Non-transgenic Counterparts Research Articles

Effects of transgenic glufosinate-tolerant rapeseed (Brassica napus L.) and the associated herbicide application on rhizospheric bacterial communities

Effects of transgenic crops on soil microbial communities are important constituents in ecological risk assessment. The Illumina MiSeq sequencing method was employed to examine the effects of transgenic glufosinate-resistant rapeseed and the associated herbicide application on rhizospheric bacterial communities at different growth stages. Our results showed that both transgenic rapeseed and the application of herbicide Basta (active ingredient: glufosinate) had no significant influence on alpha-diversity of rhizospheric bacterial communities. The dominant phyla across all samples were Proteobacteria, Bacteroidetes, Acidobacteria, Gemmatimonadetes and Actinobacteria. These dominant phyla show no remarkable differences under transgenic rapeseed, transgenic rapeseed with glufosinate applied, and non-transgenic acceptor counterpart rapeseed. Nevertheless, they respond differently at different growth stages. These results indicate that community variation was strongly dependent on the growth stage. Therefore, growing glufosinate-resistant transgenic rapeseed line Z7B10 and application of glufosinate herbicide had no adverse effects on the rhizospheric bacterial community composition.

Global metabolite profiling based on GC\u2013MS and LC\u2013MS/MS analyses in ABF3-overexpressing soybean with enhanced drought tolerance

Abscisic acid (ABA) is a phytohormone that plays an important role in the adaptive responses to abiotic stresses. We examined the metabolic changes in transgenic soybean that over-expressed Arabidopsis ABA responsive element-binding factor 3 (ABF3), which participates in drought tolerance. Transgenic and non-transgenic plants were exposed to a water deficit, and their metabolic differences were verified by untargeted GC‒MS and LC‒MS/MS analyses. A total of 64 and 476 primary and secondary metabolites from leaf extracts were identified based on GC‒MS and LC‒MS/MS platforms, respectively. Principal component analysis derived from both GC‒MS and LC‒MS/MS data showed a clearly greater separation in the metabolite profiles among three different degrees of drought stress. However, no discrimination of metabolites between transgenic and non-transgenic plants was apparent. Furthermore, except for some free amino acids, quantitative differences in relative levels of those metabolites were less than 50% between genotypes. These results suggest that, during periods of drought, overexpression of ABF3 in transgenic soybean might result in a negligible variance in primary and secondary metabolism when compared with its non-transgenic counterpart.

Atrial fibrillation and electrophysiology in transgenic mice with cardiac-restricted overexpression of FKBP12.

Cardiomyocyte-restricted overexpression of FK506-binding protein 12 transgenic (αMyHC-FKBP12) mice develop spontaneous atrial fibrillation (AF). The aim of the present study is to explore the mechanisms underlying the occurrence of AF in αMyHC-FKBP12 mice. Spontaneous AF was documented by telemetry in vivo and Langendorff-perfused hearts of αMyHC-FKBP12 and littermate control mice in vitro. Atrial conduction velocity was evaluated by optical mapping. The patch-clamp technique was applied to determine the potentially altered electrophysiology in atrial myocytes. Channel protein expression levels were evaluated by Western blot analyses. Spontaneous AF was recorded in four of seven αMyHC-FKBP12 mice but in none of eight nontransgenic (NTG) controls. Atrial conduction velocity was significantly reduced in αMyHC-FKBP12 hearts compared with NTG hearts. Interestingly, the mean action potential duration at 50% but not 90% was significantly prolonged in αMyHC-FKBP12 atrial myocytes compared with their NTG counterparts. Consistent with decreased conduction velocity, average peak Na+ current ( INa) density was dramatically reduced and the INa inactivation curve was shifted by approximately +7 mV in αMyHC-FKBP12 atrial myocytes, whereas the activation and recovery curves were unaltered. The Nav1.5 expression level was significantly reduced in αMyHC-FKBP12 atria. Furthermore, we found increases in atrial Cav1.2 protein levels and peak L-type Ca2+ current density and increased levels of fibrosis in αMyHC-FKBP12 atria. In summary, cardiomyocyte-restricted overexpression of FKBP12 reduces the atrial Nav1.5 expression level and mean peak INa, which is associated with increased peak L-type Ca2+ current and interstitial fibrosis in atria. The combined electrophysiological and structural changes facilitated the development of local conduction block and altered action potential duration and spontaneous AF. NEW & NOTEWORTHY This study addresses a long-standing riddle regarding the role of FK506-binding protein 12 in cardiac physiology. The work provides further evidence that FK506-binding protein 12 is a critical component for regulating voltage-gated sodium current and in so doing has an important role in arrhythmogenic physiology, such as atrial fibrillation.

