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J.D. Abbott and L. T. Thetford

Cyromazine is a triazine molecule with insect growth regulator properties being developed for control of Colorado potato beetle (Leptinotarsa decemlineata Say) (CPB) in vegetables. Research presented focuses primarily on results with potato (Solanum tuberosum L.), however, crop safety has been observed in other crops within the Solanaceae. Several trials were conducted in PA and NY during 1991 to examine the rates and timing necessary to control CPB in potatoes. Data from replicated small plot trials and non-replicated large block trials are included. Rates examined ranged 70 to 560 g ai na-1 applied alone or in combination with a pyrethroid or Bt. Comparisons were made with insecticides presently registered for CPB control in potatoes and cyromazine compared quite favorably. Two applications per CPB generation were made, the first at the beginning of CPB egg hatch and a second 7-16 days later for each generation. This application schedule provided excellent (90%) control of CPB larvae. The reduction in larvae also resulted in a reduction in adult CPB and potato leaf area damaged through insect feeding. In the test conducted in PA, an increase in size and number of tubers was observed when plants were treated with cyromazine. -These increases resulted in a 23-28% increase in total yield compared to that obtained from the untreated check plots.

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Benolt Lacasse, C. Laguë, S. Yelle, P.M. Roy, and M. Khelifi

A front-mounted prototype designed to pneumatically remove Colorado potato beetles (CPB) from potato plants was tested in the field. Effects of different combinations of airflow velocities, nozzle widths, and travel speeds were investigated. Results showed that capture and dislodging of CPBs were better for adults and big larvae (L3 & L4). On the other hand, neither the airflow width and velocity nor the travel speed affected significantly the dislodging and the collection of small larvae. Field trials on the removal of larvae under the effect of different travel speeds showed that, the slower the prototype moved, the better was the collection of L3-L4 larvae. This study demonstrates the potential of pneumatic control of adult and L3-L4 CPBs.

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D.S. Douches, T.J. Kisha, J.J. Coombs, W. Li, W.L. Pett, and E.J. Grafius

The Colorado potato beetle, Leptinotarsa decemlineata (Say), is the most serious insect pest of potatoes throughout the eastern and north central United States. Host plant resistance to the Colorado potato beetle has been identified in wild Solanum species and Bt-transgenic potato lines. Detached-leaf bioassays (72 h) were conducted on insecticide-resistant, first instar Colorado potato beetles to study the effectiveness of individual and combined host plant resistance traits in potato. Potato lines tested include non-transgenic cultivars (`Russet Burbank', `Lemhi Russet', and `Spunta'), a line with glandular trichomes (NYL235-4), a line with high foliar leptines (USDA8380-1), and transgenic lines expressing either codon-modified Bt-cry3A or Bt-cry5 (Bt-cry1Ia1). Bt-cry3A transgenic lines, foliar leptine line, and foliar leptine lines with Bt-cry5 had reduced feeding compared to non-transgenic cultivars. Glandular trichome lines and glandular trichome lines with Bt-cry5 did not reduce feeding in this no-choice feeding study. Some Bt-cry5 transgenic lines, using either the constitutive promoters CaMV35s or (ocs)3mas (Gelvin super promoter), were moderately effective in reducing larval feeding. Feeding on Bt-cry5 transgenic lines with the tuber-specific patatin promoter was not significantly different than or greater than feeding on the susceptible cultivars. Mortality of first instars was highest when fed on the Bt-cry3A lines (68% to 70%) and intermediate (38%) on the Bt-cry5 `Spunta' line SPG3 where the gus reporter gene was not included in the gene construct. Host plant resistance from foliar leptines is a candidate mechanism to pyramid with either Bt-cry3A or Bt-cry5 expression in potato foliage against Colorado potato beetle. Without multiple sources of host plant resistance, long-term sustainability is questionable for a highly adaptable insect like the Colorado potato beetle.

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D.W.A. Hunt, A. Liptay, and C.F. Drury

Host plant selection by Colorado potato beetle [Leptinotarsa decemlineata (Say)] was examined on tomato [Lycopersicon esculentum Mill.] transplants fertilized with varying N, P, and K concentrations during greenhouse production. In choice tests conducted with beetles in the field and the greenhouse, the insect preference for plants increased with increasing leaf tissue N concentration, but P and K concentrations had no effect. Five-day, seedling acclimatization to outdoor spring temperatures before planting reduced the insect preference for plants.

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Shelley Jansky, Sandra Austin-Phillips, and Corine McCarthy

The Colorado potato beetle (CPB) is a major insect pest that is controlled mainly through the use of pesticides. Development of potato clones with multiple forms of host plant resistance may provide a stable alternative or supplemental form of CPB control. Tetraploid hybrids were developed by somatic fusion of diploid interspecific Solanum clones with different forms of resistance to CPB. Hybrids were created between a clone containing leptine glycoalkaloids and four clones producing glandular trichomes. One fusion produced vigorous hybrids that were analyzed for CPB resistance traits. Somaclonal variation among hybrids was detected for trichome density and resistance to feeding by adult and larval beetles. Somatic hybrids were less resistant than the parents in adult feeding preference trials, but several were more resistant than either parent in larval feeding trials. Future studies are needed to determine whether clones producing both glandular trichomes and leptines express resistance that is more stable than that of clones with only one resistance factor.

