Thermal biology and overwintering behavior of the red sunflower seed weevil (Coleoptera: Curculionidae).

Most natural mortality of the red sunflower seed weevil, Smicronyx fulvus LeConte (Coleoptera: Curculionidae), occurs while larvae overwinter in the soil. To test the hypothesis that S. fulvus mortality is related to low temperatures, experiments were used to (i) evaluate the temperature at which larvae freeze (= supercooling point [SCP]), (ii) assess possible vertical movement between entry into the soil in fall and adult emergence in summer, and (iii) determine if realistic soil temperatures could explain patterns of overwintering mortality. Mean SCP for groups of S. fulvus larvae differed between years and months, but only ranged from -20.93 to -22.68 °C. Most overwintering larvae were found within 6 cm of the soil surface, but larvae appeared to move 1-2 cm deeper between pairs of successive sample dates (September to January, January to April). Significant larval mortality that occurred between January and April 2021 was tentatively attributed to a period in February where daily minimum soil temperatures ranged from -8 to -12 °C. When overwintering under control conditions (constant 4 °C) was interrupted with week-long exposure to -4, -8, or -12 °C in a cold bath, significant S. fulvus mortality was seen for temperatures at or below -8 °C. Combined results suggest that mortality of overwintering S. fulvus is likely caused by continuous exposure to low temperatures that may not be cold enough to freeze larvae. Additionally, the shallow overwintering by S. fulvus supports the idea that routine farm management, including tillage and herbicide incorporation, may help limit populations of this sunflower pest.

Vitrification of Lepidopteran Embryos-A Simple Protocol to Cryopreserve the Embryos of the Sunflower Moth, Homoeosoma electellum.

Embryos of the sunflower moth, Homoeosoma electellum (Hulst), were cryopreserved after modification to the method that was previously described for Pectinophora gossipiella. The workflow to develop the protocol consisted of methods to weaken the embryonic chorion followed by the application of various methods to disrupt the sub-chorionic wax layer. These steps were necessary to render the embryos permeable to water and cryoprotectants. Initially, the embryos were incubated at 21° and 24 °C, and the development of the double pigment spots/eyespot and eclosion were tracked every two hours. The embryos at 24 °C showed eyespots as early as 30 h, while in the case of the embryos that were incubated at 21 °C, there was a developmental delay of approximately 20 h. The embryos at 24 °C showed peak eclosion between 55 and 70 h, and the embryos at 21 °C eclosed between 80 and 100 h of development. Estimating this range is crucial for the purposes of stage selection and treatment initiation for cryopreservation protocol development for the embryos. The control hatch percentage at either developmental temperature was >90%, and the sodium hypochloride, 2-propanol and alkane-based treatments reduced the embryo hatchability to <10%. Hence, a modified surfactant-hypochlorite mixture-was used to destabilize the chorion and solubilize the hydrophobic lipid layers. Water permeability assessments using the dye-uptake method show that polysorbate 80 in combination with sodium hypochlorite alone is capable of permeabilizing the embryo as efficiently as sequential hypochlorite-alkane treatments, but with significantly higher hatch rates. A vitrification medium consisting of ethane diol and trehalose was used to dehydrate and load the embryos with the cryoprotective agent. The median hatch rates after vitrification were 10%, and maximum was 23%.

Genetic mapping of a pollinator preference trait: Nectar volume in sunflower (Helianthus annuus L.).

Although high pollinator visitation is crucial to ensure the yields of pollinator-dependent crops, the quantitative trait loci (QTL) controlling nectar volume in sunflower (Helianthus annuus L.), a pollinator preference trait, have yet to be identified. To address this, a recombinant inbred line mapping population, derived from lines with contrasting nectar volume, was used to identify loci responsible for the phenotype. As a result, linkage mapping and QTL analysis discovered major loci on chromosomes 2 and 16 that are associated with variation in nectar volume in sunflower. Increased nectar volume is also associated with increased sugars and total energy available per floret. The regions on chromosomes 2 and 16 associated with the nectar phenotype exhibit indications of chromosome structural variation, such that the phenotype is associated with rearrangements affecting regions containing hundreds of genes. Candidate genes underlying QTL on chromosomes 9 and 16 are homologous to genes with nectary function in Arabidopsis. These results have implications for sunflower breeding, to enhance pollination efficiency in sunflower, as well as current and future studies on sunflower evolution.