Induction of Lrp5 HBM-causing mutations in Cathepsin-K expressing cells alters bone metabolism.

High-bone-mass (HBM)-causing missense mutations in the low density lipoprotein receptor-related protein-5 (Lrp5) are associated with increased osteoanabolic action and protection from disuse- and ovariectomy-induced osteopenia. These mutations (e.g., A214V and G171V) confer resistance to endogenous secreted Lrp5/6 inhibitors, such as sclerostin (SOST) and Dickkopf homolog-1 (DKK1). Cells in the osteoblast lineage are responsive to canonical Wnt stimulation, but recent work has indicated that osteoclasts exhibit both indirect and direct responsiveness to canonical Wnt. Whether Lrp5-HBM receptors, expressed in osteoclasts, might alter osteoclast differentiation, activity, and consequent net bone balance in the skeleton, is not known. To address this, we bred mice harboring heterozygous Lrp5 HBM-causing conditional knock-in alleles to Ctsk-Cre transgenic mice and studied the phenotype using DXA, μCT, histomorphometry, serum assays, and primary cell culture. Mice with HBM alleles induced in Ctsk-expressing cells (TG) exhibited higher bone mass and architectural properties compared to non-transgenic (NTG) counterparts. In vivo and in vitro measurements of osteoclast activity, population density, and differentiation yielded significant reductions in osteoclast-related parameters in female but not male TG mice. Droplet digital PCR performed on osteocyte enriched cortical bone tubes from TG and NTG mice revealed that ~8-17% of the osteocyte population (depending on sex) underwent recombination of the conditional Lrp5 allele in the presence of Ctsk-Cre. Further, bone formation parameters in the midshaft femur cortex show a small but significant increase in anabolic action on the endocortical but not periosteal surface. These findings suggest that Wnt/Lrp5 signaling in osteoclasts affects osteoclastogenesis and activity in female mice, but also that some of the changes in bone mass in TG mice might be due to Cre expression in the osteocyte population.

Systematic miRNome profiling reveals differential microRNAs in transgenic maize metabolism

BackgroundWhile some genetically modified organisms (GMOs) are created to produce new double-stranded RNA molecules (dsRNA), in others, such molecules may occur as an unintended effect of the genetic engineering process. Furthermore, GMOs might produce naturally occurring dsRNA molecules in higher or lower quantities than its non-transgenic counterpart. This study is the first to use high-throughput technology to characterize the miRNome of commercialized GM maize events and to investigate potential alterations in miRNA regulatory networks.ResultsThirteen different conserved miRNAs were found to be dys-regulated in GM samples. The insecticide Bt GM variety had the most distinct miRNome. These miRNAs target a range of endogenous transcripts, such as transcription factors and nucleic acid binding domains, which play key molecular functions in basic genetic regulation. In addition, we have identified 20 potential novel miRNAs with target transcripts involved in lipid metabolism in maize. isomiRs were also found in 96 conserved miRNAs sequences, as well as potential transgenic miRNA sequences, which both can be a source of potential off-target effects in the plant genome. We have also provided information on technical limitations and when to carry on additional in vivo experimental testing.ConclusionsThese findings do not reveal hazards per se but show that robust and reproducible miRNA profiling technique can strengthen the assessment of risk by detecting any new intended and unintended dsRNA molecules, regardless of the outcome, at any stage of GMO development.

Early Impairments of Hippocampal Neurogenesis in 5xFAD Mouse Model of Alzheimer's Disease Are Associated with Altered Expression of SOXB Transcription Factors.

