Molecular characterization and expression patterns of a ryanodine receptor in soybean looper, Chrysodeixis includens.

Diamide insecticides, such as chlorantraniliprole, have been widely used to control insect pests by targeting the insect ryanodine receptor (RyR). Due to the efficacious insecticidal activity of diamides, as well as an increasing number of resistance cases, the molecular structure of RyR has been studied in many economically important insects. However, no research has been conducted on diamide resistance and RyR in the soybean looper, Chrysodeixis includens, a significant crop pest. In this study, we found moderate resistance to chlorantraniliprole in a field population from Puerto Rico and sequenced the full-length cDNA of the C. includens RyR gene, which encodes a 5124 amino acid-long protein. Genomic analysis revealed that the CincRyR gene consists of 113 exons, one of the largest exon numbers reported for RyR. Alternative splicing sites were detected in the cytosolic region. The protein sequence showed high similarity to other noctuid RyRs. Conserved structural features included the selectivity filter motif critical for ryanodine binding and ion conduction, as well as various domains involved in ion transport. Two mutation sites associated with diamide resistance in other insects were screened but not found in the Puerto Rico field populations or in the susceptible lab strain. Gene expression analysis indicated high expression of RyR in the third instar larval stage, particularly in muscle-containing tissues. Furthermore, exposure to a sublethal dose of chlorantraniliprole reduced RyR expression levels after 96 h. This study provides a molecular basis for understanding RyR structure and sheds light on potential mechanisms of diamide resistance in C. includens.

Standardized Field Trials in Cotton and Bioassays to Evaluate Resistance of Tobacco Thrips (Thysanoptera: Thripidae) to Insecticides in the Southern United States.

Foliar-applied insecticide treatments may be necessary to manage thrips in cotton (Gossypium hirsutum L.) under severe infestations or when at-planting insecticide seed treatments do not provide satisfactory protection. The most common foliar-applied insecticide is acephate. Field observations in Tennessee suggest that the performance of acephate has declined. Thus, the first objective was to perform leaf-dip bioassays to assess if tobacco thrips, Frankliniella fusca (Hinds) (Thysanoptera: Thripidae), in cotton production regions have evolved resistance to foliar-applied insecticides. A second objective was to assess the performance of commonly applied foliar insecticides for managing thrips in standardized field trials in Arkansas, Tennessee, Mississippi, and Texas. For both objectives, several insecticides were evaluated including acephate, dicrotophos, dimethoate, lambda-cyhalothrin, imidacloprid, and spinetoram. Field trials and bioassays were completed from 2018 to 2021. Dose-response bioassays with acephate were performed on tobacco thrips field populations and a susceptible laboratory population. Bioassay results suggest that tobacco thrips have developed resistance to acephate and other organophosphate insecticides; however, this resistance seems to be most severe in Arkansas, Tennessee, and the Delta region of Mississippi. Resistance to other classes of insecticides were perhaps even more evident in these bioassays. The performance of these insecticides in field trials was variable, with tobacco thrips only showing consistent signs of resistance to lambda-cyhalothrin. However, it is evident that many populations of tobacco thrips are resistant to multiple classes of insecticides. Further research is needed to determine heritability and resistance mechanism(s).

Sublethal Impacts of Novaluron on Tarnished Plant Bug (Hemiptera: Miridae) Adults.

Tarnished plant bug, Lygus lineolaris Palisot de Beauvois (Hemiptera: Miridae), has become a primary pest of cotton in the Midsouthern United States. Insect growth regulators such as novaluron are an important part of L. lineolaris management. While novaluron is lethal to nymphs, it does not kill adults, so it has been used when nymphs are the primary stage present. However, cotton yield protection was observed from an application of novaluron when adults were the predominant stage present. To explain this, a series of studies were conducted to examine sublethal impacts of novaluron to L. lineolaris adults. Novaluron ingestion by adults reduced hatch rate and sometimes reduced oviposition rate. Ingestion by either males or females reduced hatch rates, but the reduction was greater from female exposure. Contact exposure of adults with novaluron residues within 1 d of application reduced hatch rate by about 50%, but the impact on oviposition was inconsistent. A field study showed reduced hatch rate from contact exposure to mixed-age natural populations, but the overall net reproductive rate was not reduced. Surface exposure of eggs to novaluron did not reduce hatch rate. Overall, exposure of tarnished plant bug adults to novaluron, regardless of adult age or exposure route, reduced egg viability. However, the impact on oviposition rate and net reproductive rate varied with adult age and exposure route. This understanding of sublethal impacts of novaluron, in addition to lethal impacts on nymphs, should be considered when choosing application times to maximize effects on L. lineolaris populations.

Resistance monitoring to four insecticides and mechanisms of resistance in Lygus lineolaris Palisot de Beauvois (Hemiptera: Miridae) populations of southeastern USA cotton.

