The Effect of Threecornered Alfalfa Hopper Populations on Alfalfa Growth and the Development of Host Plant Resistance Screening Techniques.

This dissertation research was designed with three main objectives: to evaluate the effects of threecornered alfalfa hopper, Spissistilus festinus (Say), feeding on alfalfa, Medicago sativa L., yield, regrowth, root carbohydrate composition, and disease incidence; to screen alfalfa cultivars for resistance to the threecornered alfalfa hopper and develop effective screening techniques; and to determine the mechanism(s) of this resistance. Greenhouse studies determined the effects of threecornered alfalfa hopper infestations on alfalfa growth and their relationship with Fusarium crown-rot. Significant interaction effects between insect population and the presence of Fusarium were found for number of harvestable stems and acid detergent fiber concentration. The main effect of inoculation did not affect any of the plant parameters studied. Insect populations increased plant chlorosis and accelerated plant maturity. Insect populations also reduced plant height, root carbohydrate concentration, stem regrowth, and forage quality. Threecornered alfalfa hoppers also caused an increase in Fusarium crown-rot severity. Choice and no-choice tests were conducted under greenhouse conditions to determine the presence of alfalfa antibiosis or antixenosis to the threecornered alfalfa hopper. Adult threecornered alfalfa hoppers were released into cages containing six alfalfa cultivars. Fewer threecornered alfalfa hoppers and girdles were found on cultivars \u27Cimarron VR\u27 and \u27Zia\u27. In the no-choice test, one newly hatched nymph was placed on each plant and development was monitored daily. Nymphal duration was longest when nymphs developed on \u27Florida 77\u27 and shortest on \u27Zia\u27. Adult weights were reduced when nymphs developed on \u27Dona Ana\u27 and \u27Cimarron\u27. Screening techniques were developed to evaluate alfalfa resistance under field conditions. Resistance was characterized by adult populations, girdle damage, and stand persistence. \u27Cimarron VR\u27, \u27Dona Ana\u27, and \u27GA Plains\u27 exhibited resistance to the threecornered alfalfa hopper by having less girdle damage and greater stand persistence. \u27Ladak\u27 showed the highest level of susceptibility. Adult population monitoring was not an effective screening criterion. In addition, multiple girdle counts must be taken to accurately categorize alfalfa cultivars

Cross-Resistance of Cry1Ab-Selected \u3ci\u3eOstrinia nubilalis\u3c/i\u3e (Lepidoptera: Crambidae) to \u3ci\u3eBacillus thuringiensis\u3c/i\u3e δ -Endotoxins

Corn plants expressing the toxin from Bacillus thuringiensis (Berliner) have proven to be effective in controlling lepidopteran pests such as the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae). Several Bt toxins are being tested and incorporated into crop genomes, although tests for cross-resistance among different toxins have been limited by a lack of resistant colonies. Four different colonies of O. nubilalis selected with full-length Cry1Ab incorporated into artificial diet developed significant levels of resistance (2.0- to 10-fold) within 10 generations. Additionally, selection with Cry1Ab resulted in decreased susceptibility to a number of other toxins to which the selected colonies were not previously exposed. Significantly, levels of resistance were highest to Cry1Ac with resistance ratios up to 51.0-fold. Low levels (less than five-fold) of cross-resistance were detected with Cry1F. In contrast, Cry9C susceptibility was unaffected by selection with Cry1Ab. These results indicate that the availability of multiple toxins could improve resistance management strategies, provided cross-resistance among toxins is not a factor

Cross-Resistance of Cry1Ab-Selected \u3ci\u3eOstrinia nubilalis\u3c/i\u3e (Lepidoptera: Crambidae) to \u3ci\u3eBacillus thuringiensis\u3c/i\u3e δ -Endotoxins

