Clip Cages Research Articles

Transgenic cotton expressing Allium sativum leaf agglutinin exhibits resistance to whiteflies and aphids without negative effects on ladybugs

Cotton (Gossypium hirsutum L.) is of great economic importance as a cultivated crop in many parts of the world. In addition to being a pillar of the textile industry, cotton and its byproducts are used for livestock feed, seed oil, and other products. Bacillus thuringiensis crystal toxin (Bt) expression in cotton provides effective protection against chewing insects but does not defend plants from piercing/sucking insect pests. With the aim to create transgenic plants with resistance against piercing/sucking pests, we used Agrobacterium-mediated genetic transformation of cotton cultivar Coker 312 to express the Allium sativum leaf agglutinin (ASLA) gene from the phloem-specific rolC promoter. The ASLA transgene was stably inherited and showed Mendelian segregation in the T1 generation. Transgenic lines, expressing the ASLA gene, showed explicit resistance against major sap-sucking pests. Green peach aphid (Myzus persicae Sulzer) choice assays showed that 75% of aphids preferred untransformed cotton plants relative to those expressing the ASLA gene. In detached leaf bioassays, plants expressing ASLA caused 82% aphid mortality and 44–53% reduction in fecundity. Clip cage bioassays with whiteflies (Bemisia tabaci Gennadius) showed 74–82% mortality and 44–60% decrease in fecundity due to ASLA gene expression. In whole plant bioassays, whiteflies showed 77% mortality and a 54% decrease in fecundity on ASLA transgenics. Importantly, we did not observe a negative effect of the ASLA gene on ladybugs (Coccinella septempunctata) that consumed these whiteflies. Together, our findings demonstrate the potential of ASLA-transgenic cotton for providing protection against two devastating insect pests, whiteflies and aphids. The ASLA-transgenic cotton appears promising for direct commercial cultivation besides serving as a potential genetic resource in recombination breeding.

Using Environmental DNA to Detect and Identify Sweetpotato Whitefly Bemisia argentifolii and Twospotted Spider Mite Tetranychus urticae in Greenhouse‐Grown Tomato Plants

ABSTRACTEnvironmental DNA (eDNA) consists of genetic material shed by living organisms, including those that are deceased, offering a unique opportunity to detect and identify terrestrial insect pests without requiring visual identification. The sweetpotato whitefly, Bemisia argentifolii, and the twospotted spider mite, Tetranychus urticae, are notorious for causing crop losses through virus transmission and direct feeding. Our study aimed to: (1) assess the effectiveness of B. argentifolii literature‐based PCR primers compared to newly developed primers for eDNA amplification, (2) evaluate the sensitivity of conventional PCR (cPCR) and real‐time quantitative PCR (qPCR) for detecting eDNA of B. argentifolii and T. urticae, (3) establish a rapid eDNA processing methodology using the LGC Biosearch Technologies QuickExtract DNA extraction kit and the Qiagen DNeasy Blood and Tissue kit, and (4) test the specificity of the developed primers against non‐target species. B. argentifolii and T. urticae were confined to tomato leaves (Solanum lycopersicum) using clip cages for 24 h, after which eDNA was collected from leaf surfaces using a water spray method, filtered, and processed for DNA amplification. While literature‐based primers showed sufficient sensitivity, their specificity for eDNA applications was inadequate, prompting the design of novel PCR primers for both pest species. Positive eDNA detection was achieved with both amplification methods, with qPCR proving more reliable than cPCR due to the latter's inconsistent performance with positive control samples. We also introduced a rapid eDNA processing approach using the QuickExtract DNA extraction kit, contrasting it with the more conventional Qiagen DNeasy Blood and Tissue kit. We believe that our findings are the first step toward the practical use of eDNA as a highly sensitive, early detection technique.

