Evaluation of Cotton Fleahopper (Pseudatomoscelis seriatus (Reuter)) Feeding on Mpp51Aa2-Traited Cotton Utilizing Electrical Penetration Graph (EPG) Waveforms.

Prior to the recent implementation of the Mpp51Aa2 pesticidal protein (ThryvOn), transgenic cotton cultivars have historically offered no control of the cotton fleahopper (Pseudatomocelis seriatus (Reuter)). To evaluate the feeding behavior of cotton fleahoppers on ThryvOn cotton, electropenetrography (EPG) using a Giga-8 DC instrument was used to monitor the probing activity of fourth- and fifth-instar cotton fleahopper nymphs on both ThryvOn and non-ThryvOn cotton squares. Nymphs were individually placed on an excised cotton square for 8 h of EPG recording, after which resulting waveforms were classified as non-probing, cell rupturing, or ingestion. Although there were significantly more cell rupturing events per insect on ThryvOn (mean ± SEM, 14.8 ± 1.7) than on non-ThryvOn squares (mean ± SEM, 10.3 ± 1.6), there was no difference attributable to ThryvOn in the average number of ingestion events per insect. However, the average duration of ingestion events was significantly shorter on squares with ThryvOn (mean ± SEM, 509 ± 148 s) than on squares without (mean ± SEM, 914 ± 135 s). This suggests that cotton fleahoppers continued to probe despite their inability to sustain ingestion. These results provide conclusive evidence that the Mpp51Aa2 pesticidal protein affects the feeding behavior of cotton fleahopper nymphs.

Field Evaluation of Cotton Expressing Mpp51Aa2 as a Management Tool for Cotton Fleahoppers, Pseudatomoscelis seriatus (Reuter).

The cotton fleahopper (Pseudatomoscelis seriatus Reuter) is considered a highly economically damaging pest of cotton (Gossypium hirsutum L.) in Texas and Oklahoma. Current control methods rely heavily on the use of foliar-applied chemical insecticides, but considering the cost of insecticides and the critical timeliness of applications, chemical control methods are often not optimized to reduce potential yield losses from this pest. The Bacillus thuringiensis (Bt) Mpp51Aa2 (formerly Cry51Aa2.834_16) protein has proven effective against thrips and plant bugs with piercing and sucking feeding behaviors, but the impact of this toxin on cotton fleahoppers has not been investigated. To evaluate the Mpp51Aa2 trait effectiveness towards the cotton fleahopper, field trials were conducted in 2019, 2020, and 2021, comparing a cotton cultivar containing the Mpp51Aa2 trait to a non-traited isoline cultivar under insecticide-treated and untreated conditions. Populations of cotton fleahopper nymphs and adults were estimated weekly by visually inspecting cotton terminals. Square retention was also assessed during the first week of bloom to provide some insight on how the Bt trait may influence yield. While cotton fleahopper population differences between the traited and non-traited plants were not consistently noted during the pre-bloom squaring period, there was a consistent increase in square retention in cotton expressing Mpp51Aa2 relative to non-traited cotton. Additionally, cotton expressing Mpp51Aa2 offered similar square protection relative to non-traited cotton treated with insecticides for the cotton fleahopper. These findings indicate that the Mpp51Aa2 protein should provide benefits of delayed nymphal growth, population suppression, and increased square retention.

Intercropping Cover Crops for a Vital Ecosystem Service: A Review of the Biocontrol of Insect Pests in Tea Agroecosystems.