Dysregulation of neurogenesis in the subgranular zone (SGZ) of the hippocampal dentate gyrus has been related to cognitive deficits and memory loss in neurodegenerative diseases, such as Alzheimer's disease (AD). Members of the B group of SOX transcription factors play critical roles in regulating neurogenesis in the embryonic and adult nervous system, including maintaining the multipotency, renewal, and cell fate decision of neural stem/progenitor cells. The aim of the present study was to evaluate the expression patterns of selected SOXB proteins in the SGZ, of 8-week-old male and female 5xFAD mice, which represent a transgenic model of AD with a severe and very early development of amyloid pathology. Immunohistochemical analysis showed a significant decrease in the number of cells expressing SOX1, SOX2, and SOX21 transcription factors within the SGZ of 5xFAD mice in comparison to their non-transgenic counterparts which coincidences with reduced number of doublecortin immunoreactive immature neurons found in Tg males. Despite observed changes in expressional pattern of examined SOXB proteins, the proliferative capacity evaluated by the number of Ki-67 immunoreactive cells remained unaffected in transgenic mice of both genders. Based on our results, we suggest that SOXB proteins might be considered as new biomarkers for the detection of early impairments in adult neurogenesis in different animal models or/and new targets in human regenerative medicine.

Improved quantification of γ-aminobutyric acid in rice using stable isotope dilution gas chromatography–mass spectrometry

An accurate method for the analysis of γ-aminobutyric acid (GABA) in rice grain was developed using trimethylsilyl (TMS) derivatization and stable isotope dilution gas chromatography-mass spectrometry. When this method was used with GABA‑d6 as an internal standard (IS), the observed GABA concentration was maintained at 100% of the initial concentration with increasing storage time of the vial in the autosampler. In contrast, when using ribitol as an IS and multiple injections from one vial or single injections from different vials, the observed GABA concentration was 85 and 113% of the initial concentration upon increased storage time, respectively. The improved method recoveries at two different spike levels were between 93.3 and 97.8%, with relative standard deviations of less than 3.3%. The GABA content of resveratrol-enriched transgenic rice was compared with that of its non-transgenic counterpart from two field sites, and statistically non-significant differences were observed between the two grains.

Comparative compositional analysis of transgenic potato resistant to potato tuber moth (PTM) and its non-transformed counterpart.

In this study, the compositions of transgenic potatoes (TPs) resistant to potato tuber moth (Phthorimaea operculella) were compared with those of its non-transgenic (NTP) counterparts. The light inducible promoter, phosphoenolpyruvate carboxylase led to the expression of Cry1Ab only in the leaves and light-treated tubers of the TPs. No significant differences were found in the moisture, ash, dry weight, total soluble protein, carbohydrate, starch, fiber, ascorbate, cations, anions, fatty acids, and glycoalkaloids contents of TP and NTP. Moreover, light treatment significantly affected the contents of ascorbate, acetate and nitrite anions, palmitic, stearic and linolenic fatty acids, α-haconine and α-solanine glycoalkaloids in TP and NTP tubers. While, significant differences were observed in the amino acid contents in light-treated tubers of TPs than the NTP ones. Although, light treatment in potato tubers resulted in marked metabolic changes, all the variations observed in the metabolites compositions were found to be within the desired reference ranges for potato plants. In conclusion, the results indicated that the TPs were substantially and nutritionally equivalent to the NTP counterparts.

Repressible Transgenic Sterilization in Channel Catfish, Ictalurus punctatus, by Knockdown of Primordial Germ Cell Genes with Copper-Sensitive Constructs.

Repressible knockdown approaches were investigated to manipulate for transgenic sterilization in channel catfish, Ictalurus punctatus. Two primordial germ cell (PGC) marker genes, nanos and dead end, were targeted for knockdown and an off-target gene, vasa, was monitored. Two potentially copper-sensitive repressible promoters, yeast ctr3 (M) and ctr3-reduced (Mctr), were coupled with four knockdown strategies separately including: ds-sh RNA targeting the 5' end (N1) or 3' end (N2) of channel catfish nanos, full-length cDNA sequence of channel catfish nanos for overexpression (cDNA), and ds-sh RNA-targeting channel catfish dead end (DND). Each construct had an untreated group and treated group with copper sulfate as the repressor compound. Spawning rates of full-sibling P1 fish exposed or not exposed to the constructs as treated and untreated embryos were 85 and 54%, respectively, indicating potential sterilization of fish and repression of the constructs. In F1 fish, mRNA expressions of PGC marker genes for most constructs were downregulated in the untreated group and the knockdown was repressed in the treated group. Gonad development in transgenic, untreated F1 channel catfish was reduced compared to non-transgenic fish for MctrN2, MN1, MN2, and MDND. For 3-year-old adults, gonad size in the transgenic untreated group was 93.4% smaller than the non-transgenic group for females and 92.3% for males. However, mean body weight of transgenic females (781.8g) and males (883.8g) was smaller than of non-transgenic counterparts (984.2 and 1254.3g) at 3years of age, a 25.8 and 41.9% difference for females and males, respectively. The results indicate that repressible transgenic sterilization is feasible for reproductive control of fish, but negative pleiotropic effects can result.

