Distribution of Pfiesteria piscicida cyst populations in sediments of the Delaware Inland Bays, USA

The toxic dinoflagellate, Pfiesteria piscicida, is a common constituent of the phytoplankton community in the Delaware Inland Bays, USA. In this study, molecular methods were used to investigate the distributions of benthic stages (cysts) of P. piscicida in sediment cores from the Delaware Inland Bays. Cores from 35 sites were partitioned into nephloid and anoxic layers and analyzed for P. piscicida by nested amplification of the 18S rDNA gene using P. piscicida-specific primers. The presence of inhibitory substances in the PCR reaction was evaluated by inclusion of an exogenous control DNA in the extraction buffer, thus eliminating samples that may yield false-negative results. Our results indicate a patchy distribution of P. piscicida in sediments of the Delaware Inland Bays, with distinct differences between each of the three bays. Overall, P. piscicida was found more frequently in sediments from Rehoboth Bay compared to Indian River and Little Assawoman Bays. These differences suggest (i) that populations of P. piscicida may be more widely distributed in Rehoboth Bay, (ii) that populations of P. piscicida may have been introduced to Rehoboth Bay at an earlier time, (iii) that past blooms of P. piscicida in Rehoboth Bay estuaries may have seeded the sediments with higher numbers of cysts, and/or (iv) that Rehoboth Bay sediments may be more resistant to clearing due to storm turbulence.     

Use of quantitative real-time PCR to monitor population dynamics of ammonia-oxidizing bacteria in batch process

A quantitative real-time PCR (QPCR) assay with the TaqMan system was used to quantify 16S rRNA genes of β-proteobacterial ammonia-oxidizing bacteria (AOB) in a batch nitrification bioreactor. Five different sets of primers, together with a TaqMan probe, were used to quantify the 16S rRNA genes of β-proteobacterial AOB belonging to the Nitrosomonas europaea, Nitrosococcus mobilis, Nitrosomonas nitrosa, and Nitrosomonas cryotolerans clusters, and the genus Nitrosospira. We also used PCR followed by denaturing gradient gel electrophoresis (DGGE), cloning, and sequencing of their 16S rRNA genes to identify the AOB species. Seed sludge from an industrial wastewater treatment process controlling high-strength nitrogen wastewater (500 mg/L NH₄ ⁺–N) was used as the inoculum for subsequent batch experiment. The Nitrosomonas nitrosa cluster was the predominant AOB (2.3 × 10⁵ copies/mL) in the start-up period of the batch experiment. However, from the exponential growth period, the Nitrosomonas europaea cluster was the most abundant AOB, and its 16S rRNA gene copy number increased to 8.9 × 10⁶ copies/mL. The competitive dominance between the two AOB clusters is consistent with observed differences in ammonia tolerance and substrate affinity. Analysis of the DGGE results indicated the presence of Nitrosomonas europaea ATCC19718 and Nitrosomonas nitrosa Nm90, consistent with the QPCR results.     

Pelagic-benthic transition of the harmful alga, Heterosigma akashiwo: Changes in swimming and implications for benthic cell distributions

Many harmful algal blooming (HAB) species transition between a vegetative, motile phase in the water column and a dormant, non-motile resting phase in the sediments. These life history transitions potentially regulate the timing, location and persistence of bloom events. Motility promotes aggregation and influences vertical distributions in the water column. However, the contribution of this behavior to benthic distributions of resting cells is currently unknown. We used video-tracking techniques to test the hypothesis that algal cells use active down-swimming during pelagic-benthic transition to favorably influence benthic distributions. In an experimental water column, we monitored cell swimming trajectories of Heterosigma akashiwo for 14 days after cells were signaled to enter the benthic resting stage. Using the statistical characteristics of individual cell trajectories, we developed a video-based motion assay to assign each tracked Heterosigma cell to one of three cell states known to occur during pelagic-benthic transition: induced motile, transitional and resting. The primary swimming characteristic influencing benthic distribution, net vertical velocity, was essentially the same for all three cell states. Hence, we found no evidence that active down-swimming influences benthic distributions. Our data suggest that benthic distributions of Heterosigma resting cells are similar to distributions of slowly sedimenting passive particles. These observations suggest that Heterosigma benthic resting cell distributions can be predicted by modeling the effects of cell sedimentation rates combined with geophysical flow patterns.     

