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.

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.

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.

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.

Genetically Modified Doubts Imported GM soybeans impact China’s local farmers and create food safety concerns

The cooking oil made from the imported soybeans does not contain transgenic proteins. So there will be no food safety threat.

基于GM(1,1)模型与遗传算法的蔬菜定价与补货策略研究

合理订购和销售蔬菜类商品是商家实现收益最大化的重要研究课题。计算Spearman相关系数发现,蔬菜各品类销售量之间有着较强的正相关性;利用高斯函数进行回归拟合发现,蔬菜各品类销售总量与销售价格大致呈负相关。使用灰色预测GM(1,1)模型预测蔬菜各品类未来7日的定价和销售量,同时构建基于多目标规划与遗传算法的商家利润优化模型来研究并给出商家未来7日的单品补货量和定价策略。

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.

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.

Estimation of genetic parameters and their sampling variances for quantitative traits in the type 2 modified augmented design

The type 2 modified augmented design(MAD2) is an efficient unreplicated experimental design used for evaluating large numbers of lines in plant breeding and for assessing genetic variation in a population. Statistical methods and data adjustment for soil heterogeneity have been previously described for this design. In the absence of replicated test genotypes in MAD2, their total variance cannot be partitioned into genetic and error components as required to estimate heritability and genetic correlation of quantitative traits, the two conventional genetic parameters used for breeding selection. We propose a method of estimating the error variance of unreplicated genotypes that uses replicated controls, and then of estimating the genetic parameters. Using the Delta method, we also derived formulas for estimating the sampling variances of the genetic parameters.Computer simulations indicated that the proposed method for estimating genetic parameters and their sampling variances was feasible and the reliability of the estimates was positively associated with the level of heritability of the trait. A case study of estimating the genetic parameters of three quantitative traits, iodine value, oil content, and linolenic acid content, in a biparental recombinant inbred line population of flax with 243 individuals, was conducted using our statistical models. A joint analysis of data over multiple years and sites was suggested for genetic parameter estimation. A pipeline module using SAS and Perl was developed to facilitate data analysis and appended to the previously developed MAD data analysis pipeline(http://probes.pw.usda.gov/bioinformatics_ tools/MADPipeline/index.html).

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.

Genetically modified human umbilical cord blood cells as a promising strategy for treatment of spinal cord injury

Spinal cord injury （SCI） continues to be a pressing health and social problem. The injury leads to neuronal and glial cell death accompanied by degeneration of nerve fibers. There are currently no particularly effective treatments. SCI causes profound disabil- ity of people affected and has attracted increased attention in the international field of neuroregeneration. For the past two decades, much hope has been placed in cell therapies for the restoration of both structure and function of the injured spinal cord. Embryonic and neural stem cells, olfactory ensheathing cells, microglia-like cells, Schwann cells, mesenchymal stem cells.

Blood-testis barrier and spermatogenesis： lessons From genetically-modified mice

The blood-testis barrier （BTB） is found between adjacent Sertoli cells in the testis where it creates a unique microenvironment for the development and maturation of meiotic and postmeiotic germ cells in seminiferous tubes. It is a compound proteinous structure, composed of several types of cell junctions including tight junctions （TJs）, adhesion junctions and gap junctions （GJs）. Some of the junctional proteins function as structural proteins of BTB and some have regulatory roles. The deletion or functional silencing of genes encoding these proteins may disrupt the BTB, which may cause immunological or other damages to meiotic and postmeiotic cells and ultimately lead to spermatogenic arrest and infertility. In this review, we will summarize the findings on the BTB structure and function from genetically-modified mouse models and discuss the future perspectives.

人血型抗体IgG、IgM对基因修饰猪外周血PBMC和RBC杀伤作用初步研究

目的:探讨人血型抗体IgG、IgM对基因修饰猪外周血PBMC和RBC的结合和杀伤差异,为猪-人异种肾移植临床应用的供体猪血型选择提供理论依据。方法:抽取20例健康正常人、20例终末肾病患者和20例脑死亡器官捐献者血清,并根据ABO血型分成4组(A血型:n=20;B血型:n=17;AB血型:n=7;O血型:n=16)。用流式细胞术检测4组人血清分别与O血型野生型(WT)、α1、3-半乳糖基转移酶基因敲除(GTKO)、胞苷酸N-乙酰神经氨酸羟化酶基因敲除(GTKO/β4GalNT2KO)、β1、4-N-乙酰氨基半乳糖转移酶2基因敲除(GTKO/CMAHKO)和3种基因敲除(TKO/hCD55)猪外周血单个核细胞(PBMC)和红细胞(RBC)的抗体结合或补体依赖的细胞毒性试验(CDC)。结果:A血型、B血型、AB血型和O血型人血清抗体IgM和IgG分别对WT猪PBMC,GTKO猪PBMC,GTKO/β4GalNT2KO猪PBMC,GTKO/CMAHKO猪PBMC,TKO/hCD55猪PBMC的结合和杀伤无明显差异。A血型、B血型、AB血型和O血型人血清抗体分别对WT猪RBC,GTKO猪RBC,GTKO/β4GalNT2KO猪RBC,GTKO/CMAHKO猪RBC,TKO/hCD55猪RBC的结合无明显差异。结论:O型血的TKO猪可能是目前最佳基因型供猪,且受者可能不需要考虑血型因素。

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.

Georgian Farmers＇ Attitudes Towards Genetically Modified Crops

Although, genetically engineered products （GM） have to be a broadly debated topic in different countries, there has been much less attention devoted to farmers＇ attitudes towards GM crops. This paper attempts to research farmers＇ insights on GM crops in Georgia through February-March 2014. An in-depth survey of 611 farmers revealed that respondents lack sufficient knowledge about genetic engineering. They tend to have a negative attitude towards GM crops and are strongly against of import and adoption of GM seeds. An empirical examination based on analysis of variance and Pearson＇s correlation coefficient verified that both education and age were significant determinants of awareness of farmers about genetically engineered crops, while income used to have no significant influence on the farmers＇ decision to adopt GM crops. In addition, relationship between awareness about genetic engineering and farmers＇ decision to adopt GM crops has to be insignificant, as well.

Genetically-Modified Organisms in United States Agriculture: Mandate for Food Labeling

The production of foods with genetically modified organisms (GMOs) has risen rapidly over the past three decades to comprise nearly 90% of crops grown in the United States today. Currently, there are no mandates for labeling foods containing GMOs. GMO agricultural crops contain the insertion of genes encoding for pesticides, pesticide resistance, growth factors, or other substances not normally present. In addition to the foreign genes that are inserted, hundreds to thousands of mutations disrupt normal genes in GMO plants. Recently, animal studies have demonstrated toxicity of GMO foods causing organ failure, infertility, carcinomas and death. The FDA requirement of ingredients added to foods be labeled on the product is not applied to GMO foods, precluding the consumer’s right to know. GMOs provide an economic incentive to companies because the seeds can be patented, driving up costs and creating the potential for monopolies. Herbicide-resistance conferred by GMOs has resulted in higher pesticide applications, which correlate with higher human cancer rates, and the emergence of pesticide-resistant weeds and insects. GMO toxins are spreading into to non-target insects, waterways and aquatic organisms, with toxicity to non-target organisms and resultant contamination of disparate ecosystems in the food chain. The appropriateness of mandatory GMO labeling of foods in the United States is discussed.