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Simon Lachance and Conrad Cloutier

Predators and parasitoids used for biological control must possess good dispersal potential in order to ensure spatially uniform and cost-effective control. The rate of dispersal of Perillus bioculatus (F.) (Hemiptera: Pentatomidae), a predator of the Colorado potato beetle (Leptinotarsa decemlineata), was measured following central release in 0.025-ha potato plots. Factors influencing predator dispersal were also studied under controlled conditions in plant growth chambers. Temperature, predator size as affected by instar, and physiological age with respect to the completion of feeding during the intermolt stage were found to be significant factors. Predator density was also evaluated because of the strong tendency for this species to aggregate, thereby influencing dispersal. Results can be used to develop predictive models for inundative releases of P. bioculatus.

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Jim Mooney and Shelley H. Jansky

Resistance to the Colorado potato beetle (CPB) and green peach aphid (GPA) would be valuable if it could be effectively transferred from wild potato species to the cultivated potato. Eighteen diploid interspecific hybrids have been developed using Solanum tuberosum Gp. Tuberosum haploids (2n = 2x = 24) and the diploid wild species S. berthaultii (ber), S. chacoense (CHC), S. jamesii (jam), and S. tarijense (tar). Twenty-five genotypes per family were screened for resistance to CPB and GPA. Feeding trials were carried out on intact leaves. The degree of resistance to CPB was determined by the stage of instar development and weight of larvae after a four day feeding period; resistance to GPA was evaluated by aphid reproduction and survival after a fifteen day feeding period. Highly CPB or GPA resistant clones, compared to `Norgold Russet',, have been identified thus far. Some clones express high levels of resistance to both CPB and GPA. Crosses between resistant clones and S. tuberosum will be carried out at the diploid level in an attempt to combine resistance with good tuberization qualities.

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Anusuya Rangarajan, A. Raymond Miller, and Richard Veilleux

Leptine (LP) glycoalkaloids have been demonstrated to confer natural resistance to the Colorado potato beetle (CPB) in Solanum chacoense (chc). Development of cultivated potatoes with natural resistance to CPB has the potential to reduce both costs and environmental impacts of production by reducing pesticide use. To introgress the genes conferring leptine production from chc into S. tuberosum (tbr), clones of chc have been crossed with clones of S. phureja. Leaf disks from eight hybrids were subjected to a CPB second instar feeding bioassay to determine if extent of feeding was related to LP levels. Most hybrids contained leptinidine (LD, the aglycone of LP) levels intermediate to chc and tbr, and insect feeding was suppressed 30% to 50% in hybrids containing >10 mg·g–1 DW LD. One hybrid displaying feeding suppression contained a very low level of LD, whereas another hybrid that contained higher levels of LD had higher feeding rates. The presence of LD at “threshold” levels in these hybrids will suppress feeding of CPB, but other factors affecting resistance are also present and need to be explored.

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G. Jelenkovic, S. Billings, Q. Chen, J. Lashomb, and G. Ghidiu

A chimeric construct, containing the synthetic cryIIIA (Btt) gene, the NPTII selectable marker and the uidA reporter gene, was incorporated via Agrobacterium tumefaciens into eggplant, variety Hibush. The synthetic cryIIIA gene, altered at the nucleotide level without changing the amino acids of the toxic protein by J. Kemp of New Mexico State Univ., Las Cruces, is adapted for high expression in plant cells. To verify the transgenic status, GUS assays were performed on over 300 plants, from which 185 were confirmed to be transgenic. Physical incorporation of the chimeric construct was further confirmed by Southern analysis of about 30 transgenic plants; both single and multiple site incorporation of the Btt gene were found. Resistance to Colorado potato beetle (CPB) was assessed by: a) placing egg masses of CPB on leaves of plants grown in the growth chamber; b) placing first-instar larvae on detached leaves; c) observing 173 transgenic plants under field conditions. About 60% of the transgenic plants displayed a very high level of resistance to CPB. No larvae survived on the resistant plants longer than 50–60 hours after hatching. Upon selfing, the transgenic plants with a single construct segregate in the S1 generation in a Mendelian fashion.

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Nazareno Acciarri, Gabriele Vitelli, Salvatore Arpaia, Giuseppe Mennella, Francesco Sunseri, and Giuseppe L. Rotino

Colorado potato beetle (CPB; Leptinotarsa decemlineata-Say) is a serious pest because it has developed resistance against insecticides. Three transgenic eggplant (Solanum melongena L.) lines bearing a mutagenized Bacillus thuringiensis Berl. gene coding for the Cry3B toxin, and the nontransformed control DR2-line were tested in field trials to assess their insect resistance. The transgenic lines 3-2, 6-1, and 9-8 were tested at two different locations in a randomized complete-block design. Samples were taken biweekly to assess the level of CPB and the presence of other insects. At harvest, total yield and fruit number per plot were recorded. Two transgenic lines showed high levels of resistance at both locations, as measured by CPB abundance and yield. Fruit production was almost twice as great in the highly resistant lines (3-2 and 9-8) as in the nontransformed control. The 6-1 transgenic line showed an intermediate level of resistance; it was similar to the control under heavy CPB pressure and was comparable to the other transgenic lines under milder infestations. Analysis by double antibody sandwich–enzyme linked immunosorbent assay (DAS–ELISA), performed on different tissues, revealed a lower amount of Cry3B protein in the 6-1 transgenic line than in lines 3-2 and 9-8. No detrimental effects on nontarget arthropods (including the chrysomelid Altica) were evident. Field observations confirmed that Bt may be able to control CPB infestation in eggplant, representing a potential effective and environmentally safe means of pest control.