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Propylea japonica (Thunberg) (Coleoptera: Coccinellidae) is a dominant natural enemy of insect pests in farmland ecosystems. It also serves as an important non-target insect for environmental safety evaluations of transgenic crops. Widespread planting of transgenic crops may result in direct or indirect exposure of P. japonica to recombinant Bacillus thuringiensis (Bt) protein, which may in turn affect the biological performance of this natural enemy by affecting the P. japonica microflora. However, the effects of Bt proteins (such as Cry1B) on the P. japonica microbiota are currently unclear. Here, we used a high-throughput sequencing method to investigate differences in the P. japonica microbiota resulting from treatment with Cry1B compared to a sucrose control. The results demonstrated that the P. japonica microbiome was dominated by Firmicutes at the phylum level and by Staphylococcus at the genus level. Within-sample (α) diversity indices demonstrated a high degree of consistency between the microbial communities of P. japonica treated with the sucrose control and those treated with 0.25 or 0.5 mg/mL Cry1B. Furthermore, there were no significant differences in the abundance of any taxa after treatment with 0.25 mg/mL Cry1B for 24 or 48 h, and treatment with 0.5 mg/mL Cry1B for 24 or 48 h led to changes only in Staphylococcus, a member of the phylum Firmicutes. Treatment with a high Cry1B concentration (1.0 mg/mL) for 24 or 48 h caused significant changes in the abundance of specific taxa (e.g., Gemmatimonades, Patescibacteria, Thauera, and Microbacterium). However, compared with the control, most taxa remained unchanged. The statistically significant differences may have been due to the stimulatory effects of treatment with a high concentration of Cry1B. Overall, the results showed that Cry1B protein could alter endophytic bacterial community abundance, but not composition, in P. japonica. The effects of Bt proteins on endophytes and other parameters in non-target insects require further study. This study provides data support for the safety evaluation of transgenic plants. [ABSTRACT FROM AUTHOR]

Fall armyworm (FAW), Spodoptera frugiperda, invaded the south of China in December 2018 and has since posed a huge threat to crop production in China. However, transgenic Bacillus thuringiensis (Bt) corn can efficiently control the damage caused by FAWs. In fact, the Chinese government has issued biosafety certificates for several Bt corn hybrids expressing any one of four Bt proteins, Vip3A, Cry1F, Cry1Ab, and Cry2Ab, or combinations thereof, to control FAWs. These Bt corn events are soon to be commercialized in China. Therefore, it is necessary to monitor and evaluate whether the FAW has developed resistance to any of the Bt corn hybrids planted in fields in China. To address this issue, we collected 11 geographical populations of FAWs and determined the sensitivity of each to the aforementioned four purified Bt proteins as assessed by diet surface overlay bioassays. The ranges for the 50% lethal concentration (LC50) of the four Bt proteins to all FAW populations were as follows: 11.42–88.33 ng/cm2 (for Vip3A), 111.21–517.33 (Cry1F), 135.76–1108.47 (Cry1Ab), and 994.42–5492.50 (Cry2Ab). The corresponding ranges for the 50% growth inhibition concentrations (GIC50) were 1.43–14.86, 2.35–138.97, 1.58–464.86, and 25.01–1266.07 ng/cm2. The lethal effects and growth inhibition effects of the four Bt proteins on FAW were in the same order of Vip3A > Cry1F > Cry1Ab > Cry2Ab. A comparison with published LC50 values of Bt proteins towards sensitive FAW populations revealed that all 11 FAW populations in this study were sensitive to Vip3A, Cry1F, and Cry1Ab. This study provides foundational data for monitoring and controlling the resistance of Bt corn to FAW in China. [ABSTRACT FROM AUTHOR]

[Objectives] To determine the biological safety of BT protein from Bacillus thuringiensis (Bt) fermentation broth to mammals at high doses. [Methods] Healthy mice were randomly divided into 4 groups with 10 mice in each group. The experimental groups were fed with Bt fermentation supernatant at 0.2, 0.6 and 1.0 mL/kg, respectively, once a day for 7 consecutive days. The blank control group was fed normally without intragastric administration. [Results] There was no significant difference in blood routine and blood biochemical analysis between the experimental group and the control group. After intragastric administration, the mice were dissected, and no obvious pathological changes were found; the heart, liver, spleen, lung and kidney were taken to make tissue sections, and no pathological changes were found by microscopic observation. [Conclusions] Mice can tolerate high doses of BT protein from B. thuringiensis fermentation broth. [ABSTRACT FROM AUTHOR]

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• Bt protein levels in transgenic rice vary significantly across different growth stages and organs. • Bt protein levels higher in cry2A* than cry1C* transgenic early japonica rice. • Bt protein levels are highest in leaves, followed by stem sheaths and young panicles. • Expression levels of Bt proteins in brown rice are the lowest among all growth stages and organs. • Correlation between Bt gene transcription and protein levels is significant in various rice tissues. Transgenic insect-resistant rice offers an environmentally friendly approach to mitigate yield losses caused by lepidopteran pests, such as stem borers. Bt (Bacillus thuringiensis) genes encode insecticidal proteins and are widely used to confer insect resistance to genetically modified crops. This study investigated the integration, inheritance, and expression characteristics of codon-optimised synthetic Bt genes, cry1C* and cry2A*, in transgenic early japonica rice lines. The early japonica rice cultivar, Songgeng 9 (Oryza sativa), was transformed with cry1C* or cry2A*, which are driven by the ubi promoter via Agrobacterium tumefaciens-mediated transformation. Molecular analyses, including quantitative PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and Southern blot analysis were performed to confirm transgene integration, inheritance, transcriptional levels, and protein expression patterns across different tissues and developmental stages. Stable transgenic early japonica lines exhibiting single-copy transgene integration were established. Transcriptional analysis revealed variations in Bt gene expression among lines, tissues, and growth stages, with higher expression levels observed in leaves than in other organs. Notably, cry2A* exhibited consistently higher mRNA and protein levels than cry1C* across all examined tissues and developmental time points. Bt protein accumulation followed the trend of leaves > stem sheaths > young panicles > brown rice, with peak expression during the filling stage in the vegetative tissues. Synthetic cry2A* displayed markedly elevated transcription and translation compared to cry1C* in the transgenic early japonica rice lines examined. Distinct spatiotemporal patterns of Bt gene expression were elucidated, providing insights into the potential insect resistance conferred by these genes in rice. These findings will contribute to the development of insect-resistant japonica rice varieties and facilitate the rational deployment of Bt crops. [ABSTRACT FROM AUTHOR]