Evaluation of Squash (Cucurbita pepo L.) Genotypes for Resistance to Cucurbit Chlorotic Yellows Virus

Cucurbit chlorotic yellows virus (CCYV), a Crinivirus transmitted by whiteflies, poses a significant threat to cucurbit crops globally. Summer squash (Cucurbita pepo L.), an important vegetable crop in the Southeastern United States, is particularly affected. The absence of commercially available resistant summer squash cultivars necessitates the exploration of resistant sources. Cucurbita germplasms with potential resistance to CCYV were previously identified through field screening. In this study, we describe the controlled greenhouse screening of these germplasm lines aimed at validating resistance to CCYV infection. The susceptible cultivar Gentry used as control exhibited early and severe symptoms in response to CCYV infection. In contrast, all the PI accessions tested, including PI 512749, PI 615141, PI 136448, PI 442312, PI 458731, and PI 420328, displayed delayed and less severe symptoms. Nevertheless, CCYV RNA accumulated in all the PI accessions. Lower symptom severity while harboring a considerable amount of CCYV indicates their inherent tolerance to the yellowing disease induced by CCYV. When comparing CCYV RNA accumulation in PI accessions with the commercial cultivar ‘Gentry’, lower virus titers were observed across all tested accessions. Specifically, PI 420328 and PI 458731 exhibited significantly reduced CCYV titers compared to the susceptible cultivar in both mass exposure and clip cage experiments. These accessions, displaying reduced symptoms and lower virus titers, hold promise as sources of resistance to CCYV in breeding programs. This study also highlights the importance of utilizing a reliable method to assay the resistance or tolerance of selected germplasm to infection by CCYV.

The clip cage conundrum: Assessing the interplay of confinement method and aphid genotype in fitness studies.

Behavior and fitness are important ecological traits frequently measured in insect bioassays. A common method to measure them in soft-bodied herbivorous insects involves confining individuals to plant leaves using clip cages. Although studies have previously highlighted the negative effects of clip cages on leaf physiology, little is known about the impact that using this confinement method has on insect fitness. The responses of different aphid genotypes/clones to different containment methods have not previously been investigated. Here we measured key fitness traits (intrinsic rate of natural increase, mean relative growth rate, time to reach reproductive adulthood and population doubling time) in the potato aphid, Macrosiphum euphorbiae Thomas (Hemiptera: Aphididae), when confined to plants using two methods: (1) clip cages to confine aphids to individual strawberry leaves and (2) a mesh bag to confine aphids to whole strawberry plants. Our study identified a strong negative impact on all the measured aphid fitness traits when using clip cages instead of mesh bags. We also identified genotype-specific differences in response to confinement method, where clip cage confinement differentially affected the fitness of a given aphid genotype compared to the same genotype on whole plants. These results suggest that clip cage use should be carefully considered when experiments seek to quantify insect fitness and that whole plants should be used wherever possible. Given the prevalence of clip cage use in insect bioassays, our results highlight the need for caution when interpreting the existing literature as confinement method significantly impacts aphid fitness depending on their genotype.

Clinical and Molecular Epidemiology of Hemorrhagic Fever with Renal Syndrome Caused by Orthohantaviruses in Xiangyun County, Dali Prefecture, Yunnan Province, China.

