Roundup ReadyTM Soybean Research Articles

Development and Testing of an Individual Kernel Detection System for Genetically Modified Soybean Events in Non-identity-preserved Soybean Samples.

A genetically modified (GM) soybean kernel detection system using combination of DNA preparation from individual soybean kernels and event-specific real-time PCR was developed to simultaneously identify GM soybean events authorized for food after safety assessments in Japan. Over 100 kernels in the non-identity-preserved soybean samples imported from the United States of America (two U.S.A. lots) and Brazil (one lot) were randomly selected and examined. In total, 98 and 96% of the two independent U.S.A. lots, and 100% of the Brazilian lot contained GM soybean kernels. Herbicide-tolerant events, MON89788 (trade name Genuity® Roundup Ready 2 Yield™), GTS 40-3-2 (trade name Roundup Ready™ soybean) and A2704-12 (trade name Liberty Link® soybean), were detected similarly in both U.S.A. lots. In the Brazilian lot, in addition to GTS 40-3-2, a stacked GM event, MON87701 × MON89788, having insect-resistance and herbicide-tolerance, was detected. There were no unauthorized GM soybeans comingled, and the ratio of GM soybean events detected was consistent with statistical reports on the cultivated GM soybean events in both countries.

Detection of genetically modified soybeans by PCR method and immunoassay kits

The suitability of detecting genetically modified (GM) soybeans by polymerase chain reaction (PCR) and immunoassay kits is determined. Primers specific for inserted genes in the Roundup Ready™ soybean (RRS, Monsanto company) were applied. Four pairs of primers, namely, 35S (35S-promoter, originated from cauliflower mosaic virus), NOS (nopaline synthase-terminator, derived from Agrobacterium tumefaciens), EPSPS (5-enolpyruvylshikimate-3-phosphate synthase, obtained from A. tumefaciens strain CP4) and LE (endogenous gene lectin) were used to identify the GM soybeans. The detection limit of PCR was 0.1% (w/w) GM soybeans when primers 35S and EPSPS were used, and 1% when primers NOS were used. All soybean samples were evidenced by LE primer-PCR as soybean products. Results of Immunoassays by two kinds of kits, strip and ELISA were matched with PCR's, and feasibility of quantitation detecting GM soybeans is determined among 0∼2.5%. The data further revealed that the PCR method and immunoassay kits can sufficiently differentiate GM soybeans from non-GM products.

Chronoamperometric magnetogenosensing for simultaneous detection of two Roundup Ready™ soybean lines: GTS 40-3-2 and MON89788

Development of expeditious analytical methods for the detection of genetically modified organisms (GMOs) is increasingly necessary, not only to verify compliance with labelling, but also to help industry to efficiently control the reception of raw materials. On the basis of this, a disposable electrochemical magnetogenoassay is proposed for simultaneous detection of two Roundup Ready (RR) soybean lines GTS 40-3-2 and MON89788, using gold-coated magnetic nanoparticles (Fe3O4@Au) as nanosupport. To perform this magnetogenoassay, a sandwich-type hybridization assay was used with different enzymatic labelling systems (fluorescein isothiocyanate and digoxigenin) and dual screen-printed carbon electrodes (SPdCEs), which allowed the simultaneous readout of each target. A linear relationship ranging from 0.1 to 2.5 nM and from 0.1 to 1.0 nM was achieved for GTS 40-3-2 and MON89788 events, respectively, and both assays showed a similar detection limit of about 0.1 nM. Furthermore, a good performance in terms of precision and selectivity was achieved. The proposed approach is a step forward for event-specific multiplex detection.

Tracing two Roundup Ready™ soybean lines (GTS 40-3-2 and MON89788) in foods commercialised in Portugal

Soybean (Glycine max L.) is the most important genetically modified (GM) crop, from which the glyphosate tolerant soybean, also known as Roundup Ready (RR), is one of the most representative. The present work intends to assess the presence of two RR soybean lines, namely GTS 40-3-2 and MON89788, in a wide range of processed foods commercialised in Portugal. For this purpose, two quantitative real-time PCR approaches were proposed using available and newly designed primers. The results showed that, from 90 food samples, 73 confirmed the presence of soybean, while 15 others suggest their incompliance with labelled soybean. RR soybean was detected in 9 samples, from which 5 were identified as being the GTS-40-3-2 event, 3 the MON89788 event and one contained both events. The developed real-time PCR methods were successfully validated through the participation in an interlaboratory proficiency program and applied to processed foods. The estimated RR soybean contents of 8 samples ranged between 0.01 and 0.39%, but the sample containing both events accounted for 23.9% of GM ingredients. Such high level above the threshold for labelling regulations suggests the need of more strict control of GMO in foods.