Fall armyworm (FAW), Spodoptera frugiperda, invaded the south of China in December 2018 and has since posed a huge threat to crop production in China. However, transgenic Bacillus thuringiensis (Bt) corn can efficiently control the damage caused by FAWs. In fact, the Chinese government has issued biosafety certificates for several Bt corn hybrids expressing any one of four Bt proteins, Vip3A, Cry1F, Cry1Ab, and Cry2Ab, or combinations thereof, to control FAWs. These Bt corn events are soon to be commercialized in China. Therefore, it is necessary to monitor and evaluate whether the FAW has developed resistance to any of the Bt corn hybrids planted in fields in China. To address this issue, we collected 11 geographical populations of FAWs and determined the sensitivity of each to the aforementioned four purified Bt proteins as assessed by diet surface overlay bioassays. The ranges for the 50% lethal concentration (LC50) of the four Bt proteins to all FAW populations were as follows: 11.42–88.33 ng/cm2 (for Vip3A), 111.21–517.33 (Cry1F), 135.76–1108.47 (Cry1Ab), and 994.42–5492.50 (Cry2Ab). The corresponding ranges for the 50% growth inhibition concentrations (GIC50) were 1.43–14.86, 2.35–138.97, 1.58–464.86, and 25.01–1266.07 ng/cm2. The lethal effects and growth inhibition effects of the four Bt proteins on FAW were in the same order of Vip3A > Cry1F > Cry1Ab > Cry2Ab. A comparison with published LC50 values of Bt proteins towards sensitive FAW populations revealed that all 11 FAW populations in this study were sensitive to Vip3A, Cry1F, and Cry1Ab. This study provides foundational data for monitoring and controlling the resistance of Bt corn to FAW in China.

The lady beetle Propylea japonica is a dominant natural predator of insect pests in farmland ecosystems and an important non-target indicator insect for the environmental safety assessment of GM crops. The commercial cultivation of GM crops may cause P. japonica to frequently be exposed to the Bt protein environment. In this study, the biological characteristics, enzyme activity, and expression levels of detoxification and metabolism in P. japonica were studied after Cry1B protein treatment. No significant differences were observed in developmental duration, emergence rate, or body weight at different ages after feeding larvae 0.5 mg/mL of Cry1B protein compared with the control. Furthermore, there were no significant differences in the activities of glutathione S-transferase (GST), catalase (CAT), and peroxidase (POD) after feeding 0.25 mg/mL and 0.5 mg/mL Cry1B protein. However, when the concentration of Cry1B protein increased to 1.0 mg/mL, the activities of the GST, CAT, and POD increased significantly. Compared with the control group, there were no significant differences in the expression levels of most detoxification metabolism related genes; only a few genes had changed expression levels at the individual concentrations (CYP345B1, CYP4Q2, CYP9F2, GST, and microsomal GST). Overall, these results suggest that Cry1B protein has little or no effect on the biological characteristics of P. japonica. Genes related to enzyme activity and detoxification are differentially expressed at high concentration stimulation. Therefore, this research suggests that the potential risks of Cry1B for the predator P. japonica are negligible.

Bt (Bacillus thuringiensis) corn is one of the top three large-scale commercialized anti-insect transgenic crops around the world. In the present study, we tested the Bt protein content, defense chemicals contents, and defense enzyme activities in both the leaves and roots of Bt corn varieties 5422Bt1 and 5422CBCL, as well as their conventional corn 5422 seedlings, with two fully expanded leaves which had been treated with 2.5 mM exogenous salicylic acid (SA) to the aboveground part for 24 h. The result showed that the SA treatment to the aboveground part could significantly increase the polyphenol oxidase activity of conventional corn 5422, the Bt protein content, and peroxidase activities of Bt corn 5422Bt1, as well as the polyphenol oxidase and peroxidase activity of Bt corn 5422CBCL in the leaves. In the roots, the polyphenol oxidase and peroxidase activity of conventional corn 5422, the polyphenol oxidase and superoxide dismutase activities of Bt corn 5422Bt1, the DIMBOA (2,4-dihydroxy-7-methoxy-2H, 1, 4-benzoxazin-3 (4H)-one) content, and four defense enzymes activities of Bt corn 5422CBCL were systematically increased. These findings suggest that the direct effect of SA application to aboveground part on the leaf defense responses in Bt corn 5422CBCL is stronger than that in non-Bt corn. Meanwhile, the systemic effect of SA on the root defense responses in Bt corn 5422CBCL is stronger than that in conventional corn 5422 and Bt corn 5422Bt1. It can be concluded that the Bt gene introduction and endogenous chemical defense responses of corns act synergistically during the SA-induced defense processes to the aboveground part. Different transformation events affected the root defense response when the SA treatment was applied to the aboveground part. [ABSTRACT FROM AUTHOR]

This study reported the functionality integration of zeolitic imidazolate framework-8 (ZIF-8) with horseradish peroxidase (HRP) and streptavidin (SA) for the synthesis of a HRP&SA/ZIF-8 nanocomposite through one-pot coprecipitation. The synthesized HRP&SA/ZIF-8 nanocomposite was then employed as the ideal signal tag for application in the enzyme-linked immunosorbent assay (ELISA) and exhibited excellent sensitivity, selectivity and accuracy in the detection of insecticidal crystalline (Cry) protein Cry1Ab as a transgenic biomarker with a detection limit of 4.8 pg/mL. This proposed method provides a new way for the detection of transgenic biomarkers in food and may inspire further integration of a variety of biomolecules into ZIF-8 for applications ranging from biosensing, biomedicine, and catalysis to energy. [ABSTRACT FROM AUTHOR]

Transgenic rice with Bacillus thuringiensis (Bt) genes has been successfully cultivated in recent years. The stable and sustainable expression of Bt protein is the guarantee of effective resistance within these strains against target pests. It is well known that the synthesis of Bt protein in Bt crops needs nitrogen (N). Therefore, how is the efficacy of transgenic Bt rice against target pests in response to N application? In our study, we found that enhanced nitrogen inputs increased soluble protein content (SPC) and glutamine synthetase (GS), but decreased amino acid content (AAC); it is evident that the protein synthesis was enhanced, and degradation was reduced with increasing nitrogen application. Thus, both increased protein synthesis and decreased protein degradation contributed to the increased Bt protein content (BTC) in transgenic Bt rice. Responses of the three Bt rice lines to N proportions and N sources were different. All these differences can be explained by the changes of nitrogen metabolism of different genotypes. The BTC in leaves was higher than that in other organs, and the fully expanded green leaf had higher BTC than the senescent leaf and tender leaf. Despite the difference of Bt protein expression, all transgenic rice lines showed 100% resistance to target pests under different N conditions. Our results indicated that the response of Bt protein expression in transgenic Bt rice with different genotypes to N fertilizer was inconsistent; moreover, there exist obvious spatial-temporal differences. Therefore, in order to guarantee the stability and sustainability resistance of Bt rice, it is necessary to adopt appropriate N management practice according to the expression pattern of Bt protein of transgenic Bt rice with different genotypes. [ABSTRACT FROM AUTHOR]