Pathology and immune response of the blue mussel (Mytilus edulis L.) after an exposure to the harmful dinoflagellate Prorocentrum minimum

The harmful dinoflagellate Prorocentrum minimum has different effects upon various species of grazing bivalves, and these effects also vary with life-history stage. Possible effects of this dinoflagellate upon mussels have not been reported; therefore, experiments exposing adult blue mussels, Mytilus edulis, to P. minimum were conducted. Mussels were exposed to cultures of toxic P. minimum or benign Rhodomonas sp. in glass aquaria. After a short period of acclimation, samples were collected on day 0 (before the exposure) and after 3, 6, and 9 days of continuous-exposure experiment. Hemolymph was extracted for flow-cytometric analyses of hemocyte, immune-response functions, and soft tissues were excised for histopathology. Mussels responded to P. minimum exposure with diapedesis of hemocytes into the intestine, presumably to isolate P. minimum cells within the gut, thereby minimizing damage to other tissues. This immune response appeared to have been sustained throughout the 9-day exposure period, as circulating hemocytes retained hematological and functional properties. Bacteria proliferated in the intestines of the P. minimum-exposed mussels. Hemocytes within the intestine appeared to be either overwhelmed by the large number of bacteria or fully occupied in the encapsulating response to P. minimum cells; when hemocytes reached the intestine lumina, they underwent apoptosis and bacterial degradation. This experiment demonstrated that M. edulis is affected by ingestion of toxic P. minimum; however, the specific responses observed in the blue mussel differed from those reported for other bivalve species. This finding highlights the need to study effects of HABs on different bivalve species, rather than inferring that results from one species reflect the exposure responses of all bivalves.     

Prorocentrum minimum(clone Exuv) is nutritionally insufficient, but not toxic to the copepod Acartia tonsa

This study tested whether the dinoflagellate Prorocentrum minimum is nutritionally insufficient or toxic to the copepod Acartia tonsa. Experiments were carried out with adult female A. tonsa and the P. minimum clone Exuv, both isolated from Long Island Sound. Initially, the functional and numerical responses of A. tonsa feeding on exponentially growing P. minimum cells were characterized. These experiments revealed that A. tonsa readily ingested P. minimum cells, up to the equivalent of 200% of body carbon day⁻¹, but egg production was relatively low, with a maximum egg production rate of 22% of body carbon day⁻¹. Hence, the egg production efficiency (egg carbon produced versus cell carbon ingested) was low (10%). In a separate experiment, ingestion and egg production rates were measured as a function of food concentration with cells in different growth stages (early-exponential, late-exponential/early-stationary, and late-stationary growth phase) to simulate conditions during a bloom. There was no indication that cells in the stationary phase resulted in lower ingestion or egg production rates relative to actively growing cells. Egg hatching success remained high (>80%) and independent of the cell growth phase. In a third experiment specifically designed to test the hypothesis that P. minimum is toxic, ingestion, egg production and egg hatching success were measured when females were fed mixtures of P. minimum and the diatom Thalassiosira weissflogii, but in which total food concentration was held constant and the proportion of P. minimum in the mixed diet varied. A. tonsa readily ingested P. minimum when it was offered in the mixed diet, with no detrimental effects on egg production or egg hatching observed. Supplementing P. minimum with T. weissflogii increased both the egg production rate and the egg production efficiency. It is concluded that P. minimum is nutritionally insufficient, but not toxic to A. tonsa. Finally, it is estimated that in the field grazing by A. tonsa is approximately equivalent to 30% of the maximum daily growth rate of P. minimum. Hence, copepod grazing cannot be ignored in field and modeling studies of the population dynamics of P. minimum.     

