Enumerative and binomial sequential sampling plans for soybean aphid (Homoptera: Aphididae) in soybean

Since the discovery of the soybean aphid, Aphis glycines Matsumura, in midwestern U.S. soybean, Glycine max L., in 2000, the aphid has become a significant economic pest. Basic information about estimating population density within fields is unknown. Therefore, we developed two sampling plans to efficiently characterize A. glycines densities. Enumerative and binomial sequential plans were developed using 89 data sets collected from 10 commercial fields sampled during 2001-2003. Re-sampling software was used to validate the enumerative plan on whole plant counts, based on Taylor's power law parameters (a = 9.157 and b = 1.543). For research applications, the enumerative plan was modified to provide an actual precision level of 0.10 (SE/mean), which resulted in an average sample number of 310 individual plants. For integrated pest management (IPM) purposes, we developed an enumerative plan with an actual precision of 0.25, which resulted in an average sample number of 38 individual plants. For IPM applications, the binomial plan will likely be more practical. Binomial plans were developed using two tally thresholds at five action thresholds. Final analysis of the operating characteristic curve for each plan indicated that the tally threshold of > or = 40 aphids per plant, and an action threshold of 0.837 (84% of the plants infested) provided the most correct treat (4%) and no-treat (95%) decisions, with very low incorrect treat (0.5%) and no-treat (0.5%) decisions. A tally threshold of > or = 40 aphids per plant and action thresholds of 84% of plants infested is equivalent to a mean density of 250 aphids per plant, a recently recommended economic threshold. Using this threshold, the minimum required sample number for the binomial plan was 11 plants.     

Channeling of carbon monoxide during anaerobic carbon dioxide fixation

Carbon monoxide is an intermediate in carbon dioxide fixation by diverse microbes that inhabit anaerobic environments including the human colon. These organisms fix CO(2) by the Wood-Ljungdahl pathway of acetyl-CoA biosynthesis. The bifunctional CO dehydrogenase/acetyl-CoA synthase (CODH/ACS) catalyzes several key steps in this pathway. CO(2) is reduced to CO at a nickel iron-sulfur cluster called cluster C located in the CODH subunit. Then, CO is condensed with a methyl group and coenzyme A at cluster A, another nickel iron-sulfur cluster in the ACS subunit. Spectroscopic studies indicate that clusters A and C are at least 10-15 A apart. To gain a better understanding of how CO production and utilization are coordinated, we have studied an isotopic exchange reaction between labeled CO(2) and the carbonyl group of acetyl-CoA with the CODH/ACS from Clostridium thermoaceticum. When solution CO is provided at saturating levels, only CO(2)-derived CO is incorporated into the carbonyl group of acetyl-CoA. Furthermore, when high levels of hemoglobin or myoglobin are added to remove CO from solution, there is only partial inhibition of the incorporation of CO(2)-derived CO into acetyl-CoA. These results provide strong evidence for the existence of a CO channel between cluster C in the CODH subunit and cluster A in the ACS subunit. The existence of such a channel would tightly couple CO production and utilization and help explain why high levels of this toxic gas do not escape into the environment. Instead, microbes sequester this energy-rich carbon source for metabolic reactions.     

Two New Species of Pristionchus (Rhabditida: Diplogastridae): P. fissidentatus n. sp. from Nepal and La Réunion Island and P. elegans n. sp. from Japan

Pristionchus fissidentatus n. sp., isolated from soil in Nepal, and P. elegans n. sp., isolated from Phelotrupes auratus (Coleoptera: Scarabaeidae) in Japan, are described. The two new species are recognized as basal within the genus and thus occupy an important position for macroevolutionary studies that center on the model P. pacificus. Pristionchus fissidentatus n. sp. is distinguished by its unique stegostomatal morphology: in the stenostomatous form, the right subventral ridge has three prominent cusps and the left subventral sector has, in addition to a plate with two cusps, a prominent denticle slightly left of ventral; in the eurystomatous form, the right subventral stegostomatal sector shows both a tooth and a ridge with several cusps. Diagnostic of P. elegans n. sp. is the structure of the stenostomatous cheilostom, which bulges medially and is underlain by a large vacuolated ring. No eurystomatous form has been observed in P. elegans n. sp. Reproductive modes of P. fissidentatus n. sp. and P. elegans n. sp. are hermaphroditic and gonochoristic, respectively. The additional isolation of P. fissidentatus n. sp. from soil and two species of scarab beetle on La Réunion Island in the Indian Ocean suggests a broad geographic range for this species.     