An experiment was executed at field condition to evaluate ecological impact of Bt cotton (Mech 162 + Bt) on soil microbes and Bt protein behavior in soil. Root exudates of Bt cotton plant containing Bt protein enter into soil which persisted in soil for atleast 235 days after sowing (DAS). During vegetative and flowering stage, Bt protein was not detectable in both qualitative and quantitative assay but, during onset of boll formation at 150 DAS, a level of 0.0013 μg/g of Cry 1Ac protein was quantified in rhizospheric soil which further increased exponentially up to 0.0033 μg/g at 195 DAS and then maintained at the same level till 235 DAS with Cry1Ac expression level of 0.0029 μg/g soil. Rhizospheric soil of Bt cotton had significantly higher (p < 0.05) abundance of Fatty Acid Methyl Esters (FAME) of gram positive bacteria. Abundance of polyunsaturated FAMEs that are indicators of fungi, were slightly higher in Bt as compared to non-Bt cotton soils. The metabolic profiling, also called community level physiological profiling (CLPP) showed more microbial functional diversity (Shannon–Wiener Diversity) in soil samples of different growth stages of Bt cotton than its corresponding non-Bt variety. Our results showed that Cry 1AC Bt protein persists in clay loamy soil even after harvesting and significantly altered microbial community composition and functional diversity in rhizospheric soil of Bt cotton than non-Bt. Any possible and unintentional changes in composition of root exudes of Bt cotton could regulate the selection of different microbial communities and thus can alter functional responses. [ABSTRACT FROM AUTHOR]

Bacillus thuringiensis Cry and Vip proteins are highly effective at controlling agricultural pests and could be used in pyramided transgenic crops. However, the molecular mechanism underlying the Cry1Ah and Vip3Aa19 synergistic interaction has never been investigated at the molecular level in Yellow peach moth (YPM) Conogethes punctiferalis. Binding affinity and synergism of Cry1Ah and Vip3Aa19 proteins with ABC transporter subfamily C receptors ABCC1, ABCC2 and ABCC3 proteins from the midgut of YPM larva by using surface plasmon resonance (SPR) and pull-down assays. Both assays revealed that Cry1Ah could interact with ABCC1, ABCC2, and ABCC3, whereas Vip3Aa19 only interacts with ABCC1 and ABCC3, but not with ABCC2. Hence, when compared to the Vip3Aa19 protein, Cry1Ah had a higher binding affinity for ABCC1, ABCC2, and ABCC3. Furthermore, competitive binding assay between Cry1Ah and Vip3Aa19 protein with ABC transporter subfamily C receptors resulted in the final eluted protein samples displaying vibrant blue bands of Cry1Ah and very faint bands of Vip3Aa19. Suggesting that Cry and Vip proteins could deliver a synergistic effect after cleaving the midgut proteases. Therefore, this finding indicated that the Cry1Ah and Vip3Aa19 do not compete for interacting with midgut receptors and thus provide strong synergism against YPM. SPR and pull-down results revealed that Cry1Ah could interact with ABCC1, ABCC2, and ABCC3, whereas Vip3Aa19 only interacts with ABCC1 and ABCC3, but not with ABCC2. The competitive binding assay indicated that Cry1Ah and Vip3Aa19 do not compete for interacting with midgut receptors and thus provide strong synergism against YPM. [Display omitted] • Cry1Ah had a higher binding affinity for ABCC1, ABCC2, and ABCC3 than Vip3Aa19. • Cry1Ah and Vip3Aa19 does not compete for interacting with midgut receptors of Conogethes punctiferalis. • Cry1Ah and Vip3Aa19 combination provides strong synergism against C. punctiferalis. [ABSTRACT FROM AUTHOR]

Midgut juice plays an important role in food digestion and detoxification in insects. In order to understand the potential of midgut juice of Cnaphalocrocis medinalis (Guenée) to degrade Bt proteins, the enzymatic activity of midgut juice and its degradation of Bt proteins (Cry2A, Cry1C, Cry1Aa, and Cry1Ac) were evaluated in this study through protease inhibitor treatments. The activities of total protease in midgut juices were significantly inhibited by phenylmethylsulfonyl fluoride (PMSF), tosyl-L-lysine chloromethyl ketone (TLCK), pepstatin A and leupeptin. The enzymatic activity of chymotrypsin was significantly inhibited by PMSF, and enzymatic activity of trypsin was significantly inhibited by ethylenediaminetetraacetic acid (EDTA), PMSF, tosyl phenylalanine chloromethyl ketone (TPCK), TLCK and trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64). EDTA could significantly inhibit the degradation of Cry2A by C. medinalis. EDTA, PMSF, TPCK, and TLCK could inhibit the degradation of Cry1C and Cry1Aa. EDTA, PMSF, TPCK, TLCK, and E-64 could inhibit the degradation of Cry1Ac. Our results indicated that some protease inhibitors hindered various enzymatic activities in the larval midgut of C. medinalis, which may reduce the insect's ability to degrade Bt toxins. These findings may aid the application of protease inhibitors in the management of this insect pest in the future.

Since 1998, genetically engineered Bt maize varieties expressing the insecticidal Cry1Ab protein (i.e. event MON 810) have been grown in the European Union (EU), mainly in Spain. These varieties confer resistance against the European and Mediterranean corn borer (ECB and MCB), which are the major lepidopteran maize pests in the EU, particularly in Mediterranean areas. However, widespread, repeated and exclusive use of Bt maize is anticipated to increase the risk of Cry1Ab resistance to evolve in corn borer populations. To delay resistance evolution, typically, refuges of non-Bt maize are planted near or adjacent to, or within Bt maize fields. Moreover, changes in Cry1Ab susceptibility in field populations of corn borers and unexpected damage to maize MON 810, due to corn borers, are monitored on an annual basis. After two decades of Bt maize cultivation in Spain, neither resistant corn borer populations nor farmer complaints on unexpected field damage have been reported. However, whether the resistance monitoring strategy followed in Spain, currently based on discriminating concentration bioassays, is sufficiently sensitive to timely detect early warning signs of resistance in the field remains a point of contention. Moreover, the Cry1Ab resistance allele frequency to Bt maize, which has recently been estimated in MCB populations from north-eastern Spain, might exceed that recommended for successful resistance management. To ensure Bt maize durability in Spain, it is key that adequate resistance management approaches, including monitoring of resistance and farmer compliance with refuge requirements, continue to be implemented and are incorporated in integrated pest management schemes. [ABSTRACT FROM AUTHOR]

To learn the spatial and temporal distribution of Bt proteins in Bt maize leaves, Bt protein levels of Bt maize leaves and their corresponding non-transgenic lines were detected and analysed by the ELISA method. The results showed that Bt proteins were expressed in the leaves of Bt maize varieties during all periods of their growth, while no such Bt proteins were detected in the leaves of non-transgenic maize varieties. The range of leaf Bt proteins was 1.08∼5.81 µg.g−1 FW, which could effectively protect Bt maizes from the corn borer pest during their growth periods. Furthermore, the younger leaves of Bt maizes had significantly lower levels of Bt proteins than their older leaves. In most periods, Bt protein content of Bt maize leaves under drought stress was lower than that under normal growth conditions, but Bt protein content under nutrition stress was higher than that under normal growth conditions. The results provided a full pattern of the spatial and temporal distribution of Bt proteins in Bt maize leaves.

