Suppression of female melon fly,Zeugodacus cucurbitae,with cue‐lure and fipronil bait stations through horizontal insecticide transfer

Melon fly, Zeugodacus cucurbitae (Coquillett) (Diptera: Tephritidae), is an important quarantine tephritid fruit fly with resident populations established in Hawai'i, USA. In the male‐annihilation approach, male flies are targeted using dispensers with cue‐lure (C‐L) and insecticides, typically organophosphates. The efficacy of the male annihilation approach is thought to be limited to individual male flies, contacting the lure and the pesticide, after which they die. Alternative classes of insecticides, such as fipronil, have been investigated for use in male‐annihilation. We hypothesized that ingestion of fipronil by male flies could lead to horizontal transfer and mortality in female flies. Horizontal insecticide transfer extends pesticide control beyond the individual contacting the toxicant through indirect contact via food sharing or other mechanisms. We tested the possibility for horizontal transfer of fipronil from male to female Z. cucurbitae through field and laboratory studies. Two repeated field trials were conducted to compare the numbers of female flies collected in fields treated with Amulet C‐L (0.34% fipronil active ingredient) bait stations, sanitation, and spot treatments of GF‐120 Fruit Fly Bait to numbers collected in fields where sanitation and spot‐treatments were used without Amulet C‐L. In fields with Amulet C‐L bait stations in conjunction with sanitation and weekly protein bait spot treatments of GF‐120 Fruit Fly Bait, female captures were significantly lower than those in field plots treated with weekly protein bait spot treatments and sanitation. In subsequent laboratory studies, all females died within 6 h after direct exposure to male flies that had access to Amulet C‐L for 1–4 min. The possibility that male regurgitant could be a mechanism for horizontal transfer and subsequent female mortality was determined by collecting regurgitated droplets from fipronil‐fed male flies and feeding them to males and females. Both male and female flies exposed to regurgitant from fipronil‐fed male flies or droplets containing fipronil had higher mortality than the male and female flies that were exposed to regurgitant or droplets with only the C‐L compound or sugar solution. Thus, female flies do experience mortality from exposure to regurgitant from males that have fed on fipronil laced solutions. This provides evidence of at least one mechanism of horizontal transfer of insecticide in tephritid fruit flies. These findings are discussed in the context of Z. cucurbitae integrated pest management programs in Hawai'i.     

Recent discoveries and applications of Anoxybacillus

The Bacillaceae family members are a good source of bacteria for bioprocessing and biotransformation involving whole cells or enzymes. In contrast to Bacillus and Geobacillus, Anoxybacillus is a relatively new genus that was proposed in the year 2000. Because these bacteria are alkali-tolerant thermophiles, they are suitable for many industrial applications. More than a decade after the first report of Anoxybacillus, knowledge accumulated from fundamental and applied studies suggests that this genus can serve as a good alternative in many applications related to starch and lignocellulosic biomasses, environmental waste treatment, enzyme technology, and possibly bioenergy production. This current review provides the first summary of past and recent discoveries regarding the isolation of Anoxybacillus, its medium requirements, its proteins that have been characterized and cloned, bioremediation applications, metabolic studies, and genomic analysis. Comparisons to some other members of Bacillaceae and possible future applications of Anoxybacillus are also discussed.     

Diversity and Patch-Size Distributions of Biological Soil Crusts Regulate Dryland Ecosystem Multifunctionality

Recent studies report that multifunctionality—the simultaneous provision of multiple ecosystem functions—in drylands depends on biodiversity. Others report that specific size distributions of vegetation patches indicate overall ecosystem health and function. Using a biocrust (micro-vegetation of mosses, lichens, and cyanobacteria) model system, and multivariate modeling, we determined the relative importance of biodiversity, patch-size distribution, and total abundance to nutrient cycling and multifunctionality. In most cases we explained at least 20%, and up to 65%, of the variation in ecosystem functions, and 42% of the variation in multifunctionality. Species richness was the most important determinant of C cycling, constituting an uncommonly clear link between diversity and function in a non-experimental field setting. Regarding C cycling in gypsiferous soils, we found that patch size distributions with a greater frequency of small to medium patches, as opposed to very small patches, were more highly functional. Nitrogen cycling was largely a function of biocrust cover in two soil types, whereas in gypsiferous soils, more central-tending patch size distributions were less functional with regards to N cycling. All three community properties were about equally important to multifunctionality. Our results highlight the functional role of biotic attributes other than biodiversity, and indicate that high cover and diversity, together with a particular patch-size distribution, must be attained simultaneously to maximize multifunctionality. The results also agree with trends observed with other terrestrial and aquatic communities that more biodiversity is needed to sustain multifunctionality compared to single functions considered independently.     

An In-Depth Comparison of Latent HIV-1 Reactivation in Multiple Cell Model Systems and Resting CD4+ T Cells from Aviremic Patients

The possibility of HIV-1 eradication has been limited by the existence of latently infected cellular reservoirs. Studies to examine control of HIV latency and potential reactivation have been hindered by the small numbers of latently infected cells found in vivo. Major conceptual leaps have been facilitated by the use of latently infected T cell lines and primary cells. However, notable differences exist among cell model systems. Furthermore, screening efforts in specific cell models have identified drug candidates for “anti-latency” therapy, which often fail to reactivate HIV uniformly across different models. Therefore, the activity of a given drug candidate, demonstrated in a particular cellular model, cannot reliably predict its activity in other cell model systems or in infected patient cells, tested ex vivo. This situation represents a critical knowledge gap that adversely affects our ability to identify promising treatment compounds and hinders the advancement of drug testing into relevant animal models and clinical trials. To begin to understand the biological characteristics that are inherent to each HIV-1 latency model, we compared the response properties of five primary T cell models, four J-Lat cell models and those obtained with a viral outgrowth assay using patient-derived infected cells. A panel of thirteen stimuli that are known to reactivate HIV by defined mechanisms of action was selected and tested in parallel in all models. Our results indicate that no single in vitro cell model alone is able to capture accurately the ex vivo response characteristics of latently infected T cells from patients. Most cell models demonstrated that sensitivity to HIV reactivation was skewed toward or against specific drug classes. Protein kinase C agonists and PHA reactivated latent HIV uniformly across models, although drugs in most other classes did not.     

