Composition of forage and grain from second-generation insect-protected corn MON 89034 is equivalent to that of conventional corn (Zea mays L.)

Insect-protected corn hybrids containing Cry insecticidal proteins derived from Bacillus thuringiensis have protection from target pests and provide effective management of insect resistance. MON 89034 hybrids have been developed that produce both the Cry1A.105 and Cry2Ab2 proteins, which provide two independent modes of insecticidal action against the European corn borer ( Ostrinia nubilalis ) and other lepidopteran insect pests of corn. The composition of MON 89034 corn was compared to conventional corn by measuring proximates, fiber, and minerals in forage and by measuring proximates, fiber, amino acids, fatty acids, vitamins, minerals, antinutrients, and secondary metabolites in grain collected from 10 replicated field sites across the United States and Argentina during the 2004-2005 growing seasons. Analyses established that the forage and grain from MON 89034 are compositionally comparable to the control corn hybrid and conventional corn reference hybrids. These findings support the conclusion that MON 89034 is compositionally equivalent to conventional corn hybrids.     

Comparison of the nutritional profile of glyphosate-tolerant corn event NK603 with that of conventional corn (Zea mays L.)

The composition of glyphosate-tolerant (Roundup Ready) corn event NK603 was compared with that of conventional corn grown in the United States in 1998 and in the European Union in 1999 to assess compositional equivalence. Grain and forage samples were collected from both replicated and nonreplicated field trials, and compositional analyses were performed to measure proximates, fiber, amino acids, fatty acids, vitamin E, nine minerals, phytic acid, trypsin inhibitor, and secondary metabolites in grain as well as proximates and fiber in forage. Statistical analysis of the data was conducted to assess statistical significance at the p < 0.05 level. The values for all of the biochemical components assessed for corn event NK603 were similar to those of the nontransgenic control or were within the published range observed for nontransgenic commercial corn hybrids. In addition, the compositional profile of Roundup Ready corn event NK603 was compared with that of traditional corn hybrids grown in Europe by calculating a 99% tolerance interval to describe compositional variability in the population of traditional corn varieties in the marketplace. These comparisons, together with the history of the safe use of corn as a common component of animal feed and human food, support the conclusion that Roundup Ready corn event NK603 is compositionally equivalent to, and as safe and nutritious as, conventional corn hybrids grown commercially today.     

Comparison of the forage and grain composition from insect-protected and glyphosate-tolerant MON 88017 corn to conventional corn (Zea mays L.)

The next generation of biotechnology-derived products with the combined benefit of herbicide tolerance and insect protection (MON 88017) was developed to withstand feeding damage caused by the coleopteran pest corn rootworm and over-the-top applications of glyphosate, the active ingredient in Roundup herbicides. As a part of a larger safety and characterization assessment, MON 88017 was grown under field conditions at geographically diverse locations within the United States and Argentina during the 2002 and 2003-2004 field seasons, respectively, along with a near-isogenic control and other conventional corn hybrids for compositional assessment. Field trials were conducted using a randomized complete block design with three replication blocks at each site. Corn forage samples were harvested at the late dough/early dent stage, ground, and analyzed for the concentration of proximate constituents, fibers, and minerals. Samples of mature grain were harvested, ground, and analyzed for the concentration of proximate constituents, fiber, minerals, amino acids, fatty acids, vitamins, antinutrients, and secondary metabolites. The results showed that the forage and grain from MON 88017 are compositionally equivalent to forage and grain from control and conventional corn hybrids.     

Glyphosate-tolerant corn: the composition and feeding value of grain from glyphosate-tolerant corn is equivalent to that of conventional corn (Zea mays L.)

Glyphosate-tolerant (Roundup Ready) corn line GA21 has been developed by genetic modification to tolerate glyphosate, the active ingredient in Roundup herbicide. The purpose of this study was to evaluate the compositional and nutritional safety of corn line GA21 compared to that of conventional corn. Compositional analyses were conducted to measure proximate, fiber, amino acid, fatty acid, and mineral contents of grain and proximate, fiber, and mineral contents of forage collected from 16 field sites over two growing seasons. The nutritional safety of corn line GA21 was evaluated in a poultry feeding study conducted with 2-day old, rapidly growing broiler chickens, at a dietary concentration of 50-60% w/w. Compositional analysis results showed that, except for a few minor differences that are unlikely to be of biological significance, the grain and forage of GA21 corn were comparable in their composition to that of the control corn line and to conventional corn. Results from the poultry feeding study showed that there were no differences in growth, feed efficiency, adjusted feed efficiency, and fat pad weights between chickens fed with GA21 grain or with parental control grain. These data taken together demonstrate that Roundup Ready corn is as safe and nutritious as conventional corn for food and feed use.     