Nitrogen utilization by the raphidophyte Heterosigma akashiwo: Growth and uptake kinetics in laboratory cultures

The nitrogen uptake and growth capabilities of the potentially harmful, raphidophycean flagellate Heterosigma akashiwo (Hada) Sournia were examined in unialgal batch cultures (strain CCMP 1912). Growth rates as a function of three nitrogen substrates (ammonium, nitrate and urea) were determined at saturating and sub-saturating photosynthetic photon flux densities (PPFDs). At saturating PPFD (110 μE m⁻² s⁻¹), the growth rate of H. akashiwo was slightly greater for cells grown on NH₄⁺ (0.89 d⁻¹) compared to cells grown on NO₃⁻ or urea, which had identical growth rates (0.82 d⁻¹). At sub-saturating PPFD (40 μE m⁻² s⁻¹), both urea- and NH₄⁺-grown cells grew faster than NO₃⁻-grown cells (0.61, 0.57 and 0.46 d⁻¹, respectively). The N uptake kinetic parameters were investigated using exponentially growing batch cultures of H. akashiwo and the ¹⁵N-tracer technique. Maximum specific uptake rates (V_max) for unialgal cultures grown at 15 °C and saturating PPFD (110 μE m⁻² s⁻¹) were 28.0, 18.0 and 2.89 × 10⁻³ h⁻¹ for NH₄⁺, NO₃⁻ and urea, respectively. The traditional measure of nutrient affinity—the half saturation constants (K_s) were similar for NH₄⁺ and NO₃⁻ (1.44 and 1.47 μg-at N L⁻¹), but substantially lower for urea (0.42 μg-at N L⁻¹). Whereas the α parameter (α = V_max/K_s), which is considered a more robust indicator for substrate affinity when substrate concentrations are low (<K_s), were 19.4, 12.2 and 6.88 × 10⁻³ h⁻¹/(μg-at N L⁻¹) for NH₄⁺, NO₃⁻ and urea, respectively. These laboratory results demonstrate that at both saturating and sub-saturating N concentrations, N uptake preference follows the order: NH₄⁺ > NO₃⁻ > urea, and suggests that natural blooms of H. akashiwo may be initiated or maintained by any of the three nitrogen substrates examined.     

Impacts and potential effects due to Prorocentrum minimum blooms in Chesapeake Bay

The dinoflagellate Prorocentrum minimum (P. minimum) can be found in all seasons and over a broad range of habitat conditions in the Chesapeake Bay and its tributaries. Blooms (>3000 cells ml⁻¹), locally referred to as ‘mahagony tides’, were restricted to salinities of 4.5–12.8 psu, water temperatures of 12–28 °C, and occurred most frequently in April and May. P. minimum blooms have been detected at routine water quality monitoring stations located in the main channel of the Bay and tidal tributaries. Nearshore investigations of bloom events, however, have accounted for the majority of events recorded in excess of 10⁵ cells ml⁻¹. Mahogany tides were associated with widespread harmful impacts including anoxic/hypoxic events, finfish kills, aquaculture shellfish kills and submerged aquatic vegetation losses. We summarize the state of knowledge regarding physical and chemical factors related to P. minimum blooms, their abundance, distribution and frequency, and ecological effects in Chesapeake Bay.     

Reference gene selection for quantitative real-time PCR normalization in tomato subjected to nitrogen, cold, and light stress

We examined eight putative consistently expressed genes—actin (ACT), β-tubulin, elongation factor 1α (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK), ribosomal protein L2 (RPL2), ubiquitin (UBI), and a catalytic subunit of protein phosphatase 2A (PP2Acs)—for their potential as references for the normalization of gene expression in tomato leaves. Expression stability of candidate reference genes was tested during growth conditions of nitrogen (N) starvation, low temperature, and suboptimal light. The geNorm algorithm, using reciprocal cross-validation among a larger group of candidate references, was applied for this purpose. The widely used reference genes GAPDH and PGK were top ranked during light stress but poorly ranked during N and cold stress. In contrast, EF1 was top ranked during N and cold stress but poorly ranked during light stress. The novel references RPL2 and PP2Acs, as well as the traditional references ACT and UBI, appeared to be stably expressed when looking at the data set as a whole. No gene was identified that exhibited such a constant level of expression as to outperform the other candidates under all experimental conditions. Thus, the results highlight the need for normalizing gene expression in tomato using the geometric average of multiple carefully selected reference genes.     

