Value of Neonicotinoid Insecticide Seed Treatments in Mid-South Corn (Zea mays) Production Systems.

Neonicotinoid seed treatments are one of several effective control options used in corn, Zea mays L., production in the Mid-South for early season insect pests. An analysis was performed on 91 insecticide seed treatment trials from Arkansas, Louisiana, Mississippi, and Tennessee to determine the value of neonicotinoids in corn production systems. The analysis compared neonicotinoid insecticide treated seed plus a fungicide to seed only with the same fungicide. When analyzed by state, corn yields were significantly higher when neonicotinoid seed treatments were used compared to fungicide only treated seed in Louisiana and Mississippi. Corn seed treated with neonicotinoid seed treatments yielded 111, 1,093, 416, and 140 kg/ha, higher than fungicide only treatments for Arkansas, Louisiana, Mississippi, and Tennessee, respectively. Across all states, neonicotinoid seed treatments resulted in a 700 kg/ha advantage compared to fungicide only treated corn seed. Net returns for corn treated with neonicotinoid seed treatment were $1,446/ha compared with $1,390/ha for fungicide only treated corn seed across the Mid-South. Economic returns for neonicotinoid seed treated corn were significantly greater than fungicide-only-treated corn seed in 8 out of 14 yr. When analyzed by state, economic returns for neonicotinoid seed treatments were significantly greater than fungicide-only-treated seed in Louisiana. In some areas, dependent on year, neonicotinoid seed treatments provide significant yield and economic benefits in Mid-South corn.

Lepidoptera (Crambidae, Noctuidae, and Pyralidae) Injury to Corn Containing Single and Pyramided Bt Traits, and Blended or Block Refuge, in the Southern United States.

Fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae); corn earworm, Helicoverpa zea Boddie (Lepidoptera: Noctuidae); southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae); sugarcane borer, Diatraea saccharalis F. (Lepidoptera: Crambidae); and lesser cornstalk borer, Elasmopalpus lignosellus Zeller (Lepidoptera: Pyralidae), are lepidopteran pests of corn, Zea mays L., in the southern United States. Blended refuge for transgenic plants expressing the insecticidal protein derivative from Bacillus thuringiensis (Bt) has recently been approved as an alternative resistance management strategy in the northern United States. We conducted a two-year study with 39 experiments across 12 states in the southern United States to evaluate plant injury from these five species of Lepidoptera to corn expressing Cry1F and Cry1Ab, as both single and pyramided traits, a pyramid of Cry1Ab×Vip3Aa20, and a pyramid of Cry1F×Cry1Ab plus non-Bt in a blended refuge. Leaf injury and kernel damage from corn earworm and fall armyworm, and stalking tunneling by southwestern corn borer, were similar in Cry1F×Cry1Ab plants compared with the Cry1F×Cry1Ab plus non-Bt blended refuge averaged across five-plant clusters. When measured on an individual plant basis, leaf injury, kernel damage, stalk tunneling (southwestern corn borer), and dead or injured plants (lesser cornstalk borer) were greater in the blended non-Bt refuge plants compared to Cry1F×Cry1Ab plants in the non-Bt and pyramided Cry1F×Cry1Ab blended refuge treatment. When non-Bt blended refuge plants were compared to a structured refuge of non-Bt plants, no significant difference was detected in leaf injury, kernel damage, or stalk tunneling (southwestern corn borer). Plant stands in the non-Bt and pyramided Cry1F×Cry1Ab blended refuge treatment had more stalk tunneling from sugarcane borer and plant death from lesser cornstalk borer compared to a pyramided Cry1F×Cry1Ab structured refuge treatment. Hybrid plants containing Cry1F×Cry1Ab within the pyramided Cry1F×Cry1Ab blended refuge treatment had significantly less kernel damage than non-Bt structured refuge treatments. Both single and pyramided Bt traits were effective against southwestern corn borer, sugarcane borer, and lesser cornstalk borer.

