Insect Mortality Research Articles

Efficacy and profitability of five insecticides for the control of the stem girdler, Analeptes trifasciata (Coleoptera: Cerambycidae) on cashew

Current management practices for Analeptes trifasciata Fabricius (Coleoptera: Cerambycidae) on cashew are cultural, however, these are largely inadequate. Thus compelling the indiscriminate application of unapproved chemicals for its control in Nigeria. As part of a coordinated research effort to develop and integrate a diverse array of control options, we screened the effects of imidacloprid, dimethoate, chlorpyrifos, thiamethoxam and cypermethrin on A. trifasciata management on cashew by laboratory and field trials carried out in 2018 and 2019. The marginal rate of return for each insecticide was calculated to determine the cost-benefit ratio. In the laboratory toxicity trial, all the insecticides showed a dose dependent effect and caused significantly higher insect mortality compared to the control. Thiamethoxam 25WG significantly reduced A. trifasciata populations, improved yield and recorded stable marginal returns of 6.24% and 8.52% in the first and second year respectively. The findings and implications of this study will prove useful as a part of an integrated management system for A. trifasciata.

Development and evaluation of triple gene transgenic cotton lines expressing three genes (Cry1Ac-Cry2Ab-EPSPS) for lepidopteran insect pests and herbicide tolerance

Cotton is an international agricultural commodity and the main cash crop of Pakistan of which quality and quantity are subject to various whims of nature. Climate change, insect pest complex, and weeds are reducing its productivity. Here, we have developed triple gene cotton containing EPSPS gene along with two Bt toxin genes Cry1Ac and Cry2Ab using a strategy where all three genes are cloned in the same T-DNA, followed by successful cotton transformation via Agrobacterium-mediated transformation. This strategy has been developed to help cotton breeders in developing new cultivars by incorporating these genes into the non-transgenic or single Bt (Cry1Ac) gene cotton background where all three genes will inherit together. The expression of all three proteins was confirmed through immunostrips and was quantified through enzyme-linked immunosorbent assay (ELISA). The spatio-temporal expression of Bt protein in different parts of triple gene NIBGE cotton plants was determined. Maximum expression was found in leaves followed by seeds and boll rinds. Insect bioassays with cotton bollworms (Helicoverpa armigera), armyworms (Spodoptera litura), and pink bollworms (Pectinophora gossypiella) showed more than 90% mortality. The best performing line (NIBGE-E2) on the basis of spatiotemporal expression, glyphosate assays, and insect mortality data, was used for event characterization by using the genome sequencing approach. The event was successfully characterized and named NIBGE 20-01. A diagnostics test based on event-specific PCR was developed and its ability to distinguish NIBGE 20-01 event from other commercial transgenic cotton events was confirmed. To confirm stable expression of all three proteins in the field conditions, homozygous transgenic lines were grown in the field and the expression was confirmed through immunostrip assays. It was found that all three genes are expressed under field conditions. To show that all three genes are inherited together upon crossing with local elite cotton lines, the F1 generation was grown under glasshouse and field conditions. The expression of all three genes was confirmed under field conditions. Our results showed that transgenic cotton with three genes cloned in the same T-DNA can express all genes and can be conveniently transferred into elite cotton lines through a single cross.

Co-expression of Cocculus hirsutus trypsin inhibitor with Cry protein reduces resistant development in targeted insects along with complete mortality