Detection of genetically modified organisms in soy products sold in Turkish market

PCR-based technique for GMO detection is the most reliable choice because of its high sensitivity and specificity. As a candidate of the European Union, Turkey must comply with the rules for launching into the market, traceability, and labeling of GMOs as established by EU legislation. Therefore, the objective of this study is to assess soybean products in the Turkish market to verify compliance with legislation using qualitative Polymerase Chain Reaction (PCR) assay to detect the presence of GM soybean and to quantify its amount of GM soybean in the samples tested positive using real-time PCR. DNA extracted by the modified CTAB method was properly used for PCR amplification of food materials. The amplification of a 118 bp DNA fragment of the lectin gene from soybean by PCR was successfully achieved in all samples. The GMO screening was based on the detection of 35S promoter and NOS terminator sequences. The GM positive samples were subjected to detection of Roundup ReadyTM soybean (RR) using quantitative real-time PCR. It was found that 100% of the tested food samples contained less than 0.1 per cent of EPSPS gene.

Detection of genetically modified material in feed and foodstuffs containing soy and maize in Jordan

Food and feed products with genetically modified (GM) materials are not subject to labelling regulations in Jordan. The number of GM foods being developed to cope with growing food demand, locally and internationally, is growing. This situation may result in more consumer requests for traceability and assurance of food quality and safety with respect to GM foods. To this end, the main aim of this study was to investigate at what level Jordanian consumers are exposed to GM foods. DNA was extracted from 200 food and 80 feed samples bought in Jordanian markets, screened by polymerase chain reaction (PCR) and quantified by real-time PCR. The GM screening and quantification were based on the presence of Roundup Ready™ (RR) soybean, Bt 176 or 35S target sequence genes for the GM elements. GM positive samples were quantified by real-time PCR. Fifteen food and feed samples were found positive for maize Bt-176 or RR soybean genes, representing 5.4% of the total food tested samples. The GM elements were detected in more than 62.5% with less than 1% modification, compared to less than 37.5% containing more than 1% GM.

Relação entre densidade de plantas e genótipos de soja roundup readyTM

A presente pesquisa teve por objetivo estudar a influência de diferentes densidades de plantas e genótipos de soja Roundup ReadyTM (RR) em algumas características morfoagronômicas da soja, semeada em duas safras distintas. O experimento foi conduzido na região dos Campos de Cima da Serra, no Estado do Rio Grande do Sul, nas safras agrícolas 2006/07 e 2007/08, com os cultivares CD 213RR e BRS 255RR, nas densidades de 12, 18, 24, 30 e 36 plantas aptas por m². O delineamento experimental utilizado foi de blocos ao acaso, com quatro repetições. Para isso, foram avaliados altura de plantas, diâmetro da haste, número de ramificações por planta, número de vagens por planta, número de grãos por planta, número de grãos por vagem, massa de mil grãos e rendimento de grãos por hectare. O manejo das plantas daninhas com glyphosate em duas etapas (pré-semeadura e em V4) evitou eficientemente a matocompetição. O aumento da densidade de plantas de soja RR promoveu incremento linear da altura das plantas (0,5 a 0,9 cm por planta m-2), influenciada pelo genótipo e pelo ano de cultivo. O diâmetro da haste foi diminuído com o aumento da densidade de plantas de soja RR de forma linear (-0,03 a -0,2 mm por planta m-2). O número de ramos por planta reduziu com o aumento da densidade (-0,05 a -0,19 ramos por planta). Na segunda safra, o rendimento de grãos e a massa de mil grãos foram menores, afetados por um período de estiagem e também por uma geada no final do ciclo, mais acentuadamente para o CD 213RR. A combinação de maior número de plantas m-2 levou a uma redução do número de vagens por planta e de grãos por vagem, embora de magnitudes distintas entre as cultivares, o que foi discutido pela interação entre os fatores; no entanto, o rendimento de grãos não foi alterado nas diferentes densidades testadas, e sim sofreu influência da safra de cultivo.

Monitoring of GMO in Brazilian processed meat and soy-based products from 2007 to 2008

Roundup Ready™ (RR) soybean is the first genetically modified organism (GMO) approved in Brazil. In this country, the labeling of foodstuffs containing GMOs is mandatory if the GMO content exceeds 1% (10 g kg −1). In order to monitor and quantify the presence of RR soybean in soy-based and processed meat products on the Brazilian food market, DNA was extracted from 59 food samples and analyzed by polymerase chain reaction (PCR) to amplify the soybean lectin gene, and by nested PCR to amplify the recombinant DNA present in RR soybean. Positive samples for the presence of RR soybean were subjected to a real-time quantification of GMO using a duplex real-time PCR. The results showed that out of 59 samples, 54 were positive for lectin gene and six were positive for RR soybean, 5 samples contained less than10 g kg −1 GMO and one sample contained more than 10 g kg −1 GMO.

