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.

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.

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.