Long-term Treatment with Low-Dose Caffeine Worsens BPSD-Like Profile in 3xTg-AD Mice Model of Alzheimer's Disease and Affects Mice with Normal Aging.

Coffee or caffeine has recently been suggested as prophylaxis for dementia. Although memory problems are hallmarks of Alzheimer’s disease, this dementia is also characterized by neuropsychiatric symptoms called Behavioral and Psychological Symptoms of Dementia (BPSD). The impact of preventive/therapeutic strategies on both cognitive and non-cognitive symptoms can be addressed in the 3xTg-AD mice, since they exhibit cognitive but also BPSD-like profiles. Here, we studied the long-term effects of a low dose of caffeine in male 3xTg-AD mice and as compared to age-matched non-transgenic (NTg) counterparts with normal aging. Animals were treated (water or caffeine in drinking water) from adulthood (6 months of age) until middle-aged (13 months of age), that in 3xTg-AD mice correspond to onset of cognitive impairment and advanced stages, respectively. The low caffeine dosing used (0.3 mg/ml) was previously found to give a plasma concentration profile in mice roughly equivalent to that of a human coffee drinker. There were significant effects of caffeine on most behavioral variables, especially those related to neophobia and other anxiety-like behaviors, emotionality, and cognitive flexibility. The 3xTg-AD and NTg mice were differently influenced by caffeine. Overall, the increase of neophobia and other anxiety-related behaviors resulted in an exacerbation of BPSD-like profile in 3xTg-AD mice. Learning and memory, strongly influenced by anxiety in 3xTg-AD mice, got little benefit from caffeine, only shown after a detailed analysis of navigation strategies. The worsened pattern in NTg mice and the use of search strategies in 3xTg-AD mice make both groups more similar. Circadian motor activity showed genotype differences, which were found to be enhanced by caffeine. Selective effects of caffeine on NTg were found in the modulation of behaviors related to emotional profile and risk assessment. Caffeine normalized splenomegaly of 3xTg-AD mice, a physical indicator of their impaired peripheral immune system, and trended to increase their corticosterone levels. Our observations of adverse caffeine effects in an Alzheimer’s disease model together with previous clinical observations suggest that an exacerbation of BPSD-like symptoms may partly interfere with the beneficial cognitive effects of caffeine. These results are relevant when coffee-derived new potential treatments for dementia are to be devised and tested.

Toxicologic evaluation of chronic feeding of glyphosate-resistant transgenic soybean GTS40-3-2 meal to rats

To assess the safety of glyphosate-resistant transgenic soybean, the transgenic soybean GTS40-3-2 meal and its non-transgenic counterpart parent A5403 were fed to first- and second-generation Sprague-Dawley (SD) rats (Rattus norvegicus) for 195 days. The soybean meal made up 20% of the total weight of the feed. Body weight, food consumption, hematology, blood biochemistry, and pathological observations were monitored weekly. The results showed that rats of each group experienced good growth and development, and although results of some organ weight ratios and serum biochemistry between the groups were significantly different (P<0.05), they were of no biological significance because the variations were within the normal range. These results indicate that glyphosate-resistant transgenic soybean GTS40-3-2 meal is as safe as its non-transgenic parent soybean A5403 in a long-term SD rat feeding study. The study provides experimental evidence for the safe usage of transgenic soybean GTS40-3-2.