The intercropping of cover crops has been adopted in several agroecosystems, including tea agroecosystems, which promotes ecological intensification. Prior studies have shown that growing cover crops in tea plantations provided different ecological services, including the biocontrol of pests. Cover crops enrich soil nutrients, reduce soil erosion, suppress weeds and insect pests, and increase the abundance of natural enemies (predators and parasitoids). We have reviewed the potential cover crops that can be incorporated into the tea agroecosystem, particularly emphasizing the ecological services of cover crops in pest control. Cover crops were categorized into cereals (buckwheat, sorghum), legumes (guar, cowpea, tephrosia, hairy indigo, and sunn hemp), aromatic plants (lavender, marigold, basil, and semen cassiae), and others (maize, mountain pepper, white clover, round-leaf cassia, and creeping indigo). Legumes and aromatic plants are the most potent cover crop species that can be intercropped in monoculture tea plantations due to their exceptional benefits. These cover crop species improve crop diversity and help with atmospheric nitrogen fixation, including with the emission of functional plant volatiles, which enhances the diversity and abundance of natural enemies, thereby assisting in the biocontrol of tea insect pests. The vital ecological services rendered by cover crops to monoculture tea plantations, including regarding the prevalent natural enemies and their pivotal role in the biocontrol of insect pests in the tea plantation, have also been reviewed. Climate-resilient crops (sorghum, cowpea) and volatile blends emitting aromatic plants (semen cassiae, marigold, flemingia) are recommended as cover crops that can be intercropped in tea plantations. These recommended cover crop species attract diverse natural enemies and suppress major tea pests (tea green leaf hopper, white flies, tea aphids, and mirid bugs). It is presumed that the incorporation of cover crops within the rows of tea plantations will be a promising strategy for mitigating pest attacks via the conservation biological control, thereby increasing tea yield and conserving agrobiodiversity. Furthermore, a cropping system with intercropped cover crop species would be environmentally benign and offer the opportunity to increase natural enemy abundance, delaying pest colonization and/or preventing pest outbreaks for pest management sustainability.

Azotobacter inoculation can enhance the resistance of Bt cotton to cotton bollworm, Helicoverpa armigera.

The rising trend in the cultivation of Bacillus thuringiensis (Bt) transgenic crops may cause a destabilization of agroecosystems, thus increasing concerns about the sustainability of Bt crops as a valid pest management method. Azotobacter can be used as a biological regulator to increase environmental suitability and improve the soil nitrogen utilization efficiency of crops, especially Bt cotton. A laboratory test investigated effects on the development and food utilization of Helicoverpa armigera fed with different Cry1Ab/Cry1Ac proteins and nitrogen metabolism-related compounds from cotton (transgenic variety SCRC 37 vs non-Bt cotton cv. Yu 2067) inoculated with Azospirillum brasilense (Ab) and Azotobacter chroococcum (Ac). The findings indicate that inoculation with Azotobacter significantly decreased the partial development and food utilization indexes (pupal weight; pupation rate; adult longevity; fecundity; relative growth rate, RGR; efficiency of conversion of digested food, ECD; and efficiency of conversion of ingested food, ECI) of H. armigera fed on Bt cotton, but contrasting trends were found among these indexes in H. armigera fed on non-Bt cotton inoculated with Azotobacter, as a result of differences in Bt toxin production. Overall, the results showed that inoculation with Azotobacter had negative effects on the development and food utilization of H. armigera fed on Bt cotton, leading to enhanced target insect resistance. Presumably, Azotobacter inoculation can be used to stimulate plant soil nitrogen uptake to increase nitrogen metabolism-related compounds and promote plant growth for Bt and non-Bt cotton, simultaneously raising Bt protein expression and enhancing resistance efficacy against cotton bollworm in Bt cotton.

Influence of reduced N-fertilizer application on foliar chemicals and functional qualities of tea plants under Toxoptera aurantii infestation.

BACKGROUND: The tea aphid, Toxoptera aurantii (Boyer de Fonscolombe) is a polyphagous pest predominant in tea orchards and has become the most pernicious pest deteriorating tea quality. Nitrogen (N) is essential to plant growth improvement, and it can significantly impact plant defensive ability against aphid infestation. This study was designed to quantify the influence of reduced N-fertilizer application on foliar chemicals and functional quality parameters of tea plants against the infestation of T. aurantii. In this study, the tea seedlings (cv. Longjing43) were applied with normal level (NL) of N-fertilizer (240 kg N ha-1) along with reduced N-fertilizer levels (70%NL and 50%NL), and with and without T. aurantii infestation. RESULTS: The results showed that N-fertilizer application significantly affected plant biomass and photosynthetic indexes, foliar soluble nutrients and polyphenols, tea catechins, caffeine, essential amino acids, volatile organic compounds of tea seedlings, and the population dynamics of T. aurantii. Compared with the normal N-fertilizer level, the reduced N-fertilizer application (70%NL and 50%NL) significantly decreased all the foliar functional quality components of tea seedlings without aphid infestation, while these components were increased in tea seedlings with aphid infestation. Moreover, the transcript expression levels of foliar functional genes (including CsTCS, CsTs1, and CsGT1) were significantly higher in the NL, and significantly lower in the 50%NL for tea seedlings without aphid infestation, while the transcript expression levels were significantly higher in 50%NL in aphid inoculated tea seedlings. CONCLUSION: The results demonstrated that the reduced N-fertilizer application could enhance foliar chemicals and functional quality parameters of tea plants especially with T. aurantii infestation, which can relieve soil nitrogen pressure and reduce pesticide use for control of tea aphid infestation in tea plantations.

