GM Crop Database

Database Product Description

4114 (DP-ØØ4114-3)
Host Organism
Zea mays (Maize)
Trait

Herbicide tolerant, glufosinate ammonium; Insect resistant, Coleoptera; Insect resistant, Lepidoptera.

Trait Introduction
Agrobacterium tumefaciens-mediated plant transformation.
Proposed Use

Production for human consumption and livestock feed.

Product Developer
Pioneer Hi-Bred International Inc.

Summary of Regulatory Approvals

Country Food Feed Environment Notes
Australia 2015
Canada 2013 2013 2013
Colombia 2016
Japan 2015 2015
Korea 2014 2014
Mexico 2014 2014
New Zealand 2015
South Africa 2016 2016
Taiwan 2014 2014
United States 2013 2013 2012 View

Introduction Expand

DuPont Pioneer has developed maize line 4114 (DP-ØØ4114-3) as a “molecular stack” equivalent of the “breeding stack” containing the combined traits from maize lines 1507 (DAS-Ø15Ø7-1) and 59122 (DAS-59122-7). Maize line 4114 expresses the same insect-resistance and herbicide-tolerance traits as present in currently commercialized hybrids containing 1507 x 59122, which are marketed under the trade name Herculex® XTRA. Just like Herculex® XTRA hybrids, 4114 maize expresses the Cry1F protein, conferring resistance to certain lepidopteran insect pests (e.g., European corn borer), the Cry34Ab1 and Cry35Ab1 proteins, which together form an active binary insecticidal protein conferring resistance to corn rootworm, and the PAT enzyme, which inactivates glufosinate herbicide via acetylation.
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Unlike the 1507 x 59122 breeding stack, where the DNA insertions carrying traits derived from events 1507 and 59122, respectively, are located at two unlinked genetic loci, the genes encoding the Cry1F, Cry34Ab1, Cry35Ab1, and PAT proteins in 4114 maize are present at a single locus. This allows event 4114 to be bred more efficiently into new product offerings and to serve as a more efficient platform for the production of larger stack combinations through conventional breeding. Efficient breeding of multiple traits is becoming more important as growers demand more complex products, including multiple modes of action for insect control and herbicide-tolerance.

Summary of Introduced Genetic Elements Expand

Code Name Type Promoter, other Terminator Copies Form
pat phosphinothricin N-acetyltransferase HT

CaMV 35S

CaMV 35S poly(A) signal

1

Synthetic version of native gene

cry35Ab1 Cry35Ab1 delta-endotoxin IR

Triticum aestivum peroxidase gene root-preferred promoter

Solanum tuberosum proteinase inhibitor II (PINII)

1 functional

Altered coding sequence for optimal expression in maize

cry34Ab1 Cry34Ab1 delta-endotoxin IR

Zea mays ubiquitin gene promoter, intron and 5' UTR

Solanum tuberosum proteinase inhibitor II (PINII)

1 functional

Altered coding sequence for optimal expression in maize

cry1Fa2 cry1F delta-endotoxin IR

ubiquitin (ubi) ZM (Zea mays) promoter and the first exon and intron

3' polyadenylation signal from ORF25 (Agrobacterium tumefaciens)

One copy

Altered coding sequence for optimal expression in plant cells.

Characteristics of Zea mays (Maize) Expand

Center of Origin Reproduction Toxins Allergenicity

Mesoamerican region, now Mexico and Central America

Cross-pollination via wind-borne pollen is limited, pollen viability is about 30 minutes. Hybridization reported with teosinte species and rarely with members of the genus Tripsacum.

No endogenous toxins or significant levels of antinutritional factors.

Although some reported cases of maize allergy, protein(s) responsible have not been identified.

Donor Organism Characteristics Expand

Latin Name Gene Pathogenicity
Streptomyces viridochromogenes pat

S. viridochromogenes is ubiquitous in the soil. It exhibits very slight antimicrobial activity, is inhibited by streptomycin, and there have been no reports of adverse affects on humans, animals, or plants.