Crop losses due to various insects are a global problem. For decades, several types of research have been ongoing on this issue with limited success due to co-evolving insect pests. In this study, the modified Cocculus hirsutus trypsin inhibitor (ChTI), a serine proteinase inhibitor gene encoding a natural plant defense protein, inhibiting the digestive proteases in the midgut of phytophagous insects, was expressed alone and pyramided with the Bt-toxin Cry1Ab, to reduce resistance development against two herbivorous insects, Helicoverpa armigera and Spodoptera litura with complete insect mortality. In-silico analysis revealed the structural and conformational stability of ChTI protein. Furthermore, 80 % weight loss and reduced larval growth were observed in both insects challenged with 200 µg of the purified ChTI protein. The ChTI protein trypsin inhibitor activity was observed up to 5.5 U/mg expressed in E. coli, 3.0–3.5 U/mg in transgenic Arabidopsis plants, and up to 2.5–3.0 U/mg in transgenic tomato plants. Transgenic plants (Arabidopsis and tomato) were developed by pyramiding ChTI, and cry1Ab genes showed 100 % mortality against both insects while the transgenic plants expressing the ChTI gene alone showed up to 70 % mortality without yield penalty. Transcriptomic analysis of the transgenic plants further revealed that other pathogenesis-related defense proteins were also differently regulated.

Identification and environment-friendly biocontrol potential of five different bacteria against Aphis punicae and Aphis illinoisensis (Hemiptera: Aphididae).

The current work is aimed at isolating and identifying new Entomopathogenic bacterium (EPB) strains associated with Steinernema feltiae and assessing the EPB's biocontrol potential on Aphis punicae and Aphis illinoisensis adults in the laboratory. From S. feltiae, five bacterial isolates were isolated and molecularly characterized. Lysinibacillus xylanilyticus strain TU-2, Lysinibacillus xylanilyticus strain BN-13, Serratia liquefaciens strain TU-6, Stenotrophomonas tumulicola strain T5916-2-1b, and Pseudochrobactrum saccharolyticum strain CCUG are the strains. Pathogenicity tests demonstrated that bacterial cells were more toxic against the two aphid species than bacterial cell-free supernatants. S. tumulicola strain T5916-2-1b cells and filtrate were reported to have the strongest potential to kill A. punicae and A. illinoisensis individuals within 6 h after treatment, with 100% mortality of both insects 24 and 48 h after treatment. Based on the results of the study, it looked like endogenous Steinernema-associated EPB could be used directly as a biocontrol agent for A. punicae and A. illinoisensis.

Resporulation of Calcium Alginate Encapsulated Metarhizium anisopliae on Metham®-Fumigated Soil and Infectivity on Larvae of Tenebrio molitor.

Metarhizium anisopliae infects and kills a large range of insects and is a promising biocontrol agent to manage soil insects, such as wireworm in sweetpotato. The presence of other soil microbes, which exhibit competitive fungistasis, may inhibit the establishment of M. anisopliae in soil. Microbially depleted soil, for example, sterilized soil, has been shown to improve the resporulation of the fungus from nutrient-fortified M. anisopliae. Prior to planting, sweetpotato plant beds can be disinfected with fumigants, such as Metham®, to control soil-borne pests and weeds. Metham® is a broad-spectrum soil microbial suppressant; however, its effect on Metarhizium spp. is unclear. In the research presented here, fungal resporulation was examined in Metham®-fumigated soil and the infectivity of the resulting granule sporulation was evaluated on mealworm, as a proxy for wireworm. The fungal granules grown on different soil treatments (fumigated, field and pasteurized soil) resporulated profusely (for example, 4.14 × 107 (±2.17 × 106) conidia per granule on fumigated soil), but the resporulation was not significantly different among the three soil treatments. However, the conidial germination of the resporulated granules on fumigated soil was >80%, which was significantly higher than those on pasteurized soil or field soil. The resporulated fungal granules were highly infective, causing 100% insect mortality 9 days after the inoculation, regardless of soil treatments. The results from this research show that the fungal granules applied to soils could be an infective inoculant in sweetpotato fields in conjunction with soil fumigation. Additional field studies are required to validate these results and to demonstrate integration with current farming practices.

Responses of diamondback moth to diverse entomopathogenic fungi collected from non-agricultural habitats - Effects of dose, temperature and starvation.