Evaluation of compositional equivalence for multitrait biotechnology crops

Compositional analysis is an important tool in the evaluation of the safety and nutritional status of biotechnology-derived crops. As part of the comparative assessment of a biotechnology-derived crop, its composition is evaluated by quantitative measurement of the levels of key nutrients, antinutrients, and secondary metabolites and compared to that of conventional crops. To evaluate the effect of combining multiple biotech traits through conventional breeding, the forage and grain compositions of the double combinations MON 810 × NK603, MON 863 × MON 810, and MON 863 × NK603 and the triple combination MON 863 × NK603 × MON 810 were compared to their respective near-isogenic, conventional control hybrids. Overall, a total of 241 statistical comparisons between the multitrait biotechnology crop and its corresponding conventional controls were conducted. Of these comparisons 192 (79.7%) were not statistically significantly different (p > 0.05), and all 49 of the differences were within the 99% tolerance interval for commercial hybrids grown in the same field or related field trials. These data on combined trait biotechnology-derived products demonstrated that the forage and grain were compositionally equivalent to their conventional comparators, indicating the absence of any influence of combining insect protection and herbicide tolerance traits by conventional breeding on compositional variation.     

Composition of grain, forage, and processed fractions from second-generation glyphosate-tolerant soybean, MON 89788, is equivalent to that of conventional soybean (Glycine max L.)

Developments in biotechnology and molecular-assisted breeding have led to the development of a second-generation glyphosate-tolerant soybean product, MON 89788. The MON 89788 event was produced by direct transformation of a cp4 epsps (5-enolpyruvylshikimate-3-phosphate synthase) gene cassette derived from Agrobacterium sp. strain CP4 into an elite soybean germplasm known for its superior agronomic characteristics and high yielding property. The purpose of this work was to assess whether the nutrient and antinutrient levels in seed and forage tissues of MON 89788 are comparable to those in the conventional soybean variety, A3244, which has background genetics similar to MON 89788 but does not contain the cp4 epsps gene cassette. Additional conventional soybean varieties currently in the marketplace were also included in the analysis to establish a range of natural variability for each analyte, where the range of variability is defined by a 99% tolerance interval for that particular analyte. Compositional analyses were conducted on forage, seed and four processed fractions from soybeans grown in ten sites across both the United States and Argentina during the 2004-2005 growing seasons. Forage samples were analyzed for levels of proximates (ash, fat, moisture, and protein) and fiber. Seed samples were analyzed for proximates, fiber, antinutrients, and vitamin E. Defatted, toasted (DT) meal was analyzed for proximates, fiber, amino acids, and antinutrients. Refined, bleached, and deodorized (RBD) oil was analyzed for fatty acids and vitamin E. Protein isolate was analyzed for amino acids and moisture. Crude Lecithin was analyzed for phosphatides. Results of the comparisons indicate that MON 89788 is compositionally and nutritionally equivalent to conventional soybean varieties currently in commerce.     

The composition of grain and forage from glyphosate tolerant wheat MON 71800 is equivalent to that of conventional wheat (Triticum aestivum L.)

Glyphosate tolerant wheat MON 71800, simply referred to as MON 71800, contains a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) protein from Agrobacterium sp. strain CP4 (CP4 EPSPS) that has a reduced affinity for glyphosate as compared to the endogenous plant EPSPS enzyme. The purpose of this work was to evaluate the compositional equivalence of MON 71800 to its nontransgenic parent as well as to conventional wheat varieties. The compositional assessment evaluated the levels of proximates, amino acids, fatty acids, minerals, vitamins, secondary metabolites, and antinutrients in wheat forage and grain grown during two field seasons across a total of eight sites in the United States and Canada. These data demonstrated that with respect to these important nutritional components, the forage and grain from MON 71800 were equivalent to those of its nontransgenic parent and commercial wheat varieties. These data, together with the previously established safety of the CP4 EPSPS protein, support the conclusion that glyphosate tolerant wheat MON 71800 is as safe and nutritious as commercial wheat varieties.     