Corn plants expressing the toxin from Bacillus thuringiensis (Berliner) have proven to be effective in controlling lepidopteran pests such as the European corn borer, Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae). Several Bt toxins are being tested and incorporated into crop genomes, although tests for cross-resistance among different toxins have been limited by a lack of resistant colonies. Four different colonies of O. nubilalis selected with full-length Cry1Ab incorporated into artificial diet developed significant levels of resistance (2.0- to 10-fold) within 10 generations. Additionally, selection with Cry1Ab resulted in decreased susceptibility to a number of other toxins to which the selected colonies were not previously exposed. Significantly, levels of resistance were highest to Cry1Ac with resistance ratios up to 51.0-fold. Low levels (less than five-fold) of cross-resistance were detected with Cry1F. In contrast, Cry9C susceptibility was unaffected by selection with Cry1Ab. These results indicate that the availability of multiple toxins could improve resistance management strategies, provided cross-resistance among toxins is not a factor

Optimizing Egg Recovery From Wild Northern Corn Rootworm Beetles (Coleoptera: Chrysomelidae)

The northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), is one of the most important insect pests in the U.S. Corn Belt. Efforts to obtain eggs from wild northern corn rootworm populations using techniques developed for other rootworm species have been unsuccessful due to lack of oviposition. In 2016, we evaluated four oviposition media in choice tests within each of three female densities in 30.5 × 30.5 × 30.5 cm BugDorm cages. The number of eggs laid per female was significantly affected by female density and the interaction of female density × oviposition media, but oviposition was relatively poor in all oviposition media (1.2 eggs per female when averaging the three female densities and all oviposition media). Single females were also evaluated in nonchoice assays in 6 cm × 6 cm × 8 cm clear plastic boxes and averaged up to 108 eggs per female depending on the oviposition media. In 2017, the cumulative number of eggs laid per female in boxes with one female was not significantly different from the number of eggs laid per female in boxes with 3 females. In 2018, the cumulative number of eggs laid per female was not significantly different between female densities of 1, 3, 5, or 10 females per box. Total egg production per box therefore increased as female density increased. More than 27,000 wild northern corn rootworm eggs were collected from just 190 females when collected relatively early in the field season. We now have an efficient and robust system for obtaining eggs from wild northern corn rootworm females

Activity of gut proteinases from Cry1Ab-selected colonies of the European corn borer, \u3ci\u3eOstrinia nubilalis \u3c/i\u3e (Lepidoptera: Crambidae)

Susceptibility to the Cry1Ab protoxin and toxin from Bacillus thuringiensis (Berliner) and activity of gut proteinases were assessed in both susceptible and Cry1Ab-selected colonies of European corn borer, Ostrinia nubilalis (Hübner). Resistance in two different selected colonies was at least 6- and 15-fold for the Cry1Ab protoxin and 108- and 484-fold for the Cry1Ab toxin. Activities of trypsin-like, chymotrypsin-like and elastase-like proteinases were variable among the colonies tested and not indicative of a major contribution to Cry1Ab resistance. Activation of the 130-kDa Cry1Ab protoxin occurred rapidly in all colonies, with no apparent differences among colonies. In addition, there were no apparent changes in activated Cry1Ab processing, indicating that proteolytic degradation was not associated with resistance. These results suggest that mechanisms other than proteolytic activation of protoxin and toxin degradation, such as target site modification may be involved in the resistance to B. thuringiensis Cry1Ab in these O. nubilalis colonies

Optimizing Egg Recovery From Wild Northern Corn Rootworm Beetles (Coleoptera: Chrysomelidae)

The northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), is one of the most important insect pests in the U.S. Corn Belt. Efforts to obtain eggs from wild northern corn rootworm populations using techniques developed for other rootworm species have been unsuccessful due to lack of oviposition. In 2016, we evaluated four oviposition media in choice tests within each of three female densities in 30.5 × 30.5 × 30.5 cm BugDorm cages. The number of eggs laid per female was significantly affected by female density and the interaction of female density × oviposition media, but oviposition was relatively poor in all oviposition media (1.2 eggs per female when averaging the three female densities and all oviposition media). Single females were also evaluated in nonchoice assays in 6 cm × 6 cm × 8 cm clear plastic boxes and averaged up to 108 eggs per female depending on the oviposition media. In 2017, the cumulative number of eggs laid per female in boxes with one female was not significantly different from the number of eggs laid per female in boxes with 3 females. In 2018, the cumulative number of eggs laid per female was not significantly different between female densities of 1, 3, 5, or 10 females per box. Total egg production per box therefore increased as female density increased. More than 27,000 wild northern corn rootworm eggs were collected from just 190 females when collected relatively early in the field season. We now have an efficient and robust system for obtaining eggs from wild northern corn rootworm females

