Tanaka, Y. 2006. Flower colour and cytochromes P450. Phytochem Rev. 5: 283-291. 

Toguri, T., M. Azuma, and T. Ohtani. 1993. The cloning and characterization of a cDNA 

encoding a cytochrome P450 from the flowers of Petunia hybrida. Plant Sci. 94: 

119-126. 

USDA-APHIS. 2006. Federal Noxious Weed List as of January 26, 2010. 

http://www.aphis.usda.gov/plant_health/plant_pest_info/weeds/index.shtml.

USDA, NRCS. 2010. The PLANTS Database (http://plants.usda.gov, January 25, 2010). 

National Plant Data Center, Baton Rouge, LA 70874-4490 USA.
11

Vilanova, M., M. Santalla, and A. Masa. 2009. Environmental and genetic variation of 

phenolic compounds in grapes (Vitis vinifera) from northwest Spain. J. Agric. Sci. 

147: 683-697. 

Wood, D. W., J. C. Setubal, R. Kaul, D. E. Monks, J. P. Kitajima, V. K. Okura, Y. Zhou, 

L. Chen, G. E. Wood, N. F. Almeida Jr., L. Woo, Y. Chen, I. T. Paulsen, J. A. 

Eisen, P. D. Karp, D. Bovee Sr., P. Chapman, J. Clendenning, G. Deatherage, W. 

Gillet, C. Grant, T. Kutyavin, R. Levy, M.-J. Li, E. McClelland, A. Palmieri, C. 

Raymond, G. Rouse, C. Saenphimmachak, Z. Wu, P. Romero, D. Gordon, S. 

Zhang, H. Yoo, Y. Tao, P. Biddle, M. Jung, W. Krespan, M. Perry, B. GordonKamm, L. Liao, S. Kim, C. Hendrick, Z.-Y. Zhao, M. Dolan, f. Chumley, S. V. 

Tingey, J.-F. Tomb, M. P. Gordon, M. V. Olson, and E. W. Nester. 2001. The 

genome of the natural genetic engineer Agrobacterium tumefaciens C58. Science. 

294: 2317-2323.

Yu, O., M. Matsuno, and S. Subranmanian. 2006. Flavonoid Compounds in Flowers: 

Genetics and Biochemistry. In: Floriculture, Ornamental and Plant Biotechnology, 

Volume I. Global Science Books, UK. pp. 282-292.