Bacillus thuringiensis subsp. kurstaki cry1Ac

Although target insects are susceptible to oral doses of Bt proteins, there is no evidence of toxic effects in laboratory mammals or bird given up to 10 µg protein / g body wt. There are no significant mammalian toxins or allergens associated with the host organism.

Bacillus thuringiensis strain PS149B1 cry34Ab1 While target insects are susceptible to oral doses of Bt proteins, there is no evidence of toxic effects in laboratory mammals, in birds or in non-target arthropods.
Bacillus thuringiensis var. aizawai cry1F While target insects are susceptible to oral doses of Bt proteins, no evidence of toxic effects in laboratory mammals or birds.

Modification Method Expand

Maize line 4114 was produced by Agrobacterium tumefaciens-mediated transformation with plasmid PHP27118 containing the cry1F, cry34Ab1, cry35Ab1, and pat gene expression cassettes. These gene expression cassettes are exactly the same as used in the production of 1507 maize (cry1F, pat) and 59122 maize (cry34Ab1, cry35Ab1, pat). The first cassette contains a truncated version of the cry1F gene from Bacillus thuringiensis var. aizawai. Transcriptional control is provided by the maize polyubiquitin promoter, resulting in constitutive expression of the Cry1F protein in maize. This region also includes the 5' untranslated region (UTR) and intron associated with the native polyubiquitin promoter. The terminator for the cry1F gene is the polyadenylation signal from open reading frame 25 (ORF25) of the A. tumefaciens Ti plasmid pTi15955.
The second cassette contains the cry34Ab1 gene isolated from B. thuringiensis strain PS149B1. The expression of the cry34Ab1 gene is controlled by a second copy of the maize polyubiquitin promoter with 5' UTR and intron. The terminator for the cry34Ab1gene is the 3’ terminator sequence from the proteinase inhibitor II gene (pinIIterminator) of Solanum tuberosum.
The third gene cassette contains the cry35Ab1 gene, also isolated from B. thuringiensisstrain PS149B1. The expression of the cry35Ab1 gene is controlled by the Triticum aestivum (wheat) peroxidase promoter and leader sequence. The terminator for thecry35Ab1 gene is a second copy of the pinII terminator.
The fourth and final gene cassette contains a version of the phosphinothricin N-acetyltransferase (pat) gene from Streptomyces viridochromogenes that has been optimized for expression in plants. Expression of the pat gene is controlled by the promoter and terminator regions from the cauliflower mosaic virus (CaMV) 35S transcript.

Characteristics of the Modification Expand

Molecular Characterization:

Detailed Southern blot analysis of 4114 maize genomic DNA confirmed that the introduced DNA (T-DNA) was incorporated intact into a single site within the maize genome, without truncation or deletion of nucleotide sequences within any of the gene expression cassettes. Based on a combination of genotypic and phenotypic testing, it was determined that the introduced cry1F, cry34Ab1, cry35Ab1, and pat genes were stably inherited across multiple generations, and segregated as a single genetic locus in 4114 maize according to Mendelian rules of inheritance. 

Expressed Material:
The Cry1F, Cry34Ab1, Cry35Ab1, and PAT proteins present in 4114 maize are predicted, based on the method of genetic modification and nucleotide sequencing of the introduced DNA, to have the same amino acid sequence as the corresponding proteins in events 1507 and 59122, and by extension, the conventional breeding stack of 1507 x 59122 maize. Protein equivalence was further demonstrated via western immunoblot analysis comparing the molecular weight and immunoreactivity of the Cry1F, Cry34Ab1, Cry35Ab1, and PAT proteins produced either in 4114 maize or in 1507 x 59122 maize. Therefore, previous safety assessments of the Cry1F, Cry34Ab1, Cry35Ab1, and PAT proteins conducted during the reviews of 1507 and 59122 maize are also applicable for 4114 maize. In most cases, the concentrations of Cry1F, Cry34Ab1, Cry35Ab1, and PAT proteins in 4114 maize tissue samples were similar to, or lower than, corresponding concentrations of these proteins in tissue samples from 1507 x 59122 maize. Thus, evaluations of potential environmental exposure to Cry1F, Cry34Ab1, and Cry35Ab1 proteins conducted during the previous assessments of 1507 and 59122 maize are also informative for 4114 maize. Additionally, it is unlikely that any of the differences in the concentrations of Cry1F, Cry34Ab1, or Cry35Ab1 proteins between 4114 and 1507 x 59122 maize are biologically meaningful with respect to potential impact on non-target organisms.