Effect of Seed Blends and Soil-Insecticide on Western and Northern Corn Rootworm Emergence from mCry3A1+ eCry3.1Ab Bt Maize

Seed blends containing various ratios of transgenic Bt maize (Zea mays L.) expressing the mCry3A+eCry3.1Ab proteins and non-Bt maize (near-isoline maize) were deployed alone and in combination with a soil applied pyrethroid insecticide (Force CS) to evaluate the emergence of the western corn rootworm, Diabrotica virgifera virgifera LeConte, in a total of nine field environments across the Midwestern United States in 2010 and 2011. Northern corn rootworm, Diabrotica barberi Smith & Lawrence emergence was also evaluated in four of these environments. Both western and northern corn rootworm beetle emergence from all Bt treatments was significantly reduced when compared with beetle emergence from near-isoline treatments. Averaged across all environments, western corn rootworm beetle emergence from 95:5, 90:10, and 80:20 seed blend ratios of mCry3A+eCry3.1Ab: near-isoline were 2.6-, 4.2-, and 6.7-fold greater than that from the 100:0 ratio treatment. Northern corn rootworm emergence from the same seed blend treatments resulted in 2.8-, 3.2-, and 4.2-fold more beetles than from the 100:0 treatment. The addition of Force CS (tefluthrin) significantly reduced western corn rootworm beetle emergence for each of the three treatments to which it was applied. Force CS also significantly delayed the number of days to 50% beetle emergence in western corn rootworms. Time to 50% beetle emergence in the 100% mCry3A+eCry3.1Ab treatment with Force CS was delayed 13.7 d when compared with western corn rootworm beetle emergence on near-isoline corn. These data are discussed in terms of rootworm resistance management

Effect of Seed Blends and Soil-Insecticide on Western and Northern Corn Rootworm Emergence from mCry3A + eCry3.1Ab Bt Maize

Seed blends containing various ratios of transgenic Bt maize ( Zea mays L.) expressing the mCry3A + eCry3.1Ab proteins and non-Bt maize (near-isoline maize) were deployed alone and in combination with a soil applied pyrethroid insecticide (Force CS) to evaluate the emergence of the western corn rootworm, Diabrotica virgifera virgifera LeConte, in a total of nine field environments across the Midwestern United States in 2010 and 2011. Northern corn rootworm, Diabrotica barberi Smith & Lawrence emergence was also evaluated in four of these environments. Both western and northern corn rootworm beetle emergence from all Bt treatments was significantly reduced when compared with beetle emergence from near-isoline treatments. Averaged across all environments, western corn rootworm beetle emergence from 95:5, 90:10, and 80:20 seed blend ratios of mCry3A + eCry3.1Ab: near-isoline were 2.6-, 4.2-, and 6.7-fold greater than that from the 100:0 ratio treatment. Northern corn rootworm emergence from the same seed blend treatments resulted in 2.8-, 3.2-, and 4.2-fold more beetles than from the 100:0 treatment. The addition of Force CS (tefluthrin) significantly reduced western corn rootworm beetle emergence for each of the three treatments to which it was applied. Force CS also significantly delayed the number of days to 50% beetle emergence in western corn rootworms. Time to 50% beetle emergence in the 100% mCry3A + eCry3.1Ab treatment with Force CS was delayed 13.7 d when compared with western corn rootworm beetle emergence on near-isoline corn. These data are discussed in terms of rootworm resistance management.This article is published as Frank, Daniel L., Ryan Kurtz, Nicholas A. Tinsley, Aaron J. Gassmann, Lance J. Meinke, Daniel Moellenbeck, Michael E. Gray et al. "Effect of seed blends and soil-insecticide on western and northern corn rootworm emergence from mCry3A+ eCry3. 1Ab Bt maize." Journal of economic entomology 108, no. 3 (2015): 1260-1270. doi: 10.1093/jee/tov081. Posted with permission.</p