Elevated CO2 alters transgene methylation not only in promoterregion but also in codingregion of Bt rice under different N-fertilizer levels.

The earth has been undergoing climate change, especially in recent years, driven by increasing concentration of atmospheric carbon dioxide (CO2) and rising earth-surface temperature, which could reduce N allocation to Bt toxin for transgenic Bt crops (Bt crops), but the N fertilization is considered to be an effective method to enhance the C-N balance in Bt crops in the case of elevated CO2 in future. DNA methylation not only in promoterregion but also in codingregion of transgene plays a critical role in transgene expression regulation and silencing of transgenic crops. Recent research has emphasized the risks of increased transgene silencing of Bacillus thuringiensis (Bt) rice under elevated CO2. In this study, the effects of elevated CO2 (vs. ambient CO2) on exogenous Bt toxins and transgene expression in promoterregion and codingregion of Bt rice during tillering stage (cv. HH1 expressing fused Cry1Ab/Cry1Ac) were evaluated under three nitrogen (N) fertilizer rate (1/4, 1 and 2 N levels). The aboveground and belowground biomass, and foliar Bt protein content of Bt rice were all significantly increased with the augmentation of N-fertilizer. And elevated CO2 significantly increased belowground biomass, total soluble protein content, transgene methylation levels in promoterregion (P1), and in total of promoterregion(P1) and codingregion (P2 + P3) (i.e., P1 + P2 + P3) at 1 N level, and it also increased transgene methylation levels in codingregion (P2), and in total of promoterregion and codingregion (P1 + P2 + P3) at 2 N level. In addition, elevated CO2 decreased foliar Bt protein content at 1 N level. The transgene methylation levels in promoterregion and codingregion were negatively correlated with Bt-transgene expression level. The methylation level of cytosines located at CG sites was higher than those at CHG and CHH sites in P1, P2 and P3 fragments regardless of the CO2 or N-fertilizer level. The correlation of transgene mehtylation in promoterregion with transgene expression is even stronger than that in codingregion. These data indicate that N fertilization supply will increase the Bt toxin content in transgenic Bt rice, especially under elevated CO2.

Identification of Arylphorin interacting with the insecticidal protein PirAB from Xenorhabdus nematophila by yeast two-hybrid system.

PirAB toxin was initially found in the Photorhabdus luminescens TT01 strain and is a demonstrated binary toxin with high insecticidal activity. In this paper, we co-expressed the pirAB gene of Xenorhabdus nematophila HB310 in a prokaryotic expression system, and we found that the PirAB protein showed high hemocoel insecticidal activity against Galleria mellonella, Helicoverpa armigera and Spodoptera exigua. LD50 values were 1.562, 2.003 and 2.17 µg/larvae for G. mellonella, H. armigera, and S. exigua, respectively (p > 0.05). Additionally, PirAB-interaction proteins were identified from G. mellonella by 6 × His Protein Pulldown combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Of which, arylphorin of G. mellonella showed the highest matching rate. A protein domain conservative structure analysis indicated that arylphorin has three domains including Hemocyanin-N, Hemocyanin-M, and Hemocyanin-C. Among these protein domains, Hemocyanin-C has immune and recognition functions. Further, Hemocyanin-C domain of arylphorin was identified to interact with PirA but not PirB by Yeast two-hybrid system. These findings reveal, for the first time, new host protein interacting with PirAB. The identification of interaction protein may serve as the foundation for further study on the function and insecticidal mechanism of this binary toxin from Xenorhabdus.

Effects of elevated CO2 on foliar soluble nutrients and functional components of tea, and population dynamics of tea aphid, Toxoptera aurantii.