We evaluated the virulence of Beauveria bassiana and Metarhizium isolates from soil collected across different vegetation types in Queensland, against chlorantraniliprole-resistant and insecticide-susceptible diamondback moth (DBM) larvae. Host insecticide resistance status had no effect on susceptibility to the pathogens when conidia were topically applied to larvae in the laboratory, and one B.bassiana isolate was significantly more virulent to larvae than the others (seven days after inoculation). The influence of temperature (15, 20, 25 or 30°C): (i) at the point of host inoculation with conidia and (ii) when the pathogens had already initiated infection and were proliferating in the host haemocoel, was determined experimentally for its influence on virulence, disease progression, and sporulation. Temperature at inoculation had a greater effect on host insect mortality than it did when the fungus was already proliferating in the host haemocoel. The rearing temperature of hosts prior to inoculation had a greater effect on host susceptibility to disease than starvation of the larvae at the time of inoculation. Our results also show that each fungal isolate has its own temperature relations and that these can vary considerably across isolates, and at different points in the pathogen life cycle (germination and cuticular penetration versus growth in the host haemocoel). Temperature also had an idiosyncratic effect, across isolates and across the variables typically used to assess the potential of fungal entomopathogens as biological control agents (time to death, mortality and sporulation rates). This study demonstrates that in addition to pathogenicity and virulence, the temperature relationships of each fungal isolate when infecting insects needs to be taken into account if we are to understand their ecology and use them effectively in pest management.

Non-thermal plasma as emerging technology for Tribolium castaneum pest-management in stored grains and flours

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is a major secondary pest on wheat stored in metal bins, silo-bags and is also frequently found in wheat products such as flour. Non-thermal plasmas (NTPs) are (quasi-neutral) partially ionized gases that may be produced by a variety of electrical discharges. We propose the use of an atmospheric pressure dielectric barrier discharge (DBD) as an emerging technology in post-harvest integrated pest management. To this aim, a series of experiments were performed in order to test the lethality of such plasmas on three life stages of T. castaneum by measuring insect mortality and their impacts on physiological and biochemical parameters affecting insect fitness. The different NTP treatments were performed by increasing the time of exposure to either O2 or N2 used as carrier gases. After 24 hours, high levels of mortality (30–100%) were reached for each applied treatment, in both larval and adult populations. Mortality seems to be related to a significant water content loss and redox imbalance. Atomic force microscopy (AFM) scanning of the prothoracic surface showed that nitrogen causes more severe damage than oxygen. As a consequence of the cuticle damage, the quinone-containing secretions of the prothoracic and abdominal glands were affected. We also carried out experiments on egg-containing flours to test the ovicidal activity of NTP. The flours were evaluated at three and twelve weeks after treatments. A 3-min nitrogen treatment showed ovicidal properties, while the remaining NTPs partially killed the eggs and delayed the emergence of larvae and adults. In conclusion, we propose an inexpensive physical treatment, which controls the entire life cycle of a major grain pest, avoiding chemical residues.

Selecting thermal tolerant strains of entomopathogenic fungi to control Ceratitis capitata (Wiedeman) in tropical semi-arid conditions

Although it is usual, using constant temperature to select thermo-tolerant entomopathogen (EF) strains could discard potentially efficient biocontrol agents (BCA). The objectives of this work were to select virulent EF against C. capitata and evaluate the effect of continual and fluctuating temperature on fungal development and virulence. Initial experiments compared the effect of constant temperatures (CT) (20, 25, 30, 35, and 38 °C) and simulated intraday temperature variation (SIV) (20–38 °C) over conidial germination, mycelial growth, and insect mortality. Temperatures ≥ 35 °C in the CT experiments significantly reduced conidial germination, and the strains of Beauveria bassiana BbLCB81 and BbLCB289 showed the highest conidial germination and mycelial growth. SIV also caused a significant effect, and M. anisopliae MaLCB62 showed the highest conidial germination and mycelial growth. Applying EF strains as toxic baits in CT showed that the highest mortality occurred at 30 °C, and BbLCB62 and BbLCB289 showed the largest insect mortality at 38 °C. Survival analysis showed a slight increase in the average survival time (ST50) at 30 °C. In SIV conditions, MaLCB56 and BbLCB289 were highly virulent. In a field cage experiment at a warmer temperature, BbLCB62 was the most virulent strain according to the M-C test comparing survival curves. However, confirmed accumulated mortality on the 10th day was similar to the moderately tolerant BbLCB289 (Tukey, p < 0.05). During winter, all EF strains showed similar mortality on 10th day (Tukey, p < 0.05), but a significant difference in the survival analysis. EF strains with mild temperature tolerance can be applied in seasons with high temperatures, and BbLCB62 and BbLCB289 were selected as potential biocontrol agents against C. capitata.

