A dual-RPA based lateral flow strip for sensitive,on-site detection of CP4-EPSPS and Cry1Ab/Ac genes in genetically modified crops

Traditional transgenic detection methods require high test conditions and struggle to be both sensitive and efficient.In this study,a one-tube dual recombinase polymerase amplification(RPA)reaction system for CP4-EPSPS and Cry1Ab/Ac was proposed and combined with a lateral flow immunochromatographic assay,named“Dual-RPA-LFD”,to visualize the dual detection of genetically modified(GM)crops.In which,the herbicide tolerance gene CP4-EPSPS and the insect resistance gene Cry1Ab/Ac were selected as targets taking into account the current status of the most widespread application of insect resistance and herbicide tolerance traits and their stacked traits.Gradient diluted plasmids,transgenic standards,and actual samples were used as templates to conduct sensitivity,specificity,and practicality assays,respectively.The constructed method achieved the visual detection of plasmid at levels as low as 100 copies,demonstrating its high sensitivity.In addition,good applicability to transgenic samples was observed,with no cross-interference between two test lines and no influence from other genes.In conclusion,this strategy achieved the expected purpose of simultaneous detection of the two popular targets in GM crops within 20 min at 37°C in a rapid,equipmentfree field manner,providing a new alternative for rapid screening for transgenic assays in the field.

Trends in the global commercialization of genetically modified crops in 2023

The commercialization of genetically modified(GM)crops has increased food production,improved crop quality,reduced pesticide use,promoted changes in agricultural production methods,and become an important new production strategy for dealing with insect pests and weeds while reducing the cultivated land area.This article provides a comprehensive examination of the global distribution of GM crops in 2023.It discusses the internal factors that are driving their adoption,such as the increasing number of GM crops and the growing variety of commodities.This article also provides information support and application guidance for the new developments in global agricultural science and technology.

Research Progress of Agricultural Genetically Modified Crop Safety Evaluation

The commercial cultivation of genetically modified(GM)crops has eased the global food crisis and brought considerable economic and social benefits to countries.Because of the potential safety problems,it is necessary to make clear the molecular genetic characteristics,edible safety,planting,processing,and other aspects of the safety evaluation of GM crops.The safety problems existing in the cultivation of GM crops,safety evaluation and detection of GM crops were introduced in this paper,which provided the basis for safety evaluation and effective supervision of GM crops and their products.Commercial cultivation and reasonable supervision based on safety evaluation have far-reaching significance for ensuring consumer safety,enhancing the credibility of the national political system and enhancing citizens'confidence in the safety of GM crop products for consumption.

Understanding Saudi Public’s Awareness, Perception, and Attitudes towards Genetically Modified Foods

Introduction: Recent technological developments have enabled many scientists to produce a wide variety of foods that meet consumers’ desires for diversity and quality. Genetically modified (GM) foods have long been a controversial topic, and consumer reliance on these foods depends on their trust in biotechnology institutions due to their lack of knowledge about the risks and benefits. This study aimed to assess the awareness, attitude, and perception of GM foods among the Saudi population, as well as to identify the association between age, gender, and education level in the studied population and their awareness level towards genetically modified foods. Method: Data was collected using a structured, pretested questionnaire drawn from previous studies, after obtaining written consent. The questionnaire included three sections focusing on personal data, general knowledge of GM foods, and perception, attitudes, and beliefs towards GM foods. Our study included 500 responses. Results: The majority of the participants were female, located in the central region, and had a university education. Most participants provided a correct definition of GM foods and indicated that they had heard about them, but were unsure if they had eaten these foods in the past. Their knowledge of these foods was good. Regarding opinions and attitudes towards these foods, the majority of participants were neutral. They approved of using GM foods because it produces more food, tastes better, and can be stored for a long time. In conclusion, the study highlights the urgent necessity to raise awareness and knowledge regarding genetically modified foods among the Saudi population. It emphasizes the importance of educating consumers about the potential advantages and risks associated with consuming such foods, enabling them to make informed decisions. It is crucial to integrate awareness and education about genetically modified foods into the technological and agricultural advancements in Saudi Arabia.