Species composition and seasonal abundance of stink bugs (Hemiptera: Pentatomidae) in Louisiana soybean.

In Louisiana during the last decade, the redbanded stink bug, Piezodorus guildinii (Westwood), has become a significant and yield-limiting pest of soybean. The redbanded stink bug was previously reported in the United States in 1892, but was never considered an economically important pest until recently. Soybeans representing four maturity groups (MG) III, IV, V, and VI were sampled weekly from beginning bloom (R1) to physiological maturity (R8) during 2008-2010 at five locations across Louisiana to determine the Pentatomidae composition. In total, 13,146 stink bugs were captured and subsequently identified to species. The predominant species included the redbanded stink bug (54.2%); southern green stink bug (27.1%), Nezara viridula L.; brown stink bug (6.6%), Euschistus servus (Say); and green stink bug (5.5%), Acrosternum hilare (Say). Redbanded stink bug comprised the largest percentage of the complex collected at four of the five survey sites. Numbers exceeding action thresholds of this stink bug complex were only detected during R4 to R7 growth stages. Redbanded stink bug accounted for the largest percentage of the stink bug complex in early maturing soybean varieties (MG III [86%] and IV [60%]) and declined in later maturing soybeans (MG V [54%] and VI [50%]). The redbanded stink bug was initially identified in southern Louisiana during 2000 and had been reported in all soybean producing regions in Louisiana by 2006. This survey is the first to report the redbanded stink bug as a predominant pest of soybeans from locations within the United States.

Cotton aphid (Heteroptera: Aphididae) susceptibility to commercial and experimental insecticides in the southern United States.

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.

Evaluation of corn hybrids expressing Cry1F, cry1A.105, Cry2Ab2, Cry34Ab1/Cry35Ab1, and Cry3Bb1 against southern United States insect pests.

Studies were conducted across the southern United States to characterize the efficacy of multiple Bacillus thuringiensis (Bt) events in a field corn, Zea mays L., hybrid for control of common lepidopteran and coleopteran pests. Cry1F protein in event TC1507 and Cry1A.105 + Cry2Ab2 proteins in event MON 89034 were evaluated against pests infesting corn on above-ground plant tissue including foliage, stalks, and ears. Cry34Ab1/Cry35Ab1 proteins in event DAS-59122-7 and Cry3Bb1 in event MON 88017 were evaluated against the larvae of Mexican corn rootworm, Diabrotica virgifera zeae Krysan and Smith, which occur below-ground. Field corn hybrids containing Cry1F, Cry1A.105 + Cry2Ab2, Cry34Ab1/Cry35Ab1, and Cry3Bb1 insecticidal proteins (SmartStax) consistently demonstrated reductions in plant injury and/or reduced larval survivorship as compared with a non-Bt field corn hybrid. Efficacy provided by a field corn hybrid with multiple Bt proteins was statistically equal to or significantly better than corn hybrids containing a single event active against target pests. Single event field corn hybrids provided very high levels of control of southwestern corn borer, Diatraea grandiosella (Dyar), lesser cornstalk borer, Elasmopalpus lignosellus (Zeller), and fall armyworm, Spodoptera frugiperda (J.E. Smith), and were not significantly different than field corn hybrids with multiple events. Significant increases in efficacy were observed for a field corn hybrid with multiple Bt events for sugarcane borer, Diatraea saccharalis (F.), beet armyworm, Spodoptera exigua (Hübner), corn earworm, Helicoverpa zea (Boddie), and Mexican corn rootworm. Utilization of field corn hybrids containing multiple Bt events provides a means for managing insect resistance to Bt proteins and reduces non-Bt corn refuge requirements.

Development of a plant-based threshold for tarnished plant bug (Hemiptera: Miridae) in cotton.

