Effects of gossypol on fitness costs associated with resistance to Bt cotton in pink bollworm

Fitness costs associated with insect resistance to Bacillus thuringiensis (Bt) crops may help to delay or prevent the spread of resistance alleles, especially when refuges of non-Bt host plants are present. The potential for such delays increases as the magnitude and dominance of fitness costs increase. Here, we examined the idea that plant secondary chemicals affect expression of fitness costs associated with resistance to Bt cotton in Pectinophora gossypiella (Saunders). Specifically, we tested the hypotheses that gossypol affects the magnitude or dominance of fitness costs, by measuring performance of three independent sets of pink bollworm populations fed artificial diet with and without gossypol. Each set had an unselected susceptible population, a resistant population derived by selection from the susceptible population, and the F1 progeny of the susceptible and resistant populations. No individuals completed development on diets with gossypol in one set, suggesting that these individuals partially lost the ability to detoxify this chemical. In the other two sets, costs affecting survival did not support the hypotheses, but costs affecting pupal weight did. Adding gossypol to diet increased the magnitude and dominance of costs affecting pupal weight. In one of the two sets with survivors on diet with gossypol, costs affecting development time were less recessive when gossypol was present in diet. These results indicate that gossypol increased the magnitude and dominance of some fitness costs. Better understanding of the effects of natural plant defenses on fitness costs could improve our ability to design refuges for managing insect resistance to Bt crops.     

Real-time analysis of ternary complex on particles: direct evidence for partial agonism at the agonist-receptor-G protein complex assembly step of signal transduction

We developed a novel and generalized approach to investigate G protein-coupled receptor molecular assemblies. We solubilized a fusion protein consisting of the beta(2)-adrenergic receptor and green fluorescent protein (GFP) for bead-based flow cytometric analysis. beta(2)-Adrenergic receptor GFP bound to dihydroalprenolol-conjugated beads, providing a K(d) for the fusion protein and, in competition with beta(2)-adrenergic receptor ligands, K(d) values for agonists and antagonists. Beads displaying chelated nickel bound purified hexahistidine-tagged G protein heterotrimers and, subsequently, the binary complex of agonist with beta(2)-adrenergic receptor GFP. The dose-response curves of ternary complex formation revealed maximal assembly for ligands previously classified as full agonists and reduced assembly for ligands previously classified as partial agonists. Guanosine 5'-3-O-(thio)triphosphate-induced dissociation rates of the ternary complex were the same for full and partial agonists. Soluble G protein, competing with ternary complexes on beads provided an affinity estimate of agonist-receptor complexes to G protein. When performed simultaneously, the two assemblies discriminated between agonist, antagonist or inactive molecule in a manner appropriate for high throughput, small volume drug discovery. The assemblies can be further generalized to other G protein coupled receptor protein-protein interactions.     

Measuring conditions and trends in ecosystem services at multiple scales: the Southern African Millennium Ecosystem Assessment (SAfMA) experience

The Southern African Millennium Ecosystem Assessment (SAfMA) evaluated the relationships between ecosystem services and human well-being at multiple scales, ranging from local through to sub-continental. Trends in ecosystem services (fresh water, food, fuel-wood, cultural and biodiversity) over the period 1990-2000 were mixed across scales. Freshwater resources appear strained across the continent with large numbers of people not securing adequate supplies, especially of good quality water. This translates to high infant mortality patterns across the region. In some areas, the use of water resources for irrigated agriculture and urban-industrial expansion is taking place at considerable cost to the quality and quantity of freshwater available to ecosystems and for domestic use. Staple cereal production across the region has increased but was outstripped by population growth while protein malnutrition is on the rise. The much-anticipated wood-fuel crisis on the subcontinent has not materialized but some areas are experiencing shortages while numerous others remain vulnerable. Cultural benefits of biodiversity are considerable, though hard to quantify or track over time. Biodiversity resources remain at reasonable levels, but are declining faster than reflected in species extinction rates and appear highly sensitive to land-use decisions. The SAfMA sub-global assessment provided an opportunity to experiment with innovative ways to assess ecosystem services including the use of supply-demand surfaces, service sources and sink areas, priority areas for service provision, service 'hotspots' and trade-off assessments.     

