Transgenic Maize Research Articles

Morphological and molecular characterization of Purpureocillium lilacinum along with its biopesticidal effect against fall armyworm (Spodoptera frugiperda) in Southern Taiwan

Abstract Background Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), a key agricultural pest, impacts a broad spectrum of crop species, such as cotton, maize, rice, sorghum, and a range of vegetables. Chemical pesticides and transgenic maize are commonly used to protect crops against this major pest. However, biological control offers a safer and more sustainable solution for long-term FAW management. Entomopathogenic fungi (EPF), a substitute for synthetic insecticides, have been proven to be an effective biocontrol agent for managing pest populations. The present study objective is to isolate and identify the EPF, Purpureocillium lilacinum from soil using morphological and molecular techniques and evaluate its pathogenicity against FAW eggs and various larval stages under different spore concentrations. Results The micro-morphological characteristics fluctuated in growth behavior, appearance, and color under diverse growth media and temperatures. Molecular analysis, which involved constructing a phylogenetic tree from ITS and TEF gene regions, confirmed that PT-02 was P. lilacinum. The pathogenicity of the fungus was tested on FAW eggs, neonates, and larvae at multiple concentrations (1 × 108, 1 × 107, and 1 × 106 spores/ml), resulting in variation mortality. The egg mortality rate at 1 × 108, 1 × 107, and 1 × 106 spores/ml was 98, 98 and 96%, respectively. The neonate mortality rate was 100%, while first and second instars larvae mortality ranged from 10–36% across three concentrations, seven days after treatment (DAT). The median lethal concentration (LC50) values of P. lilacinum PT-02 against FAW eggs, first and second instars, were 3.02 × 107, 1.38 × 108, and 2.56 × 108 spore/ml, respectively. Conclusions The findings revealed that the native strain of P. lilacinum exhibited strong virulence against FAW, suggesting its potential use as an effective biocontrol agent for managing field pest populations.

Applying a computer model to evaluate the evolution of resistance by western corn rootworm to multiple Bt traits in transgenic maize.

Western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is a major pest of maize in the United States. Transgenic maize producing insecticidal toxins from the bacterium Bacillus thuringiensis (Bt) have been used to manage this pest since 2003. Refuges of non-Bt maize have been used to delay resistance to Bt maize by western corn rootworm, and are planted in conjunction with maize producing single or multiple (i.e., pyramids) Bt toxins. Two Bt toxins, Cry3Bb1 and Gpp34/Tpp35Ab1, were used individually before being combined as a pyramid, at which point resistance had already evolved to Cry3Bb1. Pyramids targeting western corn rootworm therefore contained at least one toxin to which resistance had evolved. Western corn rootworm has now evolved resistance to all four commercially available Bt toxins used for rootworm management. We used laboratory and field-generated data to parameterize a deterministic model to simulate the effectiveness of refuges and Bt pyramids to delay resistance to Bt maize in western corn rootworm. Resistance to the pyramid of Cry3Bb1 with Gpp34/Tpp35Ab1 evolved more rapidly when resistance to Cry3Bb1 was already present. This effect arose when model conditions affecting initial resistance allele frequency, inheritance of resistance, and fitness costs were varied. Generally, resistance evolved faster when initial resistance allele frequencies were higher, inheritance of resistance was nonrecessive, and fitness costs were absent, which is consistent with previous models that simulated resistance evolution. We conclude that new transgenic pyramids should pair novel, independently acting toxins with abundant refuges to minimize the risk of rapid resistance evolution.

Optimization of Plant Oxalate Quantification and Generation of Low-Oxalate Maize (Zea mays L.) through O7 Overexpression

Oxalate, the simplest dicarboxylic acid, is a prevalent antinutrient that chelates with various metals and can lead to the formation of kidney stones in humans. The accurate detection of the oxalate concentration in food and the cultivation of low-oxalate crops are important for enhancing public health. In this study, we established a high-throughput and highly sensitive technique for oxalate detection using ultra-high-performance liquid chromatographic–triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS). Additionally, we overexpressed the gene O7, which encodes oxalyl-CoA synthetase in the maize oxalate degradation pathway, resulting in O7-OE lines. By employing the UPLC-QqQ-MS/MS method to measure oxalate levels in these transgenic lines, we observed that the oxalate content in the kernels of O7-OE lines was reduced by approximately 43%, with a concurrent increase in some micronutrients such as zinc. Importantly, the transgenic maize showed normal seed storage compound accumulation or other agronomic characteristics. In summary, we developed a high-throughput detection method that advances oxalate measurement. Furthermore, by generating new maize germplasm with diminished oxalate, our work offers potential health advantages to consumers.