To learn the spatial and temporal distribution of Bt proteins in Bt maize leaves, Bt protein levels of Bt maize leaves and their corresponding non-transgenic lines were detected and analysed by the ELISA method. The results showed that Bt proteins were expressed in the leaves of Bt maize varieties during all periods of their growth, while no such Bt proteins were detected in the leaves of non-transgenic maize varieties. The range of leaf Bt proteins was 1.08∼5.81 µg.g−1 FW, which could effectively protect Bt maizes from the corn borer pest during their growth periods. Furthermore, the younger leaves of Bt maizes had significantly lower levels of Bt proteins than their older leaves. In most periods, Bt protein content of Bt maize leaves under drought stress was lower than that under normal growth conditions, but Bt protein content under nutrition stress was higher than that under normal growth conditions. The results provided a full pattern of the spatial and temporal distribution of Bt proteins in Bt maize leaves. [ABSTRACT FROM AUTHOR]

Transgenic cry1Ab/cry1Ac rice (Huahui 1) is able to control the stored pest Plodia interpunctella.Our previous study confirmed that the enzymatic activities of three major antioxidases, including POD, CAT,and SOD, could be significantly changed after short-term stressed with these Bt containing rice. This study focused on the characteristics of rice during storage, the results showed that the Bt protein content in rice husk and brown rice was 5.534 μg/g and 1.003 μg/g respectively, and after 270 days at 25 ℃, the degradation rate was 67.3% and 27.7%, respectively. Moreover, the enzymatic activities of three antioxidases in P.interpunctella long-term fed with transgenic cry1Ab/cry1Ac rice powder would be addressed by enzymatic detection assays. Our data showed that the SOD activities could be significantly inhibited by the Bt containing rice after fed with Bt-rice powder for 12 days or 5 months, However, the enzymatic activities of POD and CAT significantly increase with the concentrations of Bt toxins. In addition, the change trend of protective enzyme activity in the Indian Meal Moth fed with transgenic rice flour for 5 months was consistent with that after 12 days of feeding. Our researches find that there are differences in the trend of increase and decrease of the three major protective enzymes after fed with Bt containing rice powder for a long time,which indicates that the expression mechanisms of the three protective enzymes might be different. Our results provide a theoretical basis for the study of toxicology and related resistance mechanism of transgenic Bt crops.

This study reported the functionality integration of zeolitic imidazolate framework-8 (ZIF-8) with horseradish peroxidase (HRP) and streptavidin (SA) for the synthesis of a HRP&SA/ZIF-8 nanocomposite through one-pot coprecipitation. The synthesized HRP&SA/ZIF-8 nanocomposite was then employed as the ideal signal tag for application in the enzyme-linked immunosorbent assay (ELISA) and exhibited excellent sensitivity, selectivity and accuracy in the detection of insecticidal crystalline (Cry) protein Cry1Ab as a transgenic biomarker with a detection limit of 4.8 pg/mL. This proposed method provides a new way for the detection of transgenic biomarkers in food and may inspire further integration of a variety of biomolecules into ZIF-8 for applications ranging from biosensing, biomedicine, and catalysis to energy.

Bt (Bacillus thuringiensis) corn is one of the top three large-scale commercialized anti-insect transgenic crops around the world. In the present study, we tested the Bt protein content, defense chemicals contents, and defense enzyme activities in both the leaves and roots of Bt corn varieties 5422Bt1 and 5422CBCL, as well as their conventional corn 5422 seedlings, with two fully expanded leaves which had been treated with 2.5 mM exogenous salicylic acid (SA) to the aboveground part for 24 h. The result showed that the SA treatment to the aboveground part could significantly increase the polyphenol oxidase activity of conventional corn 5422, the Bt protein content, and peroxidase activities of Bt corn 5422Bt1, as well as the polyphenol oxidase and peroxidase activity of Bt corn 5422CBCL in the leaves. In the roots, the polyphenol oxidase and peroxidase activity of conventional corn 5422, the polyphenol oxidase and superoxide dismutase activities of Bt corn 5422Bt1, the DIMBOA (2,4-dihydroxy-7-methoxy-2H, 1, 4-benzoxazin-3 (4H)-one) content, and four defense enzymes activities of Bt corn 5422CBCL were systematically increased. These findings suggest that the direct effect of SA application to aboveground part on the leaf defense responses in Bt corn 5422CBCL is stronger than that in non-Bt corn. Meanwhile, the systemic effect of SA on the root defense responses in Bt corn 5422CBCL is stronger than that in conventional corn 5422 and Bt corn 5422Bt1. It can be concluded that the Bt gene introduction and endogenous chemical defense responses of corns act synergistically during the SA-induced defense processes to the aboveground part. Different transformation events affected the root defense response when the SA treatment was applied to the aboveground part.

Reproductive organs of Bacillus thuringiensis transgenic cotton, which contribute to cotton final yield, have low insect resistant efficacy, so it is important to improve their insect resistance. This study was conducted to find out the impact of different urea spray doses on the expression of Cry1A protein in boll shell of Bt cotton (Sikang 1 and Sikang 3), and nitrogen metabolism in this process was also studied to uncover the physiological mechanism. The experiment with six urea doses was organized during peak boll stage in 2017 and 2018. The results showed that urea spray could significantly increase boll shell insecticidal protein contents in both cultivars, with the highest Bt protein content observed at 28–32 kg ha−1 urea dose. In addition, urea spray increased the contents of soluble protein and free amino acid and the activities of GS, GOGAT, GOT, and GPT, but decreased the activities of peptidase and protease in boll shell. Correlation analysis showed that the amount of boll shell Bt protein was positively correlated with levels of soluble protein and amino acid, and activities of GS, GOGAT, GOT, and GPT, but negatively correlated with peptidase and protease activities. Thus, this study demonstrated that higher protein synthesis ability and lower proteolysis ability were related to increased Bt protein content in urea-sprayed boll shell.