Use of a real-time remote monitoring network (RTRM) and shipborne sampling to characterize a dinoflagellate bloom in the Neuse Estuary, North Carolina, USA

The spatial-temporal distribution of a dinoflagellate bloom dominated or co-dominated by Prorocentrum minimum was examined during autumn through early spring in a warm temperate, eutrophic estuary. The developing bloom was first detected from a web-based alert provided by a network of real-time remote monitoring (RTRM) platforms indicating elevated dissolved oxygen and pH levels in upper reaches of the estuary. RTRM data were used to augment shipboard sampling, allowing for an in-depth characterization of bloom initiation, development, movement, and dissipation. Prolonged drought conditions leading to elevated salinities, and relatively high nutrient concentrations from upstream inputs and other sources, likely pre-disposed the upper estuary for bloom development. Over a 7-month period (October 2001–April 2002), the bloom moved toward the northern shore of the mesohaline estuary, intensified under favorable conditions, and finally dissipated after a major storm. Bloom location and transport were influenced by prevailing wind structure and periods of elevated rainfall. Chlorophyll a within bloom areas averaged 106 ± 13 μg L⁻¹ (mean ± 1 S.E.; maximum, 803 μg L⁻¹), in comparison to 20 ± 1 μg L⁻¹ outside the bloom. There were significant positive relationships between dinoflagellate abundance and TN and TP. Ammonium, NO₃⁻, and SRP concentrations did not decrease within the main bloom, suggesting that upstream inputs and other sources provided nutrient-replete conditions. In addition, PAM fluorometric measurements (09:00–13:00 h) of maximal PSII quantum yield (F_v/F_m) were consistently 0.6–0.8 within the bloom until late March, providing little evidence of photo-physiological stress as would have been expected under nutrient-limiting conditions. Nitrogen uptake kinetics were estimated for P. minimum during the period when that species was dominant (October–December 2001), based on literature values for N uptake by an earlier P. minimum bloom (winter 1999) in the Neuse Estuary. The analysis suggests that NH₄⁺ was the major N species that supported the bloom. Considering the chlorophyll a concentrations during October and December and the estimated N uptake rates, phytoplankton biomass was estimated to have doubled once per day. Bloom displacement (January–February) coincided with higher diversity of heterotrophic dinoflagellate species as P. minimum abundance decreased. This research shows the value of RTRM in bloom detection and tracking, and advances understanding of dinoflagellate bloom dynamics in eutrophic estuaries.     

Prorocentrum minimum bloom and its possible link to a massive fish kill in Bolinao, Pangasinan, Northern Philippines

For the first time, a Prorocentrum minimum bloom at a maximum cell density of 4.7 × 10⁵ cells/L was recorded on January 31 to February 4, 2002 at Bolinao, Pangasinan, Northern Philippines where intensive and extensive aquaculture of Chanos chanos (milkfish) in fish pens and cages has been practiced for years now. The fish kill, which lasted almost simultaneously with the bloom of the organism had its peak when the organisms bloom was declining. Lack of oxygen in the cages and pens was the fundamental cause of the fish kill. Losses due to the fish kill were estimated at six million pesos (equivalent to US$ 120,000), which includes only the worth of dead cultivated fish. Lack of oxygen in the cages and pens was the fundamental cause of the fish kill, and toxicity of the Prorocentrum could not be confirmed. The cells had minute spinules equally all over the surface of valves. Intercalary striae were wide with many ridges perpendicular to valve margin. Outline of cells was rounder than typical P. minimum cells and similar to P. balticum. Recommendations for future research on the organism are incorporated together with monitoring and management interventions in order to mitigate or possibly prevent damages in similar future events.     