Effect of harvest method and feed value on the economic optimum nitrogen rate for corn

Abstract

Consideration of the harvest method and protein content of corn grain and forage may be important variables for estimating Economic Optimum Nitrogen (N) (EON) rates for corn. Corn grain yield, forage yield, grain protein and forage protein data from twenty five N response experiments conducted in Pennsylvania were analyzed to determine the effect of harvest method and a price adjustment based on grain and forage protein content on the EON rate. The value of the protein in this study was assumed to be that of substituting soybean meal in the ration for growing beef cattle or dairy cattle. There was no difference in the EON for corn harvested for grain compared to corn harvested for forage when the protein content was not considered. When price adjustments were made for the protein content of the grain and forage, the EON rates increased 10 kg/ha for grain and 22 kg/ha for forage. With the protein price adjustments, the EON was 14 kg/ha higher for forage than grain. These results support the recommendation for higher N rates for corn harvested for forage, where the protein value of the forage is considered, compared to corn harvested for grain. In situations where the amino acid composition of the grain is important, such as for poultry or swine, or where urea or anhydrous ammonia can be substituted in the ration for protein, it is unlikely that a higher EON level would result when protein content or quality is considered.     

Event specific qualitative and quantitative polymerase chain reaction detection of genetically modified MON863 maize based on the 5'-transgene integration sequence

Because of the genetically modified organisms (GMOs) labeling policies issued in many countries and areas, polymerase chain reaction (PCR) methods were developed for the execution of GMO labeling policies, such as screening, gene specific, construct specific, and event specific PCR detection methods, which have become a mainstay of GMOs detection. The event specific PCR detection method is the primary trend in GMOs detection because of its high specificity based on the flanking sequence of the exogenous integrant. This genetically modified maize, MON863, contains a Cry3Bb1 coding sequence that produces a protein with enhanced insecticidal activity against the coleopteran pest, corn rootworm. In this study, the 5'-integration junction sequence between the host plant DNA and the integrated gene construct of the genetically modified maize MON863 was revealed by means of thermal asymmetric interlaced-PCR, and the specific PCR primers and TaqMan probe were designed based upon the revealed 5'-integration junction sequence; the conventional qualitative PCR and quantitative TaqMan real-time PCR detection methods employing these primers and probes were successfully developed. In conventional qualitative PCR assay, the limit of detection (LOD) was 0.1% for MON863 in 100 ng of maize genomic DNA for one reaction. In the quantitative TaqMan real-time PCR assay, the LOD and the limit of quantification were eight and 80 haploid genome copies, respectively. In addition, three mixed maize samples with known MON863 contents were detected using the established real-time PCR systems, and the ideal results indicated that the established event specific real-time PCR detection systems were reliable, sensitive, and accurate.     

Chemical composition of glyphosate-tolerant soybean 40-3-2 grown in Europe remains equivalent with that of conventional soybean (Glycine max L.)

The composition of glyphosate-tolerant (Roundup Ready) soybean 40-3-2 was compared with that of conventional soybean grown in Romania in 2005 as part of a comparative safety assessment program. Samples were collected from replicated field trials, and compositional analyses were performed to measure proximates (moisture, fat, ash, protein, and carbohydrates by calculation), fiber, amino acids, fatty acids, isoflavones, raffinose, stachyose, phytic acid, trypsin inhibitor, and lectin in grain as well as proximates and fiber in forage. The mean values for all biochemical components assessed for Roundup Ready soybean 40-30-2 were similar to those of the conventional control and were within the published range observed for commercial soybean. The compositional profile of Roundup Ready soybean 40-3-2 was also compared to that of conventional soybean varieties grown in Romania by calculating a 99% tolerance interval to describe compositional variability in the population of traditional soybean varieties already on the marketplace. These comparisons, together with the history of the safe use of soybean as a common component of animal feed and human food, lead to the conclusion that Roundup Ready soybean 40-3-2 is compositionally equivalent to and as safe and nutritious as conventional soybean varieties grown commercially.     

Compositional equivalency of Cry1F corn event TC6275 and conventional corn (Zea mays L.)

