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Evaluations of Bollgard[R], Bollgard II[R], and WideStrike[R] technologies against beet and fall armyworm larvae (Lepidoptera: Noctuidae).

Since the first Cry1Ac Bacillus thuringiensis Berliner (Bt) cotton variety was commercialized in 1996 (Bollgard [R], Monsanto Ag. Co., St. Louis, MO), advancements for insect control with transgenic technology has occurred that offer improved efficacy against many lepidopteran pests. Current varieties can contain Cry1Ac alone or they can be stacked with Cry2Ab (Bollgard[R] II, Monsanto Ag. Co.) or Cry1F (WideStrike[R], Dow Agrosciences, Indianapolis, IN).

The beet armyworm, Spodoptera exigua (Hubner) is a secondary, but serious migratory pest of various vegetable and certain row crops in the southern part of the United States of America. Although larval feeding on cotton is primarily concentrated on foliage, larvae can cause devastating losses in yield during outbreaks (Hardee & Herzog 1997; Adamczyk et al. 1998). The fall armyworm, S. frugiperda (J. E. Smith) also is a destructive migratory pest of many crops in the Western Hemisphere, where it appears to be more common and widespread (Sparks 1979; Young 1979). Like the beet armyworm, this pest has the potential to damage both conventional and Bollgard[R] cotton bolls (Adamczyk et al. 1998).

Although certain lepidopteran pests of cotton are controlled by Bollgard[R] cotton (e.g., tobacco budworms, Heliothis virescens (F.) and pink bollworms, Pectinophora gossypiella (Saunders)), the Cry1Ac [delta]-endotoxin in Bollgard[R] cotton is less effective for controlling beet and fall armyworms (MacIntosh et al. 1990; Adamczyk et al. 1998). Consequently, outbreaks of these pests on Bollgard[R] often need full application rates of foliar insecticide treatments to keep these populations below economic injury levels (Hood 1997; Smith 1997). The addition of other Cry proteins stacked with Cry1Ac (i.e., Bollgard II[R] and WideStrike[R]) has improved the efficacy against these armyworms (Adamczyk et al. 2001; Stewart et al. 2001; Adamczyk & Gore 2004). However, differences in survivorship of beet and fall armyworm larvae feeding on Bollgard II[R] versus WideStrike[R] cottons have been suggested, but never characterized or explained. The purpose of the study was to examine the efficacy of Bollgard II[R] and WideStrike[R] against beet and fall armyworms.


Field Plots

In May 2005 transgenic cotton varieties containing the Bollgard[R], Bollgard II[R] and WideStrike[R] traits were planted in research plots in the Mississippi Delta near Stoneville, MS (Table 1). Plots consisted of 2 rows (1.0 m centers) x 10.67 m. All plots were arranged in a randomized complete block design with each variety replicated 4 times (once in each block). Only insecticides not active on Lepidoptera were applied to all plots throughout the season as dictated by local management practices. In Mar 2007 transgenic cotton varieties containing the Bollgard, Bollgard II[R] and WideStrike[R] traits were planted in strip plots in the lower Rio Grande Valley of Texas near Weslaco, TX (Table 1).


All Lepidoptera utilized in these studies were obtained from laboratory colonies maintained at the USDA, ARS located in Stoneville, MS or Weslaco, TX.

Bioassays with Larvae

In 2005 bioassays were conducted with only fall armyworm larvae. A single neonate was placed into individual 9-cm diameter petri dishes (8) that each contained a moistened filter paper and a single lower leaf obtained from all plots for a total of 32 larvae per variety. Cotton plants were at peak bloom. The plates were covered with corresponding lids. After 5 d, surviving larvae were carefully transferred with a camel-hair brush into new petri dishes containing fresh filter paper and a new leaf. This procedure continued until pupation. At 7 and 10 d, larvae were prodded with a camel-hair brush and considered alive if coordinated movement was observed. Beginning at 15 d, plates were checked daily for the presence of pupae. Percent survival was analyzed by REML-ANOVA, and means were separated according to Fisher's Protected LSD (Littell et al. 1996; PROC MIXED, SAS Institute 2001).

In 2007 leaves obtained from various sections of the plant containing various transgenic traits were assayed for bioactivity against beet and fall armyworms. Individual leaves were placed into a 50 x 9-mm Tight-Fit Lid sealing Petri dish (BD Falcon[R] #351006, VWR International). Beet armyworms (5) were placed in a dish containing a terminal (upper canopy) leaf or a mid-canopy leaf (10 dishes per variety) for a total of 50 larvae per variety. Fall armyworm bioassays were conducted identically, except only mid-canopy leaves were used. Leaves were also collected from various times during the growing season. At 5 d after exposure to cotton tissue, larvae were prodded with a camel-hair brush and considered alive if coordinated movement was observed. Percent mortality by trait was analyzed by REML-ANOVA, and means were separated according to Fisher's Protected LSD (Littell et al. 1996; PROC MIXED, SAS Institute 2001).