Environmental Safety Considerations Expand

Agronomic and phenotypic evaluations of 4114 maize were conducted in order to support a conclusion of “familiarity” based on the similarity of phenotypic characteristics with conventional and/or control maize. The phenotypic evaluations were based on laboratory experiments, to examine seed dormancy and germination, and replicated, multi-location field trials to collect data on representative characteristics that influence reproductive and survival biology. In each of these assessments, 4114 maize was compared to a near?isogenic control (i.e., ca. 98 percent similar) that did not go through the transformation process, and/or to commercial maize hybrids.

Germination rates of 4114 maize seed under warm, cold, and diurnal conditions were comparable to those of control maize under corresponding conditions. No potentially dormant seed were identified using a standard tetrazolium chloride test, which was applied to any non-germinated seed that were classified as hard or imbibed. Maize line 4114 can therefore be considered comparable to conventional maize with respect to seed germination and potential dormancy.
Comparisons of emergence and vegetative growth parameters between 4114 and control maize were based on assessments of early plant population, seedling vigour, plant height, ear height, stalk lodging, root lodging, final plant population, and stay green. There were no statistically significant differences in any of these parameters between 4114 and control maize.
Potential changes to the reproductive characteristics of 4114 maize were assessed through a combination of laboratory evaluations of seed germination and dormancy (previously described); evaluation of pollen viability, as correlated to pollen shape and colour; and collection of relevant field data on time to silking, time to pollen shed, and yield. In addition, visual estimates of foliar disease incidence and insect damage were collected for both 4114 and control maize at each location. There were no statistically significant differences in any of the parameters related to reproductive biology or disease and pest susceptibility between 4114 and control maize.

Food and/or Feed Safety Considerations Expand

For new varieties without purposefully altered nutritional properties, such as 4114 maize, the compositional assessment is part of the weight-of-evidence approach for evaluating whether there were any unanticipated consequences of the genetic modification. 

Compositional analyses were performed on samples of maize forage and kernels (grain) obtained from hybrid plants of 4114 and near-isogenic control maize grown in side-by-side trials across six locations in the United States and Canada in 2010. The near-isogenic control plants had a genetic background that was ca. 98 percent similar to that of the 4114 maize generation used, but did not go through the transformation process. 
Among the 82 compositional components that were analyzed in samples of 4114 and near-isogenic control maize grain, and nine components that were analyzed in forage samples, the only statistically significant differences observed were in concentrations of ash, phosphorus, potassium, and eicosenoic acid in grain samples. This number of statistically significant differences is within the range that would be expected purely by chance. The magnitudes of the differences in mean concentrations were small, less than 15 percent, and in every case the mean values from 4114 maize grain samples were within the ranges of natural variation as reported in the literature. Overall, no consistent patterns emerged to suggest that biologically meaningful changes in composition or nutritive value of the grain had occurred as a consequence of the genetic modification, or expression of the Cry1F, Cry34Ab1, Cry35Ab1, and PAT proteins in 4114 maize.
The conclusion based on these data was that grain and forage from 4114 maize were compositionally equivalent to these same products from unmodified near-isogenic control maize, and to other commercial maize hybrids. Processing is unlikely to alter the compositional components of maize grain, thus, products derived from 4114 grain will also be compositionally equivalent to their conventional counterparts.

Links to Further Information Expand


This record was last modified on Tuesday, September 6, 2016