Using clay to control harmful algal blooms: deposition and resuspension of clay/algal flocs

Harmful algal blooms (HABs) may be legitimate targets for direct control or mitigation, due to their impacts on commercial fisheries and public health. One promising control strategy is the rapid sedimentation of HABs through flocculation with clay. The objective of this study was to evaluate flow environments in which such a control strategy might be effective in removing harmful algae from the water column and depositing a layer of clay/algal flocs on the sea floor. We simulated the natural environment in two laboratory flumes: a straight-channel “17 m flume” in which flocs settled in a still-water column and a “racetrack flume” in which flocs settled in flow. The 17 m flume experiments were designed to estimate the critical bed shear stress for resuspension of flocs that had settled for different time periods. The racetrack flume experiments were designed to examine the deposition and repeated resuspension of flocs in a system with tidal increases in flow speed. All flume runs were conducted with the non-toxic dinoflagellate Heterocapsa triquetra and phosphatic clay (IMC-P4). We repeated the experiments with a coagulant, polyaluminum hydroxychloride (PAC), expected to enhance the removal efficiency (RE) of the clay. Our experiments indicated that at low flow speeds (≤10 cm s⁻¹), phosphatic clay was effective at removing algal cells from the water column, even after repeated resuspension. Once a layer of flocs accumulated on the bed, the consolidation, or dewatering, of the layer over time increased the critical shear stress for resuspension (i.e. decreased erodibility). Resuspension of a 2 mm thick layer that settled for 3 h in relatively low flow speeds (≤3 cm s⁻¹) would be expected at bed shear stress of 0.06–0.07 Pa, as compared to up to 0.09 Pa for a layer that was undisturbed for 9 or 24 h. For the same experimental conditions, the addition of PAC decreased the removal efficiency of algal cells in flow and increased the erodibility of flocs from the bottom. By increasing the likelihood that flocs remain in suspension, the addition of PAC in field trials of clay dispersal might have greater impact on sensitive, filter-feeding organisms. Overall, our experiments suggest that the flow environment should be considered before using clay as a control strategy for HABs in coastal waters.     

Validation of a cotton-specific gene, Sad1, used as an endogenous reference gene in qualitative and real-time quantitative PCR detection of transgenic cottons

Genetically modified (GM) cotton lines have been approved for commercialization and widely cultivated in many countries, especially in China. As a step towards the development of reliable qualitative and quantitative PCR methods for detecting GM cottons, we report here the validation of the cotton (Gossypium hirsutum) endogenous reference control gene, Sad1, using conventional and real-time (RT)-PCR methods. Both methods were tested on 15 different G. hirsutum cultivars, and identical amplicons were obtained with all of them. No amplicons were observed when DNA samples from three species of genus Gossypium, Arabidopsis thaliana, maize, and soybean and others were used as amplified templates, demonstrating that these two systems are specific for the identification and quantification of G. hirsutum. The results of Southern blot analysis also showed that the Sad1 gene was two copies in these 15 different G. hirsutum cultivars. Furthermore, one multiplex RT-quantitative PCR employing this gene as an endogenous reference gene was designed to quantify the Cry1A(c) gene modified from Bacillus thuringiensis (Bt) in the insect-resistant cottons, such as Mon531 and GK19. The quantification detection limit of the Cry1A(c) and Sad1 genes was as low as 10 pg of genomic DNA. These results indicat that the Sad1 gene can be used as an endogenous reference gene for both qualitative and quantitative PCR detection of GM cottons.     