Association between resistance to Bt cotton and cadherin genotype in pink bollworm

Two strains of pink bollworm, Pectinophora gossypiella (Saunders), each derived in 1997 from a different field population, were selected for resistance to Bacillus thuringiensis (Bt) toxin Cry1Ac in the laboratory. One strain (MOV97-R) originated from Mohave Valley in western Arizona; the other strain (SAF97-R) was from Safford in eastern Arizona. Relative to a susceptible laboratory strain, Cry1Ac resistance ratios were 1700 for MOV97-R and 520 for SAF97-R. For the two resistant strains, larval survival did not differ between non-Bt cotton and transgenic cotton producing CrylAc. In contrast, larval survival on Bt cotton was 0% for the two unselected parent strains from which the resistant strains were derived. Previously identified resistance (r) alleles of a cadherin gene (BtR) occurred in both resistant strains: r1 and r3 in MOV97-R, and r1 and r2 in SAF97-R. The frequency of individuals carrying two r alleles (rr) was 1.0 in the two resistant strains and 0.02 in each of the two unselected parent strains. Furthermore, in two hybrid strains with a mixture of susceptible (s) and r alleles at the BtR locus, all survivors on Bt cotton had two r alleles. The results show that resistance to Cry1Ac-producing Bt cotton is associated with recessive r alleles at the BtR locus in the strains of pink bollworm tested here. In conjunction with previous results from two other Bt-resistant strains of pink bollworm (APHIS-98R and AZP-R), results reported here identify the cadherin locus as the leading candidate for molecular monitoring of pink bollworm resistance to Bt cotton.     

Habitat-Specific Nitrogen Dynamics in New Zealand Streams Containing Native or Invasive Fish

Streams are important sites of nutrient transport and transformation in the landscape but little is known about the way in which individual taxa or individual habitats (riffles and pools) influence nutrient dynamics within stream reaches. We used 5-week additions of a stable isotope (¹⁵NH₄Cl) tracer to investigate nitrogen dynamics in pools and riffles of two New Zealand streams, one with native fish (Galaxias depressiceps) and the other with invasive brown trout (Salmo trutta). In New Zealand, brown trout initiate a trophic cascade leading to increased algal biomass that we predicted would lead to higher N uptake and retention. Uptake of NO₃, but not ammonium, was greater in the trout stream. Rather than causing a large increase in N demand, trout may induce a reallocation of N uptake and retention among food web compartments in different habitats. The largest differences between streams were apparent in riffles, where most uptake and retention of N occurred. In the trout stream, uptake rate by epilithon in riffles was more than six times greater than uptake rates of any other compartment. In the Galaxias stream, several compartments in both habitats had similar uptake rates. Epilithon also accounted for a larger percentage of the ¹⁵N retained in the study reach in the trout stream (51%) than the Galaxias stream (34%). Our results show that an individual predatory taxon (in our case an invader) can influence N dynamics in streams but that the magnitude and location of the impact depend on a range of abiotic and biotic factors involved in N dynamics in streams.     

A rate-independent technique for analysis of nucleic acid sequences: evolutionary parsimony

The method of evolutionary parsimony--or operator invariants--is a technique of nucleic acid sequence analysis related to parsimony analysis and explicitly designed for determining evolutionary relationships among four distantly related taxa. The method is independent of substitution rates because it is derived from consideration of the group properties of substitution operators rather than from an analysis of the probabilities of substitution in branches of a tree. In both parsimony and evolutionary parsimony, three patterns of nucleotide substitution are associated one-to-one with the three topologically linked trees for four taxa. In evolutionary parsimony, the three quantities are operator invariants. These invariants are the remnants of substitutions that have occurred in the interior branch of the tree and are analogous to the substitutions assigned to the central branch by parsimony. The two invariants associated with the incorrect trees must equal zero (statistically), whereas only the correct tree can have a nonzero invariant. The chi 2-test is used to ascertain the nonzero invariant and the statistically favored tree. Examples, obtained using data calculated with evolutionary rates and branchings designed to camouflage the true tree, show that the method accurately predicts the tree, even when substitution rates differ greatly in neighboring peripheral branches (conditions under which parsimony will consistently fail). As the number of substitutions in peripheral branches becomes fewer, the parsimony and the evolutionary-parsimony solutions converge. The method is robust and easy to use.