Hemorrhagic fever with renal syndrome (HFRS) is a zoonotic disease transmitted by several rodent species. We obtained clinical data of HFRS patients from the medical records of the People's Hospital of Xiangyun County in Dali Prefecture from July 2019 to August 2021. We collected epidemiological data of HFRS patients through interviews and investigated host animals using the night clip or night cage method. We systematically performed epidemiological analyses of patients and host animals. The differences in the presence of rodent activity at home (χ2 = 8.75, p = 0.031 < 0.05), of rodent-proof equipment in the food (χ2 = 9.19, p = 0.025 < 0.05), and of rodents or rodent excrement in the workplace (χ2 = 10.35, p = 0.014 < 0.05) were statistically different in the four clinical types, including mild, medium, severe, and critical HFRS-associated diseases. Furthermore, we conducted molecular detection of orthohantavirus in host animals. The total orthohantavirus infection rate of rodents was 2.72% (9/331); the specific infection rate of specific animal species was 6.10% (5/82) for the Apodemus chevrieri, 100% (1/1) for the Rattus nitidus, 3.77% (2/53) for the Rattus norvegicus, and 12.50% (1/8) for the Crocidura dracula. In this study, a total of 21 strains of orthohantavirus were detected in patients and rodents. The 12 orthohantavirus strains from patients showed a closer relationship with Seoul orthohantavirus (SEOOV) L0199, DLR2, and GZRn60 strains; the six orthohantavirus strains from Rattus norvegicus and Apodemus chevrieri were closely related to SEOOV GZRn60 strain. One strain (XYRn163) from Rattus norvegicus and one strain (XYR.nitidus97) from Rattus nitidus were closely related to SEOOV DLR2 strain; the orthohantavirus strain from Crocidura dracula was closely related to the Luxi orthohantavirus (LUXV) LX309 strain. In conclusion, patients with HFRS in Xuangyun County of Dali Prefecture are predominantly affected by SEOOV, with multiple genotypes of orthohantavirus in host animals, and, most importantly, these orthohantavirus strains constantly demonstrated zoonotic risk in humans.

Metabolite profile variation of chili pepper genotypes for resistance to aphids infestation

The cotton aphids (CA) were reported to reduce chili pepper production in tropical area and transmits several viruses. Using resistant cultivars can reduce the number of insecticide sprays and maintain natural enemies. The research objective was to evaluate aphid infestation in seven chili pepper genotypes and investigate secondary metabolite profiles under aphid infestation. Seven chili pepper genotypes belonging to the species Capsicum annuum L. were used in this study. The experiment was conducted using a four-replication randomized complete block design. We used a no-choice clip cage test to evaluate the chili pepper genotype. The evaluation was carried out under greenhouse conditions. Each plant was infested with one aphid using a wet brush in an insect-proof cage about five weeks after sowing. The resistance level was determined after seven days of infection (7 DAI) by counting the number of aphids. Gas chromatography-mass spectrophotometry (GC-MS) was used to perform untargeted metabolomic studies. The results showed that IPB C20 and IPB C367 genotypes had low aphid infestation, while IPB C313 showed the highest aphid infestation. Untargeted metabolomics using GC-MS detected 16 metabolites in the control plant and 27 in the treatment. Principal component analysis (PCA) (PC1 = 62% and PC2 = 23%) can used to differentiate metabolites among resistant genotypes, susceptible genotypes, and control plants. Chili pepper resistance to aphid infestation was antibiosis with numerous potential metabolite chemicals, such as fluorobenzoic acid, which strongly connected with decreased aphid colonies at 7 DAI. This study will be useful for pest control and will aid in the developing of aphid-resistant cultivars.

Construction of a Modified Clip Cage and Its Effects on the Life-History Parameters of Sitobion avenae (Fabricius) and Defense Responses of Triticum aestivum.

Simple SummaryClip cages are increasingly used in the study of plant–small insect interactions; however, the heavy weight and low light transmittance of clip cages cause damage to plants and affect insect growth and reproduction. This study aimed to develop a lightweight, transparent clip cage and investigate its effects on Sitobion avenae (Fabricius) and Triticum aestivum. The results indicate that the modified clip cages neither altered the S. avenae life-history parameters nor activated the T. aestivum defense responses and can, therefore, be used for plant–aphid interaction studies.Clip cages are commonly used to confine aphids or other small insects to a single leaf when conducting plant–small insect interaction studies; however, clip cages are usually heavy or do not efficiently transmit light, which has an impact on leaf physiology, limiting their application. Here, simple, lightweight, and transparent modified clip cages were constructed using punched clear plastic cups, cut transparent polyvinyl chloride sheets, nylon organdy mesh, and bent duck-bill clips. These cages can be clipped directly onto dicot leaves or attached to monocot leaves with bamboo skewers and elastic bands. The weight, production time, and aphid escape rates of the modified clip cages were 3.895 ± 0.004 g, less than 3 min, and 2.154 ± 0.323%, respectively. The effects of the modified clip cage on the growth, development, and reproduction of the English grain aphid (Sitobion avenae Fabricius) in comparison with the whole cage were studied. The biochemical responses of wheat (Triticum aestivum) to the cages were also investigated. No significant differences were observed in the life table parameters, nymph mortality, and adult fecundity in S. avenae confined to clip cages and whole cages, but the clip cages were more time efficient than whole cages when conducting life table studies. Moreover, the hydrogen peroxide accumulation, callose deposition, and cell necrosis in wheat leaves covered by empty clip cages and empty whole cages were similar, and significantly lower than treatments where the aphids were inside the clip cage. The results demonstrate that the modified clip cages had negligible effects on the plant and aphid physiology, suggesting that they are effective for studying plant–small insect interactions.