ZmbZIP27 regulates nitrogen-mediated leaf angle by modulating lignin deposition in maize

In grain crops such as maize (Zea mays), leaf angle (LA) is a key agronomic trait affecting light interception and thus planting density and yield. Nitrogen (N) affects LA in plants, but we lack a good understanding of how N regulates LA. Here, we report that N deficiency enhanced lignin deposition in the ligular region of maize seedlings. In situ hybridization showed that the bZIP transcription factor gene ZmbZIP27 is mainly expressed in the phloem of maize vascular bundles. Under N-sufficient conditions, transgenic maize overexpressing ZmbZIP27 showed significantly smaller LA compared with wild type (WT). By contrast, zmbzip27ems mutant showed larger LA under both N-deficient and N-sufficient conditions compared with WT. Overexpression of ZmbZIP27 enhanced lignin deposition in the ligular region of maize in the field. We further demonstrated that ZmbZIP27 could directly bind the promoters of the microRNA genes ZmMIR528a and ZmMIR528b and negatively regulate the expression levels of ZmmiR528. ZmmiR528 knockdown transgenic maize displayed erect architecture in the field by increasing lignin content in the ligular region of maize. Taken together, these results indicate that ZmbZIP27 regulates N-mediated LA size by regulating the expression of ZmmiR528 and modulating lignin deposition in maize.

Effects of transgenic modification on the bacterial communities in different niches of maize under glyphosate toxicity

Transgenic glyphosate-resistant maize has emerged as a way to expand the use of glyphosate for weed control. Studying the microbiome in the tissues and rhizosphere soil of transgenic plants is vital for understanding the glyphosate-resistant mechanism and optimizing the transgenic design of crops. In our study, the expression of a mutant cp4epsps gene in transgenic maize, which confers tolerance to glyphosate, was performed using the maize variety Xianyu 335 as the genetically modified acceptor line. This transgenic modification did not affect the initial bacterial community in the leaf, stem, or root of maize, but promoted a differential bacterial community in the rhizosphere soil. Under glyphosate application, the abundance of beneficial bacteria involved in N fixation and P solubilization in plant tissues and the rhizosphere soil of glyphosate-resistance maize were higher than those in the glyphosate-sensitive maize. In contrast, the abundance of pathogens had the opposite trend, suggesting that the enhanced health of transgenic maize prevented microbiome deterioration under glyphosate. The re-inoculation of bacterial strains isolated from glyphosate-resistance maize into the leaf and rhizosphere soil of glyphosate-sensitive maize resulted in an enhanced photosynthetic capacity in response to glyphosate, demonstrating the vital role of specific bacteria for glyphosate resistance. Our study provides important evidence of how transgenic maize tolerance to herbicides affects the bacterial communities across the maize niches under glyphosate toxicity.

Small DNA elements that act as both insulators and silencers in plants.

Insulators are cis -regulatory elements that separate transcriptional units, whereas silencers are elements that repress transcription regardless of their position. In plants, these elements remain largely uncharacterized. Here, we use the massively parallel reporter assay Plant STARR-seq with short fragments of eight large insulators to identify more than 100 fragments that block enhancer activity. The short fragments can be combined to generate more powerful insulators that abolish the capacity of the strong viral 35S enhancer to activate the 35S minimal promoter. Unexpectedly, when tested upstream of weak enhancers, these fragments act as silencers and repress transcription. Thus, these elements are capable of both insulating or repressing transcription dependent upon regulatory context. We validate our findings in stable transgenic Arabidopsis , maize, and rice plants. The short elements identified here should be useful building blocks for plant biotechnology efforts.

A simple and efficient method for betalain quantification in RUBY-expressing plant samples.