Comparative metabolic profiling of four transgenic maize lines and two non-transgenic maize lines using high-performance liquid chromatography mass spectrometry

We compared metabolic changes in transgenic maize lines (differing in genetic backgrounds and exogenous genes insertion) with non-transgenic counterparts. At the corn seedling stage, entirely expanding leaves were obtained from maize lines 1 (transgenic maize SK12-5; overexpressing cry1Ab/cry2Aj; parental maize ZD958), 2 (transgenic maize IE034; overexpressing cry1Ie; parental maize ZD958), 3 (transgenic maize Bt799; overexpressing cry1Ac-M; parental maize Z58), 4 (transgenic maize Bt799; overexpressing cry1Ac-M; parental maize ZD958), 5 (non-transgenic maize Z58), and 6 (non-transgenic maize ZD958). For each line, six biological replicates were prepared; each contained an entirely expanded leaf at the top of the plant. Metabolites were identified with high-performance liquid chromatography mass spectrometry in three comparison groups (4 transgenic maize lines vs. their corresponding controls, line 3 vs. 4, and transgenic vs. non-transgenic maize). Top 200 cationic and top 200 anionic metabolites with higher variable importance in projection value were chosen for principal component analysis, followed by pathway enrichment analysis. Annotation was possible for 227 of 400 metabolites. Twelve up- and 26 down-regulated overlapping metabolites were identified in line 1 vs. 6, line 2 vs. 6, and line 4 vs. 6 comparison groups. Altered metabolites significantly enriched in purine and glutathione metabolism pathways, but no pathways were enriched in line 3 vs. 4. Comparing transgenic and non-transgenic maize lines revealed 59 up- and 37 down-regulated metabolites, which were also significantly enriched in purine and glutathione metabolism pathways. Therefore, transgenic Bt maize differed metabolically from non-transgenic maize. Additionally, purine and glutathione metabolism pathways may be important in the transgenic maize metabolism.

Nutritional composition analysis for beta-carotene-enhanced transgenic soybeans (Glycine max L.)

Nutritional composition is important for assessing the safety of genetically modified (GM) crops for human consumption. Three beta-carotene-enhanced soybean lines were developed by introducing the β-conglycinin promoter::Phytoene synthase-2A-Carotene desaturase/t35S gene cassette into the genome of the commercial Kwangan (Glycine max L.) soybean variety. Transgenic soybeans were successfully detected on beta-carotene productions ranged from 170.47 to 213.58 µg/g. Comparative assessments of nutrition were conducted with 3 transgenic soybeans, their non-GM counterpart, and several commercial soybean varieties. Results indicated that most levels of proximate, fatty acids, amino acids, and vitamins showed non-significant differences between transgenic soybeans and their counterpart, and fit within the reference ranges established for other commercial soybeans and Organization for Economic Cooperation and Development Guidelines. However, significant differences on levels of crude fat, carbohydrate, δ-tocopherol, and oleic acid of transgenic soybeans comparing to those of non-transgenic counterpart Kwangan cannot eliminate the influences of transgene insertion. Alternations on compositions should be definite by further studies, such as transcriptome and metabolome profiling.

Different responses of transgenic Bt rice and conventional rice to elevated ozone concentration.

To assess the different sensitivity to ozone (O3) between transgenic Bt Shanyou63 (Bt-SY63) and its nontransgenic counterpart Shanyou63 (SY63), the leaf gas exchange, yield, grain elements, and antioxidant enzymes were investigated by performing a pot experiment under ambient O3 concentration (A-O3) and elevated O3 concentration (1.5×A-O3, E-O3). Under A-O3, the chlorophyll content and yield of Bt-SY63 were significantly lower than those of SY63, whereas the stomatal conductance (Gs), cellular CO2 concentration (Ci), and Fe, Zn concentration showed the opposite trends. No significant difference was detected for malondialdehyde (MDA) content between two cultivars, although the antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities in Bt-SY63 tended to be higher than those in SY63. Exposure to E-O3 resulted in significant reduction for photosynthesis (Pn), yield and all grain elements (except for N) concentration in both varieties, and the extents in Bt-SY63 were much greater than those in SY63. Meanwhile, significant increases for MDA content by 30.6 and 23.7% in Bt-SY63 and SY63, respectively, were detected under E-O3. These results demonstrated that insertion of exogenous gene could induce several unintentional changes of Bt-SY63 in physiology and growth progress, compared with SY63 under ambient O3 concentration. On the other hand, the injury of Bt-SY63 caused by elevated O3 concentration was more severe than that of SY63. This study provided valuable baseline information for the commercial release and breeding strategies of Bt-SY63 under the projected future climate.

