Agrobacterium-mediated transformation of Nicotiana attenuata, a model ecological expression system

Summary. Research into the genetic basis of the ecological sophistication of plants is hampered by the availability of transformable systems with a wealth of well-described ecological interactions. We present an Agrobacterium-mediated transformation system for the model ecological expression system, Nicotiana attenuata, a native tobacco that occupies the post-fire niche in the Great Basin Desert of North America. We describe a transformation vector and a transformation procedure that differs from the standard cultivated tobacco transformation protocols in its use of selectable markers, explants, media and cultivation conditions. We illustrate its utility in the transformations with genes coding for key enzymes in the oxylipin cascade (lipoxygenase, allene oxide synthase, hydroperoxide lyase) in antisense orientations and present high-throughput screens useful for the detection of altered phenotypes for the oxylipin cascade (green leaf volatiles and jasmonic acid after wounding). Received 14 March 2002; accepted 10 May 2002     

Conservation Implications of Host Use for Rare Parasitic Plants

Both the study and management of parasites have historically focused on the control, and even elimination, of parasite populations. In contrast, rare parasitic plants represent an uncommon challenge for conservation biologists and managers who often wish to bolster populations of these parasites. Although parasitic plants may suffer any of the maladies known to affect small populations of plants, parasitic plants may also be limited by the additional suite of factors of host availability, host quality, host resistance to parasitism, and parasite preference. We describe studies that have examined parasite growth and reproductive performance with a variety of host species to argue that consideration of the host needs of parasitic plants is necessary for successful conservation of rare species using this mode of resource acquisition. Although it is clear that parasite performance varies greatly with the availability of different host species, little is known about the host requirements of most parasitic plants, and the relative importance of particular host species may not immediately be obvious. Further, because published host lists generally do not distinguish minor hosts from those that sustain parasite populations, such lists may be misleading for conservation efforts. We argue that successful conservation and restoration of parasitic plants may necessitate the management of thoughtfully selected host populations.     

Transport of cadmium and other metals in the blood of the bivalve mollusc Mercenaria mercenaria

The distribution of metals (Ba, Cr, Cu, Fe, Mn, Ni and Zn) in the blood plasma and the circulating hemocytes was determined for a eulamellibranch bivalve, the quahog Mercenaria mercenaria (L.), collected from a relatively clean site at Falmouth, Massachusetts, USA. Whole blood volume was an exponential function of quahog length (Y=5.71×10^-5 X^3.0678; Y=ml of whole blood; X=mm length). Of this volume, 1.2±0.3% ( ) was attributable to blood cells. Total metal content (g metal, or g metal normalized per g of whole blood) was much higher in blood plasma than in hemocytes. In quahogs exposed in the laboratory to 100 ppb ¹⁰⁹Cd, 93.0% of the total accumulated blood Cd was in the plasma rather than in the circulating hemocytes (7.0%), irrespective of the length of exposure (1 h to 31 d). Less than 5% of the plasma Cd was either Cd²⁺, small inorganic Cd complexes or bound to organic molecules with a molecular weight smaller than 1 000. Cadmium was primarily bound to high molecular weight protein(s) (>60 000 daltons) within the plasma. This plasma protein-Cd complex has a low affinity constant (approximately 10⁴ M ^-1), indicating non-specific Cd binding, although the capacity for Cd-binding in the plasma is great (as high as 200 g Cd per ml of plasma). Blood plasma may be far more important in metal transport than has previously been realized.     

Reproductive dominance and differential ovicide in the communally breeding burying beetle Nicrophorus tomentosus

Communal breeding can be characterized by the division of reproduction among cooperating individuals and, if the distribution of reproduction is inequitable, by the mechanisms for achieving skewed reproductive success. The burying beetle (Nicrophorus tomentosus) is a facultative communal breeder. Unrelated adults, especially females, provide extensive parental care to broods of mixed parentage. The frequency and degree of reproductive skew between two females were examined experimentally. On medium-size carcasses, the proportion of eggs attributed to each female was not significantly different from random in 42% of the broods, skewed in 42% and not shared in 16%. Although reproduction was usually skewed in favor of the larger female, the relative sizes of the two females did not predict the degree of skew. On large carcasses, the proportion of eggs attributed to each female was not different from random in 87% of the broods and weakly skewed in 13%. Several mechanisms for biasing reproductive success were investigated. Females increase the proportion of their offspring in the brood by committing differential ovicide. Secondly, burying and preparing a carcass cooperatively stimulates ovarian development of the larger female and slows it for the smaller female, reducing or delaying oviposition by the subordinate. Thirdly, larger females are more likely to be dominant and are more fecund than smaller females. Received: 20 July 1996 / Accepted after revision: 30 November 1996     

Genotypic differences in brood rearing in honey bee colonies: context-specific?