Local and systemic effect of azadirachtin on host choice and feeding activity of Macrosiphum rosae on rose plants

Many studies in the last decades addressed the antifeedant and repellent effects of neem (Azadirachta indica)-based pesticides on arthropod herbivores with azadirachtin as the leading active ingredient. However, results for aphids regarding repellence and feeding were often contradictory and mainly based on laboratory studies with self-made neem extracts in high concentrations. Moreover, no data are available on the presence of azadirachtin in the phloem of treated plants—the tissue that most aphids feed on. We investigated how the application of the commercial neem product NeemAzal-T/S onto only one plant half (upper or lower) affects nymphs of the aphid Macrosiphum rosae in clip cages on the untreated plant half. Results for aphid mortality indicate that active ingredients of NeemAzal-T/S are translocated both upwards and downwards in rose plants and that active ingredients are transported in the phloem. Furthermore, we investigated whether NeemAzal-T/S has a repellent or antifeedant effect on M. rosae. A choice test in the greenhouse as well as the quantification of honeydew excretion and electrical penetration graph analysis do not show differences in settling or feeding on untreated and NeemAzal-T/S-treated rose plants. Collectively, our data show that the effect of approved concentrations of NeemAzal-T/S on M. rosae is based on the toxicity after feeding on treated plants, not on starvation.

Chili pepper genotypes assay approach for resistance to Aphis gossypii (Hemiptera: Aphididae)

The cotton aphid (Aphis gossypii), is one of the most devastating insect pests for chili pepper that damages the crop and transmits several viruses. Thus far, there is no commercial chili cultivar with effective resistance to cotton aphids (CA). The present study aimed to develop a reliable and practical screening protocol in chili peppers for aphids resistance in the tropical areas. Three no-choice test methods i.e., seedling cage, detached leaf, and clip cage tests have been developed. The experiment was conducted in a randomized complete block design (RCBD) with four replications in a greenhouse at the Gunung Putri, Bogor, West Java, Indonesia. Seven chili pepper genotypes belong to the species Capsicum annuum L. provided by the Department of Agronomy and Horticulture, IPB University, Bogor, Indonesia were used in this study. Chili pepper resistance to aphids was observed through the number of aphids progeny, and aphids fecundity among the genotypes. Two chili genotypes consistently showed low infestation while other genotypes showed the highest infestation with three screening methods. All the methods have a high heritability value (90%–91%), indicating that the evaluated chili genotypes could be used to determine effective screening methods. The correlation between the detached leaf and clip cage tests was significant (r = 0.84, P ≤ 0.05). Therefore, the clip cage test could be used as a reliable and practical screening test for the assay of chili peppers resistance to CA infestation. These information will be helpful in the development of aphid resistant cultivars in the future.

Histology of Damage Caused by Euschistus heros (F.) Nymphs in Soybean Pods and Seeds.

