Degradation and detection of transgenic Bacillus thuringiensis DNA and proteins in flour of three genetically modified rice events submitted to a set of thermal processes

Wang, Xiaofu; Chen, Xiaoyun; Dai, Chen; Shen, Wenbiao

doi:10.1016/j.fct.2015.08.010

Cited by 10 publications

(3 citation statements)

References 42 publications

(39 reference statements)

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“…Similar degradation results were observed for MON‐810 maize and RR soybean under autoclaving (Vijayakumar et al ., ). Our results about DNA degradation after heat treatment are in accordance with literature (van der Colff & Podivinsky, ; Wang et al ., ).…”

Section: Resultsmentioning

confidence: 97%

Applicability of quantitative polymerase chain reaction (qPCR) assays for common bean authentication in processed food

Venturelli

Bischoff

Scariot

et al. 2019

Int J of Food Sci Tech

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Food authentication by quantitative polymerase chain reaction (qPCR) methods assures food quality. The aim was to evaluate three qPCR assays for DNA quantification after heat processing of common bean grains, genus-specific FAS assay for Phaseolus, species-specific LEC assay for common bean (Phaseolus vulgaris) and genetically modified (GM) event-specific FGM assay for Embrapa 5.1 event GM common bean. FAS assay showed high stability among Phaseolus genus samples. Common bean grains were heat-treated in autoclave (at 120°C for 15-60 min) and target DNA copy number decreased as processing time increased. Even with DNA degradation, qPCR assays were capable to detect low DNA quantity, and the limit of detection was 100 copy number. Mean efficiency value of FGM assay was 92% in the presence of background DNA. Background DNA did not cause any interference, and 0.39% of GM material can be detected. These qPCR assays are able to quantify common bean in processed food. Primers and probesIn this study, we used two endogenous reference assays targeting the a-phaseolin and lectin genes of P. vulgaris common bean. The primers PvFASF/PvFASR targeting

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Section: Resultsmentioning

confidence: 97%

Applicability of quantitative polymerase chain reaction (qPCR) assays for common bean authentication in processed food

Venturelli

Bischoff

Scariot

et al. 2019

Int J of Food Sci Tech

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“…The present study is the first to detect Cry1Ac in the fat body of lepidopteran larvae ( Figure 2 ), likely because we used Bt resistant strains of H. armigera , that can survive from the 1st instar on high doses of Cry1Ac toxin, whereas susceptible insects can only survive feeding on Bt-crops for a few days. Cry1Ac in the PM content, midgut and hemolymph was shown to be present as the 62–65 kDa activated toxin fragment ( Figure 3 ), which was demonstrated to be stable in in vitro activation studies and thermal treatment experiments [ 22 , 23 , 24 ]. Therefore, we speculated that Cry1Ac in the fat body of H. armigera larvae was still in the form of the activated toxin fragment.…”

Section: Discussionmentioning

confidence: 99%

Distribution and Metabolism of Bt-Cry1Ac Toxin in Tissues and Organs of the Cotton Bollworm, Helicoverpa armigera

Zhao

Xiao

et al. 2016

Toxins

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Crystal (Cry) proteins derived from Bacillus thuringiensis (Bt) have been widely used in transgenic crops due to their toxicity against insect pests. However, the distribution and metabolism of these toxins in insect tissues and organs have remained obscure because the target insects do not ingest much toxin. In this study, several Cry1Ac-resistant strains of Helicoverpa armigera, fed artificial diets containing high doses of Cry1Ac toxin, were used to investigate the distribution and metabolism of Cry1Ac in their bodies. Cry1Ac was only detected in larvae, not in pupae or adults. Also, Cry1Ac passed through the midgut into other tissues, such as the hemolymph and fat body, but did not reach the larval integument. Metabolic tests revealed that Cry1Ac degraded most rapidly in the fat body, followed by the hemolymph, peritrophic membrane and its contents. The toxin was metabolized slowly in the midgut, but was degraded in all locations within 48 h. These findings will improve understanding of the functional mechanism of Bt toxins in target insects and the biotransfer and the bioaccumulation of Bt toxins in arthropod food webs in the Bt crop ecosystem.

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“…However, PCR methods depend on thermal cycler, which are complicated and time consuming, and fail to detect exogenous proteins ( Grel et al, 2011 ). Methods based on protein-specific expression, such as enzyme-linked immunosorbent assay (ELISA), the test strip, and Western blot ( Wang et al, 2015 ; Albright et al, 2016 ; Zeng et al, 2020 ), have also been applied to GM crops. While these methods are fast, they fail to sensitively and quantitatively detect the protein in GM crops.…”

Section: Introductionmentioning

confidence: 99%

A Label-Free Immunosensor Based on Gold Nanoparticles/Thionine for Sensitive Detection of PAT Protein in Genetically Modified Crops

et al. 2021

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Genetically modified (GM) crops containing phosphinothricin acetyltransferase (PAT) protein has been widely planted worldwide. The development of a rapid method for detecting PAT protein is of great importance to food supervision. In this study, a simple label-free electrochemical immunosensor for the ultrasensitive detection of PAT protein was constructed using thionine (Thi)/gold nanoparticles (AuNPs) as signal amplification molecules and electrochemically active substances. Under optimum conditions, the limits of detection of the sensor for soybean A2704-12 and maize BT-176 were 0.02% and 0.03%, respectively. The sensor could detect crops containing PAT protein and had no cross-reaction with other proteins. After storage at 4°C for 33 days, the sensor still retained 82.5% of the original signal, with a relative standard deviation (RSD) of 0.92%. The recoveries of the sensor for soybean A2704-12 and maize BT-176 were 85%–108% and 98%–113%, respectively. The developed PAT-target immunosensor with high sensitivity, specificity, and satisfactory reproducibility and accuracy will be a useful tool in the trace screening of GM crops. Moreover, this design concept can be extended to other proteins by simply changing the antibody.

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Degradation and detection of transgenic Bacillus thuringiensis DNA and proteins in flour of three genetically modified rice events submitted to a set of thermal processes

Cited by 10 publications

References 42 publications

Applicability of quantitative polymerase chain reaction (qPCR) assays for common bean authentication in processed food

Applicability of quantitative polymerase chain reaction (qPCR) assays for common bean authentication in processed food

Distribution and Metabolism of Bt-Cry1Ac Toxin in Tissues and Organs of the Cotton Bollworm, Helicoverpa armigera

A Label-Free Immunosensor Based on Gold Nanoparticles/Thionine for Sensitive Detection of PAT Protein in Genetically Modified Crops

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