Detection and quantification of Enterococcus gilvus in cheese by real-time PCR

The objective of this work was to investigate the occurrence of Enterococcus gilvus in cheese. For this purpose, a real-time PCR protocol using phenylalanyl-tRNA synthase (pheS) as a target gene was optimized to evaluate the presence and abundance of this microorganism in Italian artisan cheeses. The real-time assay unequivocally distinguished E. gilvus from 25 non-target LAB and non-LAB species, demonstrating its absolute specificity. The assay performed well not only with purified DNA but also with DNA extracted from cheese samples artificially contaminated with E. gilvus. The dynamic range of target determination of the method in the cheese matrix (from 10⁷ to 10⁴ cfu/ml, covering three orders of magnitude) was lower and the detection limit higher than in vitro conditions, but still high enough to obtain an excellent quantification accuracy in cheese. Twenty commercially available cheeses were analyzed by real-time PCR and approximately 40% of the cheese samples contained E. gilvus at levels ranging from 4.17±0.10 to 6.75±0.01 log cfu/g. Such levels represented 0.1–10% of the total enterococci counted on kanamycin aesculin azide agar (KAA) from the corresponding cheeses. The successful isolation of E. gilvus from cheeses containing high loads of this species, as detected by real-time PCR, provided definitive proof on both assay specificity and presence of this organism in cheeses. Despite the relatively low sensitivity in cheese (≥4 log cfu/g), the real-time PCR described here may, however, be useful to detect E. gilvus rapidly when present at (sub)dominant levels within the enterococcal cheese microflora. The assay may be helpful to detect and quantify E. gilvus strains from food, thus enabling a better understanding of technological role, ecological and safety aspects in cheeses and other fermented food products of this infrequent species.     

Entomopathogenic nematodes, phoretic Paenibacillus spp., and the use of real time quantitative PCR to explore soil food webs in Florida citrus groves

Quantitative real-time PCR (qPCR) is a powerful tool to detect and quantify species of cryptic organisms such as bacteria, fungi and nematodes from soil samples. As such, qPCR offers new opportunities to study the ecology of soil habitats by providing a single method to characterize communities of diverse organisms from a sample of DNA. Here we describe molecular tools to detect and quantify two bacteria (Paenibacillus nematophilus and Paenibacillus sp.) phoretically associated with entomopathogenic nematodes (EPNs) in the families Heterorhabditidae and Steinernematodae. We also extend the repertoire of species specific primers and TaqMan® probes for EPNs to include Heterorhabditis bacteriophora, Steinernema carpocapsae, Steinernema feltiae and Steinernema scapterisci, all widely distributed species used commercially for biological control. Primers and probes were designed from the ITS rDNA region for the EPNs and the 16S rDNA region for the bacteria. Standard curves were established using DNA from pure cultures of EPNs and plasmid DNA from the bacteria. The use of TaqMan probes in qPCR resolved the non-specificity of EPN and some bacterial primer amplifications whereas those for Paenibacillus sp. also amplified Paenibacillus thiaminolyticus and Paenibacillus popilliae, two species that are not phoretically associated with nematodes. The primer-probe sets for EPNs were able to accurately detect three infective juvenile EPNs added to nematodes recovered from soil samples. The molecular set for Paenibacillus sp. detected the bacterium attached to Steinernema diaprepesi suspended in water or added to nematodes recovered from soil samples but its detection decreased markedly in the soil samples, even when a nested PCR protocol was employed. Using qPCR we detected S. scapterisci at low levels in a citrus grove, which suggested natural long-distance spread of this exotic species, which is applied to pastures and golf courses to manage mole crickets (Scapteriscus spp.). Paenibacillus sp. (but not P. nematophilus) was detected in low quantities in the same survey but was unrelated to the spatial pattern of S. diaprepesi. The results of this research validate several new tools for studying the ecology of EPNs and their phoretic bacteria.     

Investigation of Toxoplasma gondii presence in farmed shellfish by nested-PCR and real-time PCR fluorescent amplicon generation assay (FLAG)

To evaluate the presence of Toxoplasma gondii in edible farmed shellfish, 1734 shellfish specimens i.e., 109 Crassostrea gigas (6 pools), 660 Mytilus galloprovincialis (22 pools), 804 Tapes decussatus (28 pools) and 161 Tapes philippinarum (6 pools), were collected from the Varano Lagoon (Apulia, Italy).Shellfish from 62 pools were subjected to two molecular techniques: a nested-PCR assay, and a fluorescent amplicon generation (FLAG) real-time PCR assay, both based on the multi-copy B1 target, were performed.One pooled sample of gills from C. gigas and one pooled sample of haemolymphs from T. decussatus were assessed as positive for T. gondii DNA by both techniques.The results demonstrated the presence of T. gondii in edible farmed C. gigas and T. decussatus and indicate that there may be a considerable health threat involved in eating contaminated raw shellfish.     