Testing for evolutionary change in restoration: A genomic comparison between ex situ, native, and commercial seed sources of Helianthus maximiliani.

Globally imperiled ecosystems often depend upon collection, propagation, and storage of seed material for use in restoration. However, during the restoration process demographic changes, population bottlenecks, and selection can alter the genetic composition of seed material, with potential impacts for restoration success. The evolutionary outcomes associated with these processes have been demonstrated using theoretical and experimental frameworks, but no study to date has examined their impact on the seed material maintained for conservation and restoration. In this study, we compare genomic variation across seed sources used in conservation and restoration for the perennial prairie plant Helianthus maximiliani, a key component of restorations across North American grasslands. We compare individuals sourced from contemporary wild populations, ex situ conservation collections, commercially produced restoration material, and two populations selected for agronomic traits. Overall, we observed that ex situ and contemporary wild populations exhibited similar genomic composition, while four of five commercial populations and selected lines were differentiated from each other and other seed source populations. Genomic differences across seed sources could not be explained solely by isolation by distance nor directional selection. We did find evidence of sampling effects for ex situ collections, which exhibited significantly increased coancestry relative to commercial populations, suggesting increased relatedness. Interestingly, commercially sourced seed appeared to maintain an increased number of rare alleles relative to ex situ and wild contemporary seed sources. However, while commercial seed populations were not genetically depauperate, the genomic distance between wild and commercially produced seed suggests differentiation in the genomic composition could impact restoration success. Our results point toward the importance of genetic monitoring of seed sources used for conservation and restoration as they are expected to be influenced by the evolutionary processes that contribute to divergence during the restoration process.

Fatty Acid Data and Crop Surveys Indicate Sources of Red Sunflower Seed Weevil (Coleoptera: Curculionidae), Populations and Suggest Strategies for Management.

The red sunflower seed weevil, Smicronyx fulvus L., is a univoltine seed-feeding pest of cultivated sunflower, Helianthus annuus L. Artificial infestations of S. fulvus onto sunflowers with traditional (<25% oleic acid), mid-oleic (55-75%), or high oleic (>80%) fatty acid profiles were used to test if fatty acids could be used as natural markers to estimate the proportion of weevils developing on oilseed sunflowers rather than wild Helianthus spp. and confection (non-oil) types. Oleic acid (%) in S. fulvus confirmed the fatty acid compositions of mature larvae and weevil adults reflected their diets, making primary (oleic or linoleic) fatty acids feasible as natural markers for this crop-insect combination. Oleic acid in wild S. fulvus populations in North Dakota suggests at least 84 and 90% of adults originated from mid-oleic or high oleic sunflower hybrids in 2017 and 2018, respectively. Surveys in 2017 (n = 156 fields) and 2019 (n = 120 fields) extended information provided by S. fulvus fatty acid data; no significant spatial patterns of S. fulvus damage were detected in samples, damage to oilseed sunflowers was greater than confection (non-oil) types, and the majority of damage occurred in ≈10% of surveyed fields. Combined, data suggest a few unmanaged or mismanaged oilseed sunflower fields are responsible for producing most S. fulvus in an area. Improved management seems possible with a combination of grower education and expanded use of non-insecticidal tactics, including cultural practices and S. fulvus-resistant hybrids.

Capitate Glandular Trichomes Fail to Provide Significant Resistance to Banded Sunflower Moth (Lepidoptera: Tortricidae).

Extracts from capitate glandular trichomes (CGT) of wild and cultivated sunflowers, Helianthus spp., have repellent or toxic effects on sunflower specialists and generalist herbivores less closely associated with sunflower. Though CGT have been primarily examined for their potential to provide partial resistance to the sunflower moth, Homoeosoma electellum Hulst (Lepidoptera: Pyralidae), a floret- and seed-feeding pest, the banded sunflower moth (Cochylis hospes Walsingham [Lepidoptera: Tortricidae]) is a similar species more common in the primary sunflower-producing states of North Dakota and South Dakota. Replicated field trials using partially inbred lines with low or high CGT densities were used to evaluate possible reductions to seed damage by C. hospes larvae in 2016-2017. Results failed to support the idea that CGT are a useful defense against larvae of C. hospes; the putative plant defense of high trichome density corresponded to slightly more, rather than less, insect damage. A test of a secondary explanation, that strength of sunflower hulls could help determine patterns of seed damage among tested lines, produced similarly negative results. Though timing of bloom differed between groups of most- and least-damaged lines, prior research and pheromone-trapping data suggest differences in plant maturity also cannot adequately explain the observed results. While the specific mechanisms remain unclear, significant differences in susceptibility to C. hospes exist for cultivated sunflower and limit losses from this primary insect pest.