Two New and Two Recharacterized Species from a Radiation of Pristionchus (Nematoda: Diplogastridae) in Europe

Two new species of Pristionchus, P. lucani n. sp. and P. bulgaricus n. sp., are described from France and Bulgaria, respectively. Additionally, new morphological and morphometric data are provided for two previously described species from Europe, P. brevicauda (Kotlán, 1928) Paramonov, 1952 and P. clavus (von Linstow, 1901) Sudhaus and Fürst von Lieven, 2003. A phylogeny including these four species was inferred from a dataset including 26 ribosomal protein-coding genes, sequences of which are original for P. bulgaricus n. sp. and P. clavus. Relationships support a radiation of all sequenced European Pristionchus species from a single, gonochoristic common ancestor, and current knowledge of species ranges supports “western” and “eastern” clades. Similar diagnostic morphologies reflect the close relationships among the new and recharacterized species, especially P. bulgaricus n. sp., P. brevicauda, and P. clavus, although mating tests as well as genetic and phylogenetic separation support their identities as unique species. Our results show that Pristionchus species in Europe are more diverse than typological characters suggest, and thus biological and molecular profiling will be essential for future delimitation of Pristionchus species from the region.     

Adaptive value of a predatory mouth-form in a dimorphic nematode

Polyphenisms can be adaptations to environments that are heterogeneous in space and time, but to persist they require conditional-specific advantages. The nematode Pristionchus pacificus is a facultative predator that displays an evolutionarily conserved polyphenism of its mouthparts. During development, P. pacificus irreversibly executes either a eurystomatous (Eu) or stenostomatous (St) mouth-form, which differ in the shape and number of movable teeth. The Eu form, which has an additional tooth, is more complex than the St form and is thus more highly derived relative to species lacking teeth. Here, we investigate a putative fitness trade-off for the alternative feeding-structures of P. pacificus. We show that the complex Eu form confers a greater ability to kill prey. When adults were provided with a prey diet, Eu nematodes exhibited greater fitness than St nematodes by several measures, including longevity, offspring survival and fecundity when followed by bacterial feeding. However, the two mouth-forms had similar fecundity when fed ad libitum on bacteria, a condition that would confer benefit on the more rapidly developing St form. Thus, the two forms show conditional fitness advantages in different environments. This study provides, to our knowledge, the first functional context for dimorphism in a model for the genetics of plasticity.     

Role of carbon monoxide dehydrogenase in acetate synthesis by the acetogenic bacterium, Acetobacterium woodii

Carbon monoxide dehydrogenase (CODH) plays a key role in acetate synthesis by the acetogenic bacterium, Clostridium thermoaceticum. Acetobacterium woodii, like C. thermoaceticum contains high levels of CODH. In this work we show that crude extracts of A. woodii synthesize acetate from methyl tetrahydrofolate or methyl iodide, carbon monoxide and coenzyme A (CoA). The purified CODH from A. woodii catalyzes an exchange reaction between CO and the carbonyl group of acetyl-CoA even faster than the C. thermoaceticum enzyme, indicating the CODH of A. woodii, like that of C. thermoaceticum is an acetyl-CoA synthetase. Fluorescence and EPR studies further support this postulate by demonstrating that CODH binds CoA near the CO binding site involving a tryptophan residue. The UV absorption spectra and the amino acid compositions of A. woodii and C. thermoaceticum CODHs are very similar. Evidence is presented using purified enzymes from A. woodii that the synthesis of acetyl-CoA occurs by a pathway similar to that utilized by C. thermoaceticum.     

EPR evidence for nickel-substrate interaction in carbon monoxide dehydrogenase from Clostridium thermoaceticum

Carbon monoxide dehydrogenase from Clostridiumthermoaceticum contains two different Fe₄S₄ rhombic-type EPR resonances with g-values at 2.04, 1.94, 1.90 and 2.01, 1.86, 1.75, respectively. The enzyme after reacting with CO or HCO₃^?/CO₂ also reveals in EPR signal at g = 2.07 and 2.02. This signal, readily observed at 95K, is attributed to a Ni(III) interaction with a radical species formed from CO or HCO₃^?/CO₂.     

BrdU Birth Dating Can Produce Errors in Cell Fate Specification in Chick Brain Development

Birth dating neurons with bromodeoxyuridine (BrdU) labeling is an established method widely employed by neurobiologists to study cell proliferation in embryonic, postnatal, and adult brain. Birth dating studies in the chick dorsal telencephalon and the mammalian striatum have suggested that these structures develop in a strikingly similar manner, in which neurons with the same birth date aggregate to form “isochronic clusters.” Here we show that isochronic cluster formation in the chick dorsal telencephalon is an artifact. In embryos given standardly employed doses of BrdU, we observed isochronic clusters but found that clusters were absent with BrdU doses close to the limits of detection. In addition, in situ hybridization experiments established that neurons in the clusters display errors in cell type specification: BrdU cell clusters in nidopallium adopted a mesopallial neuronal fate, mesopallial clusters were misspecified as nidopallial cells, and in some instances, the BrdU clusters failed to express neuronal differentiation markers characteristic of the dorsal telencephalon. These results demonstrate that the chick dorsal telencephalon does not develop by isochronic cluster formation and highlight the need to test the integrity of BrdU-treated tissue with gene expression markers of regional and cell type identity.