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), is an important pest of cotton, Gossypium hirsutum L., in the mid-southern United States. It is exclusively controlled with foliar insecticide applications, and sampling methods and thresholds need to be revisited. The current experiment was designed to establish a plant-based threshold during the flowering period of cotton development. Experiments were conducted in MisSissippi in 2005 and 2006, Arkansas in 2005, and Louisiana in 2005 through 2008. Treatments consisted of various combinations of thresholds based on the percentage of dirty squares that were compared with the current threshold with a drop cloth or automatic weekly applications. Dirty squares were characterized as those with yellow staining on the developing bud resulting from tarnished plant bug excrement. Treatments consisted of 5, 10, 20, and 30% dirty squares. Each plot was sampled weekly, and insecticides were applied when the mean of all replications of a particular treatment reached the designated threshold. At the end of the season, plots were harvested and lint yields were recorded. Differences were observed in the number of applications and yields among the different treatments. The 10% dirty squares threshold resulted in a similar economic return compared with the drop cloth. A threshold of 10% dirty squares resulted in a similar number of insecticide applications, yields, and economic returns compared with that observed with the drop cloth. These results suggest that a threshold of 10% dirty squares could be used to trigger insecticide applications targeting tarnished plant bugs in flowering cotton.

Susceptibility to insecticides used for control of Piezodorus guildinii (Heteroptera: Pentatomidae) in the United States and Brazil.

Limited information exists on the insecticide susceptibility of redbanded stink bug, Piezodorus guildinii (Westwood) (Heteroptera: Pentatomidae), despite its impact on soybean, Glycine max (L.) Merr., production in Brazil and the United States. Therefore, this study set out to 1) determine baseline levels of susceptibility to currently recommended pesticides using topical and vial bioassays, 2) determine the levels of esterase activity in populations in the United States and Brazil, and 3) compare control among products in field trials. In topical bioassays conducted in the United States using technical grade materials, the LC50 values of lambda-cyhalothrin, acephate, and methamidophos were 4-25, 141-295, and 40-151 ng per insect, respectively. The LC50 values of imidacloprid and thiamethoxam were 11 and 27 ng per insect, respectively. In vial bioassays conducted in the United States using technical grade materials, the LC50 values of cypermethrin, acephate, and methamidophos were 0.4-0.9, 3.8, and 1.6 microg per vial, respectively. In topical bioassays conducted in Brazil by using commercially formulated products, the LC50 values of acephate, methamidophos, endosulfan, and imidacloprid were 0.90-1.9, 0.4-0.6, 1.5-6.6, and 0.2-0.3 microg per insect, respectively. In vial bioassays conducted in Brazil using commercially formulated products, the LC50 values of endosulfan, methamidophos, and lambda-cyhalothrin were 4-32 and 2-24 microg/cm2 for thiamethoxam and imidacloprid. Esterase activity in Louisiana (United States) populations ranged from 251 to 658 nmol alpha-naphthol formed/min/mg protein. Esterase activity levels in Londrina (Brazil) populations averaged 163 nmol/min/mg. In field tests, P. guildinii in Louisiana were controlled by organophosphates thiamethoxam and imidacloprid and in Brazil, with combinations of neonicotinoids and pyrethroids.

Efficacy of Cry1Ac:Cry1F proteins in cotton leaf tissue against fall armyworm, beet armyworm, and soybean looper (Lepidoptera: Noctuidae).