Evaluating hydrologic impact from concurrent insect and fire disturbances

Study RegionRio Grande Headwaters Study FocusIn the western US wildfires are increasing in duration and frequency, insect inducted forest mortality is widespread, and both wildfires and forest mortality are exacerbated by drought. These forest disturbances have hydrologic implications that are not always evident in the post-disturbance water budget – especially at the watershed outlet. In this study we evaluate two snow dominated paired catchment systems in the forested Rio Grande headwaters – where each impacted site was severely burned by the 2013 West Fork Complex (WFC) fire, and all sites (including control sites) were severely impacted by spruce beetle induced forest mortality from approximately 2005 to 2010. A combination of observed runoff, reanalysis precipitation, and remotely sensed evapotranspiration and vegetation metrics are used to evaluate the post-fire surface water budget (runoff and evapotranspiration) in paired catchments. New hydrological insights for the regionSignificant changes in post-fire surface water budget partitioning are not detectable, which is likely the result of decreased evapotranspiration due to insect-induced vegetation change in all study catchments prior to the fire. Our findings highlight the importance of considering overlapping disturbances, especially in paired catchment studies, and in alpine and subalpine forest regions like the Rio Grande headwaters that are increasingly prone to drought, insect mortality and wildfire. Our results also illustrate that insect induced forest mortality has significant implications for post-fire hydrology studies that rely on control-impact or before-and-after comparisons.

Bioactivity of two plant products against Tribolium castaneum infesting wheat grains and their impact on some biological parameters in Japanese quail

Several experiments were carried out in the current study to assess the efficacy of Caraway seed (Carum carvi) and petitgrain lemon (Citrus aurantium) plants against the stored product insect Tri­bolium casteneum. To measure some factors, various plant preparations (powder, oil, extract, and new formulation) with concentrations of 5-10-15- and 20% oils and extracts and 5-10-20- and 40% powder and their formulations were used (biological effect, insect mortality rate). Furthermore, to assess the negative effects of tested materials on certain biochemical parameters of Japanese quail (Coturnix coturnix japonica). There have been no significant effects of oral treatment on creatinine level while significant effect were noticed on total protein, total anti-oxidants and Serum glutamic pyruvic transaminase (SGPT) levels in quail. In every treatments for quail, caraway was more effective than petitgrain lemon on some biological parameters. The determination of some biological parameters revealed that treatment after two weeks was affected in comparison to treatment four weeks later. The findings suggest that the tested plant products could be used to protect wheat grains from T. castaneum adults in Egyptian storage facilities.

Evaluation of entomopathogenic nematode (EPN) isolate, Heterorhabditis indica of Vidarbha region, against the tobacco cutworm, Spodoptera litura

