Parasite Diversity in an Endemic Region for Avian Malaria and Identification of a Parasite Causing Penguin Mortality

ABSTRACT Understanding the population structure of Plasmodium parasites is essential for malaria intervention. A survey of parasites in vectors and host infections was conducted in an area of intense mortality due to malaria in a captive penguin ( Spheniscus demersus ) colony, using a novel method for identification of Plasmodium species by amplification of ribosomal sequences in DNA or RNA. Three phylogenetically distinct groups of avian Plasmodium were detected in mosquitoes ( Culex ) collected at the study site (Baltimore Zoo, Baltimore, MD) during a period of high transmission. One of the three clades of Plasmodium was found to be prevalent in penguins monitored through the malaria transmission season and consistent with morphological identification as Plasmodium relictum. This parasite sequence was directly associated with the death of a penguin. Thus, a complete transmission cycle is defined at this site. Phylogenetic comparison of ribosomal sequences to an authenticated reference strain of Plasmodium relictum indicates that this is not the parasite causing death in the penguins, suggesting that different parasites may be morphologically indistinguishable.     

Inheritance of Anthracnose Resistance in the Common Bean Differential Cultivar G 2333 and Identification of a New Molecular Marker Linked to the Co-42 gene

The inheritance of anthracnose resistance of the common bean ( Phaseolus vulgaris L.) differential cultivar G 2333 to Colletotrichum lindemuthianum races 73 and 89 was studied in crosses with the susceptible cultivar Rudá. The segregation ratios of 15 : 1 in the F₂ and 3 : 1 in the backcrosses to Rudá indicate that for each of the races tested there are two independent resistance loci in G 2333. A random amplified polymorphic DNA (RAPD) molecular marker (OPH18_1200C) linked in resistance to race 73 was identified in a BC₃F_2:3 population derived from crosses between Rudá and G 2333. A RAPD molecular marker OPAS13_950C, previously identified as linked to gene Co-4² , was also amplified in this population. Co-segregation analyses showed that these two markers are located at 5.6 (OPH18_1200C) and 11.2 (OPAS13_950C) cM of the Co-4² gene. These markers were not present in BC₁F_2:3 plants resistant to race 89 indicating that this population carries a different resistance gene. DNA amplification of BC₁F_2:3 plants with RAPD molecular marker OPAB_450C, previously identified as linked to gene Co-5 , indicated that this gene is present in this population.     

A reciprocal transplant experiment within a climatic gradient in a semiarid shrub-steppe ecosystem: effects on bunchgrass growth and reproduction, soil carbon, and soil nitrogen

We investigated the effect of climate change on Poa secunda Presl. and soils in a shrub‐steppe ecosystem in south‐eastern Washington. Intact soil cores containing P. secunda were reciprocally transplanted between two elevations. Plants and soils were examined, respectively, 4.5 and 5 years later. The lower elevation (310 m) site is warmer (28.5 °C air average monthly maximum) and drier (224 mm yr^?1) than the upper elevation (844 m) site (23.5 °C air average monthly maximum, 272 mm yr^?1). Observations were also made on undisturbed plants at both sites. There was no effect of climate change on plant density, shoot biomass, or carbon isotope discrimination in either transplanted plant population. The cooler, wetter environment significantly reduced percent cover and leaf length, while the warmer, drier environment had no effect. Warming and drying reduced percent shoot nitrogen, while the cooler, wetter environment had no effect. Culm density was zero for the lower elevation plants transplanted to the upper site and was 10.3 culms m^?2 at the lower site. There was no effect of warming and drying on the culm density of the upper elevation plants. Culm density of in situ lower elevation plants was greater than that of the in situ upper elevation plants. Warming and drying reduced total soil carbon 32% and total soil nitrogen 40%. The cooler, wetter environment had no effect on total soil C or N. Of the C and N that was lost over time, 64% of both came from the particulate organic matter fraction (POM, > 53 µ m). There was no effect of warming and drying on the upper population of P. secunda while exposing the lower population to the cooler, wetter environment reduced reproductive effort and percent cover. With the warmer and drier conditions that may develop with climate change, total C and N of semiarid soils may decrease with the active fraction of soil C also rapidly decreasing, which may alter ecosystem diversity and function.     