The rising atmospheric CO2 concentration has shown to affect plant physiology and chemistry by altering plant primary and secondary metabolisms. Nevertheless, the impacts of elevated CO2 on plant nutrients and functional components of tea remain largely unknown, which will likely affect tea quality and taste under climate change scenario. Being sources of nutrients and secondary chemicals/metabolites for herbivorous insects, the variation in foliar soluble nutrients and functional components of tea plants resulting from CO2 enrichment will further affect the herbivorous insects' occurrence and feeding ecology. In this study, the tea aphid, Toxoptera aurantii was selected as the phloem-feeding herbivore to study the effects of elevated CO2 on foliar soluble nutrients and functional components of tea seedlings, and the population dynamics of T. aurantii. The results indicated that elevated CO2 enhanced the photosynthetic ability and improved the plant growth of tea seedlings compared with ambient CO2, with significant increases in net photosynthetic rate (+20%), intercellular CO2 concentration (+15.74%), leaf biomass (+15.04%) and root-to-shoot ratio (+8.08%), and significant decreases in stomatal conductance (-5.52%) and transpiration rate (-9.40%) of tea seedlings. Moreover, elevated CO2 significantly increased the foliar content of soluble sugars (+4.74%), theanine (+3.66%) and polyphenols (+12.01%) and reduced the foliar content of free amino acids (-9.09%) and caffeine (-3.38%) of tea seedlings compared with ambient CO2. Furthermore, the relative transcript levels of the genes of theanine synthetase (+18.64%), phenylalanine ammonia lyase (+49.50%), s'-adenosine methionine synthetase (+143.03%) and chalcone synthase (+61.86%) were up-regulated, and that of caffeine synthase (-56.91%) was down-regulated for the tea seedlings grown under elevated CO2 relative to ambient CO2. In addition, the foliar contents of jasmonic acid (+98.6%) and salicylic acid (+155.6%) also increased for the tea seedlings grown under elevated CO2 in contrast to ambient CO2. Also, significant increases in the population abundance of T. aurantii (+4.24%-41.17%) were observed when they fed on tea seedlings grown under elevated CO2 compared to ambient CO2. It is presumed that the tea quality and taste will be improved owing to the enhanced foliar soluble nutrients and functional components of tea seedlings under the climate change scenario, especially on account of the rising atmospheric CO2 concentration, while the climate change may exacerbate the occurrence of tea aphid, T. aurantii, despite the enhanced secondary defensive chemicals manifested by the CO2 enrichment.

Interactive Effects of [CO2] and Temperature on Plant Chemistry of Transgenic Bt Rice and Population Dynamics of a Non-Target Planthopper, Nilaparvata lugens (Stål) under Different Levels of Soil Nitrogen.

Gaining a better understanding of the interactive effect of projected atmospheric CO2 level increase and the Earth's rising temperature on plant chemistry (nutritional and defensive characteristics) of transgenic crops is essential when attempting to forecast the responses of target and non-target insects to climate change. In this study, effects of carbon dioxide (CO2; elevated versus ambient), temperature (T; high versus low), and their interactions on leaf nitrogen content (N%) and C:N ratio of transgenic Bt rice and its non-Bt isoline grown under low- and high-N fertilizer were systematically analyzed together with the resulting insect population dynamics of a non-target planthopper Nilaparvata lugens (Stâl) in open-top-chamber experiments. The results indicated that under low-N treatment, elevated CO2 at low T (i.e., eCO2) (compared to ambient CO2 at low T, i.e., CK) significantly decreased N% and Bt-toxin content and significantly increased C:N ratio in leaf sheath and leaf of Bt rice, especially during the tillering stage, whereas inverse effects of high T were shown on the plant chemistry of Bt rice, especially during heading stage. The combination of elevated CO2 and high T (i.e., Combined) (in contrast to CK) significantly increased N% and decreased C:N ratio in leaf sheath of Bt rice during the heading stage under low-N fertilizer, while significantly decreased N% and increased C:N ratio in leaf of Bt rice during the tillering stage, regardless of fertilizer-N level, and significantly increased Bt-toxin content in leaf sheath and leaf during the tillering stage under both low- and high-N. Moreover, no discernable relationships between Bt-toxin content and N% or leaf C:N ratio were observed at any CO2 or N levels evaluated. Furthermore, transgenic treatment, temperature and fertilizer-N level interactions, and CO2 and fertilizer-N level interactions all significantly affected the population dynamics of N. lugens. Specifically, high-N significantly enhanced the population dynamics of N. lugens fed on non-Bt rice grown under eTemp and Bt cultivar significantly reduced the population dynamics of N. lugens under eCO2 regardless of N fertilizer levels. The study demonstrates that the planting of transgenic Bt rice would not increase the risk of increased N. lugens severity under the combined condition of elevated CO2 and increased temperature, particularly under moderate level of N fertility.