The present study on pathogenicity of entomopathogenic nematodes against Spodoptera litura in laboratory conditions was undertaken during 2020-21, with the aim to ascertain the effectiveness of entomopathogenic nematodes, against an obnoxious cosmopolitan pest S. litura. Experiments were conducted by using entomopathogenic nematode (EPN) isolate Heterorhabditis indica (CICR-Guava), on filter paper, against Galleria mellonella and S. litura at the treatment dose of 10, 20, 30, 40, 60, 80 and 100 IJs/100µl along with control (Sterile distilled water). The results of our study revealed that, EPN isolate H. indica (CICR-Guava) caused 100% mortality at the treatment dose of 40 IJs/100µl within 72 h of infection in 5th instar larvae of G. mellonella and in case of S. litura, 100% mortality was recorded within 72 h of infection at the treatment dose of 100 IJs/100µl in 3rd instar larvae, which was found more susceptible. The median lethal concentration of H. indica (CICR-Guava) for 5th instar larvae was 2.29 IJs/100µl. The result of reproductive potential of isolates of entomopathogenic nematodes revealed that the highest yield was obtained from 5th instar larvae of G. mellonella at treatment dose of 100 IJs/100µl 278667 IJs per larva. In case of S. litura, the highest yield obtained was 152533 IJs. It could be concluded that, there was a positive correlation between nematode treatment concentration, time of exposure and the insect mortality of the tobacco cut worm and multiplication rate of IJs increased with increase of exposure time and size of larvae. This EPN isolate, H. indica (CICR-Guava) can be suggested as biocontrol agents for the control of S. litura in the Vidarbha region.

Root Colonization by Fungal Entomopathogen Systemically Primes Belowground Plant Defense against Cabbage Root Fly.

Entomopathogenic fungi infect insects via spores but also live inside plant tissues as endophytes. Frequently, colonization by entomopathogens provides plants with increased resistance against insects, but the mechanisms are little understood. This study investigated direct, local, and systemic root-mediated interactions between isolates of the fungus Metarhizium brunneum and larvae of the cabbage root fly (CRF) Delia radicum attacking Brassica napus plants. All fungal isolates infected CRF when conidia were present in the soil, leading to 43–93% mortality. Locally, root-associated M. brunneum isolates reduced herbivore damage by 10–20% and in three out of five isolates caused significant insect mortality due to plant-mediated and/or direct effects. A split-root experiment with isolate Gd12 also demonstrated systemic plant resistance with significantly reduced root collar damage by CRF. LC-MS analyses showed that fungal root colonization did not induce changes in phytohormones, while herbivory increased jasmonic acid (JA) and glucosinolate concentrations. Proteinase inhibitor gene expression was also increased. Fungal colonization, however, primed herbivore-induced JA and the expression of the JA-responsive plant defensin 1.2 (PDF1.2) gene. We conclude that root-associated M. brunneum benefits plant health through multiple mechanisms, such as the direct infection of insects, as well as the local and systemic priming of the JA pathway.

Population growth of stored product insects on wheat containing wheat bugs

In the present study we examined the effect of the presence of wheat bugs (Eurygaster spp. and Aelia spp.) on wheat on the mortality, progeny production and infestation patterns of three stored product insects, Trogoderma granarium Everts, Sitophilus oryzae (L.) and Rhyzopertha dominica (F.). The bioassays were carried out under laboratory conditions (25 ± 1 °C, 60 ± 5% relative humidity (r.h.) and continuous darkness), and 20 adults of each stored product insect species were placed in plastic vials (3 cm in diameter, 8 cm in height) with wheat that contained different numbers of wheat bugs (1, 2, 3 or 5 adults). In general, stored product insect mortality was not affected by the presence of wheat bugs, in most of the combinations examined. In most of the cases, mortality of T. granarium was higher on wheat which contained Eurygaster that wheat containing Aelia, while progeny production of S. oryzae was also affected by the presence of wheat bugs. Frass production for all combinations was similar, but greater frass production occurred by all combinations of R. dominica with Aelia. Overall, the presence of wheat bugs could affect the survival and the progeny production of both S. oryzae and R. dominica. In general, our results showed that beetle longevity and fecundity can be affected by the presence of wheat bugs that are likely to be present in stored grains after harvest.Graphical abstract