Cotton, Gossypium hirsutum L., plants expressing Cry1Ac and Cry1F insecticidal crystal proteins of Bacillus thuringiensis Berliner (Bt) were evaluated against selected lepidopteran pests including fall armyworm, Spodoptera frugiperda (J. E. Smith), beet armyworm, Spodoptera exigua (Hübner), and soybean looper, Pseudoplusia includens (Walker). Studies were conducted in a range of environments, challenging various cotton tissue types from several varieties containing a combination of Cry1Ac and Cry1F proteins. In fresh tissue bioassays of mature leaves and squares (flower buds) and in artificial field infestations of white flowers, plants containing Cry1Ac:Cry1F significantly reduced levels of damage (leaf defoliation, bract feeding, penetrated squares and bolls, and boll abscission) and induced significantly greater mortality (90-100%) of fall armyworm compared with that on non-Bt cotton plants. Plants containing Cry1Ac:Cry1F conferred high levels (100%) of soybean looper mortality and low levels (0.2%) of leaf defoliation compared with non-Bt cotton. Beet armyworm was relatively less sensitive to Cry1Ac:Cry1F cotton plants compared with fall armyworm and soybean looper. However, beet armyworm larval development was delayed 21 d after infestation (DAI), and ingestion of plant tissue was inhibited (14 and 21 DAI) on the Cry1Ac:Cry1F plants compared with that on non-Bt cotton plants. These results show Cry1Ac:Cry1F cotton varieties can be an effective component in a management program for these lepidopteran pest species. Differential susceptibility of fall armyworm, beet armyworm, and soybean looper larvae to Cry1Ac:Cry1F cotton reinforces the need to sample during plant development and respond with a foliar insecticide if local action thresholds are exceeded.

Quantification of Cry1Ac and Cry1F Bacillus thuringiensis insecticidal proteins in selected transgenic cotton plant tissue types.

Quantitative enzyme-linked immunosorbent assays were used to characterize the geographical (locations) and temporal (through 6 wk) expression of CrylAc, from Bacillus thuringiensis variety kurstaki, and Cry1F, from B. thuringiensis variety aizawai, in transgenic cotton, Gossypium hirsutum L., plant structures. Terminal leaves, squares (flower buds), flowers, bolls (fruit), and mature leaves located five and eight nodes below the terminal apex were sampled during weeks 2, 4, and 6 after the initiation of anthesis. The effect of location (environment) significantly influenced protein expression levels, although similar trends were observed across locations. Cry1F was expressed at levels greater (1.1-29.0-fold) than that for CrylAc in all structures with exception to flowers. In contrast, the level of CrylAc in flowers was generally greater than Cry1F. Within each sampling period, concentrations of Cry1F in mature leaves (five and eight node) were greater than that for other structures. Expression was also greater for older, eight-node mature leaves than younger, five-node mature leaves. CrylAc expression in bolls was lowest compared with terminal leaves, squares, flowers, and mature leaves, which expressed at similar concentrations. Cry1F expression increased through time for mature leaves and terminal leaves; whereas, a decline in Cry1Ac protein concentration was observed for terminal leaves and bolls. The data presented here provides a means to understand observed levels of efficacy (patterns of insect damage) by comparing the spatial and temporal dynamics of expression for Cry1Ac and Cry1F in PhytoGen 440W transgenic cotton.

Survivorship of Helicoverpa zea and Heliothis virescens on cotton plant structures expressing a Bacillus thuringiensis vegetative insecticidal protein.

A series of tests quantified bollworm, Helicoverpa zea (Boddie), and tobacco budworm, Heliothis virescens (F.), larval survival on plant structures of a nontransgenic cotton (Gossypium hirsutum L.), 'Coker 312', and two transgenic cottons expressing Vip3A protein or both Vip3A + CrylAb proteins (VipCot). Vegetative and reproductive structures including terminal leaves, flower bud (square) bracts, whole debracted squares, flower petals, flower anthers, and intact capsules (bolls) were harvested from plants in field plots. Each structure was infested with 2-d-old larvae from one of the two heliothine species. Larvae were allowed to feed for 96 h on fresh tissue. Survivorship at 96 h after infestation was significantly lower on all structures of Vip3A and VipCot cotton lines compared with similar structures of Coker 312. VipCot plant structures generally resulted in lower larval survivorship compared with similar structures of the Vip3A cotton line. H. zea survivorship ranged from 4 to 28% and from 1 to 18% on Vip3A and VipCot plant structures, respectively. H. virescens survivorship ranged from 10 to 43% and from 2 to 12% on Vip3A and VipCot plant structures, respectively. H. virescens survivorship was higher on VIP3A plant structures compared with that for H. zea on similar structures. These differences between species were not observed on plants from the cotton line expressing VipCot proteins. The results for these plant structures demonstrate that the combination of proteins expressed in VipCot cotton lines are more effective than Vip3A cotton lines against this heliothine complex.