Plant Response and Economic Injury Levels for a Boll-Feeding Sucking Bug Complex on Cotton.

Whole-plant cage field experiments were conducted in 2014, 2015, and 2016 to characterize cotton injury from a species complex of boll-feeding sucking bugs represented by the verde plant bug, Creontiades signatus (Distant) (Hemiptera: Miridae), brown stink bug, Euschistus servus (Say), green stink bug, Acrosternum hilare (Say), and redbanded stink bug, Piezodorus guildinii (Westwood) (Hemiptera: Pentatomidae). Field-collected adult bugs were used to infest cotton plants previously maintained free of insect injury. Plants caged in groups of four were infested at mid-bloom and late-bloom for 7 d with four insect densities: 0 (control), 0.25 bugs per plant, 1 bug per plant, and 2 bugs per plant. Species and water stress conditions varied across years, allowing selective within-year comparisons. Response to feeding resulted in boll injury in the form of lint deterioration and cotton boll rot at mid- and late-bloom stages, and in water limiting and non-water limiting conditions. Although plant injury was apparent across a wide range of conditions, subsequent yield decline attributed to insect feeding was seen primarily under water limiting conditions when plants were infested at mid-bloom. For these conditions, significant yield-insect density relationships were used to calculate economic injury levels (EILs) for each species. EILs expressed as bugs per plant from lowest to highest were the brown stink bug (0.29-0.31 bugs per plant), redbanded stink bug (0.33), verde plant bug (0.49), and green stink bug (0.50). Given the variability observed among species, species-specific EILs may be used where the injurious species is known and combined for stink bugs (a common EIL of 0.34 bugs per plant) where multiple species occur. Verde plant bug was less damaging and can be considered separately, but its EIL was generally within a range of values for the boll-feeders encountered.

Effects of seed mixture sowing with transgenic Bt rice and its parental line on the population dynamics of target stemborers and leafrollers, and non-target planthoppers.

The widespread planting of insect-resistant crops has caused a dramatic shift in agricultural landscapes, thus raising concerns about the potential impacts on both target and non-target pests. In this study, we examined the potential effects of intra-specific seed mixture sowing with transgenic Bt rice (Bt) and its parental non-transgenic line (Nt) (100% Bt rice [Bt100 ], 5% Nt+95% Bt [Nt05 Bt95 ], 10% Nt+90% Bt [Nt10 Bt90 ], 20% Nt+80% Bt [Nt20 Bt80 ], 40% Nt+60% Bt [Nt40 Bt60 ] and 100% Nt rice [Nt100 ]) on target and non-target pests in a 2-year field trial in southern China. The occurrence of target pests, Sesamia inferens, Chilo suppressalis and Cnaphalocrocis medinalis, decreased with the increased ratio of Bt rice, and the mixture ratios with more than 90% Bt rice (Bt100 and Nt05 Bt95 ) significantly increased the pest suppression efficiency, with the lowest occurrences of non-target planthoppers, Nilaparvata lugens and Sogatella furcifera in Nt100 and Nt05 Bt95 . Furthermore, there were no significant differences in 1000-grain dry weight and grain dry weight per 100 plants between Bt100 and Nt05 Bt95 . Seed mixture sowing of Bt rice with ≤10% (especially 5%) of its parent line was sufficient to overcome potential compliance issues that exist with the use of block or structured refuge to provide most effective control of both target and non-target pests without compromising the grain yield. It is also expected that the strategy of seed mixture sowing with transgenic Bt rice and the non-transgenic parental line would provide rice yield stability while decreasing the insecticide use frequency in rice production.

Molecular Evidence for the Fitness of Cotton Aphid, Aphis gossypii in Response to Elevated CO2 From the Perspective of Feeding Behavior Analysis.