Reproductive organs of Bacillus thuringiensis transgenic cotton, which contribute to cotton final yield, have low insect resistant efficacy, so it is important to improve their insect resistance. This study was conducted to find out the impact of different urea spray doses on the expression of Cry1A protein in boll shell of Bt cotton (Sikang 1 and Sikang 3), and nitrogen metabolism in this process was also studied to uncover the physiological mechanism. The experiment with six urea doses was organized during peak boll stage in 2017 and 2018. The results showed that urea spray could significantly increase boll shell insecticidal protein contents in both cultivars, with the highest Bt protein content observed at 28–32 kg ha−1 urea dose. In addition, urea spray increased the contents of soluble protein and free amino acid and the activities of GS, GOGAT, GOT, and GPT, but decreased the activities of peptidase and protease in boll shell. Correlation analysis showed that the amount of boll shell Bt protein was positively correlated with levels of soluble protein and amino acid, and activities of GS, GOGAT, GOT, and GPT, but negatively correlated with peptidase and protease activities. Thus, this study demonstrated that higher protein synthesis ability and lower proteolysis ability were related to increased Bt protein content in urea-sprayed boll shell. [ABSTRACT FROM AUTHOR]

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[Objective] To study the effect of abiotic stress on the expression of Bt protein in transgenic insect-resistant cotton. [Method] The seedlings of two transgenic insect-resistant cotton varieties Zhongmian 45 (ZGK9822) and Ekang insect cotton 1 (GK19) were treated with low temperature (4℃), high temperature (37℃), 0.1, 0.2, 0.5 mol· L -1 NaCl, mild drought, moderate drought, and severe drought 8 abiotic stress treatments. [Results] Low temperature and high temperature stress caused the Bt protein content in the roots, stems and leaves of the two insect-resistant cotton varieties to decrease to varying degrees. Among them, the Bt protein content of each tissue under low temperature stress was higher than that of high temperature stress; Compared with CK, the content of Bt protein in stems and leaves showed a trend of first decreasing and then increasing. In roots, stems and leaves of different degrees of drought stress, the change trend of Bt protein content was consistent with salt stress. [Conclusion] Abiotic stress seriously affects the expression of Bt protein in transgenic insect-resistant cotton, resulting in dynamic changes in the expression of Bt protein. When planting transgenic insect-resistant cotton in the field, attention should be paid to the impact of environmental factors. [ABSTRACT FROM AUTHOR]

Fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith), is the main pest of maize in Brazil, attacking plants from emergence to reproductive stages. Here, we conducted studies to evaluate the efficacy of two seed treatments (chlorantraniliprole alone and imidacloprid combined with thiodicarb) on Bt and non-Bt maize in laboratory bioassays with distinct FAW strains that are susceptible, selected for resistance to Bt-maize single (Cry1F) or pyramided (Cry1A.105 + Cry2Ab2) events and F1 hybrids of the selected and susceptible strains (heterozygotes), and in the field against a natural infestation. In the laboratory, leaf-discs from seed treated Bt-maize plants at 7 d after emergence (DAE) increased the mortality of FAW resistant, heterozygote, and susceptible strains up to 24.8%, when compared with the respective maize grown without a seed treatment. In the field against natural infestations of FAW, Bt maize with a seed treatment had ~30% less FAW damage than non-Bt maize with the same seed treatment at 7 and 14 DAE. No differences in FAW damage was observed between Bt and non-Bt maize grown with and without a seed treatment at 21 DAE. Maize seeds treated with chlorantraniliprole alone or imidacloprid and thiodicarb combined presented limited protection against early infestations of FAW strains under laboratory and field studies. [ABSTRACT FROM AUTHOR]

Copyright of Acta Agronomica Sinica is the property of Crop Science Society of China and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is currently the most important maize pest in Mexico. Its control is mainly based on the use of conventional insecticides. Additionally, Bt-maize expressing Cry1F protein represents an alternative to control this pest. We estimated the baseline susceptibility in Mexican populations of S. frugiperda to Cry1F protein. Twenty-eight geographical populations were field collected from Baja California Sur, Chihuahua, Coahuila, Durango, Sinaloa, Sonora, and Tamaulipas states. The F 1 neonate larvae of each population were subjected to diet-overlay bioassay. After 7 d of Cry1F exposure, the percent mortality and the percent growth inhibition with respect to the untreated control were recorded (S-LAB). The LC50 ranged from 14.4 (6.3−24.0) (Cajeme 1, Sonora) to 161.8 ng/cm2 (92.0–320) (Ahumada 2, Chihuahua), while the LC95 was between 207.1 (145–363) (Obregón, Sonora) and 1,217 ng/cm2 (510.8–7,390.0) (Río Bravo 2, Tamaulipas). The sensitivity ratios at 50% mortality, (LC50 field/LC50 S-Lab) and 95% mortality were ≤6.45 and ≤5.05-fold, respectively. The 50% growth inhibition (GI50) ranged from 2.8 (0.008–9.3) (Obregón, Sonora) to 42.4 ng/cm2 (3.6–147.0) (Cajeme 1, Sonora). The GI95 was between 75.4 (San Luis Río Colorado, Sonora) to 1,198 ng/cm2 (Cajeme 1, Sonora). The relative inhibition at 50% of the growth, (RI50 = GI50 field /GI50 S-LAB) was ≤3.5 and at 95% (RI95) was ≤1.91-fold. These results indicated susceptibility to Cry1F protein in the evaluated populations of S. frugiperda. [ABSTRACT FROM AUTHOR]

Abstract Background Pardosa pseudoannulata is a prevailing spider species, and has been regarded as an important bio-control agent of insect pests in farmland of China. However, the available genomic and transcriptomic databases of P. pseudoannulata and their venom are limited, which severely hampers functional genomic analysis of P. pseudoannulata. Recently high-throughput sequencing technology has been proved to be an efficient tool for profiling the transcriptome of relevant non-target organisms exposed to Bacillus thuringiensis (Bt) protein through food webs. Results In this study, the transcriptome of the venom apparatus was analyzed. A total of 113,358 non-redundant unigenes were yielded, among which 34,041 unigenes with complete or various length encoding regions were assigned biological function annotations and annotated with gene ontology and karyotic orthologous group terms. In addition, 3726 unigenes involved in response to stimulus and 720 unigenes associated with immune-response pathways were identified. Furthermore, we investigated transcriptomic changes in the venom apparatus using tag-based DGE technique. A total of 1724 differentially expressed genes (DEGs) were detected, while 75 and 372 DEGs were functionally annotated with KEGG pathways and GO terms, respectively. qPCR analyses were performed to verify the DEGs directly or indirectly related to immune and stress responses, including genes encoding heat shock protein, toll-like receptor, GST and NADH dehydrogenase. Conclusion This is the first study conducted to specifically investigate the venom apparatus of P. pseudoannulata in response to Bt protein exposure through tritrophic chain. A substantial fraction of transcript sequences was generated by high-throughput sequencing of the venom apparatus of P. pseudoannulata. Then a comparative transcriptome analysis showing a large number of candidate genes involved in immune response were identified by the tag-based DGE technology. This transcriptome dataset will provide a comprehensive sequence resource for furture molecular genetic research of the venom apparatus of P. pseudoannulata.