The Properties of an Enzyme System Degrading Endogenous Phospholipids of Tetrahymena pyriformis

SYNOPSIS. Tetrahymena pyriformis contains a potent system of lipolytic enzymes which is activated by disruption of the cells. The extent to which endogenous lipids of the homogenate are degraded depends upon the age of the culture and the strain of T. pyriformis used. In their most active form, the enzymes can hydrolyze nearly 60% of the endogenous phospholipids within one hour at room temperature. The products of this reaction are probably responsible for the frequently reported instability of Tetrahymena enzyme systems in vitro. The use of inhibitors and the careful choice of culture conditions can reduce lipid degradation to a negligible level.     

RNA and Protein Synthesis in Germinating Pine Pollen

The results of autoradiographic experiments demonstrate that,as with the pollen of most other species, both the generativeand vegetative nuclei of Loblolly Pine (Pinus taeda) activelyengage in RNA synthesis from the very early stages of pollengermination. Unlike most other species, however, this newlysynthesized RNA includes rRNA. Evidence is provided for theimportance of this newly synthesized RNA in the process of continuedpollen tube growth. One and two-dimensional gel electrophoretic analysis revealsa number of both qualitative and quantitative differences amongthe proteins synthesized during the early stages of germinationand the later stages of pollen tube growth. One of the mostnotable of these is a 36 kD protein, the synthesis of whichpredominates during the later stages of pollen germination.A similar pattern of 36 kD protein synthesis is observed whenmRNA extracted from pollen at each of these stages is translatedin vitro. Key words: Pinus, pollen tube growth     

The Ontogeny of Osmoregulation and Its Neurosecretory Control In the Decapod Crustacean, Rhithropanopeus harrisii (Gould)

Laboratory-hatched larvae of this estuarine crab were rearedat 25°C in seawater of 25 salinity for 18 days coveringzoeal Stages I to IV and a megalops. Three-day periods betweenzoeal stages represent intermolt stages of circadean metecdysis,diecdysis, and proecdysis.Larvae were exposed to either a seriesof seawater salinities from 5-40 in 5 increments or of 10-40in 10 increments for one hour during each day of their development.The osmoconcentrations of 20-80 nanoliter hemolymph samplesfrom each of four larvae were measured separately by determinationsof freezing point depression. Eyestalkless larvae in metecdysis of zoeal Stage II were exposedto the same osmoconcentrations as unoperated controls to testfor osmoregulation by eyestalk nerve tissue. Larvae tend to be isosmotic with seawater of 30-40 salinity(S) and to hyperregulate in more dilute media except for larvaein their first diecdysis which remain isosmotic. Larvae in thelast few hours of proecdysis hyperregulate against 40 S as well,presumably to insure inflow of water to establish a greaterbody volume during hardening of the exoskeleton. They are consequentlyisosmotic in the very early metecdysis. The presence of eyestalks at the first metecdysis (Stage II)keeps zoeas hyperosmotic to 5-30 S, but prevents them from hyperregulationagainst 40 S. Eyestalkless zoeas become isosmotic with 5-30S and hyperregulate against 40 S like late proecdysal larvae.Lack of eyestalks makes diecdysal animals hyperregulate againsta medium with which normal animals are isosmotic. The eyestalkinfluence affects second metecdysal (Stage III) larvae in away similar to those in first metecdysis except that it apparentlyalso prevents a curious tendency to hyporegulate in 5-30 S.Similarly, in this stage, the eyestalks prevent hyperosmosityin 40 S seawater as they do during the first day of zoeal StageII. Eyestalk nerve tissue reduces the degree to which diecdysallarvae of this stage remain hyperosmotic to media of 10 S and20 S and apparently causes larvae to be hypoosmotic at 40 S. Preliminary data indicate that removal of eyestalks has littleeffect on proecdysal larval osmoregulation or on regulationof Stage IV zoeas. In other experiments ablation of eyestalks caused Stage II larvaeto lose the ability to osmoregulate against 10-30 S seawaterwithin two hours after the operation. The same zoeas did nothyperregulate against 40 seawater until four hours after removalof eyestalks.     