Quantification of Roundup Ready™ soybean in Brazilian soy‐derived foods by real‐time PCR

SummaryA significant increase in the growth of genetically modified organisms has been observed in recent years. As a result of the market introduction of these genetically modified (GM) crops in foods, legislation worldwide was prepared to deal with the question of the use and labelling requirements on GM crops and their derivatives. The aim of this work was to detect and to quantify GM soybean in soy‐derived food products sold commercially in Florianópolis, Brazil, by polymerase chain reaction‐based analysis. In total sixty‐two samples of texturized soy protein and powdered soymilk were analysed and forty samples have been shown to contain Roundup Ready (RR) soy. Quantitative analysis revealed that only two out of these forty samples presented RR soybean content above 1%. These findings confirm the presence of RR soybean in Brazilian soy‐derived food products and that the amount of samples with % RR content above 1% decreases when compared with previously published data.

Recombinant DNA in meat additives: Specific detection of Roundup Ready™ soybean by nested PCR

AbstractSoybean proteins are widely used by the meat industry as technological coadjutor when producing processed products such as emulsified and ground meat products. Since regulations for the use and labeling of GMOs and derived ingredients are in force in Brazil, a PCR‐based method capable of detecting Roundup Ready™ (RR) soybean was employed for meat additives. Thirty‐two samples of meat additives containing soy proteins were tested for the presence of soybean amplifiable DNA and RR soybean DNA. Twenty‐five samples gave a positive signal for the lectin gene, confirming the presence of soybean amplifiable DNA and 15 samples returned a positive signal for specific RR detection confirming the presence of genetically modified soy. These results demonstrate for the first time the presence of RR soybean in meat additives. This method may be useful for meat industries interested in controlling the presence of RR soybean in additives used for meat products manufacture. Copyright © 2007 Society of Chemical Industry

Monitoring of Roundup™ Ready soybean in Guangdong province in China

This study was aimed at monitoring the presence of Roundup™ Ready soybean (RRS) in the market (660 samples) and fields (630 samples) in Guangdong Province in China using polymerase chain reaction (PCR) method. According to the gene sequence transgened to Roundup Ready™ soybean, four primer pairs, aiming at cp4-epsps, lectin, CaMV 35S promoter and nos terminator were designed and PCR method to detect the GM soybean has been established. The detection results demonstrate the presence of one Roundup™ Ready soybean seed sample from 60 samples in Shenzhen City. No soybean plant samples from Guangdong Province were found.

Nested PCR detection of genetically modified soybean in soybean flour, infant formula and soymilk

Due to the Brazilian market introduction of the genetically modified (GM) crop Roundup Ready™ (RR) soybean, the ability to detect GM crops has become a legal necessity. In order to detect the presence of RR soybean, a polymerase chain reaction (PCR) amplification method was evaluated for the detection of RR in soybean mixtures and commercially available soy flour, infant formula and soymilk powder. To detect the presence of RR soybean, a nested PCR resulted in an amplicon of 169 bp, present for all soybean mixed samples containing 0.01–10% GM soybean and absent for 0% GM soybean. None of the analysed infant formulas showed a positive signal after the nested PCR; four out of six soy flour samples and 15 out of 25 soymilk powder samples were positive for the presence of RR soybean. Results show that the nested PCR method used is adequate to determine the presence of GM soybean in the presented products.

PCR-based detection of genetically modified soybean and maize in raw and highly processed foodstuffs.

The PCR method has proved to be an invaluable tool for the specific and sensitive detection of genetically modified material (e.g., Roundup Ready Soybean and Bt-176 "Maximizer" Maize) in foodstuffs. The first step in the procedure, namely the purification of nucleic acids from the sample, is often the deciding factor in the production of meaningful results. In this study, we present two procedures that enable an efficient isolation of trace amounts of genetic material from both raw and highly processed foodstuffs. We show that for optimal, PCR-ready DNA purification from highly processed foodstuffs and PCR inhibitor-rich substances--such as cocoa-containing products--adapted protocols for the QIAGEN QIAamp DNA Stool Mini Kit can be utilized. For complete DNA isolation from raw foodstuffs, a protocol using the DNeasy Plant Mini Kit is presented.

Quantitative competitive PCR for the detection of genetically modified organisms in food

Present polymerase chain reaction (PCR) detection methods only allow the qualitative detection of GMO in food without quantitation of the GMO content. Clearly, the availability of quantitative detection methods for GMO analysis is an important prerequisite for the introduction of threshold limits for GMOs in food. PCR is well known to be quantitative if internal DNA standards are co-amplified together with the target DNA. This quantitative competitive (QC) PCR was first described in the early nineties and is widely used nowadays. We have developed and evaluated QC–PCR systems for the quantitative detection of Roundup Ready TM soybean (RRS) and Maximizer maize (MM) in food samples. Three DNA fragments differing from the GMO specific sequences by DNA insertions were constructed and used as internal standards in QC–PCR. These standards were calibrated by co-amplifying with mixtures containing defined amounts of RRS DNA and MM DNA, respectively. The calibrated QC–PCR systems were applied to several commercial food samples containing RRS and to three certified RRS flour mixtures (Fluka standards). Recently, quantitative methods for the detection of RRS were successfully tested in a collaborative study involving twelve European control laboratories. Thus, QC–PCR methods will allow to survey “de minimis thresholds” of GMOs in food.