Efficacy of transgenic cotton expressing Cry1Ac and Cry1F insecticidal protein against heliothines (Lepidoptera: Noctuidae).

Cotton, Cossypium hirsutum L, plants expressing Cry1Ac and Cry1F (Phytogen 440W) insecticidal crystal proteins of Bacillus thuringiensis (Bt) Berliner, were evaluated against natural populations of tobacco budworm, Heliothis virescens (F.), and bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), across 13 southern U.S. locations that sustained low, moderate, and high infestations. The intrinsic activity of Phytogen 440W was compared with nontreated non-Bt cotton (PSC355) and with management strategies in which supplemental insecticides targeting heliothines were applied to Phytogen 440W and to PSC355 cotton. Infestations were composed primarily of bollworm, which is the least sensitive of the heliothine complex to Cry toxins. Therefore, damage recorded in these studies was primarily due to bollworm. Greater than 75% of all test sites sustained heliothine infestations categorized as moderate to high (10.6-64.0% peak damaged bolls in nontreated PSC355). Phytogen 440W, alone or managed with supplemental insecticide applications, reduced heliothine-damaged plant terminals, squares (flower buds), flowers, and bolls equal to or better (1.0-79.0-fold) than managing a non-Bt cotton variety with foliar insecticides across all infestation environments. Rarely (frequency of < or = 11% averaged across structures), sprayed Phytogen 440W reduced damaged structures compared with nontreated Phytogen 440W. Protection against heliothine-induced plant damage was similar across the three levels of infestation for each viable management strategy, with exception to damaged squares for nontreated Phytogen 440W. In situations of moderate to high heliothine infestations, cotton plants expressing Cry1Ac and Cry1F may sustain higher levels of damage compared with that same variety in low infestations. No significant difference in yield was observed among heliothine management strategies within each infestation level, indicating cotton plants may compensate for those levels of plant damage. These findings indicate Phytogen 440W containing Cry1Ac and Cry1F provided consistent control of heliothines across a range of environments and infestation levels.

Influence of Nezara viridula feeding on cotton yield, fiber quality, and seed germination.

The influence of southern green stink bug, Nezara viridula (L.), adults (males and females) and fourth to fifth instars on cotton, Gossypium hirsutum L., boll abscission, seedcotton yield, fiber quality, and seed viability was evaluated in field studies conducted during 2004 and 2005. Cotton bolls representing several age classes ranging from 0-600 heat units were individually infested with a specific gender or life stage of southern green stink bug. Adults and nymphs induced abscission of bolls that accumulated 0-280 heat units after anthesis. Seedcotton yield was significantly lower in bolls infested with adults (males and females) and late instars through approximately 500 heat units after anthesis. Southern green stink bug feeding on bolls significantly affected the physical fiber properties of micronaire (measure of fiber fineness or maturity), strength, uniformity, and fiber length. Discolored cotton lint in the stink bug-infested bolls was more common than in noninfested bolls. Seed germination and development of normal seedlings for seed harvested from stink bug-infested bolls that accumulated < or =500 heat units beyond anthesis were significantly lower compared with noninfested bolls. No significant differences in boll abscission, yield, fiber quality, and seed germination were detected between southern green stink bug males and females or between adults and fourth to fifth instars.

F2 screen for resistance to a Bacillus thuringiensis-maize hybrid in the sugarcane borer (Lepidoptera: Crambidae).