Biocidal potential of indigenous isolates of Entomopathogenic Nematodes (EPNs) against tobacco cutworm, Spodoptera litura Fabricius (Lepidoptera: Noctuidae)

BackgroundEntomopathogenic nematodes (EPNs) have been regarded as the most convenient strategy for insect pest management. The native strains of EPNs: Heterorhabditis bacteriophora EUPT-SD, H. bacteriophora EUPT-R, H. bacteriophora EUPT-KN, H. bacteriophora EUPT-K and H. bacteriophora EUPT-H isolated from mid-Himalayan region of Himachal Pradesh were tested in laboratory for their multiplication and virulence against 3rd and 4th larval instars of the tobacco cutworm, Spodoptera litura Fabricius (Lepidoptera: Noctuidae), the serious polyphagous pest affecting a wide range of agricultural crops worldwide.ResultsAll the EPN strains were effective against 3rd and 4th larval instars of S. litura. Insect mortality reached 90–96% after 96 h at nematode concentrations of 150 infective juveniles (IJs)/ml. The insect mortality was also recorded at low concentrations of IJs, but the most exposure period was required. High virulence was shown by H. bacteriophora EUPT-SD 96 and 94%, followed by H. bacteriophora EUPT-R 92 and 90%, H. bacteriophora EUPT-KN 92 and 90%, H. bacteriophora EUPT-K 92 and 90% and H. bacteriophora EUPT-H 92 and 90%, respectively, against 3rd and 4th larval instars in terms of reproductive potential and killing. All the insects were alive in the absolute control.ConclusionUtilization of EPNs for the management of S. litura may be the best method to overcome the insect resistance problems and to manage the population of this insect pest. It may be an effective method and may be a partial substitute of synthetic insecticides, thus minimizing the excessive use of synthetic chemicals. The results demonstrated the potential of indigenous EPNs isolates against S. litura, but before further recommendation, multiplication field trials need to be conducted to confirm their efficacy at farm level.

Efficacy of Metarhizium anisopliae and (E)-2-hexenal combination using autodissemination technology for the management of the adult greenhouse whitefly, Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae).

The efficiency of an autodissemination technique in controlling adult whiteflies, Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae) on tomato, Solunum lycopersicum was investigated with previously identified potent fungal isolates of Metarhizium anisopliae ICIPE 18, ICIPE 62 and ICIPE 69 under screenhouse or semi-field conditions. The autodissemination device was inoculated with dry conidia of the M. anisopliae isolates, while control insects were exposed to a fungus-free device. Sampling for conidia uptake, conidial viability and persistence, and insect mortality was done at 1, 2, 3, 5 and 8 days post-exposure, and collected insects were monitored for mortality over ten days. Overall, mortality was higher in insects exposed to ICIPE 18 (62.8%) and ICIPE 69 (61.8%) than in those exposed to ICIPE 62 (42.6%), with median lethal times, (LT50) ranging between 6.73-8.54 days. The control group recorded the lowest mortality rates (18.9%). A general linear reduction in conidial viability with exposure time was observed, although this was more pronounced with M. anisopliae ICIPE 62. Insects exposed to M. anisopliae ICIPE 69 also recorded the highest conidia uptake, hence selected for further evaluation with a T. vaporariorum attractant volatile organic compound, (E)-2-hexenal. The volatile inhibited fungal germination in laboratory compatibility tests, therefore, spatial separation of M. anisopliae ICIPE 69 and (E)-2-hexenal in the autodissemination device was conducted. The inhibitory effects of the volatile were significantly reduced by spatial separation at a distance of 5 cm between the fungus and the volatile, which was found to be more suitable and chosen for the subsequent experiments. Results showed that (E)-2-hexenal did not influence conidia uptake by the insects, while fungal viability and the subsequent mortality variations were more related to duration of exposure. The fungus-volatile compatibility demonstrated with spatial separation provides a basis for the optimisation of the volatile formulation to achieve better T. vaporariorum suppression with an excellent autodissemination efficiency when used in the management of whiteflies under screenhouse conditions.