The Neotropical brown stink bug, Euschistus heros (F.), feeds on stems, leaves, pods, and seeds of soybean, Glycine max (L.) Merrill. Knowledge of the damage that nymphs at different instars can cause to soybean pods and seeds, as well as efficient histological techniques for locating the salivary sheath are sparse. This study developed a new double-staining method to facilitate distinguishing the salivary sheath from plant tissues and to anatomically evaluate the damage caused by nymphs of different instars as they feed on soybean pods and seeds. Five insects from each of the analyzed instars (1st, 2nd, 3rd, 4th, and 5th) per pod at the R6 stage (full pod-filling) were kept in clip cages for 48h of feeding. The salivary sheath was analyzed to localize the damage (pod, vascular bundle, and seed) and the depth reached by the damage (categorized tissue). Double staining with xylidine ponceau and toluidine blue provided the best differentiation between the salivary sheath and watery sheath (proteins stained red) and the plant tissues (stained blue). First instar nymphs do not feed. Second instar and older nymphs caused damage to seeds, which became more severe with later developmental stages. The damage consists of coalescence of protein bodies and degradation and breakdown of the cell wall, marked by darkened regions in the embryo tissue of seeds. The information generated will contribute to new studies on feeding habits and emphasizes the need to control E. heros in early development stages.

Life cycle and morphometry of Rugose spiraling whitefly, Aleyrodicus rugioperculatus Martin (Hemiptera: Aleyrodidae) on coconut

The present study investigated the biology and morphometric analysis of rugose spiralling whitefly (RSW), Aleyrodicus rugioperculatus on coconut under mini net house condition at Department of Agricultural Entomology, Tamil Nadu Agricultural University-Coimbatore during 2019-2020. The biology of rugose spiralling whitefly was studied by clip cage method and morphometrics were done using Leica image analyser. Total lifecycle of rugose spiraling whitefly was 56.23 ± 2.20 days. Developmental period of egg, nymphal, pupal and adult period was 8.47 ± 0.26, 17.46 ± 0.76, 10.30 ± 0.29 and 20.00 ± 1.00 days, respectively. In morphometrics, Length and width of egg (0.31 ± 0.01 mm and 0.11± 0.02 mm), nymphal (0.94 ± 0.01 mm and 0.82 ± 0.01 mm), pupal (1.23 ± 0.01 mm and 1.00 ± 0.01 mm) was recorded. A nymphal parasitoid, Encarsia guadeloupae can be potential natural enemy for effective management of rugose spiraling whitefly.

Artificial microRNA-Based RNA Interference and Specific Gene Silencing for Developing Insect Resistance in Solanum lycopersicum

RNA Interference (RNAi), which works against invading nucleic acids or modulates the expression of endogenous genes, is a natural eukaryotic regulating system, and it works by noncoding smaller RNA molecules. Plant-mediated gene silencing through RNAi can be used to develop plants with insect tolerance at transcriptional or post-transcriptional levels. In this study, we selected Myzus persicae’s acetylcholinesterase 1 gene (Ace 1) as a silencing target to develop transgenic Solanum lycopersicum L. plants’ resistance to aphids. An RNAi plasmid vector containing an artificial microRNA (amiRNA) sequence was engineered and successfully transformed into Jamila and Tomaland, two elite tomato cultivars. A northern blot analysis and PCR were carried out to check the efficacy of Agrobacterium-mediated transformation in T0 transgenic plants. The quantitative PCR data showed a substantial downregulation of the Ace 1 gene in aphids fed in clip cages on T1 transgenic plants. Furthermore, there was a substantial drop in aphid colonies that were fed on T1 transgenic plants of both the cultivars. These findings strongly suggest that transgenic plants that express amiRNA could be an important tool for engineering plants resistant to aphids and possibly for the prevention of viral disease in other plant-infested pests.

Proteome changes in pepper (Capsicum annuum L.) leaves induced by the green peach aphid (Myzus persicae Sulzer)

