David L. Kerns

Cry1F Resistance in Fall Armyworm Spodoptera frugiperda: Single Gene versus Pyramided Bt Maize

Evolution of insect resistance to transgenic crops containing Bacillus thuringiensis (Bt) genes is a serious threat to the sustainability of this technology. However, field resistance related to the reduced efficacy of Bt maize has not been documented in any lepidopteran pest in the mainland U.S. after 18 years of intensive Bt maize planting. Here we report compelling evidence of field resistance in the fall armyworm, Spodoptera frugiperda (J.E. Smith), to Cry1F maize (TC 3507) in the southeastern region of the U.S. An F2 screen showed a surprisingly high (0.293) Cry1F resistance allele frequency in a population collected in 2011 from non-Bt maize in south Florida. Field populations from non-Bt maize in 2012–2013 exhibited 18.8-fold to >85.4-fold resistance to purified Cry1F protein and those collected from unexpectedly damaged Bt maize plants at several locations in Florida and North Carolina had >85.4-fold resistance. In addition, reduced efficacy and control failure of Cry1F maize against natural populations of S. frugiperda were documented in field trials using Cry1F-based and pyramided Bt maize products in south Florida. The Cry1F-resistant S. frugiperda also showed a low level of cross-resistance to Cry1A.105 and related maize products, but not to Cry2Ab2 or Vip3A. The occurrence of Cry1F resistance in the U.S. mainland populations of S. frugiperda likely represents migration of insects from Puerto Rico, indicating the great challenges faced in achieving effective resistance management for long-distance migratory pests like S. frugiperda.

Sublethal effects of insecticides on the intrinsic rate of increase of cotton aphid

Insecticides are often implicated in causing outbreaks of the cotton aphid, Aphis gossypii (Glover) (Homoptera: Aphididae), through stimulation of reproduction. In this study we report the sublethal effects of dosages of bifenthrin, acephate, carbofuran or pyriproxifen on cotton aphid reproduction. We could not detect any increase or decrease in the intrinsic rate of increase of cotton aphids exposed to bifenthrin, acephate or carbofuran. However, we did detect some increases in the net reproductive rate of aphids treated with bifenthrin justifying further investigation of the effect on reproduction by this insecticide. Trends based on simple linear regression models suggest that sublethal dosages of bifenthrin or carbofuran have a negative impact on aphid population growth as dosages increase. These data suggest that stimulation of reproduction by these insecticides probably does not play a major role in cotton aphid outbreaks or resurgence. Pyriproxyfen is a juvenoid currently used for control of whiteflies in cotton. It demonstrated significant activity towards cotton aphid reared on treated cotton in our bioassays. Pyriproxyfen caused sterility in most aphids exposed to dosages exceeding 1 ppm, and reduced aphid longevity by approximately 50%. However, it did not appear to greatly influence the reproductive potential or longevity of reproductively mature aphids. A field study indicates that pyriproxyfen affects cotton aphid population structure and may have potential in managing cotton aphid outbreaks. Modifying aphid population structure and growth through the use of juvenoids such as pyriproxifen may prove to be an effective proactive approach to pest control without adversely impacting beneficial organisms or causing pest resurgence.

Spatial and Temporal Variation in Fungal Endophyte Communities Isolated from Cultivated Cotton (Gossypium hirsutum)

Studies of fungi in upland cotton (Gossypium hirsutum) cultivated in the United States have largely focused on monitoring and controlling plant pathogens. Given increasing interest in asymptomatic fungal endophytes as potential biological control agents, surveys are needed to better characterize their diversity, distribution patterns and possible applications in integrated pest management. We sampled multiple varieties of cotton in Texas, USA and tested for temporal and spatial variation in fungal endophyte diversity and community composition, as well as for differences associated with organic and conventional farming practices. Fungal isolates were identified by morphological and DNA identification methods. We found members of the genera Alternaria, Colletotrichum and Phomopsis, previously isolated as endophytes from other plant species. Other recovered species such as Drechslerella dactyloides (formerly Arthrobotrys dactyloides) and Exserohilum rostratum have not, to our knowledge, been previously reported as endophytes in cotton. We also isolated many latent pathogens, but some species such as Alternaria tennuissima, Epicoccum nigrum, Acremonium alternatum, Cladosporium cladosporioides, Chaetomium globosum and Paecilomyces sp., are known to be antagonists against plant pathogens, insects and nematode pests. We found no differences in endophyte species richness or diversity among different cotton varieties, but did detect differences over time and in different plant tissues. No consistent patterns of community similarity associated with variety, region, farming practice, time of the season or tissue type were observed regardless of the ecological community similarity measurements used. Results indicated that local fungal endophyte communities may be affected by both time of the year and plant tissue, but the specific community composition varies across sites. In addition to providing insights into fungal endophyte community structure, our survey provides candidates for further evaluation as potential management tools against a variety of pests and diseases when present as endophytes in cotton and other plants.