The RUBY reporter system has demonstrated great potential as a visible marker to monitor gene expression in both transiently and stably transformed plant tissues. Ectopic expression of the RUBY reporter leads to bright red pigmentation in plant tissues that do not naturally accumulate betalain. Unlike traditional visual markers such as β-glucuronidase (GUS), luciferase (LUC), and various fluorescent proteins, the RUBY reporter system does not require sample sacrifice or special equipment for visualizing the gene expression. However, a robust quantitative analysis method for betalain content has been lacking, limiting accurate comparative analyses. In this work, we present a simple and rapid protocol for quantitative evaluation of RUBY expression in transgenic plant tissues. Using this method, we demonstrate that differential RUBY expression can be quantified in transiently transformed leaf tissues, such as agroinfiltrated Nicotiana benthamiana leaves, and in stable transgenic maize tissues, including seeds, leaves, and roots. We found that grinding fresh tissues with a hand grinder and plastic pestle, without the use of liquid nitrogen, is an effective method for rapid betalain extraction. Betalain contents estimated by spectrophotometric and High-Performance Liquid Chromatography (HPLC) analyses were highly consistent, validating that our rapid betalain extraction and quantification method is suitable for comparative analysis. In addition, betalain content was strongly correlated with RUBY expression level in agroinfiltrated N. benthamiana leaves, suggesting that our method can be useful for monitoring transient transformation efficiency in plants. Using our rapid protocol, we quantified varying levels of betalain pigment in N. benthamiana leaves, ranging from 110 to 1066 mg/kg of tissue, and in maize samples, ranging from 15.3 to 1028.7 mg/kg of tissue. This method is expected to streamline comparative studies in plants, providing valuable insights into the effectiveness of various promoters, enhancers, or other regulatory elements used in transgenic constructs.

Investigation of the toxicity and safety concerns of transgenic maize seeds expressing immunogenic F and HN protein genes against Newcastle disease virus.

The presented study investigated the potential toxicity and safety concerns associated with transgenic maize seeds expressing immunogenic F and HN protein genes against Newcastle disease virus (NDV). The experiment involved feeding Sprague-Dawley rats with transgenic maize seeds formulated into standard diets at levels of 30% (w/w) for a duration of 90days. The rats were divided into three groups, with 10 rats per group. We assessed various parameters including overall appearance, feed consumption, body weight, organ weight, hematological parameters, serum chemistry, and histopathology. The results of these assessments were compared between the control group and the treatment groups. The study findings revealed that there were no significant differences between the control and treatment groups in terms of overall appearance, feed consumption, body weight, organ weight, hematological parameters, serum chemistry, microscopic histopathology, and gross appearance of tissues. These observations suggest that the consumption of transgenic maize seeds did not lead to any treatment-related adverse effects or toxicological issues. Furthermore, the transgenic maize seeds were found to be nutritionally equivalent to their non-transgenic counterpart. Overall, no physiological, pathological, or molecular toxicity was observed in the Rats fed with transgenic feed.However, it is important to note that this study focused specifically on the parameters measured and the outcomes observed in Sprague-Dawley rats, and further research and studies are necessary to fully evaluate the safety and potential applications of transgenic edible vaccines in humans or other animals.

Impact of Transgenic Maize Ruifeng125 on Diversity and Dynamics of Bacterial Community in Rhizosphere Soil.

With the development of commercialized planting of genetically modified crops, their ecological security risks remain a key topic of public concern. Insect-resistant genetically modified maize, Ruifeng125, which expresses a fusion Bt protein (Cry1Ab-Cry2Aj), has obtained the application safety certificate issued by the Chinese government. To determine the effects of Ruifeng125 on the diversity and dynamics of bacterial communities, the accumulation and degradation pattern of the fusion Bt protein in the rhizosphere soil of transgenic maize were detected. Results showed that the contents of Bt protein varied significantly at different developmental stages, but after straw was returned to the field, over 97% of Bt proteins were degraded quickly at the early stages (≤10 d) and then they were degraded at a relatively slow rate. In addition, the variations in bacterial community diversity in the rhizosphere soil were detected by 16S ribosomal RNA (Rrna) high-throughput sequencing technology. A total of 44 phyla, 435 families, and 842 genera were obtained by 16S rRNA sequencing, among which Proteobacteria, Actinobacia, Acidobacter Acidobacterium, and Chloroflexi were the dominant taxa. At the same developmental stage, no significant differences in soil bacterial diversity were detected between Ruifeng125 and its non-transgenic control variety. Further analysis revealed that developmental stage, rather than the transgenic event, made the greatest contribution to the changes in soil microbial diversity. This research provides important information for evaluating the impacts of Bt crops on the soil microbiome and establishes a theoretical foundation for their environmental safety assessment.