Bioassays with Egg Masses

Inoculations of beet and fall armyworm egg masses to leaves located in various sections of the plant were conducted in 2007. In the laboratory, egg masses were deposited on nylon cloth placed on the top of adult rearing cages (3.79-L cardboard containers). For each inoculation, an egg mass of equal size (ca. 100-300 eggs per 2.54 [cm.sup.2] cloth sample) was pinned to the underside of a leaf for all traits and covered with a cage that consisted of a condiment cup (118 mL) (Solo Co., Highland Park, IL) coupled with a hard plastic lid (Fig. 1). Five d after inoculations, the infested leaf and corresponding cages were harvested and transported to the laboratory. Leaf damage (0-5) was estimated with a categorical rating scale where 0% indicated no leaf damage, while 80-100% of leaf consumption was given a value of 5. Damage ratings were analyzed by non-parametric statistics (SAS Institute 2001).


Bioassays with Larvae

Both Bollgard II[R] and WideStrike[R] had significantly higher efficacy against fall armyworms than Bollgard[R] (Fig. 2 and Fig. 3); however, in both 2005 and 2007, WideStrike[R] had typically higher efficacy compared to Bollgard II[R]. It is interesting to note that larval development to pupation was observed for fall armyworms when fed Bollgard[R] or Bollgard II[R], but not WideStrike[R]. In WideStrike[R] isolines containing only Cry1F or Cry1Ac, Adamczyk & Gore (2004) showed that the Cry1F protein provided greater efficacy compared to CrylAc against this pest. In addition, previous studies have shown little efficacy of Cry1Ac against fall armyworms (Adamczyk et al. 1998). We believe that the greater efficacy of WideStrike[R] compared to Bollgard II[R] against fall armyworms is primarily due to the action of the Cry1F protein. Although we did not evaluate isolines containing only Cry1F or Cry2Ab, our data suggests greater efficacy of Cry1F versus Cry2Ab against this pest.


The expression of the Cry1F protein affects beet armyworm survival both temporally and spatially. When using mid-canopy leaves sampled throughout the season, no significant differences (P > 0.05) were observed between the survival of beet armyworms fed Bollgard II[R] and WideStrike[R] (Fig. 4). However late in the season when beet armyworms were fed terminal leaves located in the upper part of the plant (i.e., upper-canopy leaves), survival of larvae on WideStrike[R] was very high (>%60) (Fig. 5). Siebert et al. (2009) reported that expression of Cry1F protein is greatest in mature leaves as compared to other structures such as terminal leaves, squares, flowers, and bolls, and that protein levels in mature leaves increases with age. This expression profile for Cry1F is very different than what was reported for Cry2Ab in Bollgard II[R], where expression in all tissue appears to remain relatively constant over time Adamczyk et al. (2001). In addition, mortality at >109 DAP of beet armyworms on WideStrike[R] terminal leaves was similar to what was observed with Bollgard[R] which also contains a Cry1Ac-like transgene that provides little efficacy against both beet and fall armyworms (Stewart et al. 2001; Adamczyk et al. 1998; Adamczyk & Gore 2004). We believe that low levels of Cry1F protein reported for new foliage (e.g., young terminal leaves) as compared to mature, fully expanded leaves may partially explain the low mortality observed against beet armyworms. Furthermore, our field studies with leaves collected from different portions of the plant supports these temporal and spatial conclusions. More damage was observed from beet armyworms feeding on WideStrike[R] leaves compared to Bollgard II[R] leaves in the upper canopy compared to the middle canopy (Fig. 6). Even with the presumed greater efficacy of Cry1F compared to Cry2Ab against the fall armyworm, this same trend was observed (Fig. 7). Regardless of transgenic trait, it is interesting to note that more damage was observed for both armyworm species caged on leaves located in the upper canopy. This may be partially explained by greater amounts of secondary plant compounds, as well as a thicker leaf cuticle, for leaves located lower in the plant canopy. In some situations, we believe that beet armyworms may need supplemental foliar insecticides for controlling outbreak populations that may be feeding on young tissues especially late in the season where Cry1F levels do not have the time to build to provide adequate control.





Various transgenes that are used to produce cotton with the WideStrike[R] trait may partially explain the observed protein expression profile of Cry1F. In particular, the cry1F trait in WideStrike[R] is regulated by the (4ocs) DeltaMas 2' Promoter. Previous studies have shown that the dual promoter of Agrobacterium tumefaciens mannopine synthase (mas) genes is regulated by plant growth hormones, and that the activity of the mas dual promoters increases basipetally in developing plants; it has also been reported that the apical meristem contains a factor that inhibits stimulation of mas promoter activity (Langridge et al. 1989). However, it is important to note that WideStrike[R] traits contain enhancers that make the promoter more constitutive, whereby losing sensitivity to hormones (US Patent: 5,955,646). Thus, further research is needed to determine the various plant mechanisms that confer protein expression for the WideStrike[R] traits.