Abstract The transgenic rice line T1C-19 provides high resistance to lepidopteran pests because of the synthesis of the Bacillus thuringiensis (Bt) insecticidal protein Cry1C. It thus shows good prospect for commercial planting in China. Species of Cladocera, an order of aquatic arthropods commonly found in aquatic ecosystems such as rice paddies, might be exposed to the insecticidal protein released from Bt -transgenic rice-straw residues. For the study reported herein, we used Daphnia magna (water flea) as a representative of Cladocera to evaluate whether aquatic arthropods are adversely affected when exposed to Bt rice-straw leachates. We exposed D. magna to M4 medium containing various volume percentages of medium that had been incubated with T1C-19 rice straw or rice straw from its non-transformed near-isoline Minghui 63 (MH63) for 21 days. Compared with pure M4 medium (control), the fitness and developmental and reproduction parameters of D. magna decreased significantly when exposed to rice-straw leachates; conversely, no significant differences between the T1C-19 and MH63 rice-straw leachate treatments were observed, indicating that the Bt rice straw leachate did not adversely affect this non-target species. Highlights • Rice-straw leachate affects water quality and different life-table parameters of Daphnia magna. • D. magna is not affected by rice-straw leachate from Bt rice expressing Cry1C when compared to its corresponding non-Bt rice. • Catalase activity in D. magna exposed to rice-straw leachate was significantly higher than that of blank control. [ABSTRACT FROM AUTHOR]

Abstract Background: To know the effects of transgenic crops on soil microorganisms, animals and other expected hazards due to the introduction of GM crops into the environment is critical both scientifically and environmentally. The work was conducted to study the effect of insecticidal Bt protein on Rats and Earthworms. Methods: For this purpose, animals like rat and soil organisms like Earthworm were selected. Rats were selected on the basis of its 95% homology on genomic, cellular and enzymatic level with human while earthworm were preferred on the basis of their direct contact with soil to evaluate the impact of Bt (Cry1AC) crop field soil on earthworm, secreted by root exudates of Bt cotton. Several physical, molecular, biochemical and histological analyses were performed on both Rats/Earthworms fed on standard diet (control group) as well containing Bt protein (experimental group). Results: Molecular analyses such as immune Dot blot, SDS-PAGE, ELISA and PCR, confirmed the absence of Cry1Ac protein in blood and urine samples of rats, which were fed with Bt protein in their diet. Furthermore, histological studies showed that there was no difference in cellular architecture in liver, heart, kidney and intestine of Bt and non-Bt diet fed rats. To see the effect of Bt on earthworm two different groups were studied, one with transgenic plant field soil supplemented with grinded leaves of cotton and second group with non-Bt field soil. Conclusions: No lethal effects of transgenic Bt protein on the survival of earthworm and rats were observed. Bradford assay, Dipstick assay ELISA demonstrated the absence of Cry1Ac protein in the mid-gut epithelial tissue of earthworm. The results of present study will be helpful in successful deployment and commercial release of genetically modified crop in Pakistan.

Monsanto's Bt-cotton NC 33B, planted in northern China for more than one decade, effectively controls cotton bollworms; however the understanding of its potential effects on soil microorganisms is limited. The dynamics of eubacterial, fungal and actinomycetes population sizes in rhizosphere soil of the Bt cotton were analysed by real-time PCR (qPCR) at the different growth stages under field conditions during 2009 to 2011. Results showed that the population sizes (microbial rDNA gene copies) of eubacteria, fungi and actinomycetes in rhizosphere soil were markedly affected by natural variations in the environment related to the year, cotton growth and cultivar. However, there was no significant difference in eubacterial, fungal and actinomycetes population size in rhizosphere soil between the Bt-cotton NC 33B and its near-isogenic comparator DP 5415. In general, the Bt-cotton NC 33B did not show evident effects on the population sizes of eubacteria, fungi and actinomycetes in rhizosphere soil under field conditions after three-year cultivation.

Higher boll worm survival rates were detected after high temperature presented during square period in Bt cotton. The objective of this study was to investigate the effects of high temperature level on the Bt efficacy of two different types of Bt cotton cultivars at squaring stage. During the 2011 to 2013 cotton growth seasons, high temperature treatments ranged from 34 to 44°C in climate chambers, and field experiments under high temperature weather with various temperature levels were conducted to investigate the effects of the high temperature level on square Bt protein concentration and nitrogen metabolism. The climate chamber experiments showed that the square insecticidal protein contents reduced after 24 h elevated temperature treatments for both cultivars, whereas significant declines of the square insecticidal protein contents were detected at temperature >38°C, and only slightly numerical reductions were observed when temperature below 38°C. Similar high temperature responses were also observed at the two field experimental sites in 2013. Correspondingly, high temperature below 38°C seems have little effect on the square amino acid concentrations, soluble protein contents, glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) activities as well as protease and peptidase activities; however, when the temperature was above 38°C, reduced soluble protein contents, enhanced amino acid concentrations, decreased GPT and GOT activities, bolstered protease and peptidase activities in square were detected. In general, the higher the temperature is (>38°C), the larger the changes for the above compound contents and key enzymes activities of the square protein cycle. The findings indicated that the unstable insect resistance of the square was related to high temperature level during square stage.

To clarify the effects of soil salinity on the insect-resistance of flower buds in transgenic Bt cotton of Xinmian 33B (salt-sensitive) and Zhong 07 (salt-tolerant), five levels (0, 0.15%, 0.30%, 0.45% and 0.60%) of soil salinity were set to investigate the impacts of soil salinity on Bt protein content, relative expression of Bt gene, activities of nitrogen metabolism-related enzymes and substances in flower buds during flowering stage. The results showed that Bt protein contents of flower buds decreased with increasing soil salinity. The Bt protein contents of flower buds decreased significantly when the soil salinity level was above 0.30% in both cotton cutivars. Greater reduction of Bt protein content occurred under severe than moderate soil salinity stress. However, the relative expression of Cry1Ac gene increased in flower buds of both cultivars when the stress level increased. Greater decreases of Bt protein content of flower buds was observed in Xinmian 33B compared to Zhong 07 under the same salinity level. The cultivars with greater reduction in Bt protein contents of flower buds also had greater reduction in the soluble protein content, glutamate pyruvate transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) activities, and greater increments in free amino acid contents, protease activities and peptidase activities. Therefore, the decrease of Bt protein content was caused by decreased synthesis and increased decomposition of protein in flower buds of Bt cotton under soil salinity stress. [ABSTRACT FROM AUTHOR]

In order to clarify the effects of soil salinity on the insect-resistance of boll in transgenic Bt cotton, potted plants of two Bt cotton cultivars Xinmian 33B (salt-sensitive) and Zhong 07 (salt-tolerant) were exposed to five levels of soil salinity (0, 0.15%, 0.30%, 0.45% and 0.60%). The results showed that Bt protein content of boll shell decreased with increasing soil salinity. Compared with the control (0% soil salinity level), the Bt protein content of boll shell decreased significantly when the soil salinity level was above 0.15% for Xinmian 33B and above 0.30% for Zhong 07. The reduction extent of Bt protein content of boll shell at 30 days post anthesis (DPA) was greater than that at 10 DPA under the same soil salinity level. Significant reductions of soluble protein contents, nitrate reductase (NR), and glutamate pyruvate transaminase (GPT) activities were observed when the boll shell Bt protein content was significantly reduced. The content of free amino acid, protease, and peptidase activity of boll shell significantly increased when the soil salinity level was above 0.30%. In conclusion, soil salinity affected boll shell nitrogen metabolism and reduced Bt protein synthesis. Middle and high soil salinity levels could enhance decomposition of Bt protein, which further decreases the expression level of insecticidal protein. [ABSTRACT FROM AUTHOR]