Resistance of Transgenic Maize Cultivars to Mycotoxin Production-Systematic Review and Meta-Analysis.

Approximately 25% of cereal grains present with contamination caused by fungi and the presence of mycotoxins that may cause severe adverse effects when consumed. Maize has been genetically engineered to present different traits, such as fungal or insect resistance and herbicide tolerance. This systematic review compared the observable quantities, via meta-analysis, of four mycotoxins (aflatoxins-AFL, fumonisins-FUM, deoxynivalenol-DON, zearalenone-ZEA) between genetically modified (GM) and conventional maize kernels. This study was conducted following the PRISMA guidelines, with searches performed using PubMed, Web of Science, Scopus, Google Scholar, and CAPES journals databases. Analyses were conducted using RevMan v.5.4 software. Transgenic maize showed a 58% reduction in total mycotoxins (p < 0.001) compared to conventional maize. FUM were the most impacted, with a 59% reduction (p < 0.001) in GM maize. AFL and ZEA levels were also lower in GM maize by 49% (p = 0.02) and 51% (p < 0.001), respectively. On the other hand, DON levels increased by 6% (p < 0.001) in GM maize compared to conventional maize. However, results for ZEA and DON were inconclusive due to the limited research and sample sizes. We conclude that transgenic maize reduces total mycotoxins by over 50%, primarily fumonisin and aflatoxin. Most studies presented maize varieties that were resistant to insects or herbicides, not fungal pathogens, showing a positive collateral effect of these genetic alterations. Therefore, transgenic maize appears to be a safer product for animal and human consumption from a toxicological point of view. Further studies with larger sample sizes are needed to confirm our findings for ZEA and DON in transgenic maize.

ZmDST44 Gene Is a Positive Regulator in Plant Drought Stress Tolerance.

Improving drought tolerance in plants is essential for increasing crop yields under water-limited conditions. In this study, we investigated the functional role of the maize gene ZmDST44, which is targeted by the miRNA ZmmiR139. Our results indicate that ZmmiR139 regulates ZmDST44 by cleaving its mRNA, as confirmed by inverse expression patterns and 5'-RACE analysis. Overexpression of ZmDST44 in Arabidopsis, rice, and maize resulted in significant enhancements in drought tolerance. Transgenic plants exhibited reduced malondialdehyde (MDA) levels, increased proline accumulation, and upregulation of drought-responsive genes compared to wild-type plants. Transgenic Arabidopsis and rice showed improved drought resistance and higher post-drought recovery rates, and transgenic maize displayed lower sensitivity to drought stress. These findings suggest that ZmDST44 acts as a positive regulator of drought tolerance across different plant species and holds promise for developing drought-resistant crops through genetic engineering.

Control Efficacy of the Bt Maize Event DBN3601T Expressing Cry1Ab and Vip3Aa Proteins against Beet Armyworm, Spodoptera exigua (Hübner), in China.

The beet armyworm, Spodoptera exigua (Hübner), is a major pest of maize, cotton, soybean, and many other crops globally. Despite the widespread deployment of Bt transgenic maize for pest control worldwide, the efficacy of Bt lepidopteran-resistant transgenic maize in managing S. exigua remains rarely studied. In this study, we quantified the expression level of pyramided Cry1Ab and Vip3Aa toxins in Bt maize (event DBN3601T) and evaluated their control efficiency against S. exigua under both laboratory and field conditions. The enzyme-linked immunosorbent assay (ELISA) results showed that the expression levels of Cry1Ab and Vip3Aa proteins in DBN3601T maize tissues followed a decreasing order as follows: V5-leaf > V8-leaf > VT-tassel > R2-kernel > R1-silk. Diet-overlay assay results showed that the LC50 values of Cry1Ab and Vip3Aa proteins against S. exigua larvae were 11.66 ng/cm2 and 27.74 ng/cm2, respectively, with corresponding GIC50 values at 1.59 ng/cm2 and 7.93 ng/cm2. Bioassay using various tissues of the DBN3601T maize indicated that after 7 days of infestation, mortality rates of neonates and third-instar larvae ranged from 86% to 100% and 58% to 100%, respectively. Mortality was highest on V5 and V8 leaves, followed by R2-kernel, VT-tassel, and R1-silk. Field trials demonstrated that DBN3601T maize exhibited significantly lower larval density, damage rate, and leaf damage score compared to non-Bt maize. Field cage trial showed that the control efficacy of DBN3601T maize at the vegetative stage could reach 98%. These findings provide a theoretical basis for utilizing Bt transgenic maize to enhance the sustainable management of S. exigua in Asia.