BackgroundAphid attack induces defense responses in plants activating several signaling cascades that led to the production of toxic, repellent or antinutritive compounds and the consequent reorganization of the plant primary metabolism. Pepper (Capsicum annuum L.) leaf proteomic response against Myzus persicae (Sulzer) has been investigated and analyzed by LC-MS/MS coupled with bioinformatics tools.ResultsInfestation with an initially low density (20 aphids/plant) of aphids restricted to a single leaf taking advantage of clip cages resulted in 6 differentially expressed proteins relative to control leaves (3 proteins at 2 days post-infestation and 3 proteins at 4 days post-infestation). Conversely, when plants were infested with a high density of infestation (200 aphids/plant) 140 proteins resulted differentially expressed relative to control leaves (97 proteins at 2 days post-infestation, 112 proteins at 4 days post-infestation and 105 proteins at 7 days post-infestation). The majority of proteins altered by aphid attack were involved in photosynthesis and photorespiration, oxidative stress, translation, protein folding and degradation and amino acid metabolism. Other proteins identified were involved in lipid, carbohydrate and hormone metabolism, transcription, transport, energy production and cell organization. However proteins directly involved in defense were scarce and were mostly downregulated in response to aphids.ConclusionsThe unexpectedly very low number of regulated proteins found in the experiment with a low aphid density suggests an active mitigation of plant defensive response by aphids or alternatively an aphid strategy to remain undetected by the plant.Under a high density of aphids, pepper leaf proteome however changed significantly revealing nearly all routes of plant primary metabolism being altered. Photosynthesis was so far the process with the highest number of proteins being regulated by the presence of aphids.In general, at short times of infestation (2 days) most of the altered proteins were upregulated. However, at longer times of infestation (7 days) the protein downregulation prevailed.Proteins involved in plant defense and in hormone signaling were scarce and mostly downregulated.

Comparison of Toxicological Bioassays for Whiteflies.

Simple SummaryInsecticides are commonly used to manage whiteflies in many crops including vegetables, but frequent use can cause these pests to become resistant to insecticides. Resistance can lead to control failure and severe crop damage, thus the need for insecticide efficacy testing and insecticide resistance monitoring. A study was conducted to determine whether any current methods of toxicity assays are better than others for testing whiteflies for insecticide resistance and efficacy for better information to make effective pest control decisions.Two Bemisia tabaci populations from Georgia and Florida, USA, were tested for their response to insecticides across different toxicological bioassay methods. Five insecticides in four Insecticide Resistance Action Committee (IRAC) groups (imidacloprid (4A), dinotefuran (4A), flupyradifurone (4D), pyriproxyfen (7C) and cyantraniliprole (28)), were evaluated against a water check. The routes of application to the plant used were either leaf drench or (systemic) root drench. The four different whitefly bioassay methodologies tested were two published IRAC methods, a clip cage method, and a new tube method. A split–split experimental design was used to assess any interactions between application route, bioassay method and insecticide treatment. Application route had no significant effect on efficacy. However, bioassay method affected overall whitefly mortality, with the dish method having reduced mortality compared to other methods, except for the clip cage method. High rates of cyantraniliprole, dinotefuran and flupyradifurone insecticides resulted in the highest incidence of adult whitefly mortality. Significant interactions relative to percent adult mortality were found between the insecticide and bioassay method for both populations assayed. The clip cage method was more sensitive in terms of dose mortality response followed by the cup and tube methods. The dish method was the least responsive to insecticide dose. Other interactions are discussed.

A standardised bioassay method using a bench-top spray tower to evaluate entomopathogenic fungi for control of the greenhouse whitefly, Trialeurodes vaporariorum.

BACKGROUNDBioassays evaluating entomopathogenic fungi (EPF) isolates for effective microbial control of whitefly are a fundamental part of the screening process for bioprotectants, but development of repeatable, robust bioassays is not straightforward. Currently, there is no readily available standardised method to test the efficacy of EPF on whitefly.Here, we describe the calibration and use of a spray tower to deliver a standardised protocol to assess EPF activity; the method was validated using 18 EPF from four genera in tests against greenhouse whitefly, Trialeurodes vaporariorum (Westwood).RESULTSAt 138 kPa, the sprayer delivered 0.062 mL mm−2 (620 L ha−1) and an even deposition of spray across the central 1590 mm2 of the spray area.Average conidial deposition for all EPF was 252 conidia mm−2 and equivalent to 2.5 × 1012 conidia ha−1 at an application concentration of 1 × 107 conidia mL−1. Conidial deposition of a test Beauveria bassiana suspension increased with increasing application concentration.Egg laying by T. vaporariorum adults was restricted to 177 mm2 using clip cages specifically designed to ensure that third‐instar T. vaporariorum received a uniform spray coverage. Nymphs occupied 373 ± 5 mm2 of the leaf after migrating during the first instar.Average T. vaporariorum mortality totaled 8–89% 14 days after application of 1 × 107 conidia mL−1 of each EPF isolate.CONCLUSIONCombining the calibrated sprayer and bioassay method provides a reliable, standardised approach to test the virulence of EPF against whitefly nymphs. This laboratory‐based assay is affordable, replicable and allows the user to alter the dose of conidia applied to the target.