A Challenge for the Seed Mixture Refuge Strategy in Bt Maize: Impact of Cross-Pollination on an Ear-Feeding Pest, Corn Earworm

To counter the threat of insect resistance, Bacillus thuringiensis (Bt) maize growers in the U.S. are required to plant structured non-Bt maize refuges. Concerns with refuge compliance led to the introduction of seed mixtures, also called RIB (refuge-in-the-bag), as an alternative approach for implementing refuge for Bt maize products in the U.S. Maize Belt. A major concern in RIB is cross-pollination of maize hybrids that can cause Bt proteins to be present in refuge maize kernels and negatively affect refuge insects. Here we show that a mixed planting of 5% nonBt and 95% Bt maize containing the SmartStax traits expressing Cry1A.105, Cry2Ab2 and Cry1F did not provide an effective refuge for an important above-ground ear-feeding pest, the corn earworm, Helicoverpa zea (Boddie). Cross-pollination in RIB caused a majority (>90%) of refuge kernels to express ≥ one Bt protein. The contamination of Bt proteins in the refuge ears reduced neonate-to-adult survivorship of H. zea to only 4.6%, a reduction of 88.1% relative to larvae feeding on ears of pure non-Bt maize plantings. In addition, the limited survivors on refuge ears had lower pupal mass and took longer to develop to adults.

Cotton Aphid (Heteroptera: Aphididae) Susceptibility to Commercial and Experimental Insecticides in the Southern United States

ABSTRACT Cotton aphid, Aphis gossypii Glover, has a history of developing resistance to novel insecticides. A program is needed to monitor cotton aphid susceptibility to new insecticides. Concentration-mortality bioassays were conducted from 2008 to 2011 to monitor the susceptibility of cotton aphids from fields across the midsouthern United States to thiamethoxam and sulfoxaflor. Flonicamid was included in 2010 and 2011. Bioassays followed the procedures described by the Insecticide Resistance Action Committee for testing neonicotinoids against cotton aphid. Mortality was rated at 48 and 72 h. These bioassays suggest that high levels of resistance to thiamethoxam occur in cotton aphid throughout the midsouthern United States. Resistance ratios ranged from 0.9 to 562.6 at 48 h, and from 0.9 to 29.1 at 72 h. Aphid colonies tested were considered susceptible to flonicamid and sulfoxaflor. The LC50 values ranged from 1.43 to 6.60 ppm for flonicamid. The LC50 values for sulfoxaflor ranged from 1.01 to 5.85 ppm and 0.92–4.13 ppm at 48 and 72 h, respectively. These values represent the baseline variability of the susceptibility of cotton aphid to flonicamid and sulfoxaflor. The moderate level of variability observed combined with the high level of efficacy at low rates and the high reproductive rate of cotton aphid suggests that an effective resistance management plan needs to be devised for these insecticides. Flonicamid and sulfoxaflor should provide effective control of cotton aphid in areas where thiamethoxam resistance occurs. However, these insecticides need to be incorporated into a rotation strategy to preserve their efficacy against cotton aphid.

Performance and cross-crop resistance of Cry1F-maize selected Spodoptera frugiperda on transgenic Bt cotton: implications for resistance management.

Transgenic crops producing Bacillus thuringiensis (Bt) proteins have become a primary tool in pest management. Due to the intensive use of Bt crops, resistance of the fall armyworm, Spodoptera frugiperda, to Cry1F maize has occurred in Puerto Rico, Brazil, and some areas of the southeastern U.S. The sustainability of Bt crops faces a great challenge because the Cry1F-maize resistant S. frugiperda may also infest other Bt crops in multiple cropping ecosystems. Here we examined the survival and plant injury of a S. frugiperda population selected with Cry1F maize on three single-gene and five pyramided Bt cotton products. Larvae of Cry1F-susceptible (SS), -heterozygous (RS), and -resistant (RR) genotypes of S. frugiperda were all susceptible to the pyramided cotton containing Cry1Ac/Cry2Ab, Cry1Ac/Cry1F/Vip3A, Cry1Ab/Cry2Ae, or Cry1Ab/Cry2Ae/Vip3A, and the single-gene Cry2Ae cotton. Pyramided cotton containing Cry1Ac/Cry1F was effective against SS and RS, but not for RR. These findings show that the Cry1F-maize selected S. frugiperda can cause cross-crop resistance to other Bt crops expressing similar insecticidal proteins. Resistance management and pest management programs that utilize diversify mortality factors must be implemented to ensure the sustainability of Bt crops. This is especially important in areas where resistance to single-gene Bt crops is already widespread.

Mating disruption of beet armyworm (Lepidoptera: Noctuidae) in vegetables by a synthetic pheromone.

