Late blight, caused by the pathogen Phytophthora infestans, is a devastating disease affecting potato production globally, with adverse effects in Africa where limited access to fungicides exacerbates its impact. Outbreaks of late blight lead to reduced yields and substantial economic losses to potato farmers and agricultural systems. The development of resistant potato varieties, tailored to African agroecological conditions, offers a viable solution in mitigating the devastating effects of late blight on potato cultivation. Leading to this study, two consumer-preferred varieties, Victoria and Shangi, with high susceptibility to late blight were targeted for conferring late blight resistance through genetic engineering. This was achieved by inserting R genes from wild relatives of potato displaying resistance to the disease. The intended effect of conferring resistance to the late blight disease has been consistently observed over twenty experimental field trials spanning 8 years at three locations in Uganda and Kenya. In this study, we assessed whether the genetic transformation has led to any significant unintended effects on the nutritional and anti-nutritional composition of potato tubers compared to the non-transgenic controls grown under the same agroecological conditions. The compositional assessments were conducted on commercial-size potato tubers harvested from regulatory trials at three locations in Uganda and Kenya. Statistical analysis was conducted using two-way analysis of variance comparing transgenic and non-transgenic samples. Overall, the results showed that the transgenic and non-transgenic samples exhibited similar levels of nutritional and antinutritional components. Variations detected in the levels of the analysed components fell within the expected ranges as documented in existing literature and potato composition databases. Thus, we conclude that there are no biologically significant differences in the nutritional and anti-nutritional composition of transgenic and non-transgenic potato tubers engineered for resistance to late blight.

Multigene cotransformation has been widely used in the study of genetic improvement in crops and trees. However, little is known about the unintended effects and causes of multigene cotransformation in poplars. To gain insight into the unintended effects of T-DNA integration during multigene cotransformation in field stands, here, three lines (A1-A3) of Populus × euramericana cv. Neva (PEN) carrying Cry1Ac-Cry3A-BADH genes and three lines (B1-B3) of PEN carrying Cry1Ac-Cry3A-NTHK1 genes were used as research objects, with non-transgenic PEN as the control. Experimental stands were established at three common gardens in three locations and next generation sequencing (NGS) was used to identify the insertion sites of exogenous genes in six transgenic lines. We compared the growth data of the transgenic and control lines for four consecutive years. The results demonstrated that the tree height and diameter at breast height (DBH) of transgenic lines were significantly lower than those of the control, and the adaptability of transgenic lines in different locations varied significantly. The genotype and the experimental environment showed an interaction effect. A total of seven insertion sites were detected in the six transgenic lines, with B3 having a double-site insertion and the other lines having single copies. There are four insertion sites in the gene region and three insertion sites in the intergenic region. Analysis of the bases near the insertion sites showed that AT content was higher than the average chromosome content in four of the seven insertion sites within 1000 bp. Transcriptome analysis suggested that the differential expression of genes related to plant hormone transduction and lignin synthesis might be responsible for the slow development of plant height and DBH in transgenic lines. This study provides an integrated analysis of the unintended effects of transgenic poplar, which will benefit the safety assessment and reasonable application of genetically modified trees.

The extent of molecular diversity and differentially expressed proteins (DEPs) in transgenic lines provide valuable information to understand the phenotypic performance of transgenic crops compared with their parents. Here, we compared the differences in the phenotypic variation of twelve agronomic and end-use quality traits, the extent of microsatellite diversity, and DEPs of a recurrent parent line with three transgenic rice restorer lines carrying either CRY1C gene on chromosome 11 or CRY2A gene on chromosome 12 or both genes. The three transgenic lines had significantly smaller stem borer infestation than the recurrent parent without showing significant differences among most agronomic traits, yield components, and end-use quality traits. Using 512 microsatellite markers, the three transgenic lines inherited 2.9-4.3% of the Minghui 63 donor genome and 96.3-97.1% of the CH891 recurrent parent genome. As compared with the recurrent parent, the number of upregulated and down-regulated proteins in the three transgenic lines varied from 169 to 239 and from 131 to 199, respectively. Most DEPs were associated with the secondary metabolites biosynthesis transport and catabolism, carbohydrate transport and metabolism, post-translational modification, and signal transduction mechanisms. Although several differentially expressed proteins were observed between transgenic rice and its recurrent parent, the differences may not have been associated with grain yield and most other phenotypic traits in transgenic rice.