Effects of combining soil-applied insecticide and Bt corn for integrated pest management and resistance management of western corn rootworm (Coleoptera: Chrysomelidae).

The western corn rootworm, (Diabrotica virgifera virgifera LeConte, Coleoptera: Chrysomelidae), is a serious pest of corn (Zea mays Linnaeus, Cyperales: Poaceae) in the midwestern United States. Management practices for corn rootworm larvae include crop rotation, transgenic corn producing insecticidal toxins from the bacterium Bacillus thuringiensis Berliner (Bacillales: Bacillaceae) (Bt), and soil-applied insecticides. The extent to which combining soil-applied insecticide with Bt corn would be beneficial from the perspective of insect resistance management (IRM) or integrated pest management (IPM) remains uncertain. We conducted a 3-yr field study to characterize the implications of combining a soil-applied insecticide and Bt corn for IRM and IPM of western corn rootworm. Experimental treatments were Bt corn, a soil-applied insecticide, the combination of these factors, and an experimental control in which both factors were absent. Data were collected on root injury to corn by rootworm, survival to adulthood, adult size, and emergence time for western corn rootworm. We found that mortality caused by the soil-applied insecticide was insufficient to delay resistance to Bt corn. While combining Bt corn and a soil-applied insecticide may provide a short-term economic benefit, additional research is needed to determine appropriate economic thresholds for combining these tactics. Additionally, combining a soil-applied insecticide and Bt corn would not be sustainable over multiple growing seasons because of its potential to rapidly select for Bt resistance. In general, a more sustainable IRM strategy for rootworm management would include using crop rotation and alternating between non-Bt corn with soil-applied insecticide and Bt corn without soil-applied insecticide.

A novel member of miR169 family negatively regulates maize resistance against Bipolaris maydis.

MicroRNAs (miRNAs) have been confirmed to play important roles in plant defense response. However, the key maize miRNAs involved in the defense response against Bipolaris maydis are very limited. In this study, a novel member of the miR169 family in response to B. maydis, named zma-miR169s, was discovered and investigated. The expression levels of pre-miR169s and zma-miR169s were significantly repressed during B. maydis infection. CRISPR/Cas9-induced zma-miR169s mutant exhibited more resistance against B. maydis, whereas overexpression zma-miR169s enhanced susceptibility, supporting that zma-miR169s might play a negative role in maize resistance. Moreover, RNA-seq and GO analysis showed that differentially expressed genes were highly enriched in the oxidation-reduction process and plant hormone pathway. Hence, reactive oxygen species (ROS) and plant hormone levels were further investigated. ROS detection confirmed that zma-miR169s mutant accumulated more ROS, while less ROS was detected in transgenic maize OE-miR169s. Furthermore, more remarkable changes in PR-1 expression levels and salicylic acid (SA) contents were detected in zma-miR169s mutant compared to wild-type and transgenic maize during B. maydis infection. Additionally, nuclear transcription factors (NF-YA1 and NF-YA13) were identified as targets regulated by zma-miR169s through the agrobacterium-mediated transient expression method. Overexpression of ZmNF-YA13 enhanced Arabidopsis resistance to Pseudomonas syringae pv. tomato DC3000. Taken together, our results suggest that zma-miR169s negatively regulate maize defense responses by influencing ROS accumulation and the SA-dependent signaling pathway.

CRISPR/Cas9-mediated genome editing technique to control fall armyworm (Spodoptera frugiperda) in crop plants with special reference to maize.