Transgenic Winter Wheat Expressing the Sucrose Transporter HvSUT1 from Barley does not Affect Aphid Performance.

Winter wheat expressing the sucrose transporter HvSUT1 from barley (HOSUT) has an increased yield potential. Genetic engineering should improve cultivars without increasing susceptibility to biotic stresses or causing negative impacts on ecosystem services. We studied the effects of HOSUT wheat on cereal aphids that feed on the sugar-rich phloem sap. Three HOSUT winter wheat lines, their conventional parental cultivar Certo, and three conventional cultivars were used. Clip cage experiments in the greenhouse showed no differences in life-table parameters of Rhopalosiphum padi and Sitobion avenae (Hemiptera: Aphididae) on transgenic lines compared to Certo, except higher fecundity of S. avenae on one HOSUT line. Population development of both aphid species over three weeks on caged flowering tillers did not reveal differences between the HOSUT lines and Certo. When aphids were monitored in a Swiss field study over two years, no differences between HOSUT lines and Certo were observed. We conclude that HOSUT wheat did not have consistent effects on aphids compared to the parental cultivar and measured parameters were generally in the range observed for the conventional winter wheat cultivars. Thus, HOSUT wheat is unlikely to suffer from increased aphid damage.

Endophytic Beauveria bassiana in maize affects survival and fecundity of the aphid Sitobion avenae

The entomopathogenic fungus Beauveria bassiana is able to colonize and establish a symbiotic relationship with plants as an endophyte, naturally as well as through artificial inoculation. This study aims at implementing a reliable and efficient method to establish endophytic B. bassiana in maize plants. The level of the endophytic establishment was evaluated and the survival and fecundity of the English grain aphid (Sitobion avenae) was investigated. Eight-days-old maize plants were sprayed with a fungal spore suspension, while control plants were treated only with a Tween solution. Two weeks after inoculation, seven clip cages per plant, each containing an adult aphid, were attached to different leaves and left for seven days. All inoculated plants contained endophytic B. bassiana as confirmed by plating surface disinfected leaf pieces on selective plates. None of the control plants showed growth of B. bassiana. The highest colonization levels were 61% in older inoculated leaves and 19% in younger non-inoculated leaves. The survival of S. avenae on inoculated plants was reduced up to 49% as compared to control plants. Dead aphids did not show any signs of mycosis. The number of nymphs produced by a single aphid was reduced up to 70% and 74% on inoculated and non-inoculated leaves, respectively. Presence of endophytic B. bassiana in non-inoculated younger leaves implies the movement of the endophyte inside plants. The negative effect on aphid fecundity appears to be systemic as implied by the almost equal reduction of mean numbers of nymphs produced on older and younger leaves on the inoculated plants, irrespective of the level of colonization by endophytic B. bassiana.

Efficacy of Chlorantraniliprole Seed Treatments Against Armyworm (Mythimna unipuncta [Lepidoptera: Noctuidae]) Larvae on Corn (Zea mays).