Seed and floret size parameters of sunflower are determined by partially overlapping sets of quantitative trait loci with epistatic interactions.

KEY MESSAGE: Floret and seed traits are moderately correlated phenotypically in modern sunflower cultivars, but the underlying genetics are mostly independent. Seed traits in particular are governed in part by epistatic effects among quantitative trait loci. Seed size is an important quality component in marketing commercial sunflower (Helianthus annuus L.), particularly for the in-shell confectionery market, where long and broad seed types are preferred as a directly consumed snack food globally. Floret size is also important because corolla tube length was previously shown to be inversely correlated with pollinator visitation, impacting bee foraging potential and pollinator services to the plant. Commercial sunflower production benefits from pollinator visits, despite being self-compatible, and bees are required in hybrid seed production, where "female" and "male" inbred lines are crossed at field scale. Issues with pollination of long-seed confectionery sunflower suggest that there may be an unfavorable correlation between seed and floret traits; thus, our objective was to determine the strength of the correlation between seed and floret traits, and confirm any co-localization of seed and floret trait loci using genome-wide association analysis in the SAM diversity panel of sunflower. Our results indicate that phenotypic correlations between seed and floret traits are generally low to moderate, regardless of market class, a component of population substructure. Association mapping results mirror the correlations: while a few loci overlap, many loci for the two traits are not overlapping or even adjacent. The genetics of these traits, while modestly quantitative and influenced by epistatic effects, are not a barrier to simultaneous improvement of seed length and pollinator-friendly floret traits. We conclude that breeding for large seed size, which is required for the confectionery seed market, is possible without producing florets too long for efficient use by pollinators, which promotes bee foraging and associated pollination services.

Native Solitary Bees Provide Economically Significant Pollination Services to Confection Sunflowers (Helianthus annuus L.) (Asterales: Asteraceae) Grown Across the Northern Great Plains.

The benefits of insect pollination to crop yields depend on genetic and environmental factors including plant self-fertility, pollinator visitation rates, and pollinator efficacy. While many crops benefit from insect pollination, such variation in pollinator benefits across both plant cultivars and growing regions is not well documented. In this study, across three states in the northern Great Plains, United States, from 2016 to 2017, we evaluated the pollinator-mediated yield increases for 10 varieties of confection sunflowers, Helianthus annuus L. (Asterales: Asteraceae), a plant that is naturally pollinator-dependent but was bred for self-fertility. We additionally measured pollinator visitation rates and compared per-visit seed set across pollinator taxa in order to determine the most efficacious sunflower pollinators. Across all locations and hybrids, insect pollination increased sunflower yields by 45%, which is a regional economic value of over $40 million and a national value of over $56 million. There was, however, some variation in the extent of pollinator benefits across locations and plant genotypes, and such variation was significantly related to pollinator visitation rates, further highlighting the value of pollinators for confection sunflowers. Female Andrena helianthi Robertson (Hymenoptera: Andrenidae) and Melissodes spp. (Hymenoptera: Apidae) were the most common and effective pollinators, while other bees including managed honey bees (Hymenoptera: Halictidae), Apis mellifera L. (Hymenoptera: Apidae), small-bodied sweat bees (Hymenoptera: Halictidae), bumble bees Bombus spp. (Hymenoptera: Apidae), and male bees were either infrequent or less effective on a per-visit basis. Our results illustrate that wild bees, in particular the sunflower specialists A. helianthi and Melissodes spp., provide significant economic benefits to confection sunflower production.

Using Nectar-Related Traits to Enhance Crop-Pollinator Interactions.

Floral nectar and other reward facilitate crop pollination, and in so doing, increase the amount and breadth of food available for humans. Though abundance and diversity of pollinators (particularly bees) have declined over the past several decades, a concomitant increase in reliance on pollinators presents a challenge to food production. Development of crop varieties with specific nectar or nectar-related traits to attract and retain pollinating insects is an appealing strategy to help address needs of agriculture and pollinators for several reasons. First, many crops have specific traits which have been identified to enhance crop-pollinator interactions. Also, an improved understanding of mechanisms that govern nectar-related traits suggest simplified phenotyping and breeding are possible. Finally, the use of nectar-related traits to enhance crop pollination should complement other measures promoting pollinators and will not limit options for crop production or require any changes by growers (other than planting varieties that are more attractive or rewarding to pollinators). In this article, we review the rationale for improving crop-pollinator interactions, the effects of specific plant traits on pollinator species, and use cultivated sunflowers as a case study. Recent research in sunflower has (i) associated variation in bee visitation with specific floral traits, (ii) quantified benefits of pollinators to hybrid yields, and (iii) used genetic resources in sunflower and other plants to find markers associated with key floral traits. Forthcoming work to increase pollinator rewards should enable sunflower to act as a model for using nectar-related traits to enhance crop-pollinator interactions.