A novel F2 screening technique was developed for detecting resistance in sugarcane borer, Diatraea saccharalis (F.), to transgenic Bacillus thuringiensis (Bt)-maize expressing the Cry1Ab insecticidal protein. The F2 screening method involved (i) collecting larvae from maize fields; (ii) establishing two-parent families; (iii) screening F2 neonates for survival on Bt-maize leaf tissues; and (iv) confirming resistance on commercial Bt-maize plants. With the F2 screening method, 213 iso-line families of D. saccharalis were established from field collections in northeast Louisiana, USA and were screened for Bt resistance. One family was confirmed to carry a major Bt resistance allele(s). In a laboratory bioassay, larval mortality of the Bt-resistant D. saccharalis on Bt-maize leaf tissues was significantly lower than that of a Bt-susceptible strain. This Bt-resistant D. saccharalis population is the first corn stalk borer species that has completed larval development on commercial Bt-maize. The F2 screening protocol developed in this study could be modified for detecting Bt resistance alleles in other similar corn stalk borers, such as the European corn borer, Ostrinia nubilalis (Hübner), and the southwestern corn borer, D. grandiosella Dyar.

Sugarcane borer (Lepidoptera: Crambidae) management threshold assessment on four sugarcane cultivars.

This research assesses the potential for using different economic injury thresholds in management of a key insect pest on susceptible and resistant commercially produced cultivars of sugarcane (Saccharum spp. hybrids). In a 2-yr sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), study involving four commercially produced sugarcane cultivars and four insecticide treatment thresholds, 'LCP 85-384' and 'HoCP 91-555' were the most susceptible based on percentage of bored internodes compared with the more resistant 'HoCP 85-845' and 'CP 70-321'. In 2001, the 10% infested stalks threshold was not as effective as the 5% early season-10% late season and 5% full season for HoCP 91-555. Based on D. saccharalis injury under natural infestation conditions, susceptible cultivars seem to require a lower infestation threshold than the more resistant cultivars to achieve adequate injury reduction. Among yield components, only the theoretical recoverable sugar per stalk was significantly increased by applying insecticides. With the resistant HoCP 85-845, differences were not detected for percentage of bored internodes among treated versus untreated management regimes. The resistant HoCP 85-845 had higher levels of fiber in our study; however, no clear pattern on resistance mechanisms was established, because the resistant cultivar CP 70-321 had comparatively low levels of fiber. The development of cultivar-specific thresholds is expected to lower the amount of insecticide used for D. saccharalis management in the sugarcane industry, reduce selection pressure, and delay the development of insecticide resistance.

Cotton boll age influences feeding preference by brown stink bug (Heteroptera: Pentatomidae).

Cotton plants were infested with brown stink bug, Euschistus servus (Say), to define cotton boll age classes (based on heat unit accumulation beyond anthesis) that are most frequently injured during each of the initial 5 wk of flowering. Bolls from each week were grouped into discrete age classes and evaluated for the presence of stink bug injury. Brown stink bug injured significantly more bolls of age class B (approximately 165-336 heat units), age class C (approximately 330-504 heat units), and age class D (approximately 495-672 heat units) during the initial 3 wk in both years and in week 5 in 2002 compared with other boll ages. Generally, the frequency of injured bolls was lowest in age class A (< or = 168 heat units) during these periods. The preference by brown stink bug for boll age classes B, C, and D within a week was similar when ages were combined across all 5 wk. Based on these data, bolls that have accumulated 165.2 through 672 heat units beyond anthesis (approximately 7-27-d-old) are more frequently injured by brown stink bug when a range of boll ages are available. The boll ages in our studies corresponded to a boll diameter of 1.161-3.586 cm with a mid-range of 2.375 cm. A general protocol for initiating treatments against stink bugs is to sample bolls for evidence of injury as an indicator of presence of infestations in cotton. Sampling bolls within a defined range, which is most likely to be injured, should improve the precision of this method in detecting economic stink bug infestations in cotton.

Injury to preflowering and flowering cotton by brown stink bug and southern green stink bug.