The Nucellus, Embryo Sac, Endosperm, and Embryo of Aesculus and their Interdependence during Growth

Immature fruits of Aesculus yield powerful stimuli to growthand cell division. Therefore, the developing fruit of Aesculuswoerlitzensis Koehne has been investigated from pollinationto maturity. The fluid, or liquid endosperm, which containsthe growth-promoting substances is produced in a large vesiclewhich forms at the chalazal tip of the embryo sac. As the vesiclegrows, it encroaches upon the nucellus and, when the embryodevelops, one of its cotyledons penetrates into the vesicleof the embryo sac where it grows and absorbs the contents. Theembryo, which has only a vestigial suspensor, reaches the vesicleby growing along the neck of the long curved embryo sac. Thecotyledon which first penetrates the vesicle grows into a massivestructure; the other remains small. The tip of the cotyledonseems to function as an absorbing surface, for the endospermwith which it comes into contact disorganizes. Fertilizationand the presence of a viable embryo at the micropylar end ofthe embryo sac therefore sets in train a number of other events.These are the extensive development of the nucellus at the chalazalend of the embryo sac, the swelling of the vesicle and the formationof a free nuclear and some cellular endosperm, and the disorganizationof the nucellus as it is encroached upon by the vesiculate embryosac. Attention is directed to the organization of the nucellusin the vicinity of the embryo sac. Files or richly protoplasmicnucellar cells(hypostase) which converge upon the chalazal tipof the embryo sac suggest a principal route by which the vesiclemay be nourished. Special attention is drawn to the very differentsizes of cells, their nuclei and nucleoli, in the differentparts of the nucellus. The growth and development of the embryohas also been traced from the zygote to the mature seed. Thenutritive role of the veaiculate embryo sac, and the supplyof growthstimulating substances, through the function of a cotyledonas an absorbing organ, are now seen as important features ofthe development of the Aesulus embryo in the ovule. Many outstandingproblems still remain. The sequential events that follow fertilizationin the different interdependent regions (nucellus, embryo sac,cotyledon, &c.) are here described, but not casually explained.     

Reduction and Maintenance of a White-Tailed Deer Herd in Central Massachusetts

Abstract: Controlled public hunts have been used in a variety of settings to reduce overabundant white-tailed deer (Odocoileus virginianus) herds. We present the results of a large-scale (160 km²) controlled hunt at Quabbin Reservation (QR) in central Massachusetts, USA. The QR was divided into 5 hunt zones. Hunting was initiated in each zone from 1991 to 1994 and continued through 2004. The management goal was to achieve posthunt deer densities of4 deer/km². Initial estimated deer densities in each zone ranged from 11.4 deer/km² to 27.6 deer/km². The management goals were reached in each zone after 2-4 years of hunting. Posthunt populations were maintained at or below the goal even though total hunter effort was reduced. Hunters were not required to harvest antlerless deer, but antlerless deer comprised 55-83% of the harvest each year. We simulated the effects of 5 years without hunting on deer populations. The simulated deer population exceeded management goals after 2 years. Our results demonstrate that controlled public deer hunts can effectively reduce deer populations and maintain them at desired levels over large areas with minimal hunter restrictions. Managers should prepare stakeholders during the hunt planning process for the need to continue overall harvest rates of >30% during the maintenance phase of a deer management program.     

Functional analysis of the Alternaria brassicicola non-ribosomal peptide synthetase gene AbNPS2 reveals a role in conidial cell wall construction

Alternaria brassicicola is a necrotrophic pathogen causing black spot disease on virtually all cultivated Brassica crops worldwide. In many plant pathosystems fungal secondary metabolites derived from non-ribosomal peptide synthetases (NPSs) are phytotoxic virulence factors or are antibiotics thought to be important for niche competition with other micro-organisms. However, many of the functions of NPS genes and their products are largely unknown. In this study, we investigated the function of one of the A. brassicicola NPS genes, AbNPS2 . The predicted amino acid sequence of AbNPS2 showed high sequence similarity with A. brassicae , AbrePsy1, Cochliobolus heterostrophus , NPS4 and a Stagonospora nodorum NPS. The AbNPS2 open reading frame was predicted to be 22 kb in length and encodes a large protein (7195 amino acids) showing typical NPS modular organization. Gene expression analysis of AbNPS2 in wild-type fungus indicated that it is expressed almost exclusively in conidia and conidiophores, broadly in the reproductive developmental phase. AbNPS2 gene disruption mutants showed abnormal spore cell wall morphology and a decreased hydrophobicity phenotype. Conidia of abnps2 mutants displayed an aberrantly inflated cell wall and an increase in lipid bodies compared with wild-type. Further phenotypic analyses of abnps2 mutants showed decreased spore germination rates both in vitro and in vivo , and a marked reduction in sporulation in vivo compared with wild-type fungus. Moreover, virulence tests on Brassicas with abnps2 mutants revealed a significant reduction in lesion size compared with wild-type but only when aged spores were used in experiments. Collectively, these results indicate that AbNPS2 plays an important role in development and virulence.