Micropropagation of Daylily(Hemerocallis fulva)from Crown-Tip Explants and Assessment of Somaclonal Variation of in Vitro-Propagated Plants Using SCoT Markers

Determination of the somaclonal variation of in vitro-propagated plants is crucial to determine the appropriate micropropagation protocol and growth regulators for commercial scale multiplication.In this research,nine multiplication media(MM)augmented with different concentrations of 6-benzyl adenine(BA),Kinetin(Kin),and Thidiazuron(TDZ),Three rooting media(RM)supplemented with three levels ofα-naphthalene acetic acid(NAA)and three types of soil mixtures(v/v);Coco peat/Vermiculite/Sand(CVS),Peat moss/Perlite/Sand(PPS)and Peat moss/Perlite(PP)were used in the micropropagation protocol of daylily plants.MM2 showed the maximum shoot length and the number of leaves,while MM9 showed the maximum number of shoots.The RM1 showed the maximum root length and the number of roots.During acclimatization,CVS,PPS,and PP soil mixture showed similar performance except the CVS mixture showed lower performance regarding plant height and diameter.The genetic fidelity of micropropagated plants was evaluated using Start Codon Targeted(SCoT)Markers.Six SCoT primers amplified 51 scorable bands with an approximate range from 146 bp to 1598 bp size.Thirty one out of 51 loci were presented in the mother plants.40 loci were polymorphic,11 were monomorphic and 7 were unique.The amplification patterns of the micropropagated plants demonstrated genetic integrity to the mother plant ranging from 84.32 to 47.06 and somaclonal variations ranging from 52.94 with 5 mg/l BA pathway to 15.68 with 1mg/l TDZ pathway,thus demonstrating that the homogeneity and the variation of the micropropagated plants affected by the type and the quantity of the plant growth regulator used during multiplication subcultures.This research can be successfully used for other ornamental and medicinal plants’bulk multiplication,germplasm conservation,and future genetic improvement.

Comparison of the Evolution of Genetically Modified Food Safety Policies between the United States and the European Union

With the research on and development of Genetically Modified Food （GMF）, people＇s attitude toward GMF may fall into two divergent categories, typical- ly represented by the United States（US） and the European Union（EU）, respectively. The former follows a ＂sound science principle＂ and firmly objects to the precautionary principle, namely a permissive policy of positive support of and voluntary labelling on GMF; while the latter adopts a cautiously precautionary principle, requiring mandatory labelling and traceability. From the standpoint of regulatory principles, together with corresponding supervisory measures and relevant provisions, this paper compared the execution of directives and provisions on GMF from the initial policies enacted by the US and EU to current situation combined with the track and analysis of latest polic ies issued.

Breeding of High Yield and Genetically Modified Hybrid Cotton-Sumian 29

Sumian 29, a genetically modified cotton variety, was approved by Autho- rized Committee of Crop Varieties of Jiangsu Province in 2013. Yield performance, cultivation characteristics of Sumian 29, and its selection and breeding process were introduced in the paper. Regional tests from 2010 to 2011 in Jiangsu Province showed that seed cotton yield and lint yield averaged 4 185 and 1 737 kg/hm2, and increased by 10.6% and 8.5% respectively, when compared with control （Siza 3）. In production test, seed cotton yield and lint yield of Sumian 29 averaged 4 176 and 1 744.5 kg/hm2, respectively. Sumian 29 had high resistance to cotton bollworm, and also resistance to Fusarium wilt and Verticillium wilt of cotton. All of its fiber qualities achieved National Standard III and above. Sumian 29 has good application prospects.

Uncertainty in Measuring Construct-specific Fragments of Genetically Modified Maize MON863 by Real Time Quantitative PCR

[Objective] The study aimed at evaluating the uncertainty in measuring the construct-specific fragments of genetically modified maize MON863 by real time quantitative PCR.[Method] The content of construct-specific fragments in MON863 samples was determined by real time quantitative PCR,and then the uncertainty of measurement result was evaluated according to the sources of uncertainty like the PCR system,the data processing and the micropipette.[Result] Type A evaluation of uncertainty（uA） in the measurement was 1.7×10^-2;Type B evaluation of uncertainty（uB） was 9.0×10^-4;the combined standard uncertainty（uC） was 1.7×10^-2;the expanded uncertainty（U95） was 0.036 and the finally measured result was 1.08%±0.036.[Conclusion] The main uncertainty of the result measured by real time quantitative PCR came from the randomizing effect in the experimental process.