Fall Armyworm imposes a major risk to agricultural losses. Insecticides have historically been used to manage its infestations, but it eventually becomes resistant to them. To combat the pest, a more recent strategy based on the use of transgenic maize that expresses Bt proteins such as Cry1F from the bacteria has been used. Nonetheless, there have been numerous reports of Cry1F maize resistance in FAW populations. Nowadays, the more effective and less time-consuming genome editing method known as CRISPR/Cas9 technology has gradually supplanted these various breeding techniques. This method successfully edits the genomes of various insects, including Spodoptera frugiperda. On the other hand, this new technique can change an insect's DNA to overcome its tolerance to specific insecticides or to generate a gene drive. The production of plant cultivars resistant to fall armyworms holds great potential for the sustainable management of this pest, given the swift advancement of CRISPR/Cas9 technology and its varied uses. Thus, this review article discussed and critically assessed the use of CRISPR/Cas9 genome-editing technology in long-term fall armyworm pest management. However, this review study focuses primarily on the mechanism of the CRISPR-Cas9 system in both crop plants and insects for FAW management.

Impact of glyphosate on the rhizosphere microbial communities of a double-transgenic maize line D105

Plant roots shape the rhizosphere microbiome, recruiting microbes with beneficial functions. While genetically engineered crops offer yield advantages, their impacts on rhizosphere microbial communities remain understudied. This study evaluated the effects of transgenic maize, alongside a non-transgenic counterpart, on rhizosphere bacterial and fungal community composition using 16S rRNA and ITS amplicon sequencing. Additionally, glyphosate was used to evaluate its impact on microbial assembly and the magnitude of its effect at various maize growth stages. The results showed that transgenic maize D105 line significantly increased bacterial alpha diversity but not fungal diversity. Beta diversity analysis showed clear separation between bacterial and fungal communities at higher glyphosate treatment. Specific bacterial taxa such as Pseudomonas and Sphingomonas were enriched, while fungal taxa such as Ascomycota, Lasiosphaeriaceae, Verticillium were differentially abundant in glyphosate treatments. LEfSe analysis identified distinct enrichment patterns of bacterial (Proteobacteria and Actinobacteria) and fungal taxa (Verticillium and Guehomyces) associated with the transgenic line and glyphosate levels. KEGG functional analysis suggested potential impacts on bacterial metabolic pathways and shifts in fungal trophic modes (saprotrophs, pathogens) within the rhizosphere microbiome. This research provides insights into the classification, functional relationships, and underlying mechanisms shaping microbial communities carrying insect resistance and glyphosate resistance traits.

Heterologous synthesis of poly-γ-glutamic acid enhanced drought resistance in maize (Zea mays L.)

Drought stress is the main factor restricting maize yield. Poly-γ-glutamic acid (γ-PGA), as a water-retaining agent and fertilizer synergist, could significantly improve the drought resistance and yield of many crops. However, its high production costs and unclear long-term impact on soil ecology limit its large-scale application. In this study, an environmentally friendly green material γ-PGA was heterologous synthesized in maize for the first time using the synthetic biology method. The genes (PgsA, PgsB, PgsC) participated in γ-PGA synthesis were cloned from Bacillus licheniformis and transformed into maize to produce γ-PGA for the first time. Under drought stress, transgenic maize significantly increased the ear length, ear weight and grain weight by 50 % compared to the control, whereas the yield characteristic of ear weight, grain number per ear, grain weight per ear and 100-grain weight increased by 1.67 %–2.33 %, 3.78 %–13.06 %, 8.41 %–22.06 %, 6.03 %–19.28 %, and 11.85 %–18.36 %, respectively under normal growth conditions. γ-PGA was mainly expressed in the mesophyll cells of maize leaf rosette structure and improved drought resistance and yield by protecting and increasing the expression of genes for the photosynthetic and carbon fixation. This study is an important exploration for maize drought stress molecular breeding and building resource-saving agriculture.