FAR knockout significantly inhibits Chilo suppressalis survival and transgene expression of double-stranded FAR in rice exhibits strong pest resistance.

Chilo suppressalis is one of the most prevalent and damaging rice pests, causing significant economic losses each year. Chemical control is currently the primary method of controlling C. suppressalis. However, the indiscriminate use of chemical insecticides increases pest resistance, pollutes the environment and poses a significant health threat to humans and livestock, highlighting the need to find safer, more pest-specific and more effective alternatives to pest control. Plant-mediated RNA interference (RNAi) is a promising agricultural pest control method that is highly pest-specific and has less of an impact on the environment. Using multi-sgRNAs/Cas9 technology to delete Fatty acyl-CoA reductase (FAR) of C. suppressalis in the G0 generation, we show that downregulating FAR transcription may significantly increase the mortality rate and darken the epidermis of C. suppressalis compared with the control. Subsequently, we developed dsFAR transgenic rice lines using Agrobacterium-mediated genetic transformation and then screened three strains expressing dsFAR at high levels using transcriptional level analysis. Using transgenic rice stems, a laboratory feeding bioassay indicated that at least one line (L#10) displayed a particularly high level of insect resistance, with an insect mortality rate of more than 80%. In the field trials, dsFAR transgenic rice displayed high levels of resistance to C. suppressalis damage. Collectively, these results suggest the potential of a new environment-friendly, species-specific strategy for rice pest management.

Whitefly-tolerant transgenic common bean (Phaseolus vulgaris) line.

Common bean (Phaseolus vulgaris L.) is a staple food in Brazil with both nutritional and socioeconomic importance. As an orphan crop, it has not received as much research attention as the commodity crops. Crop losses are strongly related to virus diseases transmitted by the whitefly Bemisia tabaci, one of the most important agricultural pests in the world. The main method of managing whitefly-transmitted viruses has been the application of insecticides to reduce vector populations. Compared to chemical vector control, a more sustainable strategy for managing insect-borne viruses is the development of resistant/tolerant cultivars. RNA interference has been applied to develop plant lines resistant to the whitefly in other species, such as tomato, lettuce and tobacco. Still, no whitefly-resistant plant has been made commercially available to date. Common bean is a recalcitrant species to in vitro regeneration; therefore, stable genetic transformation of this plant has been achieved only at low frequencies (<1%) using particle bombardment. In the present work, two transgenic common bean lines were obtained with an intron-hairpin construct to induce post-transcriptional gene silencing against the B. tabaci vATPase (Bt-vATPase) gene, with stable expression of siRNA. Northern blot analysis revealed the presence of bands of expected size for siRNA in leaf samples of the line Bt-22.5, while in the other line (11.5), the amount of siRNA produced was significantly smaller. Bioassays were conducted with both lines, but only the line Bt-22.5 was associated with significant mortality of adult insects (97% when insects were fed on detached leaves and 59% on the whole plant). The expression of the Bt-vATPase gene was 50% lower (p < 0.05) in insects that fed on the transgenic line Bt-22.5, when compared to non-transgenic controls. The transgenic line did not affect the virus transmission ability of the insects. Moreover, no effect was observed on the reproduction of non-target organisms, such as the black aphid Aphis craccivora, the leafminer Liriomyza sp. and the whitefly parasitoid Encarsia formosa. The results presented here serve as a basis for the development of whitefly-tolerant transgenic elite common bean cultivars, with potential to contribute to the management of the whitefly and virus diseases.

Changes in immune responses, gene expression, and life table parameters of Helicoverpa armigera Hübner fed on a diet containing the saponin of tea plant, Camellia sinensis.