Abstract: Resistances of newly bred Bacillus thuringiensis (Bt) crops have been dramatically improved because of the effective and high expression of Bt protein in the plant. However, poorer adaptabilities to environmental stresses were observed in some Bt crops compared to their non‐Bt counterparts. The biological reasons for the poorer adaptabilities were still unclear. A nitrogen (N) deficiency experiment was conducted to investigate variations in growth and physiology characteristics of a newly bred Bt rice [Oryza sativa L. line MH63 (Cry2A*)] compared to its non‐Bt counterpart MH63. MH63 (Cry2A*) showed lower grain yields and lower biomass under low N levels compared to MH63. Earlier leaf senescence associated with disorder in protein metabolism was observed in MH63 (Cry2A*) when the N concentration was lower than 13.50 mg g−1 in MH63 (Cry2A*) leaves and the ratio of Bt protein to soluble protein (BT : SP) was higher than 2203 μg g−1 in MH63 (Cry2A*) leaves. The lower grain yield, the lower biomass and the earlier leaf senescence associated with disorder in protein metabolism in MH63 (Cry2A*) were correlated to the high BT : SP in MH63 (Cry2A*) leaves. The results suggest that MH63 (Cry2A*) has a poorer adaptability to N deficiency compared to its non‐Bt counterpart MH63. This poorer adaptability might be related to the high Bt protein expression in MH63 (Cry2A*). [ABSTRACT FROM AUTHOR]

High expression of Bacillus thuringiensis ( Bt ) protein in transgenic Bt crops has dramatically improved the resistance of crops to some insect pests. However, some Bt crops were reported to have poorer adaptabilities to abiotic stress environments compared to their non- Bt counterparts. The biological reasons for poorer adaptabilities were still unclear. A two-year field experiment was conducted to investigate yield and physiological response of newly bred Bt rice lines with different Bt protein expression levels [MH63 ( Cry1C* ), MH63 ( Cry2A* ) and MH63 ( Cry1Ab/Ac )] and their non- Bt counterpart MH63 to drought stress. MH63 ( Cry1C* ), MH63 ( Cry2A* ) and MH63 ( Cry1Ab/Ac ) all showed lower grain yields and lower biomass under drought treatment compared to MH63. The lower grain yields and lower biomass of Bt -MH63 were mainly due to the earlier leaf senescence associated with disorder in protein metabolism under drought treatment. These variations in Bt -MH63 were found to be correlated to the BT/SP (ratio of Bt protein to soluble protein) in Bt -MH63 leaves. The impact of drought on traits of Bt -MH63 was aggravated with increase of the Bt protein expression level in the plant. These results suggested that Bt -MH63 had poorer adaptabilities to drought stress compared to their non- Bt counterpart MH63. “Bio-burden” caused by the high expression of Bt protein in the plant was proposed to be one possible reason for the poorer adaptability, and BT/SP was estimated to be an indicator for the assessment of the “bio-burden”. [ABSTRACT FROM AUTHOR]

Cry1C is a Bacillus thuringiensis ( Bt ) insecticidal protein and it can be produced by transgenic rice lines developed in China. Cladocera species are common aquatic arthropods that may be exposed to insecticidal proteins produced in Bt -transgenic plants through ingestion of pollen or crop residues in water. As the cladoceran Daphnia magna plays an important role in the aquatic food chain, it is important to assess the possible effects of Bt crops to this species. To evaluate the safety of the Cry1C protein for D. magna , individuals were exposed to different concentrations of purified Cry1C protein in M4 medium for 21 days. Potassium dichromate (K 2 Cr 2 O 7 ), a known toxicant to D. magna , was added to M4 medium as a positive control treatment, and pure M4 medium was used as a negative control. Our results show that developmental, reproductive, and biochemical parameters of D. magna were not significantly different between Cry1C and negative control treatments but were significantly inhibited by the positive control. We thus conclude that D. magna is insensitive to Cry1C. [ABSTRACT FROM AUTHOR]

Expression of insecticidal protein for transgenic Bacillus thuringiensis (Bt) cotton is unstable and related to nitrogen metabolism. The objective of this study was to investigate the relationship between leaf carbon nitrogen ratio (C/N) and insecticidal efficacy of two Bt cotton cultivars. C/N ratio and Bt protein content were both measured at peak square period and peak boll period respectively under 5-7 d high temperature and different nitrogen fertilizer rates on the Yangzhou University Farm and the Ludong Cotton Farm, China. All plants were grown in field. The results showed that the C/N ratio enhanced slightly and the Bt protein content remained stable at peak square period, but significant increases for the C/N ratio and decreases markedly for the leaf Bt protein concentration were detected at the peak boll period. The similar patterns at the two growth periods were found for the leaf C/N ratio and Bt protein content by different N fertilizer treatments. When nitrogen rate was from 0 to 600 kg ha−1, the C/N ratio was reduced by 0.017 and 0.006 for Sikang 1 and Sikang 3 at peak square period, compared to the 1.350 to 1.143 reduction for Sikang 1 and Sikang 3 at peak boll period, respectively. Correspondingly, the leaf Bt protein contents were bolstered by 2.6-11.8 and 26.9-36.9% at the two different growth periods, respectively. The results suggested that enhanced C/N ratio by high temperature and nitrogen application may result in the reduction of insectiocidal efficacy in Bt cotton, especially in peak boll period.

