Quantitative trait locus mapping and functional genomics of an organophosphate resistance trait in the western corn rootworm, Diabrotica virgifera virgifera.

The western corn rootworm, Diabrotica virgifera virgifera, is an insect pest of corn and population suppression with chemical insecticides is an important management tool. Traits conferring organophosphate insecticide resistance have increased in frequency amongst D. v. virgifera populations, resulting in the reduced efficacy in many corn-growing regions of the USA. We used comparative functional genomic and quantitative trait locus (QTL) mapping approaches to investigate the genetic basis of D. v. virgifera resistance to the organophosphate methyl-parathion. RNA from adult methyl-parathion resistant and susceptible adults was hybridized to 8331 microarray probes. The results predicted that 11 transcripts were significantly up-regulated in resistant phenotypes, with the most significant (fold increases ≥ 2.43) being an α-esterase-like transcript. Differential expression was validated only for the α-esterase (ST020027A20C03), with 11- to 13-fold greater expression in methyl-parathion resistant adults (P < 0.05). Progeny with a segregating methyl-parathion resistance trait were obtained from a reciprocal backcross design. QTL analyses of high-throughput single nucleotide polymorphism genotype data predicted involvement of a single genome interval. These data suggest that a specific carboyxesterase may function in field-evolved corn rootworm resistance to organophosphates, even though direct linkage between the QTL and this locus could not be established.

Impact of the Bt Corn Proteins Cry34/35Ab1 and Cry3Bb1, Alone or Pyramided, on Western Corn Rootworm (Coleoptera: Chrysomelidae) Beetle Emergence in the Field.

Western corn rootworm, Diabrotica virgifera virgifera LeConte, is a major pest of corn, Zea mays L. The effect of the Bt proteins Cry34/35Ab1 and Cry3Bb1, alone or pyramided in corn hybrids on D. v. virgifera adult emergence was evaluated in field experiments for 3 yr. Experiments were infested artificially with 2,500 viable D. v. virgifera eggs per row meter of corn. The reduction in beetle emergence compared with non-Bt controls, from Cry34/35Ab1, Cry3Bb1, and the pyramided hybrids ranged from 64.3 to 97.4%, 91.1 to 95.2%, and 98.1 to 99.6%, respectively. The sex ratio of emerged beetles was usually female-biased from the Cry3Bb1 and pyramided treatments, but not from Cry34/35Ab1 treatment alone. Emergence from all Bt hybrids was delayed compared with the control, with the delay longest from the pyramided hybrid. In 2013, three egg infestation levels were tested, with density-dependent mortality observed at 1,250 viable eggs per row meter. The effect of Bt proteins on the emergence timing and sex ratio of D. v. virgifera may impact the suitability of resistance management plans, specifically the effectiveness of the refuge strategy. Susceptible males emerging from refuge might not be synchronized to mate with potentially resistant females emerging later from Bt corn hybrids.

Phenotypic trait changes in laboratory--reared colonies of the maize herbivore, Diabrotica virgifera virgifera.

The North American and European maize pest Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) was used to assess whether conditions of the natal field, subsequent laboratory rearing, or genetic population origin affect phenotypic traits of fitness, activity, or morphometrics. Standardized laboratory bioassays with large sample sizes revealed that none of the 16 tested traits, except crawling behaviours, appeared consistently stable across all seven tested colonies. Environmental conditions in the natal field of the F 0 generation affected trait averages of the subsequently reared F 1 generation in laboratory in ca. 47% of cases, and trait variability in 67% of cases. This was apparent for fitness and morphometrics, but less obvious for activity traits. Early generation laboratory rearing affected trait averages in ca. 56% of cases: morphometrics changed; fecundity and egg survival increased from F 1 to F 2. Trait variability increased or decreased in 38% of cases. Laboratory rearing for over more than 190 generations affected the trait averages in 60% of cases, reflected by decreases in flight activity and increases in body size, weight, and fecundity to some extent. It had little effect on trait variability, especially so for morphometric variability. The genetic population origin affected average levels of 55% and variability of 63% of phenotypic traits. A comparison among D. v. virgifera studies might be difficult if they use different populations or laboratory colonies. It is advised to consider possible effects of original field conditions, laboratory rearing, and population genetics when planning comparative studies targeting fitness, activity, or morphometric questions regarding Diabrotica species.

Abundance of Coleomegilla maculata (Coleoptera: Coccinellidae) in corn rootworm-resistant Cry3Bb1 maize.

Lady beetles (Coleoptera: Coccinellidae) are important polyphagous predators in maize, Zea mays L., fields. Transgenic Cry3Bb1 maize hybrids express a coleopteran-specific insecticidal protein derived from Bacillus thuringiensis (Berliner) subsp. kumamotoensis that is targeted at corn rootworm larvae. This study evaluated impacts of Cry3Bb1 protein-expressing maize, tefluthrin-treated maize, and untreated controls on lady beetle abundance at preanthesis, anthesis, and postanthesis maize-developmental periods near Brookings in eastern South Dakota during 2001 and 2002. The dominant lady beetle species captured on Pherocon AM sticky traps was Coleomegilla maculata De Geer. It comprised 73.5 and 69.9% of all adult Coccinellidae caught in 2001 and 2002, respectively. Numbers of C. maculata captured in Cry3Bb1 maize were not significantly different from those in untreated plots during preanthesis, and adults were more abundant in Cry3Bb1 maize than in tefluthrin-treated and untreated plots during anthesis and postanthesis. Whole-plant sampling confirmed C. maculata predominance with the species representing 89.2 and 91.4% of all adult lady beetles observed in 2001 and 2002, respectively. Whole-plant sampling also indicated a lack of negative effects from Cry3Bb1 maize on abundance of lady beetle eggs, larvae, pupae, or adults. Overall, these findings indicate that Cry3Bb1-expressing hybrids are not likely to impose harmful effects on C. maculata, a species common to maize production systems in the northern Great Plains. This research further suggests that Cry3Bb1 maize has the potential for conservation of these beneficial coccinellids in maize production systems.