We thank the efforts of Alex Gomezplata, Carlos Gracia, Jay Alejandro Frank Garza, and Chuy Caballero for helping with rearing insects and conducting bioassays. Mention of a trademark, warranty, proprietary product or vendor does not constitute a guarantee by the USDA and does not imply approval or recommendation of the product to the exclusion of others that may be suitable.



ADAMCZYK, JR., J. J., AND J. GORE. 2004. Laboratory and field performance of cotton containing Cry1Ac, Cry1F, and both Cry1Ac and Cry1F (WideStrike[R]) against beet armyworm and fall armyworm larvae (Lepidoptera: Noctuidae). Florida Entomol. 87: 427-432.

ADAMCZYK, JR., J. J., L. C. ADAMS, AND D. D. HARDEE. 2001. Field efficacy and seasonal expression profiles for terminal leases of single and double Bt toxin cotton genotypes. J. Econ. Entomol. 94: 1589-1593.

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LANGRIDE, W. H. R., K. J. FITZGERALD, C. KONCZ, J. SCHELL, AND A. A. SZALAY 1989. Dual promoter of Agrobacterium tumefaciens mannopine synthase genes is regulated by plant growth hormones. Proc. Natl. Acad. Sci. USA 86: 3219-3223.

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J. J. ADAMCZYK, JR., S. GREENBERG, J. S. ARMSTRONG, W. J. MULLINS (1), L. B. BRAXTON (2), R. B. LASSITER (2) AND M. W. SIEBERT (2) USDA, ARS, KSARC, Beneficial Insects Research Unit, 2413 E. Hwy 83, Weslaco, TX 78501

(1) Monsanto Co., St. Louis, MO (2) Dow AgroSciences LLC, Indianapolis, IN

Year Location Bt Trait Variety

2005 Stoneville, MS Bollgard St 4691B
2005 Stoneville, MS Bollgard II DP 424B11/RR
2005 Stoneville, MS Non-Bt DP 5415
2005 Stoneville, MS Non-Bt SG 521 RR
2005 Stoneville, MS Non-Bt St 474
2005 Stoneville, MS WideStrike Phy 470WR
2007 Weslaco, TX Bollgard DPL 444 BG/RR
2007 Weslaco, TX Bollgard II Americot 1532 BGII/RR
2007 Weslaco, TX Bollgard II St 4357 BGII/RRF
2007 Weslaco, TX Bollgard II DPL 424 BGII/RR
2007 Weslaco, TX Bollgard II DPL 143 BGII/RR
2007 Weslaco, TX Non-Bt Americot 262R
2007 Weslaco, TX Non-Bt Phy 425 RF
2007 Weslaco, TX Non-Bt DPL 5415 RR
2007 Weslaco, TX WideStrike Phy 485 WRF

Year Bt Endotoxins Owner of Bt Trait

2005 CrylAc Monsanto
2005 CrylAc + Cry2Ab Monsanto
2005 None None
2005 None None
2005 None None
2005 CrylAc+ CryiF DowAgrosciences
2007 CrylAc Monsanto
2007 CrylAc + Cry2Ab Monsanto
2007 CrylAc + Cry2Ab Monsanto
2007 CrylAc + Cry2Ab Monsanto
2007 CrylAc + Cry2Ab Monsanto
2007 None None
2007 None None
2007 None None
2007 CrylAc + CryiF Dow Agrosciences

Year Owner of Variety

2005 Stoneville (Bayer Cropscience)
2005 Delta & Pineland (Monsanto)
2005 Delta & Pineland (Monsanto)
2005 Delta & Pineland (Monsanto)
2005 Stoneville (Bayer Cropscience)
2005 Phytogen (Dow Agrosciences)
2007 Delta & Pineland (Monsanto)
2007 Americot
2007 Stoneville Seed Co. (Bayer Cropscience)
2007 Delta & Pineland (Monsanto)
2007 Delta & Pineland (Monsanto)
2007 Americot
2007 Dow Agrosciences
2007 Delta & Pineland (Monsanto)
2007 Dow Agrosciences
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Article Details
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Author:Adamczyk, J.J., Jr.; Greenberg, S.; Armstrong, J.S.; Mullins, W.J.; Braxton, L.B.; Lassiter, R.B.; S
Publication:Florida Entomologist
Article Type:Report
Geographic Code:1USA
Date:Dec 1, 2008
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