Structure of chitinase D from Serratia proteamaculans reveals the structural basis of its dual action of hydrolysis and transglycosylation

Chitinases are known to hydrolyze chitin polymers into smaller chitooligosaccharides. Chitinase from bacterium Serratia proteamaculans (SpChiD) is found to exhibit both hydrolysis and transglycosylation activities. SpChiD belongs to family 18 of glycosyl hydrolases (GH-18). The recombinant SpChiD was crystallized and its three-dimensional structure was determined at 1.49 Å resolution. The structure was refined to an R-factor of 16.2%. SpChiD consists of 406 amino acid residues. The polypeptide chain of SpChiD adopts a (β/α)₈ triosephosphate isomerase (TIM) barrel structure. SpChiD contains three acidic residues, Asp149, Asp151 and Glu153 as part of its catalytic scheme. While both Asp149 and Glu153 adopt single conformations, Asp151 is observed in two conformations. The substrate binding cleft is partially obstructed by a protruding loop, Asn30 - Asp42 causing a considerable reduction in the number of available subsites in the substrate binding site. The positioning of loop, Asn30 - Asp42 appears to be responsible for the transglycosylation activity. The structure determination indicated the presence of sulfone Met89 (SMet89). The sulfone methionine residue is located on the surface of the protein at a site where extra domain is attached in other chitinases. This is the first structure of a single domain chitinase with hydrolytic and transglycosylation activities.     

Monitoring of diamondback moth (Lepidoptera: Plutellidae) resistance to spinosad, indoxacarb, and emamectin benzoate

Six to nine populations of the diamondback moth, Plutella xylostella (L.), were collected annually from fields of crucifer vegetables in the United States and Mexico from 2001 to 2004 for baseline susceptibility tests and resistance monitoring to spinosad, indoxacarb, and emamectin benzoate. A discriminating concentration for resistance monitoring to indoxacarb and emamectin benzoate was determined based on baseline data in 2001 and was used in the diagnostic assay for each population in 2002-2004 together with a discriminating concentration for spinosad determined previously. Most populations were susceptible to all three insecticides, but a population from Hawaii in 2003 showed high levels of resistance to indoxacarb. Instances of resistance to spinosad occurred in Hawaii (2000), Georgia (2001), and California (2002) as a consequence of a few years of extensive applications in each region. The collaborative monitoring program between university and industry scientists we discuss in this article has provided useful information to both parties as well as growers who use the products. These studies provide a baseline for developing a more effective resistance management program for diamondback moth.     

Cytochrome P450 oxygenases of monoterpene metabolism

The cytochrome P450 monoterpene oxygenases are largely responsible for imparting structural and functional diversity to this family of natural products. In most cases, cytochrome P450-mediated allylic hydroxylation of a parental monoterpene olefin leads to a series of redox transformations and conjugation reactions which yield a family of structurally related derivatives and isomers. An overview is provided of the extant monoterpene oxygenases, with examples mainly from the mint (Lamiaceae) family of essential oil plants, and, where possible, information on the structure, mechanism, localization and regulation of these enzymes is described. The review concludes with a brief assessment of biotechnological applications and a view to future research in this area.     

On the origin and evolution of apomixis in Boechera

The genetic mechanisms causing seed development by gametophytic apomixis in plants are predominantly unknown. As apomixis is consistently associated with hybridity and polyploidy, these confounding factors may either (a) be the underlying mechanism for the expression of apomixis, or (b) obscure the genetic factors which cause apomixis. To distinguish between these hypotheses, we analyzed the population genetic patterns of diploid and triploid apomictic lineages and their sexual progenitors in the genus Boechera (Brassicaceae). We find that while triploid apomixis is associated with hybridization, the majority of diploid apomictic lineages are likely the product of intra-specific crosses. We then show that these diploid apomicts are more likely to sire triploid apomictic lineages than conspecific sexuals. Combined with flow cytometric seed screen phenotyping for male and female components of apomixis, our analyses demonstrate that hybridization is an indirect correlate of apomixis in Boechera.     

Biotransformation of podophyllotoxin by Hordeum vulgare cell suspension cultures

Hordeum vulgare cell suspension cultures were used to modify podophyllotoxin (1) One major product (1a) and one minor product (1b) were detected in both the culture medium and cells. To optimize the yield of compound 1a, we showed that: (1) the optimal concentration of added podophyllotoxin (1) was 33 mg L^-1; higher concentrations caused cell toxicity; (2) the stage of the cell cycle (lag/log/stationary) at which podophyllotoxin was added only marginally affected the yield of compound 1a; the optimal addition time was after lag phase, in which the yield of compound 1a reached ca. 76%, and (3) biotransformation of podophyllotoxin (1) was relatively slow; podophyllotoxin fed at 4 days after subculture resulted in yields of compound 1a of ca. 56, 64 and 76% after an additional 3, 6 and 10 days of incubation, respectively. Product 1a was purified and identified as isopicropodophyllone (1a) based on MS and NMR data.