The use of omics techniques to analyze the differences between genetic engineering organisms and their parents can identify unintended effects and explore whether such unintended effects will have negative consequences. In order to evaluate whether genetic engineering will cause changes in crops beyond the changes introduced by conventional plant breeding, we compared the extent of transcriptome and metabolome modification in the leaves of three lines developed by RNA interference (RNAi)-based genetic engineering and three lines developed by conventional breeding. The results showed that both types of plant breeding methods can manifest changes at the short interfering RNA (siRNA), transcriptomic, and metabolic levels. Relative expression analysis of potential off-target gene revealed that there was no broad gene decline in the three RNAi-based genetic engineering lines. We found that the number of DEGs and DAMs between RNAi-based genetic engineering lines and the parental line was less than that between conventional breeding lines. These unique DEGs and DAMs between RNAi-based genetic engineering lines and the parental lines were not enriched in detrimental metabolic pathways. The results suggest that RNAi-based genetic engineering do not cause unintended effects beyond those found in conventional breeding in maize.

UNASSIGNED: RNAi-based genetically modified maize resistant to Monolepta hieroglyphica (Motschulsky) was demonstrated with negligible transcriptome and metabolome alterations compared to its unmodified equivalent. As one of the most prevalent insect pests afflicting various crops, Monolepta hieroglyphica (Motschulsky) causes severe loss of agricultural and economic productivity for many years in China. In an effort to reduce damages, in this study, an RNA interference (RNAi)-based genetically modified (GM) maize was developed. It was engineered to produce MhSnf7 double-stranded RNAs (dsRNAs), which can suppress the Snf7 gene expression and then lead M. hieroglyphica to death. Field trail analysis confirmed the robustly insecticidal ability of the MhSnf7 GM maize to resist damages by M. hieroglyphica. RNA sequencing analysis identified that only one gene was differentially expressed in the MhSnf7 GM maize compared to non-GM maize, indicating that the transcriptome in MhSnf7 GM maize is principally unaffected by the introduction of the MhSnf7 dsRNA expression vector. Likewise, metabolomics analysis identified that only 8 out of 5787 metabolites were significantly changed. Hence, the integration of transcriptomics and metabolomics demonstrates that there are negligible differences between MhSnf7 GM maize and its unmodified equivalent. This study not only presents a comprehensive assessment of cellular alteration in terms of gene transcription and metabolite abundance in RNAi-based GM maize, but also could be used as a reference for evaluating the unintended effect of GM crops.

Advancements in -omics techniques provide powerful tools to assess the potential effects in composition of a plant at the RNA, protein and metabolite levels. These technologies can thus be deployed to assess whether genetic engineering (GE) causes changes in plants that go beyond the changes introduced by conventional plant breeding. Here, we compare the extent of transcriptome and metabolome modification occurring in leaves of four GE rice lines expressing Bacillus thuringiensis genes developed by GE and seven rice lines developed by conventional cross-breeding. The results showed that both types of crop breeding methods can bring changes at transcriptomic and metabolic levels, but the differences were comparable between the two methods, and were less than those between conventional non-GE lines were. Metabolome profiling analysis found several new metabolites in GE rice lines when compared with the closest non-GE parental lines, but these compounds were also found in several of the conventionally bred rice lines. Functional analyses suggest that the differentially expressed genes and metabolites caused by both GE and conventional cross-breeding do not involve detrimental metabolic pathways. The study successfully employed RNA-sequencing and high-performance liquid chromatography mass spectrometry technology to assess the unintended changes in new rice varieties, and the results suggest that GE does not cause unintended effects that go beyond conventional cross-breeding in rice.