Summary Three experiments were performed to determine whether brood care in honey bee colonies is influenced by colony genetic structure and by social context. In experiment 1, there were significant genotypic biases in the relative likelihood of rearing queens or workers, based on observations of individually labeled workers of known age belonging to two visually distinguishable subfamilies. In experiment 2, no genotypic biases in the relative likelihood of rearing drones or workers was detected, in the same colonies that were used in experiment 1. In experiment 3, there again were significant genotypic differences in the likelihood of rearing queens or workers, based on electrophoretic analyses of workers from a set of colonies with allozyme subfamily markers. There also was an overall significant trend for colonies to show greater subfamily differences in queen rearing when the queens were sisters (half- and super-sisters) rather than unrelated, but these differences were not consistent from trial to trial for some colonies. Results of experiments 1 and 3 demonstrate genotypic differences in queen rearing, which has been reported previously based on more limited behavioral observations. Results from all three experiments suggest that genotypic differences in brood care are influenced by social context and may be more pronounced when workers have a theoretical opportunity to practice nepotism. Finally, we failed to detect persistent interindividual differences in bees from either subfamily in the tendency to rear queen brood, using two different statistical tests. This indicates that the probability of queen rearing was influenced by genotypic differences but not by the effect of prior queen-rearing experience. These results suggest that subfamilies within a colony can specialize on a particular task, such as queen rearing, without individual workers performing that task for extended periods of time.     

New Proof-of-Concept in Viral Inactivation: Virucidal Efficacy of 405 nm Light Against Feline Calicivirus as a Model for Norovirus Decontamination

The requirement for novel decontamination technologies for use in hospitals is ever present. One such system uses 405 nm visible light to inactivate microorganisms via ROS-generated oxidative damage. Although effective for bacterial and fungal inactivation, little is known about the virucidal effects of 405 nm light. Norovirus (NoV) gastroenteritis outbreaks often occur in the clinical setting, and this study was designed to investigate potential inactivation effects of 405 nm light on the NoV surrogate, feline calicivirus (FCV). FCV was exposed to 405 nm light whilst suspended in minimal and organically-rich media to establish the virucidal efficacy and the effect biologically-relevant material may play in viral susceptibility. Antiviral activity was successfully demonstrated with a 4 Log₁₀ (99.99%) reduction in infectivity when suspended in minimal media evident after a dose of 2.8 kJ cm^?2. FCV exposed in artificial faeces, artificial saliva, blood plasma and other organically rich media exhibited an equivalent level of inactivation using between 50–85% less dose of the light, indicating enhanced inactivation when the virus is present in organically-rich biologically-relevant media. Further research in this area could aid in the development of 405 nm light technology for effective NoV decontamination within the hospital environment.     

Long-Term Changes in the Avifauna of Barro Colorado Island, Panama, a Tropical Forest Isolate

Since its isolation from the mainland more than 85 years ago, 65 bird species have disappeared from Barro Colorado Island, Panama. Because of these extinctions the island is often used as a model for the study of faunal relaxation, or loss of species through time. The most recent survey of the island's bird community was completed in 1970. Between June 1994 and May 1996 I surveyed the island and observed 218 species, including 5 species never before recorded. Three relatively sedentary species have experienced severe population declines since Willis's 1970 survey and may be on the brink of disappearing from the island. Willis estimated 500 Slate-colored Grosbeaks ( Pitylus grossus ), whereas I found only two pairs of this conspicuous midstory-dwelling finch. Two cotingas—Speckled Mourner ( Laniocera rufescens ) and Rufous Piha ( Lipaugus unirufus )—have declined by 85–95%. I did not locate any individuals of 36 other species that were present during the 1970s. Most (21) of these are uncommon aquatic or nocturnal species, which I may have missed during my surveys, or are rare to uncommon edge species that probably move frequently to and from the island. Seven species, however, are primarily inhabitants of second-growth forest and have been lost probably because of continuing successional maturation of the forest, including changes in land use around the laboratory clearing. Seven forest-dwelling species disappeared during the 1970s and have not recolonized. I detected only one sedentary forest-dwelling species, Great Currasow ( Crax rubra ), that previously had been reported as missing from the island. The nearly complete lack of recolonizations by such forest-interior species suggests that local extinction from tropical forest isolates may be extremely persistent. Tropical forest reserves as small as Barro Colorado Island (1600 ha) may not preserve high levels of regional avian diversity over long periods of time.     

Flooding impacts on responses of a riparian consumer to cross-ecosystem subsidies

Landscape-driven processes impact the magnitude and direction of cross-ecosystem resource subsidies, but they may also control consumers' numerical and functional responses by altering habitat availability. We investigated effects of the interaction between habitat availability and subsidy level on populations of a riparian fishing spider, Dolomedes aquaticus, using a flood disturbance gradient in the Waimakariri River catchment, New Zealand. D. aquaticus predominantly eat aquatic prey as they hunt from the water surface. However, D. aquaticus biomass peaked at rivers with intermediate flood disturbance, rather than at less flood-prone rivers where the biomass of aquatic insect prey was markedly higher. Flooding positively influenced spider habitat quality, and an experimental manipulation at stable rivers indicated that unembedded cobbles, preferred D. aquaticus habitat, were a limiting factor, preventing response to the increased prey resource at stable sites. Potential terrestrial prey abundance was low, did not vary across the disturbance gradient, and is likely to have been a much smaller component of the fishing spiders' diet than aquatic insect prey. Thus landscape-driven factors not only controlled the magnitude of resource subsidies, but also influenced the ability of consumers to respond to them by altering the physical nature of the ecosystem boundary.