Mythimna unipuncta (Haworth) (Lepidoptera: Noctuidae) is an important insect pest of corn (Zea mays L.) in North America and can cause severe damage during outbreaks. Insecticides are the main control method; however, development of insecticide resistance poses management challenges and necessitates the use of novel insecticides. A synthetic insecticide, chlorantraniliprole, belonging to the anthranilic diamides, targets insect ryanodine receptors and is a potential alternative to conventional insecticides for management of M. unipuncta. We determined the efficacy of seed treatment with chlorantraniliprole alone compared with a positive control consisting of a neonicitinoid seed treatment of thiamethoxam and combinations of both compounds against M. unipuncta larvae in laboratory bioassays. Bioassays were conducted on two different growth stages of M. unipuncta larvae (instars 3 and 5) and two plant growth stages (V1 and V2 corn) in clip cages and whole plant experiments. Larval mortality, head capsule width, and feeding injury were measured. The chlorantraniliprole seed treatment alone or in combination with thimethoxam at different doses affected survival of M. unipuncta larvae. In all bioassays except one, larval mortality occurred earlier when a combination of chlorantraniliprole and thimethoxam seed treatment was used when compared with a thimethoxam alone seed treatment. Mythimna unipuncta larvae developed faster on the untreated control corn plants compared with corn with insecticide seed treatments. Foliar injury was low in both chlorantraniliprole alone and chlorantraniliprole + thimethoxam treatments compared with the control. Chlorantraniliprole thus offers potential alternative to conventionally used insecticides in the management of M. unipuncta in corn.

Effects of crowding and host plant quality on morph determination in the soybean aphid,Aphis glycines

AbstractPolyphenism constitutes a key example of the adaptability of a genotype to a shifting environment. In aphids, the relative production of apterous and alate individuals is influenced by cues resulting from increases in population density, changes in plant quality, and natural enemies. The ability to respond to multiple environmental cues offers aphids an effective way to optimize their fitness. Understanding the mechanisms behind alate morph production is particularly important in aphid species that are agricultural pests. We tested the effects of crowding and host plant quality cues on morph determination inAphis glycinesMatsumura (Hemiptera: Aphididae). We exposedA. glycinesto pre‐natal and post‐natal crowding under laboratory conditions. In the field, we reared nymphs ofA. glycinesat high‐density vs. low‐density conditions in clip cages set on the top vs. bottom nodes of infested and uninfested soybean plants,Glycine max(L.) Merrill (Fabaceae). Apterous adult body size was used as an estimate of aphid performance, and the proportion of morphs developing under each condition was determined. Both pre‐ and post‐natal crowding induced the production of asexual alate individuals byA. glycines. In the field, we observed additive negative effects of lower overall plant quality (infested vs. uninfested plants), within‐plant differences in nutritional quality (bottom vs. top nodes), and high vs. low levels of crowding on aphid size. These three factors also interacted to increase the proportionA. glycinesnymphs that developed wings. Our results suggest two levels in the production of asexual alate individuals byA. glycines: (1) a moderate level in response to aggregation, triggered by crowding cues, and (2) a high level in response to plant quality deterioration, triggered by both plant quality and crowding cues. These two processes ensure a flexible production of asexual alate individuals, allowing aphids to adapt to changing environmental conditions.

Soybean aphid, Aphis glycines (Hemiptera: Aphididae), developmental and reproductive capacity on white clover, Trifolium repens (Rosales: Leguminosae), in northeast China

Nymphs of Aphis glycines Matsumura were individually reared to adults in the laboratory on detached leaf discs of Trifolium repens L. (white clover) mounted on agar medium. Adults of A. glycines were fed T. repens within small clip cages in the field. Development, reproduction and intrinsic rates of increase of A. glycines were studied. These data were compared to those of controls fed known host plants including cultivated soybean Glycine max (L.) Merr. and the wild soybean species Glycine soja Sieb & Zucc. The results demonstrated that nymphs of A. glycines successfully developed into adults and reproduced efficiently when reared on T. repens in the laboratory. The lower development temperature threshold for nymphs fed T. repens was estimated as 8.27 °C, and the effective cumulative temperature for A. glycines development from nymph to adult was 90.91 degree-days. Adults of A. glycines could also survive on T. repens in the field, but only a few nymphs were produced.