Maize (Zea mays L.) plants have been transformed to express a Cry1F insecticidal crystal protein originally isolated from Bacillus thuringiensis Berliner. This protein controls lepidopteran pests of maize, including the European corn borer, Ostrinia nubilalis (Hübner). As part of the safety assessment for crops containing transgenes, a compositional analysis of the food and feed is conducted. This analysis is designed to detect unintended changes in the nutrient and antinutrient content of the raw commodities produced by the crop due to the insertion of the genes into the genomic DNA of the plant (pleotropic effects). Samples of transgenic and nontransgenic maize forage and grain were collected from six field sites located in the U.S. and Canada. Forage samples were analyzed for proximates and minerals, and grain was further analyzed for fatty acids, amino acids, vitamins, secondary metabolites, and antinutrients. Results demonstrated that maize expressing the Cry1F protein was equivalent to nontransgenic maize with respect to these important components. Comparison of the variability within the nontransgenic and transgenic hybrid, as compared to composition values reported in the literature, suggest that factors other than transgenes may contribute more substantially to the composition of crops.     

分层破胚剥皮玉米不同部位营养成分富集特征

为明确玉米籽粒营养成分的分布差异及不同部位富集特征,应用快速缓苏、微量着水半湿法分层破胚剥皮技术,结合靶向代谢组学方法,对郑单958玉米不同部位的营养成分及基础代谢物质进行分析与比较。结果表明玉米籽粒不同部位的淀粉、脂肪、矿物元素和膳食纤维等营养物质含量存在显著差异（P<0.05）。该研究中的玉米内皮层可能主要由种皮、糊粉层及部分外胚乳构成,该部位营养成分的种类及含量均较为丰富,其中水溶性膳食纤维含量显著高于其他部位（P<0.05）,可作为玉米水溶性膳食纤维的提取分离来源。K、P和Mg元素是玉米中含量最高的矿物元素,主要存在于胚芽中,Fe、Zn、Mn和Cu元素在胚芽和玉米皮层中均有较多分布,精制加工会导致这些矿物元素的损失。玉米胚芽中水解氨基酸种类较其他部位丰富且含量较高（P<0.05）,甜味氨基酸占总游离氨基酸含量的24.49%,高于玉米皮层部位、显著高于胚乳部位。研究结果为玉米营养健康食品的创制、玉米精深加工及相关专用装备的研发提供参考。     

Assessment of the arbuscular mycorrhizal fungal community in roots and rhizosphere soils of Bt corn and their non-Bt isolines

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) in combination with sequencing of amplified partial 18S ribosomal DNA was used to assess the effects of transgenic Bt corn 5422Bt1 (event Bt 11) and 5422CBCL (event MON810) on the community structure of a non-target microorganism, namely the arbuscular mycorrhizal fungi (AMF) Glomus in corn roots and rhizosphere soils, relative to their non-Bt isolines 5422 (conventional parent) and 5422wx (conventional hybrid). AMF colonization in roots of different corn genotypes was also assessed using microscopic visualization. No adverse effect was detected on the indigenous AMF colonization of the roots of Bt hybrids 5422Bt1 and 5422CBCL. Two-way indicator species analysis (TWINSPAN^®) and detrended correspondence analysis (DCA) of the DGGE data from corn roots presented differences between Bt and non-Bt corn isolines (5422Bt1 vs. 5422wx, and 5422CBCL vs. 5422wx). However, differences were also recognized between the two non-Bt corn cultivars (5422 vs. 5422wx), and between the two Bt corn lines (5422Bt1 vs. 5422CBCL) in roots. Our results suggest that corn genotypes may have a greater influence on the AMF community structure of plant roots and rhizosphere soils than other factors, such as the age of the growing plants.     

Bollgard II cotton: compositional analysis and feeding studies of cottonseed from insect-protected cotton (Gossypium hirsutum L.) producing the Cry1Ac and Cry2Ab2 proteins

Bollgard II cotton event 15985 producing the Cry1Ac and Cry2Ab2 proteins has been developed by genetic modification to broaden the spectrum of insects to which the plant is tolerant and to provide an insect resistance management tool to impede the onset of resistance. The purpose of this study was to evaluate the composition and nutrition of Bollgard II cotton, relative to the use for food and animal feed, compared to that of conventional cotton varieties. Compositional analyses were conducted to measure proximate, fiber, amino acid, fatty acid, gossypol, and mineral contents of cottonseed from a total of 14 U.S. field sites over two years. Compositional analysis results showed that the cottonseed and cottonseed oil from Bollgard II cotton were comparable in their composition to those of the conventional control cotton line and other commercial varieties. The composition data are supported by nutritional safety studies conducted with dairy cows, catfish, and quail. Results from these studies showed that Bollgard II performed similarly to the conventional control cotton varieties. These data demonstrate that Bollgard II cotton is compositionally and nutritionally equivalent to conventional cotton varieties. These data support the conclusion that Bollgard II cotton is as safe and nutritious as conventional cotton for food and feed use.     