Wind-driven river plume dynamics and toxic Alexandrium tamarense blooms in the St. Lawrence estuary (Canada): A modeling study

In the lower St. Lawrence estuary (LSLE, eastern Canada), blooms of the toxic dinoflagellate Alexandrium tamarense are a recurrent phenomenon, resulting in paralytic shellfish poisoning outbreaks every summer. A first coupled physical–biological model of A. tamarense blooms was developed for this system in order to explore the interactions between cyst germination, cellular growth and water circulation and to identify the effect of physical processes on bloom development and transport across the estuary. The simulated summer (1998) was characterized by an A. tamarense red tide with concentrations reaching 2.3 × 10⁶ cells L⁻¹ along the south shore of the LSLE. The biological model was built with previously observed A. tamarense cyst distribution, cyst germination rate and timing, and A. tamarense growth limitation by temperature and salinity. The coupled model successfully reproduced the timing of the A. tamarense bloom in 1998, its coincidence with the combined plumes from the Manicouagan and Aux-Outardes (M-O) rivers on the north shore of the estuary, and the temporal variations in the north-south gradients in cell concentrations. The simulation results reveal that the interaction between cyst germination and the estuarine circulation generates a preferential inoculation of the surface waters of the M-O river plume with newly germinated cells which could partly explain the coincidence of the blooms with the freshwater plume. Furthermore, the results suggest that the spatio-temporal evolution of the bloom is dominated by alternating periods of retention and advection of the M-O plume: east or north-east winds favor the retention of the plume close to the north shore while west or north-west winds result in its advection toward the south shore. The response of the simulated freshwater plume to fluctuating wind forcing controls the delivery of the A. tamarense bloom from the northern part of the estuary to the south shore. In addition, our results suggest that a long residence time of the M-O plume and associated A. tamarense population in the LSLE during the summer 1998 contributed to the development of the red tide. We thus hypothesize that the wind-driven dynamics of the M-O plume could partly determine the success of A. tamarense blooms in the LSLE by influencing the residence time of the blooms and water column stability, which in turn affects A. tamarense vertical migrations and growth.     

Quantitative trait loci for lodging resistance,plant height and partial resistance to mycosphaerella blight in field pea (Pisum sativum L.)

With the development of genetic maps and the identification of the most-likely positions of quantitative trait loci (QTLs) on these maps, molecular markers for lodging resistance can be identified. Consequently, marker-assisted selection (MAS) has the potential to improve the efficiency of selection for lodging resistance in a breeding program. This study was conducted to identify genetic loci associated with lodging resistance, plant height and reaction to mycosphaerella blight in pea. A population consisting of 88 recombinant inbred lines (RILs) was developed from a cross between Carneval and MP1401. The RILs were evaluated in 11 environments across the provinces of Manitoba, Saskatchewan and Alberta, Canada in 1998, 1999 and 2000. One hundred and ninety two amplified fragment length polymorphism (AFLP) markers, 13 random amplified polymorphic DNA (RAPD) markers and one sequence tagged site (STS) marker were assigned to ten linkage groups (LGs) that covered 1,274 centi Morgans (cM) of the pea genome. Six of these LGs were aligned with the previous pea map. Two QTLs were identified for lodging resistance that collectively explained 58% of the total phenotypic variation in the mean environment. Three QTLs were identified each for plant height and resistance to mycosphaerella blight, which accounted for 65% and 36% of the total phenotypic variation, respectively, in the mean environment. These QTLs were relatively consistent across environments. The AFLP marker that was associated with the major locus for lodging resistance was converted into the sequence-characterized amplified-region (SCAR) marker. The presence or absence of the SCAR marker corresponded well with the lodging reaction of 50 commercial pea varieties.Communicated by H. F. Linskens