Rising atmospheric carbon dioxide (CO2) concentration is likely to influence insect-plant interactions. Aphid, as a typical phloem-feeding herbivorous insect, has shown consistently more positive responses in fitness to elevated CO2 concentrations than those seen in leaf-chewing insects. But, little is known about the mechanism of this performance. In this study, the foliar soluble constituents of cotton and the life history of the cotton aphid Aphis gossypii and its mean relative growth rate (MRGR) and feeding behavior were measured, as well as the relative transcript levels of target genes related appetite, salivary proteins, molting hormone (MH), and juvenile hormone, to investigate the fitness of A. gossypii in response to elevated CO2 (800 ppm vs. 400 ppm). The results indicated that elevated CO2 significantly stimulated the increase in concentrations of soluble proteins in the leaf and sucrose in seedlings. Significant increases in adult longevity, lifespan, fecundity, and MRGR of A. gossypii were found under elevated CO2 in contrast to ambient CO2. Furthermore, the feeding behavior of A. gossypii was significantly affected by elevated CO2, including significant shortening of the time of stylet penetration to phloem position and significant decrease in the mean frequency of xylem phase. It is presumed that the fitness of A. gossypii can be enhanced, resulting from the increases in nutrient sources and potential increase in the duration of phloem ingestion under elevated CO2 in contrast to ambient CO2. In addition, the qPCR results also demonstrated that the genes related to appetite and salivary proteins were significantly upregulated, whereas, the genes related to MH were significantly downregulated under elevated CO2 in contrast to ambient CO2, this is in accordance with the performance of A. gossypii in response to elevated CO2. In conclusion, rise in atmospheric CO2 concentration can enhance the fitness of A. gossypii by increasing their ingestion of higher quantity and higher quality of host plant tissues and by simultaneously upregulating the transcript expression of the genes related to appetite and salivary proteins, and then this may increase the control risk of A. gossypii under conditions of climate change in the future.

Influence of elevated CO2 on development and food utilization of armyworm Mythimna separata fed on transgenic Bt maize infected by nitrogen-fixing bacteria.

BACKGROUND: Bt crops will face a new ecological risk of reduced effectiveness against target-insect pests owing to the general decrease in exogenous-toxin content in Bt crops grown under elevated carbon dioxide (CO2). The method chosen to deal with this issue may affect the sustainability of transgenic crops as an effective pest management tool, especially under future atmospheric CO2 level raising. METHODS: In this study, rhizobacterias, as being one potential biological regulator to enhance nitrogen utilization efficiency of crops, was selected and the effects of Bt maize (Line IE09S034 with Cry1Ie vs. its parental line of non-Bt maize Xianyu 335) infected by Azospirillum brasilense (AB) and Azotobacter chroococcum (AC) on the development and food utilization of the target Mythimna separate under ambient and double-ambient CO2 in open-top chambers from 2016 to 2017. RESULTS: The results indicated that rhizobacteria infection significantly increased the larval life-span, pupal duration, relative consumption rate and approximate digestibility of M. separata, and significantly decreased the pupation rate, pupal weight, adult longevity, fecundity, relative growth rate, efficiency of conversion of digested food and efficiency of conversion of ingested food of M. separata fed on Bt maize, while here were opposite trends in development and food utilization of M. separata fed on non-Bt maize infected with AB and AC compared with the control buffer in 2016 and 2017 regardless of CO2 level. DISCUSSION: Simultaneously, elevated CO2 and Bt maize both had negative influence on the development and food utilization of M. separata. Presumably, CO2 concentration arising in future significantly can increase their intake of food and harm to maize crop; however, Bt maize infected with rhizobacterias can reduce the field hazards from M. separata and the application of rhizobacteria infection can enhance the resistance of Bt maize against target lepidoptera pests especially under elevated CO2.

Host-selection behavior and physiological mechanisms of the cotton aphid, Aphis gossypii, in response to rising atmospheric carbon dioxide levels.

Rising atmospheric carbon dioxide (CO2) levels can markedly affect the growth, development, reproduction and behavior of herbivorous insects, mainly by changing the primary and secondary metabolites of their host plants. However, little is known about the host-selection behavior and the respective intrinsic mechanism of sap-sucking insects in response to elevated CO2. In this experiment, the host-selection behavior, as well as the physiological mechanism based on the analysis of growth, development and energy substances, and the expression of the olfactory-related genes of the cotton aphid, Aphis gossypii, were studied under ambient (407.0 ±â€¯4.3 µl/L) and elevated (810.5 ±â€¯7.2 µl/L) CO2. The results indicated that the aphids reared under ambient and elevated CO2 did not differ in their level of preference for cotton seedlings, whatever the CO2 conditions in which the plants developed. However, aphids reared under elevated CO2 showed a greater ability to respond to the plant volatiles compared to aphids that developed under ambient CO2 (+23.3%). This suggests that rising atmospheric CO2 enhances the activity of host selection in this aphid. Compared with ambient CO2, elevated CO2 significantly increased aphid body weight (+36.7%) and the contents of glycogen (+18.9%), body fat (+14.6%), and amino acids (+16.8%) and increased the expression of odor-binding protein genes, OBP2 (+299.6%) and OBP7 (+47.4%), and chemosensory protein genes, CSP4 (+265.3%) and CSP6 (+50.9%), potentially enhancing the overall life activities and upregulating the olfactory ability of A. gossypii. We speculated that the rising atmospheric CO2 level would likely aggravate the damage caused by A. gossypii due to the higher potential host selection and increased general activity under future climate change.