The impact of brown stink bug, Euschistus servus (Say), and southern green stink bug, Nezara viridula (L.), injury was evaluated on preflowering and flowering cotton, Gossypium hirsutum L., plants in no-choice tests. Vegetative stage cotton seedlings and reproductive structures, including flower buds (square) and bolls, were infested with adults and/or nymphs of both species. There were no significant differences in height, height to node ratio, square retention, and flower initiation for cotton seedlings or plants with a match-head square between southern green stink bug adult- or brown stink bug adult-infested and noninfested treatments. Abscission for individual large squares (precandle) and multiple squares (medium and small square on the same sympodial branch) was not significantly different among infested and noninfested treatments for the following species and developmental stages: brown stink bug adults, southern green stink bug adults, and third and fourth to fifth instar southern green stink bug nymphs. In boll infestation studies, the relationship between boll maturity, expressed as heat units beyond anthesis, and boll growth, abscission, hard locked carpels, seedcotton yield, and seed germination was measured. Brown stink bug induced abscission in bolls that had accumulated > 0-350 heat units beyond anthesis. Boll growth and seedcotton yield was significantly lower for bolls infested with brown stink bug through 266.5 and 550 heat units beyond anthesis, respectively. The proportion of hard locked carpels per boll was significantly greater for the infested treatment in a cohort of bolls that accumulated from 51 to 400 heat units beyond anthesis. Seed germination in bolls infested with brown stink bug was significantly lower in bolls aged 101-600 heat units beyond anthesis.

Boll injury and yield losses in cotton associated with brown stink bug (Heteroptera: Pentatomidae) during flowering.

Brown stink bug, Euschistus servus (Say), was infested on cotton, Gossypium hirsutum L., plants during reproductive stages to determine the effects on boll injury and seedcotton yield. During each week in 2002 and 2003, significantly more bolls with > or = 1 injured locule, bolls with > or = 2 injured locules, and bolls with discolored lint were recorded on stink bug-infested plants compared with that on noninfested plants. Significantly fewer bolls displayed external injury on the boll exocarp compared with bolls with only internal locule injury. Boll injury was significantly underestimated by the presence of external symptomology. The boll population increased 6.6- and 5.1-fold from weeks 1-5 in 2002 and 2003, respectively. There was a corresponding 6.2- and 4.6-fold increase in 2002 and 2003, respectively, for total bolls injured from weeks 1-5. Percentage of boll injury ranged from 10.7 (week 4) to 27.4 (week 2) in 2002 and from 9.2 (week 3) to 16.0 (week 2) in 2003. Percentage of injury was greatest during weeks 1 and 2 in both years and also in week 5 in 2002. Brown stink bug significantly reduced seedcotton yield of bolls present on cotton plants during weeks 1, 2, and 5 in 2002 and in weeks 4 and 5 in 2003. However, total seedcotton yield, as a function of bolls exposed to brown stink bug and subsequent bolls produced on plant in the absence of stink bugs, was not significantly different for plots infested during weeks 1-4 in 2002 and weeks 1-3 in 2003. Flowering period and boll population influence the severity of stink bug injury on seedcotton yield. Infestation timing and number of bolls should be considered, in addition to insect densities, when initiating treatments against brown stink bug.

Distribution of bollworm, Helicoverpa zea (Boddie), injured reproductive structures on genetically engineered Bacillus thuringiensis var. kurstaki Berliner cotton.