Detection of Genetically Modified Crops by Combination of Multiplex PCR and Low-density DNA Microarray

Objective To develop a technique for simultaneous detection of various target genes in Roundup Ready soybean by combining multiplex PCR and low-density DNA microarray. Methods Two sets of the multiplex PCR system were used to amplify the target genes in genetically modified （GM） soybean. Seventeen capture probes （PCR products） and 17 pairs of corresponding primers were designed according to the genetic characteristics of Rroundup Ready soybean （GTS40-3-2）, maize （MonS10, Nk603, GA21）, canola （T45, MS1/RF1）, and rice （SCK） in many identified GM crops. All of the probes were categorized and identified as species-specific probes. One negative probe and one positive control probe were used to assess the efficiency of all reactions, and therefore eliminate any false positive and negative results. After multiplex PCR reaction, amplicons were adulterated with Cy5-dUTP and hybridized with DNA microarray. The array was then scanned to display the specific hybridization signals of target genes. The assay was applied to the analysis of sample of certified transgenic soybean （Roundup Ready GTS40-3-2） and canola （MS1/RF1）. Results A combination technique of multiplex PCR and DNA microarray was successfully developed to identify multi-target genes in Roundup Ready soybean and MS 1/RF1 canola with a great specificity and reliability. Reliable identification of genetic characteristics of Roundup Ready of GM soybean from genetically modified crops was achieved at 0.5% transgenic events, indicating a high sensitivity. Conclusion A combination technique of multiplex PCR and low-density DNA microarray can reliably detect and identify the genetically modified crops.

Immunotoxicological Evaluation of Wheat Genetically Modified with TaDREB4 Gene on BALB/c Mice

Objective To evaluate the immunotoxicological effects of genetically modified wheat with TaDREB4 gene in female BALB/c mice. Methods Female mice weighing 18-22 g were divided into five groups （10 mice/group）, which were set as negative control group, common wheat group, parental wheat group, genetically modified wheat group and cyclophosphamide positive control group, respectively. Mice in negative control group and positive control group were fed with AIN93G diet, mice in common wheat group, non-genetically modified parental wheat group and genetically modified wheat group were fed with feedstuffs added corresponding wheat （the proportion is 76%} for 30 days, then body weight, absolute and relative weight of spleen and thymus, white blood cell count, histological examination of immune organ, peripheral blood lymphocytes phenotyping, serum cytokine, serum immunoglobulin, antibody plaque-forming cell, serum half hemolysis value, mitogen-induced splenocyte proliferation, delayed-type hypersensitivity reaction and phagocytic activities of phagocytes were detected. Results No immunotoxicological effects related to the consumption of the genetically modified wheat were observed in BALB/c mice when compared with parental wheat group, common wheat group and negative control group. Conclusion From the immunotoxicological point of view, results from this study demonstrate that genetically modified wheat with TaDREB4 gene is as safe as the parental wheat.

Comparison of Ileal Digested Production of Parental Rice and Rice Genetically Modified With Cowpeas Trypsin Inhibitor

Objective To compare the ileal digestibility of protein and amino acids in parental rice and rice genetically modified with sck gene. Methods Six experimental swines were surgically fixed with a simple T-cannula at the terminal ileum and fed with parental rice and rice genetically modified with sck gene alternately. The ileum digesta were collected and analyzed for determination of apparent and true digestibility of protein and amino acids. Results The apparent and true digestibility of protein was similar in these two types of rice. Except for the apparent digestibility of lysine, there was no difference in the apparent and true digestibility of the other 17 amino acids. Conclusion The digestibility of protein and amino acids is not changed by the insertion of foreign gene, so it can meet the request of ＂substantial equivalence＂ in digestibility of protein and amino acids.