Effects of seed mixture sowing with resistant and susceptible rice on population dynamics of target planthoppers and non-target stemborers and leaffolders.

BACKGROUND: The widespread planting of insect-resistant crops has caused a dramatic shift in agricultural landscapes, thus raising concerns about the potential impact on both target and non-target pests worldwide. In this study, we examined the potential effects of six seed mixture ratios of insect-resistance dominance [100% (R100), 95% (S05R95), 90% (S10R90), 80% (S20R80), 60% (S40R60), and 0% (S100)] on target and non-target pests in a 2-year field trial in southern China. RESULTS: The occurrence of the target pests Nilaparvata lugens and Sogatella furcifera decreased with an increase in the ratio of resistant rice, and mixture ratios with ≥90% resistant rice significantly increased the pest suppression efficiency, with the lowest occurrences of the non-target pests Sesamia inferens, Chilo suppressalis and Cnaphalocrocis medinalis for S100 and S10R90 seed mixture ratios. Furthermore, there were no significant differences in the 1000-grain dry weight and grain yield between R100 and other treatments with ≥80% resistant seeds in the mixture (S20R80, S10R90 and S05R95). CONCLUSION: S10R90 produced a good yield and provided the most effective control of both target and non-target pests, with the potential to significantly reduce the application of chemical pesticides for integrated pest management in paddy fields. It is further presumed that the strategy of seed mixture with resistant and susceptible rice would be advantageous for rice yield stability. © 2018 Society of Chemical Industry.

Impacts of elevated CO2 on exogenous Bacillus thuringiensis toxins and transgene expression in transgenic rice under different levels of nitrogen.

Recent studies have highlighted great challenges of transgene silencing for transgenic plants facing climate change. In order to understand the impacts of elevated CO2 on exogenous Bacillus thuringiensis (Bt) toxins and transgene expression in transgenic rice under different levels of N-fertilizer supply, we investigated the biomass, exogenous Bt toxins, Bt-transgene expression and methylation status in Bt rice exposed to two levels of CO2 concentrations and nitrogen (N) supply (1/8, 1/4, 1/2, 1 and 2 N). It is elucidated that the increased levels of global atmospheric CO2 concentration will trigger up-regulation of Bt toxin expression in transgenic rice, especially with appropriate increase of N fertilizer supply, while, to some extent, the exogenous Bt-transgene expression is reduced at sub-N levels (1/4 and 1/2N), even though the total protein of plant tissues is reduced and the plant growth is restricted. The unpredictable and stochastic occurrence of transgene silencing and epigenetic alternations remains unresolved for most transgenic plants. It is expected that N fertilization supply may promote the expression of transgenic Bt toxin in transgenic Bt rice, particularly under elevated CO2.

DNA barcoding and molecular evolution of mosquito vectors of medical and veterinary importance.