<i>AtDREB2A-CA</i> Influences Root Architecture and Increases Drought Tolerance in Transgenic Cotton

Drought is a major environmental factor limiting cotton (Gossypium hirsutum L.) productivity worldwide and projected climate changes could increase their negative effects in the future. Thus, targeting the molecular mechanisms correlated with drought tolerance without reducing productivity is a challenge for plant breeding. In this way, we evaluated the effects of water deficit progress on AtDREB2A-CA transgenic cotton plant responses, driven by the stress-inducible rd29 promoter. Besides shoot and root morphometric traits, gas exchange and osmotic adjustment analyses were also included. Here, we present how altered root traits shown by transgenic plants impacted on physiological acclimation responses when submitted to severe water stress. The integration of AtDREB2A-CA into the cotton genome increased total root volume, surface area and total root length, without negatively affecting shoot morphometric growth parameters and nor phenotypic evaluated traits. Additionally, when compared to wild-type plants, transgenic plants (17-T0 plants and its progeny) highlighted a gradual pattern of phenotypic plasticity tosome photosynthetic parameters such as photosynthetic rate and stomatal conductance with water deficit progress. Transgene also promoted greater shoot development and root robustness (greater and deeper root mass) allowing roots to grow into deeper soil layers. The same morpho-physiological trend was observed in the subsequent generation (17.6-T2). Our results suggest that the altered root traits shown by transgenic plants are the major contributors to higher tolerance response, allowing the AtDRE2A-CA-cotton plants to maintain elevated stomatal conductance and assimilate rates and, consequently, reducing their metabolic costs involved in the antioxidant responses activation. These results also suggest that these morpho-physiological changes increased the number of reproductive structures retained per plant (26% higher) when compared with its non-transgenic counterpart. This is the first report of cotton plants overexpressing the AtDRE2A-CA transcription factor, demonstrating a morpho-physiological and yield advantages under drought stress, without displaying any yield penalty under irrigated conditions. The mechanisms by which the root traits influenced the acclimation of the transgenic plants to severe water deficit conditions are also discussed. These data present an opportunity to use this strategy in cotton breeding programs in order to improve drought adaptation toward better rooting features.

Proteome rebalancing in transgenic Camelina occurs within the enlarged proteome induced by β-carotene accumulation and storage protein suppression

Oilseed crops are global commodities for their oil and protein seed content. We have engineered the oilseed Camelina sativa to exhibit increased protein content with a slight decrease in oil content. The introduction of a phytoene synthase gene with an RNAi cassette directed to suppress the storage protein 2S albumin resulted in seeds with an 11-24% elevation in overall protein. The phytoene synthase cassette alone produced enhanced β-carotene content of an average 275±6.10μg/g dry seed and an overall altered seed composition of 11% less protein and comparable nontransgenic amounts of both oil and carbohydrates. Stacking an RNAi to suppress the major 2S storage protein resulted in seeds that contain elevated protein and slight decrease in oil and carbohydrate amounts showing that Camelina rebalances its proteome within an enlarged protein content genotype. In both β-carotene enhanced seeds with/without RNAi2S suppression, the seed size was noticeably enlarged compared to nontransgenic counterpart seeds. Metabolic analysis of maturing seeds indicate that the enhanced β-carotene trait had the larger effect than the RNAi2S suppression on the seed metabolome. The use of a GRAS (generally regarded as safe) β-carotene as a visual marker in a floral dip transformation system, such as Camelina, might eliminate the need for costly regulatory and controversial antibiotic resistance markers. β-carotene enhanced RNAi2S suppressed Camelina seeds could be further developed as a rapid heterologous protein production platform in a nonfood crop leveraging its enlarged protein content and visual marker.

Analysis of exogenous gene and protein digestion and absorption of SD rats (Rattus norvegicus) fed roundup ready soybean meal

Metabolism and deposition of exogenous gene and protein from transgenic glyphosate herbicide-tolerant soybean meal in SD rats were studied in the experiment. The transgenic soybean GTS40-3-2 meal and its non-transgenic counterpart (parent A5403) were fed to the generation and the second generation Sprague-Dawley (SD) rats (Rattus norvegicus). The study added the genetically modified (GM) soybean meal and its non-transgenic control soybean meal (parent A5403) in a ratio of 20% respectively to the feeds. By using qualitative, quantitative PCR and ELISA methods to detect transgenic soybean residues of metabolism ingredients in rats, the safety and influence of GM soybean were evaluated. The results showed that the intestinal fecal and cecum contents of rats were detected with residues of GM ingredients, intestinal flora and organs were not found related genes and protein. These results indicated that transgenic glyphosate herbicide-tolerant soybean GTS40-3-2 meal was as safe as its non-GM soybean meal in long-term feeding study.