Evaluation of genetically modified rice detection methods 2011/884/EU and 2008/289/EC proposed by the European Union

Increases in the number of cases of identified genetically modified （GM） rice contamination can be traced back to the first Rapid Alert System for Food and Feed （RASFF） in 2006. In response to the lack of reliable detection methods, Decision 2011/884/EU proposed that new screening methods replace Decision 2008/289/EC, to identify all possible GM rice products originating in China. However, the synergy brands （SYBR） Green real-time PCR assay proposed by Decision 2011/884/EU has been shown to lack conformity with other TaqMan methods currently in use. To evaluate the specificity and repeatability of the methods recommended in Decision 2011/884/EU and Decision 2008/289/EC, we collected 74 rice products originating from six countries or districts. The 74 rice samples were tested using the Decision 2011/884/EU and Decision 2008/289/ EC methods. The parallel use of different instruments and reagents were used for testing in parallel, and the results were analyzed statistically. To avoid the limitations of specific laboratories, eight GM organism detection laboratories in China participated in a collaborative trial. In our tests, 24.3% （18/74） of the samples tested were positive with the SYBR Green real-time PCR assay using the Decision 2011/884/EU method, but were negative with the TaqMan real-time PCR assay using the Decision 2011/884/EU and Decision 2008/289/EC methods. Sequencing the PCR-amplified CrylA（b/c） genes in three samples （6, 30 and 43） showed that the products consisted of primer dimers rather than the targeted sequence. The combined experimental results showed that testing for the nopaline synthase gene （NOS） of Agrobacterium tumefasciens terminator and CrylA（b/c） produced false-positive results when the Decision 2011/884/EU method was used. Because of the high rate of false-positive results, the Decision 2011/884/EU SYBR Green method to detect GM rice requires improvement.

Development of a Multiplex Ligation Dependent Probe Amplification (MLPA) Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events

For the official surveillance of genetically modified crops, efficient and simple detection methods need to be on hand to implement the strict requirements regarding approval, labelling and traceability determined by the European Union. Therefore, a multiplex ligation dependent probe amplification (MLPA) module was developed for simultaneous detection of five genetically modified rapeseed events (MS8, RF3, GT73, Falcon GS40/90 and T45). Probes were designed and concentrations were adapted in order to obtain high sensitivity and specificity. This MLPA module was validated using certified reference materials and its applicability was tested analyzing routine honey, mustard and rapeseed samples. The limit of detection was determined by analyzing a dilution series (n = 16 for each concentration) of respective transgenic DNA. After optimization, the MLPA revealed limits of detection between 10 to 50 copies of the transgene DNA/assay. The method proved to be sensitive and highly reproducible. When analyzing routine samples, results obtained applying the MLPA module were similar compared to real-time PCR.

Absolute quantitative detection of genetically modified soybean MON87708×MON89788 with stacked traits by digital polymerase chain reaction

The main advantage of digital PCR(dPCR) is that it facilitates absolute quantification of the target without reference to the standard/calibration curve.Crystal droplet dPCR has a three-color staining detection function,which enables multiplex PCR reaction.In this study,this technique was used to establish triple dPCR detection for the genetically modified soybean MON87708 × MON89788 with stacked traits.Specific absolute quantitative detection was accomplished for the genomic DNA extracted from the homogenized seeds of GM stack MON87708× MON89788 soybean.Our results can serve as a reference for the absolute quantitative detection of stacked events of genetically modified crops.

Defensive Role of Plant Hormones in Advancing Abiotic Stress-Resistant Rice Plants

Consistent climatic perturbations have increased global environmental concerns, especially the impacts of abiotic stresses on crop productivity. Rice is a staple food crop for the majority of the world’s population. Abiotic stresses, including salt, drought, heat, cold and heavy metals, are potential inhibitors of rice growth and yield. Abiotic stresses elicit various acclimation responses that facilitate in stress mitigation. Plant hormones play an important role in mediating the growth and development of rice plants under optimal and stressful environments by activating a multitude of signalling cascades to elicit the rice plant’s adaptive responses. The current review describes the role of plant hormone-mediated abiotic stress tolerance in rice, potential crosstalk between plant hormones involved in rice abiotic stress tolerance and significant advancements in biotechnological initiatives including genetic engineering approach to provide a step forward in making rice resistance to abiotic stress.