Copyright of Journal of Economic Entomology is the property of Oxford University Press / USA and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Agricultural Science & Technology (1008-0864) is the property of Journal of Agricultural Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Transgenic Bacillus thuringiensis ( Bt) rice have been reported to acquire effective resistance against the target pests; however, the insertion and expression of alien Bt genes may have some unintended effects on the growth characteristics of rice. A screen-house experiment was conducted and repeated twice to investigate the growth characteristics and Bt protein expressions in two Bt rice lines [MH63 ( Cry2A*) and MH63 ( Cry1Ab/Ac)], which had different Bt protein expression levels in leaves, under zero nitrogen (N0) and recommended nitrogen (NR) fertilizer applications. Compared to the counterpart MH63, MH63 ( Cry2A*) under N0 experienced accelerated leaf senescence and a lower internal N use efficiency (IE), resulting in a 23.2% decrease in grain yield and a lower accumulated biomass. These variations were revealed to be correlated to the higher ratio of the Bt protein content to the soluble protein content (BTC/SPC) with a maximum value of 4.3‰ in MH63 ( Cry2A*) leaves in the late growth stage. Under NR, no differences in growth characteristics between MH63 ( Cry2A*) and MH63 were found. The growth characteristics of MH63 ( Cry1Ab/Ac), with a lower BTC/SPC in the late growth stage compared to MH63 ( Cry2A*), were identical to those of MH63 under the two N applications. Results show that the transgenic Bt rice MH63 ( Cry2A*), with a relatively higher Bt protein expression in the late growth stage, had an inferior adaptation to nitrogen deficiency compared to its non- Bt counterpart. And this inferior adaptation was found to be correlated with the higher BTC/SPC in MH63 ( Cry2A*) leaves in the late growth stage. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Applied Ecology / Yingyong Shengtai Xuebao is the property of Chinese Journal of Applied Ecology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Bollgard ® III was developed by combining cotton events COT102 and MON 15985 through conventional breeding to improve efficacy against lepidopteran feeding damage. COT102 produces the Vip3Aa19 protein and MON 15985 produces the Cry1Ac and Cry2Ab2 proteins. COT102 × MON 15985 has also been bred with Roundup Ready Flex ® cotton (MON 88913) that confers glyphosate tolerance. This study evaluated the activity of COT102 and MON 15985 and the combined activity of COT102 and MON 15985 against the cotton bollworm (CBW, Helicoverpa zea ). COT102, MON 15985, COT102 × MON 15985 and COT102 × MON 15985 × MON 88913 have comparable Vip3Aa19 and/or Cry1Ac, Cry2Ab2 protein expression levels as determined by enzyme-linked immunosorbent assay. CBW demonstrated concentration-dependent growth inhibition after 7-days of feeding on lyophilized leaf tissue derived from COT102, MON 15985, COT102 × MON 15985 and COT102 × MON 15985 × MON 88913 incorporated into an artificial diet. Observed EC 50 values for COT102 × MON 15985 and COT102 × MON 15985 × MON 88913 were comparable (≤4% deviation) with the predicted EC 50 value under the assumption of additivity using the combined activity of COT102 and MON 15985. No interaction in biological activity between COT102 and MON 15985 is consistent with results from competition and ligand blotting assays that demonstrated that Vip3Aa does not inhibit the binding of either Cry1Ac or Cry2Ab2 and vice versa. The results from this study demonstrate that the activity of COT102 × MON 15985 against CBW is consistent with predictions of additivity. [ABSTRACT FROM AUTHOR]

Transgenic rice with Cry Bacillus thuringiensis ( Bt ) genes have been developed in the last few decades. The performance of transgenic rice, especially the yield and resistance, must be meticulously evaluated prior to commercial production. In this study, yield and resistance performance of newly bred transgenic Bt ( Cry1Ab/Ac ) rice lines with different genetic backgrounds were first investigated under pesticide-free field environment. Transgenic Bt ( Cry1Ab/Ac ) rice lines showed high insect resistances and superior agronomic traits, resulting in obvious yield advantages ranging from 9.1% to 38.0% over their respective non- Bt control lines. Bt protein content (BTC) was similar among the Bt ( Cry1Ab/Ac ) rice lines in different years, while it was much higher in leaves than in stems of the Bt ( Cry1Ab/Ac ) rice lines. Different genetic backgrounds of Bt ( Cry1Ab/Ac ) rice lines affected the yield advantage but had no influence on the resistance and BTC. The results suggested that Bt ( Cry1Ab/Ac ) rice lines have gained obvious yield advantages and high resistances regardless of genetic background under pesticide-free environment. [ABSTRACT FROM AUTHOR]

The lady beetle Propylea japonica is a dominant natural predator of insect pests in farmland ecosystems and an important non-target indicator insect for the environmental safety assessment of GM crops. The commercial cultivation of GM crops may cause P. japonica to frequently be exposed to the Bt protein environment. In this study, the biological characteristics, enzyme activity, and expression levels of detoxification and metabolism in P. japonica were studied after Cry1B protein treatment. No significant differences were observed in developmental duration, emergence rate, or body weight at different ages after feeding larvae 0.5 mg/mL of Cry1B protein compared with the control. Furthermore, there were no significant differences in the activities of glutathione S-transferase (GST), catalase (CAT), and peroxidase (POD) after feeding 0.25 mg/mL and 0.5 mg/mL Cry1B protein. However, when the concentration of Cry1B protein increased to 1.0 mg/mL, the activities of the GST, CAT, and POD increased significantly. Compared with the control group, there were no significant differences in the expression levels of most detoxification metabolism related genes; only a few genes had changed expression levels at the individual concentrations (CYP345B1 , CYP4Q2 , CYP9F2 , GST , and microsomal GST). Overall, these results suggest that Cry1B protein has little or no effect on the biological characteristics of P. japonica. Genes related to enzyme activity and detoxification are differentially expressed at high concentration stimulation. Therefore, this research suggests that the potential risks of Cry1B for the predator P. japonica are negligible. • Cry1B protein can be transmitted to the predator Propylea japonica. • Cry1B protein did not affect the development duration, emergence rate and body weight at different ages. • Cry1B protein did not affect the activities of GST, CAT and POD in Propylea japonica. • Cry1B protein does not affect the expression of genes related to detoxification metabolism of Propylea japonica. • The potential risks of Cry1B for the predator Propylea japonica are negligible. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Applied Ecology / Yingyong Shengtai Xuebao is the property of Chinese Journal of Applied Ecology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Using a transgenic Bacillus thuringiensis ( Bt ) crop is reportedly one of the most effective ways to improve insect resistance; however, the insertion of external Bt genes may affect the non-target growth characteristics of crop. Field experiments were conducted to investigate the yield performance and physiological traits in an elite restorer rice line, Minghui63 (MH63) with Cry1C * , Cry2A * or Cry1Ab/Ac genes under three fertilizer applications [zero nitrogen fertilizer (N0), zero potassium fertilizer (K0) and recommended nitrogen and potassium fertilizer (NK)]. In comparison with MH63, MH63 ( Cry1C * ) experienced declining grain yields of 15.3–31.0% under different fertilizer applications because of the decreased setting rates associated with the poorer matter transport-related traits. The grain yield of MH63 ( Cry2A * ) was averagely 18.4% lower than MH63 in only the N0 treatment due to the accelerated leaf senescence, which was correlated with the higher ratios of the Bt protein content to the soluble protein content (BTC/SPC) in the MH63 ( Cry2A * ) leaves at the filling stage (FS). The yield performance of MH63 ( Cry1Ab/Ac ) was identical to MH63. In addition, no significant differences were recorded between Bt- MH63 and MH63 regarding N and K assimilation. Hence, our results suggest that the Cry1C * and Cry2A * transgenes affect the yield performance and related traits of MH63 in response to nitrogen and/or potassium applications. [ABSTRACT FROM AUTHOR]