BACKGROUND: Economically damaging infestations of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), have become more frequent in Virginia and North Carolina cotton since 2013 and 2010, respectively. Foliar insecticide use has increased dramatically in response. Efficacy data (LC50 ) are needed to evaluate L. lineolaris susceptibility and resistance levels (RR50 ) to commonly used and recommended insecticides for managing this pest in the southeastern USA. RESULTS: Elevated resistance levels to acephate and bifenthrin were measured in L. lineolaris populations collected from wild and cultivated hosts in Virginia, North Carolina and northern Alabama when compared to a susceptible laboratory population. High levels of bifenthrin resistance were observed in 2018 and 2019. Mixed-function oxidase and esterase (EST) inhibitors, piperonyl butoxide and S,S,S-Tributyl phosphotrithioate, respectively, had a synergistic effect on bifenthrin with resistant populations of L. lineolaris. Bifenthrin-resistant L. lineolaris populations expressed elevated levels of cytochrome P450 (CYP450 ) monooxygenase and general EST activity. Results suggest that insecticide resistance is present in some locations and that CYP450 and EST activity in L. lineolaris contribute to pyrethroid resistance in the southeastern USA. CONCLUSIONS: Results can serve as a baseline for continued monitoring of L. lineolaris insecticide resistance and inform insecticide resistance management strategies that help southeastern USA cotton producers to minimize inputs and slow resistance development. © 2020 Society of Chemical Industry.

Novaluron prevents oogenesis and oviposition by inducing ultrastructural changes in ovarian tissue of young adult Lygus lineolaris.

BACKGROUND: The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), has emerged as a major pest of cotton, Gossypium hirsutum L, in the mid-southern USA. In the early 1990s L. lineolaris populations developed resistance to several classes of conventional insecticides, increasing the need for insecticides with alternative modes of action such as insect growth regulators (IGRs) for integrated pest management (IPM). The benzoylphenyl urea (BPU) class of IGRs acts by disrupting the growth and development of immature stages of insects, but little is known about its impact on adult stages. RESULTS: The effect of novaluron (Diamond™ 0.83EC), a BPU with known chitin synthesis inhibitor activity, was investigated on adult females of L. lineolaris. Treatment of 1-day-old adults with 600 ppm of novaluron in the diet prevented oviposition, while treatment of older females had no impact on oviposition. Oral novaluron exposure of adults of all ages reduced the viability of eggs laid. Novaluron treatment caused ultrastructural changes in the ovaries of 1-day-old adults (48 h post exposure), distorting the follicular epithelial cell architecture of developing oocytes. Additionally, novaluron treatment decreased the chitin content in ovarian tissue. CONCLUSION: Our results suggest that chitin or chitin-like components in the developing ovaries of adult L. lineolaris are a target of IGRs such as novaluron, but its activity is specific to a critical time during development. This enhances our understanding of the effects of BPUs on adult insects and could lead to incorporation of IGRs in IPM for controlling adult insect pest populations in the field. © 2020 Society of Chemical Industry.

Behavioral Responses of Thrips (Thysanoptera: Thripidae) and Tarnished Plant Bug (Hemiptera: Miridae) to a New Bt Toxin, Cry51Aa2.834_16 in Cotton.

Thrips (Thysanoptera: Thripidae) and tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae), are among the most important insect pests of cotton, Gosssypium hirsutum, in the mid-southern United States. These pests are currently managed primarily by insecticides; however, a new Bt toxin, Cry51Aa2.834_16 is under evaluation for control of thrips and tarnished plant bug. Experiments were conducted to evaluate the behavioral response of thrips and tarnished plant bug to Bt Cry51Aa2.834_16. Adult thrips avoided Bt Cry51Aa2.834_16 cotton in field choice tests and in separate field tests of Bt and non-Bt cotton not treated with insecticides. In a greenhouse choice test, approximately twice as many adult thrips and eggs were found on non-Bt compared with Bt Cry51Aa2.834_16 cotton. Similarly, in a field test of nontreated Bt Cry51Aa2.834_16 and non-Bt cotton, 68% of adult thrips collected were found on non-Bt cotton. In cotton that was not sprayed with insecticides, Bt Cry51Aa2.834_16 did not affect the distribution of tarnished plant bug within the canopy, although more square and flower injury was caused by tarnished plant bug in non-Bt cotton. Adult tarnished plant bug exhibited a nonpreference for diet containing lyophilized Bt Cry51Aa2.834_16 leaves and for excised Bt Cry51Aa2.834_16 squares in choice tests with non-Bt squares. The behavioral responses of these pests when exposed to this new Bt toxin will play a key role in the efficacy and potential resistance management strategies if this new technology is incorporated in an overall cotton insect pest management system.