Application of terahertz spectroscopy imaging for discrimination of transgenic rice seeds with chemometrics

Discrimination of genetically modified organisms is increasingly demanded by legislation and consumers worldwide. The feasibility of a non-destructive discrimination of transgenic rice seeds from its non-transgenic counterparts was examined by terahertz spectroscopy imaging system combined with chemometrics. Principal component analysis (PCA), least squares support vector machines (LS-SVM), PCA-back propagation neural network (PCA-BPNN), and random forest (RF) models with the first and second derivative and standard normal variate transformation (SNV) pre-treatments were applied to classify rice seeds based on genotype. The results demonstrated that differences between non-transgenic and transgenic rice seeds did exist, and an excellent classification (accuracy was 96.67% in the prediction set) could be achieved using the RF model combined with the first derivative pre-treatment. The results indicated that THz spectroscopy imaging together with chemometrics would be a promising technique to identify transgenic rice seeds with high efficiency and without any sample preparation.

A Comprehensive Assessment of the Effects of Transgenic Cry1Ac/Cry1Ab Rice Huahui 1 on Adult Micraspis discolor (Fabricius) (Coleoptera: Coccinellidae).

Micraspis discolor (Fabricius) (Coleoptera: Coccinellidae) is a widely distributed coleoptera predator in southern Asia in rice ecosystem, and adult M. discolor feed on both rice pollen and soft-bodied arthropods. Bitrophic bioassay and tritrophic bioassay were conducted to evaluate the potential impact of Cry1Ac/Cry1Ab-expressing rice Huahui 1 and its non-transgenic counterpart Minghui 63 on fitness parameters of adult M. discolor. The results showed that the survival, and fecundity of this beetle’ adults were not different when they fed on Bt rice or non-Bt rice pollen or Nilaparvata lugens (Stål) reared on Bt rice or non-Bt rice. Toxicity assessment to ensure M. discolor adults were not sensitive to Cry1Ab or Cry1Ac protein independent from the pollen background, M. discolor adults were fed with an artificial diet containing Cry1Ac, Cry1Ab or both protein approximately 10 times higher concentration than in Huahui 1 rice pollen. No difference was detected for any of the life-table parameters tested between Cry protein-containing and pure diet. Artificial diet containing E-64 (N-(trans-Epoxysuccinyl)-L-leucine 4-guanidinobutylamide) was included as a positive control. In contrast, the pre-oviposition and fecundity of M. discolor were significantly adversely affected by feeding on E-64-containing diet. In both bioassays, the uptakes of Cry protein by adult M. discolor were tested by ELISA measurements. These results indicated that adults of M. discolor are not affected by Cry1Ab- or Cry1Ac-expressing rice pollen and are not sensitive to Cry protein at concentrations exceeding the levels in rice pollen in Huahui1. This suggests that M. discolor adults would not be harmed by Cry1Ac/Cry1Ab rice if Bt rice Huahui 1 were commercialized.

A subchronic feeding study of dicamba-tolerant soybean with the dmo gene in Sprague–Dawley rats

The dicamba-tolerant soybean MON87708 expresses the dicamba mono-oxygenase (DMO) enzyme that is encoded by the dmo gene. In order to evaluate the safety of this soybean, a 90-day subchronic feeding toxicity study (13 weeks) was conducted on Sprague–Dawley rats. A total of 140 rats were divided into 7 groups (10/sex/group), including a standard commercial diet control group. The genetically modified (GM) soybean MON87708 and the near isogenic non-GM soybean A3525 were respectively processed to unhulled, full-fat, and heat-treated powder, then mixed into the diet at levels of 7.5%, 15%, and 30% (wt/wt) with the main nutrients of the various diets balanced and then fed to 6 groups. The remaining group of rats fed with a commercial rat diet served as blank control. Some isolated parameters indicated statistically significant differences in body weight, feed consumption/utilization, hematology, serum biochemistry, and relative organ weights. These differences were not consistent across gender or test-diet dose, which were attributed to incidental and biological variability. In conclusion, the results demonstrated that the transgenic soybean MON87708 containing DMO was as safe as non-transgenic isogenic counterpart with historical safe use.