Assessment of the arbuscular mycorrhizal fungal community in roots and rhizosphere soils of Bt corn and their non-Bt isolines

Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) in combination with sequencing of amplified partial 18S ribosomal DNA was used to assess the effects of transgenic Bt corn 5422Bt1 (event Bt 11) and 5422CBCL (event MON810) on the community structure of a non-target microorganism, namely the arbuscular mycorrhizal fungi (AMF) Glomus in corn roots and rhizosphere soils, relative to their non-Bt isolines 5422 (conventional parent) and 5422wx (conventional hybrid). AMF colonization in roots of different corn genotypes was also assessed using microscopic visualization. No adverse effect was detected on the indigenous AMF colonization of the roots of Bt hybrids 5422Bt1 and 5422CBCL. Two-way indicator species analysis (TWINSPAN^®) and detrended correspondence analysis (DCA) of the DGGE data from corn roots presented differences between Bt and non-Bt corn isolines (5422Bt1 vs. 5422wx, and 5422CBCL vs. 5422wx). However, differences were also recognized between the two non-Bt corn cultivars (5422 vs. 5422wx), and between the two Bt corn lines (5422Bt1 vs. 5422CBCL) in roots. Our results suggest that corn genotypes may have a greater influence on the AMF community structure of plant roots and rhizosphere soils than other factors, such as the age of the growing plants.     

Metabolite analyses of grain from maize hybrids grown in the United States under drought and watered conditions during the 2002 field season

Understanding natural variation in the composition of conventional crop germplasms is critical in establishing a baseline for comparison of biotechnology-derived crops. This is particularly relevant to such traits as tolerance to drought stress. Thus, there is both a need to understand the contribution of stress conditions to natural variation in plant nutritional components and to determine whether levels of small molecule metabolites such as osmoprotectants and stress metabolites are also affected. As a first step in developing such information for maize, seven conventional hybrids were grown under different moisture regimens and the impact of moisture on composition was assessed. The regimens included well-watered conditions, water restriction during the vegetative phase, and water restriction during grain fill. Compositional analyses of the harvested grain included assessments of the levels of proximates (moisture, protein, oil, starch) and small molecule metabolites such as fatty acids, free amino acids, organic acids, sugars, total glycerol, glycine betaine, and abscisic acid. Ranges for these analytes were determined across all moisture regimens, and the effect of the different water regimens on these analytes was also evaluated. The number and type of grain analytes that showed statistically significant differences in levels between different water regimens differed quite markedly by maize hybrid. However, the magnitude of mean differences between well-watered and water-restricted samples was typically small, and statistically significant differences for any given analyte were typically observed in only one to three of the seven maize hybrids. Only two analytes, free glutamine and free proline, showed a significant drought-induced difference in at least four maize hybrids.     

氮、磷和钾营养对优质玉米子粒产量和营养品质的影响

施用氮、磷和钾营养对优质玉米子粒产量和营养品质有明显的影响。氮磷钾营养平衡施用(NPK)较不施氮(PK)、不施磷(NK)和不施钾(NP)显著增加优质玉米产量,使高油玉米(农大115)分别增产15.9%、6.9%和12.1%,使高淀粉玉米(白玉109)依次增产20.3%、8.6%和12.7%。施氮能增加高油和高淀粉玉米子粒蛋白质、醇溶蛋白和清蛋白含量;施磷或施钾能增加高油玉米子粒蛋白质、醇溶蛋白和清蛋白含量以及氨基酸和必需氨基酸含量,而对高淀粉玉米子粒蛋白质及其组分含量基本无或较小影响。施氮使高油玉米子粒氨基酸和必需氨基酸含量提高0.83和0.41个百分点,使高淀粉玉米提高1.18和0.36个百分点;施氮、施磷和施钾增加高油和高淀粉玉米淀粉总量和支链淀粉含量,降低直链淀粉含量。施氮能增加高油和高淀粉玉米子粒油分、亚油酸和油酸含量,使高油玉米分别增加0.83、0.41和0.30个百分点,使高淀粉玉米分别增加0.34、0.18和0.13个百分点,而施磷或施钾对两品种子粒油分、亚油酸和油酸含量的影响不明显。     