Abstract The pheromone dispenser, Yotoh-con-S ® , was evaluated for its ability to inhibit mate location and subsequent infestation by beet armyworm, Spodoptera exigua (Hubner) in head lettuce and broccoli. Pheromone dispensers consistently prevented male beet armyworm moths from locating synthetic lures or virgin females in treated fields. However, the dispensers were inconsistent in reducing the number of beet armyworm larvae, appearing to be most effective when population densities were low but ineffective when populations were high. Where beet armyworm larval populations were significantly reduced by the presence of synthetic pheromone dispensers, the resultant populations were usually not reduced to the point where insecticide applications could be eliminated.

F2 screen for resistance to Bacillus thuringiensis Cry2Ab2-maize in field populations of Spodoptera frugiperda (Lepidoptera: Noctuidae) from the southern United States.

The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a target pest of transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins in both North and South America. In 2013 and 2014, a total of 215 F2 two-parent families of S. frugiperda were established using single-pair mating of field individuals collected from seven locations in four states of the southern U.S.: Texas, Louisiana, Georgia, and Florida. The objective of the investigation was to detect resistance alleles in field populations to Cry2Ab2, a common Bt protein produced in transgenic maize and cotton. For each F2 family, 128 F2 neonates were screened on leaf tissue of Cry2Ab2 maize plants in the laboratory. A conservative estimate of the frequency of major Cry2Ab2 resistance alleles in S. frugiperda from the four states was 0.0023 with a 95% credibility interval of 0.0003-0.0064. In addition, six families were considered to likely possess minor resistance alleles at a frequency of 0.0082 with a 95% credibility interval of 0.0033-0.0152. One F2 family from Georgia (GA-15) was confirmed to possess a major resistance allele to the Cry2Ab2 protein. Larvae from this family survived well on whole maize plants expressing Cry2Ab2 protein and demonstrated a significant level (>15-fold) of resistance when fed with the same protein incorporated in a meridic diet. The detection of the major resistance allele along with the relatively abundant minor resistance alleles revealed in this study may have important implications for resistance management.

Sugarcane Aphid Population Growth, Plant Injury, and Natural Enemies on Selected Grain Sorghum Hybrids in Texas and Louisiana

Abstract In response to the 2013 outbreak of sugarcane aphid, Melanaphis sacchari (Zehntner) (Hemiptera: Aphididae), on sorghum, Sorghum bicolor (L.), in North America, experiments were conducted at three southern U.S. grain sorghum production locations (Corpus Christi, TX; Winnsboro, LA; Rosenberg, TX). The objectives were to authenticate yield decline on susceptible hybrids (2014 and 2015) and to measure aphid population growth and natural enemy prevalence on susceptible and resistant hybrids with similar genetic background (2014).Yield decline on susceptible hybrids (Tx 2752/Tx430 and DKS53-67) was more substantial when aphid population growth accelerated quickly and peaked above 300 aphids per leaf (50 to nearly 100% yield decline). Location and year variation in maximum aphid density and cumulative aphid-days was high, with doubling time values on the susceptible hybrids ranging between 3.9 and 7.9 d. On resistant Tx2752/Tx2783, leaf injury and yield decline were not seen or less severe than on its paired susceptibleTx2752/Tx430. Aphids declined onTx2752/Tx2783 after initial colony establishment (Corpus Christi) or took about 60% longer to double in population size when compared with Tx2572/Tx430 (Winnsboro). The predominant natural enemy taxa were aphelinid mummies (Hymenoptera: Aphelinidae), ladybird beetles (Coleoptera: Coccinellidae), and sryphid flies (Diptera: Syrphidae), and they were more prevalent during flowering than prior to flowering.They were generally responsive to changes in aphid density of both susceptible and resistant hybrids, but variability points to need for further study. In future research, full season observations should continue as well as more detailed study of potential compatibility of sorghum resistance and biological control.

Value of Neonicotinoid Insecticide Seed Treatments in Mid-South Cotton (Gossypium hirsutum [Malvales: Malvaceae]) Production Systems

Neonicotinoid insecticides are currently one of two classes of chemicals available as a seed treatment for growers to manage early season insect pests of cotton, Gossypium hirsutum L. (Malvales: Malvaceae), and they are used on nearly 100% of cotton hectares in the midsouthern states. An analysis was performed on 100 seed-treatment trials from Arkansas, Louisiana, Mississippi, and Tennessee to determine the value of neonicotinoid seed treatments in cotton production systems. The analysis compared seed treated with neonicotinoid insecticides seed treatments plus a fungicide with seed only treated with fungicide. When analyzed by state, cotton yields were significantly greater when neonicotinoid seed treatments were used compared with fungicide-only treatments. Cotton treated with neonicotinoid seed treatments yielded 123, 142, 95, and 104 kg ha-1, higher than fungicide only treatments for Arkansas, Louisiana, Mississippi, and Tennessee, respectively. Across all states, neonicotinoid seed treatments provided an additional 115 kg lint ha-1 comparedwith fungicide only treated seed. Average net returns from cotton with a neonicotinoid seed treatment were