Genetic Architecture of Capitate Glandular Trichome Density in Florets of Domesticated Sunflower (Helianthus annuus L.).

Capitate glandular trichomes (CGT), one type of glandular trichomes, are most common in Asteraceae species. CGT can produce various secondary metabolites such as sesquiterpene lactones (STLs) and provide durable resistance to insect pests. In sunflower, CGT-based host resistance is effective to combat the specialist pest, sunflower moth. However, the genetic basis of CGT density is not well understood in sunflower. In this study, we identified two major QTL controlling CGT density in sunflower florets by using a F4 mapping population derived from the cross HA 300 × RHA 464 with a genetic linkage map constructed from genotyping-by-sequencing data and composed of 2121 SNP markers. One major QTL is located on chromosome 5, which explained 11.61% of the observed phenotypic variation, and the second QTL is located on chromosome 6, which explained 14.06% of the observed phenotypic variation. The QTL effects and the association between CGT density and QTL support interval were confirmed in a validation population which included 39 sunflower inbred lines with diverse genetic backgrounds. We also identified two strong candidate genes in the QTL support intervals, and the functions of their orthologs in other plant species suggested their potential roles in regulating capitate glandular trichome density in sunflower. Our results provide valuable information to sunflower breeding community for developing host resistance to sunflower insect pests.

Nonconstant Thermal Regimes Enhance Overwintering Success and Accelerate Diapause Development for Smicronyx fulvus (Coleoptera: Curculionidae).

Recent populations of the red sunflower seed weevil, Smicronyx fulvus LeConte (Coleoptera: Curculionidae), have been inconsistent or declining, particularly in North Dakota. Consequently, research on weevil biology, including development of resistant germplasm, has been limited. To determine whether cold storage of diapausing larvae could be improved, nonconstant temperature treatments (fluctuating thermal regime [FTR] and thermoperiod [TP]) were tested versus a constant 6°C for storage up to 365 d. Both alternate temperature treatments produced more adult weevils than constant 6°C for short (42, 91 d) storage, while all temperature treatments were good (≥60% adult emergence) at moderate term (182 d) cold storage, and FTR was best for long (365 d) periods. Excluding the 14-d storage period, which produced too few weevils for comparison, each doubling of cold storage time (e.g., from 42 to 91 d, 91 to 182 d), usually decreased the number of days to 50% relative emergence by ∼10 d. After 365 d of larval storage, emerged S. fulvus adults successfully infested sunflowers in a plant growth chamber, with damage per female similar to that observed in field trials. Compared with previous efforts to store weevil larvae, the method of collection and FTR storage is either more effective (greater adult emergence and reduced parasitism) or more time-efficient, and should permit year-round research using S. fulvus adults. Because successful emergence under FTR was >75% after 365 d, additional research would be required to determine the maximum effective duration of cold storage for S. fulvus.

Sesquiterpene Lactone Composition of Wild and Cultivated Sunflowers and Biological Activity against an Insect Pest.

Sesquiterpene lactones in sunflowers, Helianthus spp., are important to interactions with pathogens, weeds, and insects. Across a broad range of Helianthus annuus, differences in composition of sesquiterpene lactones extracted from disc florets were found between wild and cultivated sunflowers and also between distinct groups of inbreds used to produce sunflower hybrids. Discriminant function analysis showed the presence and relative abundance of argophyllone B, niveusin B, and 15-hydroxy-3-dehydrodesoxyfruticin were usually (75%) effective at classifying wild sunflowers, cultivated inbreds, and hybrids. Argophyllone B reduced the larval mass of the sunflower moth, Homeosoma electellum, by >30%, but only at a dose greater than that found in florets. Low doses of mixed extracts from cultivated florets produced a similar (≈40%) reduction in larval mass, suggesting combinations of sesquiterpene lactones act additively. Although the results support a role for sesquiterpene lactones in herbivore defense of cultivated sunflowers, additional information is needed to use these compounds purposefully in breeding.