Discrimination of maize transgenic and non-transgenic varieties by laser induced spectroscopy (LIBS) and machine learning algorithms

BackgroundIn the last decades, the production and consumption of genetically modified agricultural products have increased markedly due to the worldwide population growth and rising demand for food and feed. Consequently, genetically modified crops have been extensively produced and consumed, which required identifying and discriminating transgenic and non-transgenic products. ResultsLaser-induced breakdown spectroscopy (LIBS) combined with chemometric methods was applied to identify and discriminate two varieties of conventional (not-genetically modified, NGM) maize from four varieties of transgenic maize (genetically modified, GM). The LIBS spectra acquired under reduced pressure (100 Torr) conditions over two ranges, i.e., 175–330 nm and 275–770 nm, were subjected to Standard Normal Variation (SNV) and multivariate methods such as Principal Component Analysis (PCA) to reduce data matrices dimensionality and spectral noise. The supervised machine learning algorithms k-nearest neighbor (k-NN) and support vector machine (SVM) have been applied to discriminate among NGM and GM maize reserving 25 % of data for external validation. The training data were employed for hyperparameter optimization of classifiers using the Leave-One-Out Cross-Validation (LOOCV) method. Considering all six maize varieties simultaneously, the highest training accuracy achieved was 90.56 %, with an external validation accuracy of 88.33 %. In an alternative approach based on pairwise combinations of one GM variety against one NGM variety, the best outcome achieved was 100 % LOOCV and external validation accuracy. ConclusionsThese results showed that LIBS supported by appropriate chemometric methods represents an alternative screening technique for identifying and discriminating transgenic from non-transgenic maize.

Ca2+-independent ZmCPK2 is inhibited by Ca2+-dependent ZmCPK17 during drought response in maize.

Calcium oscillations are induced by different stresses. Calcium-dependent protein kinases (CDPKs/CPKs) are one major group of the plant calcium decoders that are involved in various processes including drought response. Some CPKs are calcium-independent. Here, we identified ZmCPK2 as a negative regulator of drought resistance by screening an overexpression transgenic maize pool. We found that ZmCPK2 does not bind calcium, and its activity is mainly inhibited during short term abscisic acid (ABA) treatment, and dynamically changed in prolonged treatment. Interestingly, ZmCPK2 interacts with and is inhibited by calcium-dependent ZmCPK17, a positive regulator of drought resistance, which is activated by ABA. ZmCPK17 could prevent the nuclear localization of ZmCPK2 through phosphorylation of ZmCPK2T60. ZmCPK2 interacts with and phosphorylates and activates ZmYAB15, a negative transcriptional factor for drought resistance. Our results suggest that drought stress-induced Ca2+ can be decoded directly by ZmCPK17 that inhibits ZmCPK2, thereby promoting plant adaptation to water deficit.

Modernizing and harmonizing regulatory data requirements for genetically modified crops-perspectives from a workshop.

Genetically modified (GM) crops that have been engineered to express transgenes have been in commercial use since 1995 and are annually grown on 200 million hectares globally. These crops have provided documented benefits to food security, rural economies, and the environment, with no substantiated case of food, feed, or environmental harm attributable to cultivation or consumption. Despite this extensive history of advantages and safety, the level of regulatory scrutiny has continually increased, placing undue burdens on regulators, developers, and society, while reinforcing consumer distrust of the technology. CropLife International held a workshop at the 16th International Society of Biosafety Research (ISBR) Symposium to examine the scientific basis for modernizing global regulatory frameworks for GM crops. Participants represented a spectrum of global stakeholders, including academic researchers, GM crop developers, regulatory consultants, and regulators. Concurrently examining the considerations of food and feed safety, along with environmental safety, for GM crops, the workshop presented recommendations for a core set of data that should always be considered, and supplementary (i.e., conditional) data that would be warranted only on a case-by-case basis to address specific plausible hypotheses of harm. Then, using a case-study involving a hypothetical GM maize event expressing two familiar traits (insect protection and herbicide tolerance), participants were asked to consider these recommendations and discuss if any additional data might be warranted to support a science-based risk assessment or for regulatory decision-making. The discussions during the workshop highlighted that the set of data to address the food, feed, and environmental safety of the hypothetical GM maize, in relation to a conventional comparator, could be modernized compared to current global regulatory requirements. If these scientific approaches to modernize data packages for GM crop regulation were adopted globally, GM crops could be commercialized in a more timely manner, thereby enabling development of more diverse GM traits to benefit growers, consumers, and the environment.