Effect of Various Acids and Sulfites in Steep Solution on Yields and Composition of Corn Fiber and Corn Fiber Oil

The addition of six acids (organic and inorganic) and four sulfite compounds (including gaseous SO₂) during the conventional corn wet‐milling steeping process of two yellow dent corn hybrids were evaluated for the effect on corn fiber yield, corn fiber oil yield, and the composition of three phytosterol compounds (ferulate phytosterol esters [FPE], free phytosterols [St], and phytosterol fatty acyl esters [St:E]) in the corn fiber oil. No significant effect of different sulfite compounds and acids were observed on corn fiber yields. However, a significant effect was observed on corn fiber oil yield and the composition of corn fiber oil for phytosterol compounds. Three of the sulfite compounds (including gaseous SO₂) caused very little effect on the levels of phytosterol compounds compared with the control sample (corn steeped with sodium metabisulfite and lactic acid). However, for one hybrid, ammonium sulfite gave a significantly higher yield of FPE and St:E and had no effect on the yield of St. For the other hybrid, it gave a significantly higher yield of FPE and had no effect on the yield of St and St:E compared with the control sample. This indicates that the effect of these sulfite compounds on yields of these phytosterol compounds in corn fiber oil is probably hybrid‐dependent. No significant effect of acids was observed on corn fiber yields, but significant effects were observed on corn fiber oil yields and yields of phytosterol compounds in the corn fiber oil. The effect also seems to be hybrid‐dependent because different acids affected the two hybrids differently. Overall, it seems that weak acids have a positive effect on increasing the individual phytosterol compounds in the corn fiber. When comparing the effect of experimental acids and sulfites on the two hybrids, acids have a more positive effect than sulfites in increasing the yield of phytosterol compounds in corn fiber oil.     

Phenolic acids, lipids, and proteins associated with purified corn fiber arabinoxylans

Corn fiber gum (CFG) is a hemicellulose (arabinoxylan)-enriched fraction obtained by the extraction of corn bran/fiber using a proprietary alkaline hydrogen peroxide process. When purified CFG prepared by this process was hydrolyzed with more concentrated base (1.5 N methanolic KOH at 70 degrees C for 1 hour), considerable amounts of hydroxycinnamic acids (up to 0.015% of mainly ferulic acid) and lipids (up to 0.43%) were released. The released phenolic acids and lipids were identified and quantified using high-performance liquid chromatography (HPLC) with detection by both UV and evaporative light-scattering detection (ELSD). During the wet milling of corn, two types of corn fiber are produced: coarse fiber, which is primarily from pericarp, and fine fiber, which is from the endosperm. The total phenolic acid content in CFGs purified from coarse corn fiber (pericarp fiber) is comparatively higher than that purified from fine corn fiber (endosperm fiber). It was also determined that the purified CFG samples contained significant amounts of strongly associated proteins, from 2 to 5% by weight. The presence of these phenolic acids, lipids, and proteins strongly associated or bound to CFG may contribute to its excellent ability to emulsify oil-in-water emulsions.     

Qualitative and quantitative polymerase chain reaction analysis for genetically modified maize MON863

Qualitative and quantitative analytical methods were developed for the new event of genetically modified (GM) maize, MON863. One specific primer pair was designed for the qualitative polymerase chain reaction (PCR) method. The specificity and sensitivity of the designed primers were confirmed. PCR was performed on genomic DNAs extracted from MON863, other GM events, and cereal crops. Single PCR product was obtained from MON863 by the designed primer pair. Eight test samples including GM maize MON863 were prepared at 0.01 approximately 10% levels and analyzed by PCR. Limit of detection of the method was 0.01% for GM maize MON863. On the other hand, another specific primer pair and probe were also designed for quantitative method using a real-time polymerase chain reaction. As a reference molecule, a plasmid was constructed from a taxon-specific DNA sequence for maize, a universal sequence for a cauliflower mosaic virus (CaMV) 35S promoter used in most genetically modified organisms, and a construct-specific DNA sequence for the MON863 event. Six test samples of 0.1, 0.5, 1.0, 3.0, 5.0 and 10.0% of GM maize MON863 were quantitated for the validation of this method. At the 3.0% level, the bias (mean vs true value) for MON863 was 3.0%, and its relative standard deviation was 5.5%. Limit of quantitation of the method was 0.5%. These results show that the developed PCR methods can be used to qualitatively and quantitatively detect GM maize MON863.