Bollworm, Helicoverpa zea (Boddie), larvae are commonly observed feeding in genetically engineered Bollgard cotton. Although no information is currently available characterizing the levels of injury bollworms cause, aproximately 25% of the Bollgard acreage in the United States receives at least one insecticide application annually targeting bollworm populations. Studies were conducted to determine the levels of fruiting form injury that can occur from bollworm larvae feeding on white flowers of two types of genetically engineered cotton. The two types of genetically engineered cotton included the original Bollgard that produces one protein (Cry1Ac) from Bacillus thuringiensis variety kurstaki Berliner and Bollgard II that produces two proteins (Cry1Ac + Cry2Ab) from B. thuringiensis kurstaki. In one study, individual larvae (24 +/- 6 h old) were placed in first position white flowers of Deltapine 5415 (non-Bollgard) and Deltapine NuCOTN 33B (Bollgard). Larval infestations were made on 50 plants for each of 5 d during 2000 and 2001. Each plant was visually examined at 3 d and every 2 d thereafter, until larvae were no longer recovered. Larvae injured a total of 46.6 fruiting forms per 50 plants on non-Bollgard cotton, compared with only 18.9 fruiting forms per 50 plants on Bollgard cotton. Mean larval injury per insect was 4.3 fruiting forms on non-Bollgard cotton compared with 2.7 fruiting forms on Bollgard cotton. In a second study, individual larvae (24 +/- 6 h old) were placed in first position white flowers of Deltapine 50 (non-Bollgard), Deltapine 50B (Bollgard), and an experimental Bollgard II line. Larval infestations were made on 10 plants per day for each of six consecutive days during 2001. Larvae injured a total of 25.0 fruiting forms per 10 plants on non-Bollgard, 11.5 on Bollgard, and 6.4 on Bollgard II cottons. Mean larval injury per insect was 6.6 fruiting forms on non-Bollgard, 3.5 on Bollgard, and 0.8 on Bollgard II cottons. These data indicate that supplemental insecticide applications may be necessary to prevent yield losses on Bollgard cotton. In contrast, injury to Bollgard II cotton was minimal and may not require additional insecticide applications for bollworms.

Behavior of bollworm (Lepidoptera: Noctuidae) larvae on genetically engineered cotton.

Reports of bollworm, Helicoverpa zea (Boddie), larvae feeding in white flowers of Bollgard cotton have been relatively common since the commercialization of this technology in 1996. Field studies were conducted in Louisiana to determine if differences in bollworm larval behavior occuron non-Bollgard (cultivar 'Deltapine 5415') and Bacillus thuringiensis (Bt), Bollgard ('NuCOTN 33B') cottons. Larvae were placed on the terminal foliage of either single cotton plants or on all plants within 1-m row micro-plots. On preflowering cotton plants, significantly more bollworms moved from the site of infestation (terminal) on Bollgard plants compared with that on non-Bollgard plants. On individual flowering plants, the number of nodes larvae moved from the terminal and the number of infested bolls was greater on Bollgard cotton plants. Similar differences between Bollgard and non-Bollgard plants in the percentage of infested terminals and squares were observed at 48-h after infestation when 1-m rows were infested. These data will be used to refine scouting protocols for bollworm larvae on Bollgard cotton.

Bollworm (Lepidoptera: Noctuidae) survival on 'Bollgard' and 'Bollgard II' cotton flower bud and flower components.

Genetically modified cotton, Gossypium hirsutum L., cultivars ('Bollgard') that produce crystalline proteins from Bacillus thuringiensis (Berliner) are valuable tools for managing lepidopteran insect pests in the United States. However, high numbers of bollworm, Helicoverpa zea (Boddie), larvae have been observed feeding in white flowers of these cultivars. Fresh tissue bioassays were conducted to investigate bollworm survival on Bollgard and 'Bollgard II' cottons. Bollworm survival was higher on square and flower anthers than on other floral structures on 'Deltapine 5415' (conventional cotton) and 'NuCOTN 33B' (Bollgard). Bollworm survival at 72 h was higher on all floral structures from Deltapine 5415 than on corresponding structures from NuCOTN 33B. ELISA tests indicated that CryIA(c) expression varied among plant parts; however, bollworm survival did not correlate with protein expression levels. Trends in bollworm survival on Bollgard II were similar to those on Bollgard and conventional cotton; however, survival was lower on all structures of Bollgard II than on corresponding structures of Bollgard and conventional cotton. These data support field observations of bollworm injury to white flowers and small bolls and provide a better understanding of larval behavior on Bollgard cotton.