Establishment of a Construct-specific Real-time PCR Method for Quantitative Detection of Genetically Modified Maize Line MIR604

In this study, a construct-specific real-time PCR method for quantitative detection of genetically modified maize line MIR604 was established with prim- ers and probes designed based on vector sequence of MIR604 under optimized reaction system and thermal cycling condition. By using the established method, six non-genetically modified crops, genetically modified maize line MIR604 and other non-target genetically modified crops were detected. According to the results, flu- orescence signal could be detected in genomie DNA of MIR604, but other non-genetically modified crops and non-target genetically modified crops exhibited no fluo- rescence signal of MIR604 molecular fragment. Certified reference materials containing 1% MIR604 were detected with the established method and the results indi- cated that the average relative content in the test samples was 1.05%. The sensitivity analysis results indicated that MIR604 nucleic acid fragment with at least five copies could be detected with the established method. In conclusion, the construct-specific real-time PCR method for quantitative detection of genetically modified maize line MIR604 is of high specificity, accuracy and sensitivity, which provides technical support for safety supervision of genetically modified organisms in China.

Consumers’ Awareness of Genetically Modified Food and Their Willingness to Buy in Yanbian Prefecture

Having a clear understanding of consumers' awareness of genetically modified food and their willingness to buy, plays a very important role in formulating the regulatory policy of genetically modified food and regulating the market of genetically modified food. This paper takes the supermarket consumers as the study object. Through on-site questionnaire survey, we find that consumers' awareness of genetically modified food is not high in Yanbian Prefecture, and their willingness to buy is also low; the prices of genetically modified food, consumers' income, educational level and so on, are the main factors that affect the willingness to buy. Based on this, we put forth the relevant recommendations: increasing publicity efforts to safeguard consumers' right to know and choose; increasing supervision efforts to improve the existing regulatory system of genetically modified food; actively organizing forces to carry out the study on safety of genetically modified food.

College Students' Cognition of Genetically Modified Foods

In order to have knowledge about the college students'awareness and acceptance of genetically modified foods and their attitudes toward the identification of genetically modified foods and the government's regulation,we conduct a questionnaire survey of 150 college students'cognition of genetically modified foods in North College Town of Zhengzhou City.The results show that the college students have a certain understanding of genetically modified foods,but the cognition level is low;electronic media has become the main channel for the current college students to know the information about genetically modified foods;for security reasons,the majority of college students are wary of genetically modified foods,and pay more attention to whether there is genetically modified component in the foods labeling;college students generally believe that the government should strengthen the supervision of genetically modified foods,and make the GM labeling system strict,to protect consumers'right to know and choice.

Establishment of a Real-time Fluorescent Quantitative PCR Assay for Detection of Genetically Modified Maize Line MON88017

In order to improve the standardized technical systems of quantitative analyses for genetically modified organisms （GMOs） and products, ensure bio-safety and reduce ecological risk in China, a real-time fluorescent quantitative PCR assay was established for detection of genetically modified maize line MON88017. The established method was evaluated based on the specificity, sensitivity, accuracy and measurement uncertainty. The results showed that the established method had strong specificity in detection of genetically modified maize line MON88017. 1.50% MON88017 sample was detected with 29 replica- tions. The average measured value （ 1. 541% ） was close to the actual value （ 1.50% ） and the relative deviation was 2.70%. The variation coefficient of the measured value was 0.110 g ; the recovery was 100.00% and the measurement uncertainty was 0. 096. The limit of detection for genetically modified maize line MON88017 with the established method was 5 copies at the 97.5% confidence level. Thus, the real-time fluorescent quantitative PCR assay established in this study exhibited high specificity, accuracy and sensitivity, which could provide technical support for the safety supervision of genetically modified organ- isms and products in China.

Histopathological Investigation of the Stomach of Rats Fed a 60% Genetically Modified Corn Diet

Genetic modification (GM) represents new opportunities for enhanced crop features such as improved insect resistance and herbicide tolerance. The technology allows for cross-species alterations, therefore potentially allowing a vast array of novel traits. Many GM crops have been developed and approved for human and animal consumption. The present study investigated a triple-stacked GM corn variety containing modifications for insect resistance (via cry1Ab and cry3Bb1 genes) and herbicide tolerance (via an EPSPS gene), which was fed to rats for six months. The study investigated the mucosa of the stomach. Alterations to tight junction apposition, gland dilatations with epithelial elongation and dysplasia in the GM-fed rats were observed. These results indicate that GM-corn may have an effect on rat stomach mucosa, which may have health implications.