The soybean aphid, Aphis glycines (Hemiptera: Aphididae), is a pest of soybeans in Asia, and in recent years has caused extensive damage to soybeans in North America. Within these agroecosystems, generalist predators form an important component of the assemblage of natural enemies, and can exert significant pressure on prey populations. These food webs are complex and molecular gut-content analyses offer nondisruptive approaches for examining trophic linkages in the field. We describe the development of a molecular detection system to examine the feeding behaviour of Orius insidiosus (Hemiptera: Anthocoridae) upon soybean aphids, an alternative prey item, Neohydatothrips variabilis (Thysanoptera: Thripidae), and an intraguild prey species, Harmonia axyridis (Coleoptera: Coccinellidae). Specific primer pairs were designed to target prey and were used to examine key trophic connections within this soybean food web. In total, 32% of O. insidiosus were found to have preyed upon A. glycines, but disproportionately high consumption occurred early in the season, when aphid densities were low. The intensity of early season predation indicates that O. insidiosus are important biological control agents of A. glycines, although data suggest that N. variabilis constitute a significant proportion of the diet of these generalist predators. No Orius were found to contain DNA of H. axyridis, suggesting intraguild predation upon these important late-season predators during 2005 was low. In their entirety, these results implicate O. insidiosus as a valuable natural enemy of A. glycines in this soybean agroecosystem.

Determining relative strengths of trophic links is critical for ranking predators for conservation biological control. Molecular gut-content analysis enables ranking by incidence of prey remains in the gut, but differential digestive rates bias such rankings toward predators with slower rates. This bias can be reduced by indexing each predator's half-life to that of the middle-most half-life in a predator complex. We demonstrate this with data from key species in the predator complex of Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), comprising adults and immatures of four taxonomically diverse species. These animals display order-of-magnitude variation in detectability half-life for the cytochrome oxidase I DNA sequence of a single CPB egg: from 7.0 h in larval Coleomegilla maculata (DeGeer) (Coleoptera: Coccinellidae) to 84.4 h in nymphal Perillus bioculatus (Fabricius) (Hemiptera: Pentatomidae). The raw species-specific incidence of L. decemlineata DNA in the guts of 351 field-collected predators ranged from 11 to 95%, ranking them as follows: C. maculata adults < Lebia grandis Hentz (Coleoptera: Carabidae) adults < Podisus maculiventris (Say) (Hemiptera: Pentatomidae) adults < P. maculiventris nymphs < P. bioculatus adults < P. bioculatus nymphs. Half-life adjustment reorders the rankings: C. maculata adults < P. bioculatus adults < P. bioculatus nymphs < P. maculiventris nymphs < L. grandis adults < P. maculiventris adults. These changes in status demonstrate the value of half-lifeadjusted molecular gut-content data for ranking predators. This is the first study to measure prey detectability half-lives for the key arthropod predators of a major insect pest, and to use them to evaluate the relative impact of all adults and immatures in this predator complex.

The time during which prey remains are detectable in the gut of a predator is an important consideration in the interpretation of molecular gut-content data, because predators with longer detectability times may appear on the basis of unweighted data to be disproportionately important agents of prey population suppression. The rate of decay in detectability, typically expressed as the half-life, depends on many variables; one that has not been explicitly examined is the manner in which the predator processes prey items. The influence of differences in feeding mode and digestive physiology on the half-life of DNA for a single prey species, the Colorado potato beetle Leptinotarsa decemlineata (Say), is examined in two predators that differ dramatically in these attributes: the pink ladybeetle, Coleomegilla maculata (DeGeer), which feeds by chewing and then ingesting the macerated material into the gut for digestion; and the spined soldier bug, Podisus maculiventris (Say), which physically and enzymatically processes the prey extra-orally before ingestion and further digestion in the gut. In order to standardize the amount of DNA consumed per predator, a single L. decemlineata egg was used as the prey item; all predators were third instars. The PCR assay yields estimated prey DNA half-lives, for animals maintained under field temperatures, of 7.0 h in C. maculata and 50.9 h in P. maculiventris. The difference in the prey DNA half-lives from these two predators underscores the need to determine detectabilities from assemblages of predators differing in feeding mode and digestive physiology, in order to weight positives properly, and hence determine the predators' relative impacts on prey population suppression.

In Escheyichia coli K-12, expression of zwf, the gene for glucose 6-phosphate dehydrogenase, is coordinated with the cellular growth rate and induced by superoxide-generating agents. To initiate the study of the molecular mechanisms regulating its expression, the gene was cloned and its DNA sequence was determined. The 5' ends of zwfmRNA isolated from cells growing in glucose and acetate minimal media were mapped. The map was complex in that transcripts mapped to -45, -52, and -62, with respect to the beginning of the coding sequence. Three analytical methods were used to search the DNA sequence for putative promoters. Only one sequence for a promoter recognized by the r70 form of RNA polymerase was found by all three search routines that could be aligned with a mapped transcript, indicating that the other transcripts arise by processing of the mRNA. A computer-assisted search did not reveal a thermodynamically stable long-range mRNA secondary structure that is capable of sequestering the translation initiation region, which suggests that growth-ratedependent regulation of glucose 6-phosphate dehydrogenase level may not be carried out by a mechanism similar to the one for the gene (gnd) for 6-phosphogluconate dehydrogenase. The DNA segment between the -10 hexamer and the start point of transcription resembles the discriminator sequence of stable RNA genes, which has been implicated in stringent control and growth-rate-dependent regulation.The oxidative branch of the pentose phosphate pathway provides ribose for nucleoside biosynthesis and NADPH for reductive biosyntheses (15,20). The two dehydrogenases of this pathway are glucose 6-phosphate dehydrogenase (G6PD; EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (6PGD; EC 1.1.1.44). In Escherichia coli, the specific activities of these enzymes increase in proportion to growth rate during steady-state growth on different carbon sources (56). Although the growth rate dependence of the level of these two enzymes resembles that of the components of the translational apparatus, they are not part of the same regulatory network. After a nutritional shiftup, the accumulation rate of ribosomal components increases immediately, whereas the accumulation rate of G6PD and 6PGD has the same kinetics as that of total protein, i.e., increasing only after a lag (16). Thus, the mechanism(s) underlying the growth-rate-dependent regulation of zwf and gnd, which encode G6PD and 6PGD, respectively, may be common to other nonribosomal proteins whose levels also increase with increasing growth rate.Studies of the control of gnd expression point to an interesting mechanism. Regulation is exerted at a posttranscriptional step and requires sequences within the 6PGD coding region (3, 4). The internal regulatory site, which lies between codons 71 and 74, is complementary to the translation initiation region on gnd mRNA (12). This internal complementary sequence (ICS) appears to function as a cis-acting antisense RNA by forming a long-range secondary structure that sequesters the ribosome bin...