Overexpression of Drosophila juvenile hormone esterase binding protein results in anti-JH effects and reduced pheromone abundance.

The titer of juvenile hormone (JH), which has wide ranging physiological effects in insects, is regulated in part by JH esterase (JHE). We show that overexpression in Drosophila melanogaster of the JHE binding protein, DmP29 results in a series of apparent anti-JH effects. We hypothesize that DmP29 functions in transport of JHE such that over- or under-expression of DmP29 results in increased or decreased JH degradation at specific sites respectively. Overexpression of DmP29 during the first or second instar was lethal, while overexpression during the third instar resulted in eclosion of small adults. Overexpression of DmP29 in newly eclosed flies reduced ovarian development and fecundity in addition to reducing the abundance of aggregation pheromone (cis-vaccenyl acetate) in males and courtship pheromone (cis,cis-7,11-heptacosadiene) in females. Both sexes also had lower levels of 23 and 25 carbon monoenes. Females exhibited reduced receptivity to mating, and males exhibited male-male courtship behavior, with both sexes being hyperactive: Male flies covered 2.7 times the distance of control flies at 2.9 times the maximum velocity. Application of the JH analog methoprene reversed impaired ovarian development, supporting a role for reduced JH in production of this phenotype. Rather than increasing lifespan as expected from a JH deficiency, overexpression of DmP29 reduced the life span of adult flies which may result from the hyperactivity of these flies. Underexpression of DmP29 resulted in reduced longevity, increased fecundity and reduced titers of pupal JHE. An alternative hypothesis, that mitochondrial dysfunction rather than JHE results in the JH-mediated phenotypes, is discussed.

Effects of insecticide exposure on movement and population size estimates of predatory ground beetles (Coleoptera: Carabidae).

BACKGROUND: Estimates of arthropod population size may paradoxically increase following insecticide applications. Research with ground beetles (Coleoptera: Carabidae) suggests that such unusual results reflect increased arthropod movement and capture in traps rather than real changes in population size. However, it is unclear whether direct (hyperactivity) or indirect (prey-mediated) mechanisms produce increased movement. RESULTS: Video tracking of Scarites quadriceps Chaudior indicated that brief exposure to lambda-cyhalothrin or tefluthrin increased total distance moved, maximum velocity and percentage of time moving. Repeated measurements on individual beetles indicated that movement decreased 240 min after initial lambda-cyhalothrin exposure, but increased again following a second exposure, suggesting hyperactivity could lead to increased trap captures in the field. Two field experiments in which ground beetles were collected after lambda-cyhalothrin or permethrin application attempted to detect increases in population size estimates as a result of hyperactivity. Field trials used mark-release-recapture methods in small plots and natural carabid populations in larger plots, but found no significant short-term (<6 day) increases in beetle trap captures. CONCLUSION: The disagreement between laboratory and field results suggests mechanisms other than hyperactivity may better explain unusual changes in population size estimates. When traps are used as a primary sampling tool, unexpected population-level effects should be interpreted carefully or with additional data less influenced by arthropod activity.

Effects of Cry1Ab-expressing corn anthers on the movement of monarch butterfly larvae.

Decreased larval feeding and weight of the monarch butterfly, Danaus plexippus L., have been detected after 4 d of exposure in the laboratory to a high density of Bacillus thuringiensis (Bt)-expressing anthers. One hypothesis is that larvae exposed to Bt anthers exhibit increased wandering, resulting in less feeding and lower weight gain. To test this hypothesis, 2-d-old monarch butterfly larvae exposed to milkweed leaf disks with no anthers, anthers that express Bt (Cry1Ab, event MON810), or other non-Bt anthers were observed using a video-tracking system. As had been shown in previous studies, larvae exposed to Bt anthers fed less and gained less weight than larvae exposed to non-Bt or no anthers, yet there was no evidence of feeding on anthers. Total distance moved, maximum displacement from release point, percentage of time spent moving or near anthers, or mean turn angle did not differ across treatments. However, larvae exposed to Bt anthers spent more time off milkweed leaf disks than those exposed to no anthers and were more likely to move off the leaf than larvae exposed to non-Bt anthers. Results suggest that larvae exposed to Bt anthers behave differently and that ingestion may not be the only way Bt can affect nontarget insects like the monarch butterfly.