Saponins cause mortality in insects by reducing food absorption and movement in the gut, which may be used to exploit the control of insect pests. In the current study, tea seed saponin (TSS) was extracted and then added to the artificial diets of Helicoverpa armigera. Pre-ovipositional period of the TSS-treated individuals increased while longevity and fecundity decreased compared to control. There was a significant reduction of the treated individuals in the life table parameters of TSS-treated Individuals including net reproduction rate (R0 ), intrinsic rate of population increase (r), finite rate of increase (λ), and gross reproduction rate (GRR). Also, we found that saponin suppressed the immune system by reducing the total hemocyte count, immune-related gene expression, and phenoloxidase activity. Our results demonstrated a lower expression of cecropin gene in the treated larvae with TSS while no significant differences were observed in attacin gene. Our results clearly showed that feeding ofH. armigera larvae in the diet containing TSS significantly reduced demographic parameters, forced insects to obtain more time to complete one generation, and caused vulnerabilities against pathogens. These discrepancies alleviated nutrient uptake of the larvae and disrupted their feeding and growth. Hence, a proper formulation with a desirable concentration would be prepared and applied in the fields suffering H. armigera damage to monitor insecticidal efficiency of TSS.

Spatial Distribution and Retention in Loblolly Pine Seedlings of Exogenous dsRNAs Applied through Roots.

Exogenously applied double-stranded RNA (dsRNA) can induce potent host specific gene knockdown and mortality in insects. The deployment of RNA-interference (RNAi) technologies for pest suppression is gaining traction in both agriculture and horticulture, but its implementation in forest systems is lagging. While numerous forest pests have demonstrated susceptibility to RNAi mediated gene silencing, including the southern pine beetle (SPB), Dendroctonus frontalis, multiple barriers stand between laboratory screening and real-world deployment. One such barrier is dsRNA delivery. One possible delivery method is through host plants, but an understanding of exogenous dsRNA movement through plant tissues is essential. Therefore, we sought to understand the translocation and persistence of dsRNAs designed for SPB throughout woody plant tissues after hydroponic exposure. Loblolly pine, Pinus taeda, seedlings were exposed to dsRNAs as a root soak, followed by destructive sampling. Total RNA was extracted from different tissue types including root, stem, crown, needle, and meristem, after which gel electrophoresis confirmed the recovery of the exogenous dsRNAs, which were further verified using Sanger sequencing. Both techniques confirmed the presence of the exogenously applied target dsRNAs in each tissue type after 1, 3, 5, and 7 d of dsRNA exposure. These findings suggest that root drench applications of exogenous dsRNAs could provide a viable delivery route for RNAi technology designed to combat tree feeding pests.

Evaluation of insecticidal activity of Lavandula coronopifolia essential oil against the Mediterranean fruit fly Ceratitis capitata Wiedemann (Diptera: Tephritidae)

The present investigation was aimed to assess the toxicity of Lavandula coronopifolia (Lamiaceae) essential oil (EO) against adult Mediterranean fruit fly. Ceratitis capitata (Wiedemann) (Diptera Tephritidae) is one of the most destructive fruit pests in Morocco.Essential oil was extracted from leaves of lavender (L. coronopifolia) by hydrodistillation and analyzed by GC and GC/MS. The main compound is carvacrol (48.9%) followed by E-caryophyllene (10.8%) and caryophyllene oxide (7.7%).The EO was examined in the laboratory for 7 days to assess their activity against Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) adults by ingestion toxicity. We also conducted oviposition bioassay in the laboratory, testing the oviposition deterrence of essential oils.The oil showed 75% mortality of adults at the concentration of 150 µl/g after 24 h of exposure. Great differences in insect mortality were observed depending on the sex of the insects, oil concentrations and exposure time.The ingestion toxicity potential of L. coronopifolia on adults was higher (LC 50 = 86.34 µl/g, LC 95 = 236.07 µl/g) and also, deterred significantly the oviposition by C. capitata, reducing the number of eggs laid by about 94% compared with the controls for the concentration of 55µl/g.Results suggested that L. coronopifolia EOs may have potential as a control agent against this major stored product insect.