Unintended effects of engineering agents and materials on the formation of undesirable disinfection byproducts (DBPs) during drinking water treatment and distribution were comprehensively reviewed. Specially, coagulants, biologically active filtration biofilms, activated carbons, nanomaterials, ion-exchange resins, membrane materials in drinking water treatment and piping materials, deposits and biofilms within drinking water distribution systems were discussed, which may serve as DBP precursors, transform DBPs into more toxic species, and/or catalyze the formation of DBPs. Speciation and quantity of DBPs generated rely heavily on the material characteristics, solution chemistry conditions, and operating factors. For example, quaternary ammonium polymer coagulants can increase concentrations of N-nitrosodimethylamine (NDMA) to above the California notification level (10 ng/L). Meanwhile, the application of strong base ion-exchange resins has been associated with the formation of N-nitrosamines and trichloronitromethane up to concentrations of 400 ng/L and 9.0 μg/L, respectively. Organic compounds leaching from membranes and plastic and rubber pipes can generate high NDMA (180-450 ng/L) and chloral hydrate (∼12.4 μg/L) upon downstream disinfection. Activated carbon and membranes preferentially remove organic precursors over bromide, resulting in a higher proportion of brominated DBPs. Copper corrosion products (CCPs) accelerate the decay of disinfectants and increase the formation of halogenated DBPs. Chlorination of high bromide waters containing CCPs can form bromate at concentrations exceeding regulatory limits. Owing to the aforementioned concern for the drinking water quality, the application of these materials and reagents during drinking water treatment and distribution should be based on the removal of pollutants with consideration for balancing DBP formation during disinfection scenarios. Overall, this review highlights situations in which the use of engineering agents and materials in drinking water treatment and distribution needs balance against deleterious impacts on DBP formation.

Gene stacking is a developing trend in agricultural biotechnology. Unintended effects in stacked transgenic plants are safety issues considered by the public and researchers. Omics techniques provide useful tools to assess unintended effects. In this paper, stacked transgenic maize 12-5×IE034 that contained insecticidal cry and glyphosate tolerance G10-epsps genes was obtained by crossing of transgenic maize varieties 12-5 and IE034. Transcriptome and metabolome analyses were performed for different maize varieties, including 12-5×IE034, 12-5, IE034, and conventional varieties collected from different provinces in China. The transcriptome results were as follows. The nine maize varieties had obvious differences in gene expression. There were 3561-5538 differentially expressed genes between 12-5×IE034 and its parents and transgenic receptor, which were far fewer than the number of differentially expressed genes in different traditional maize varieties. Cluster analysis indicated that there were close relationships between 12-5×IE034 and its parents. The metabolome results were as follows. For the nine detected maize varieties, the number of different metabolites ranged from 0 to 240. Compared with its parents, 12-5 and IE034, the hybrid variety 12-5×IE034 had 15 and 112 different metabolites, respectively. Hierarchical cluster analysis with Pearson\'s correlation analysis showed that the differences between 12-5×IE034 and its parents were fewer than those between other maize varieties. Shikimate pathway-related genes and metabolites analysis results showed that the effects of hybrid stacking are less than those from transformation and differing genotypes. Thus, the differences due to breeding stack were fewer than those due to natural variation among maize varieties. This paper provides scientific data for assessing unintended effects in stacked transgenic plants.

The key research question that this article aims to respond to is: what are the individual-level effects of wages financed by development assistance? Our hypothesis is that these effects are substantial and overall positive, depending on the level of analysis. This article theorizes about unintended wage effects at the individual level, spillover effects, and those at the macro level, yet focuses its research on individual-level effects. The empirical part consists of two case studies. One takes a sample of grants from a donor agency, the Netherlands Ministry of Foreign Affairs, as a starting point, following these grants through the aid chain to determine local staffing costs. The other case study consists of a comparative wage analysis in a developing country, the Democratic Republic of Congo. As this field of research is rather fresh, instead of answering relevant questions with respect to wage effects, this study merely aims to identify questions that merit further research.

To investigate unintended effects in genetically modified crops (GMCs), a comparative proteomic analysis between the leaves of the phytase-transgenic maize and the non-transgenic plants was performed using two-dimensional gel electrophoresis and mass spectrometry. A total of 57 differentially expressed proteins (DEPs) were successfully identified, which represents 44 unique proteins. Functional classification of the identified proteins showed that these DEPs were predominantly involved in carbohydrate transport and metabolism category, followed by post-translational modification. KEGG pathway analysis revealed that most of the DEPs participated in carbon fixation in photosynthesis. Among them, 15 proteins were found to show protein-protein interactions with each other, and these proteins were mainly participated in glycolysis and carbon fixation. Comparison of the changes in the protein and tanscript levels of the identified proteins showed that most proteins had a similar pattern of changes between proteins and transcripts. Our results suggested that although some significant differences were observed, the proteomic patterns were not substantially different between the leaves of the phytase-transgenic maize and the non-transgenic isogenic type. Moreover, none of the DEPs was identified as a new toxic protein or an allergenic protein. The differences between the leaf proteome might be attributed to both genetic modification and hybrid influence.