Mosquitoes (Diptera: Culicidae) are a key threat for millions of people worldwide, since they act as vectors for devastating pathogens and parasites. The standard method of utilisation of morphological characters becomes challenging due to various factors such as phenotypical variations. We explored the complementary approach of CO1 gene-based identification, analysing ten species of mosquito vectors belonging to three genera, Aedes, Culex and Anopheles from India. Analysed nucleotide sequences were found without pseudo genes and indels; they match with high similarity in nucleotide Basic Local Alignment Search Tool (BLASTn) search. The partial CO1 sequence of Anopheles niligricus was the first time record submitted to National Center for Biotechnology Information (NCBI). Mean intra- and interspecies divergence was found to be 1.30 and 3.83 %, respectively. The congeneric divergence was three times higher than the conspecifics. Deep intraspecific divergence was noted in three of the species, and the reason could be explained more accurately in the future by improving the sample size across different locations. The transitional and transversional substitutions were tested individually. Ts and Tv substitutions in all the 1st, 2nd and 3rd codons were estimated to be (0.44, 99.51), (40.35, 59.66) and (59.16, 40.84), respectively. Saturation of the sequences was resolved, since both the Ts and Tv exhibited a linear relationship suggesting that the sequences were not saturated. NJ and ML tree analysis showed that the individuals of the same species clustered together based on the CO1 sequence similarity, regardless of their collection site and geographic location. Overall, this study adds basic knowledge to molecular evolution of mosquito vectors of medical and veterinary importance and may be useful to improve biotechnological tools employed in Culicidae control programmes.

Intercrop movement of convergent lady beetle, Hippodamia convergens (Coleoptera: Coccinellidae), between adjacent cotton and alfalfa.

A 2-year study was conducted to characterize the intercrop movement of convergent lady beetle, Hippodamia convergens Guerin-Meneville (Coleoptera: Coccinellidae) between adjacent cotton and alfalfa. A dual protein-marking method was used to assess the intercrop movement of the lady beetles in each crop. In turns field collected lady beetles in each crop were assayed by protein specific ELISA to quantify the movement of beetles between the crops. Results indicated that a high percentage of convergent lady beetles caught in cotton (46% in 2008; 56% in 2009) and alfalfa (46% in 2008; 71% in 2009) contained a protein mark, thus indicating that convergent lady beetle movement was largely bidirectional between the adjacent crops. Although at a much lower proportion, lady beetles also showed unidirectional movement from cotton to alfalfa (5% in 2008 and 6% in 2009) and from alfalfa to cotton (9% in 2008 and 14% in 2009). The season-long bidirectional movement exhibited by the beetles was significantly higher in alfalfa than cotton during both years of the study. The total influx of lady beetles (bidirectional and unidirectional combined) was significantly higher in alfalfa compared with that in cotton for both years. While convergent lady beetles moved between adjacent cotton and alfalfa, they were more attracted to alfalfa when cotton was not flowering and/or when alfalfa offered more opportunities for prey. This study offers much needed information on intercrop movement of the convergent lady beetle that should facilitate integrated pest management decisions in cotton utilizing conservation biological control.

Nanoparticles in the fight against mosquito-borne diseases: bioactivity of Bruguiera cylindrica-synthesized nanoparticles against dengue virus DEN-2 (in vitro) and its mosquito vector Aedes aegypti (Diptera: Culicidae).

Mosquitoes are blood-feeding insects serving as the most important vectors for spreading human pathogens and parasites. Dengue is a viral disease mainly vectored through the bite of Aedes mosquitoes. Its transmission has recently increased in urban and semi-urban areas of tropical and subtropical regions worldwide, becoming a major international public health concern. There is no specific treatment for dengue. Its prevention and control solely depend on effective vector control measures. Mangrove plants have been used in Indian traditional medicine for a wide array of purposes. In this research, we proposed a method for biosynthesis of antiviral and mosquitocidal silver nanoparticles (AgNP) using the aqueous extract of Bruguiera cylindrica leaves. AgNP were characterized using a variety of biophysical analyses, including UV-visible spectrophotometry, Fourier-transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. Bruguiera cilyndrica aqueous extract and green-synthesized AgNP were tested against the primary dengue vector Aedes aegypti. AgNP were the most effective. LC50 values ranged from 8.93 ppm (larva I) to 30.69 ppm (pupa). In vitro experiments showed that 30 µg/ml of AgNP significantly inhibited the production of dengue viral envelope (E) protein in vero cells and downregulated the expression of dengue viral E gene. Concerning nontarget effects, we observed that the predation efficiency of Carassius auratus against A. aegypti was not affected by exposure at sublethal doses of AgNP. Predation in the control was 71.81 % (larva II) and 50.43 % (larva III), while in an AgNP-treated environment, predation was boosted to 90.25 and 76.81 %, respectively. Overall, this study highlights the concrete potential of green-synthesized AgNP in the fight against dengue virus. Furthermore, B. cylindrica-synthesized AgNP can be employed at low doses to reduce larval and pupal population of A. aegypti, without detrimental effects of predation rates of mosquito predators, such as C. auratus.