Characterization of Cry34Ab1 and Cry35Ab1 insecticidal crystal proteins expressed in transgenic corn plants and Pseudomonas fluorescens

Cry34Ab1 and Cry35Ab1 proteins, identified from Bacillus thuringiensis strain PS149B1, act together to control corn rootworms. Transgenic corn lines coexpressing the two proteins were developed to protect corn against rootworm damage. Large quantities of the two proteins were needed to conduct studies required for assessing the safety of this transgenic corn crop. Because it was technically infeasible to obtain sufficient quantities of high purity Cry34Ab1 and Cry35Ab1 proteins from the transgenic corn plants, the proteins were produced using a recombinant Pseudomonas fluorescens (Pf) production system. The two proteins from both the transgenic corn and the Pf were purified and characterized. The proteins from each host had the expected molecular mass and were immunoreactive to specific antibodies in enzyme-linked immunosorbent assay and Western blot analysis. Data from N-terminal sequencing, tryptic peptide mass fingerprinting, internal peptide sequencing, and biological activity provided direct evidence that the Cry34Ab1 and Cry35Ab1 proteins produced in Pf and transgenic corn were, respectively, comparable or equivalent molecules. In addition, neither protein had detectable glycosylation regardless of the host.     

Liming of Acid and Metal Contaminated Catchments for the Improvement of Drainage Water Quality

A 38 ha near-barren experimental catchment area near an abandoned Cu and Ni smelter in Sudbury, Canada was treated with 410 tons of coarse dolomitic limestone in 1994. An additional 54 tons of pelletized fine dolomite were added to 15 wetlands within the experimental catchment in 1995. The treatments significantly increased the pH and base cation concentrations in the outlet stream. Cu and Ni concentrations initially rose after the wetland treatment, but then declined to levels below those of the reference site. Bioassay tests revealed that the toxicity of the drainage water was greatly reduced by the liming, but some localized inputs of highly toxic groundwater still posed a problem for aquatic biota. The pH of surface water in the wetlands has been maintained at >6.0 for over 4 years. The wetland liming appeared to be highly effective at neutralizing drainage water, however there may be some adverse effects on wetland plant communities as a consequence.     

Japanese consumer categorisation of beef into quality grades,based on Meat Standards Australia methodology

The objective was to evaluate the sensory categorisation of beef by Japanese consumers, based on Meat Standards Australia methodology. Various cuts of beef, with a wide range of quality (from Australian and Japanese cattle) and three cooking methods (grill, yakiniku, shabu shabu), were evaluated by 1620 Japanese consumers in Tokyo and Osaka. Consumers rated each sample for four sensory attributes (tenderness, juiciness, flavor and overall satisfaction), then selected one of four grades (unsatisfactory/2‐star, good everyday/3‐star, better than everyday/4‐star, and premium quality/5‐star), based on the quality of the beef within each cooking method. Meat quality scores, denoted as MQ4 scores (weighted combination of the four sensory attributes) were calculated from the Japanese consumer test results, to describe the Japanese consumer rating of beef. The distribution of the Japanese consumer MQ4 scores showed a clear distinction between grades, with the majority of scores being included within the boundaries of each grade. The MQ4 score allocated approximately 64% of the samples to their correct consumer grades. The MQ4 score showed potential to be used as a tool in developing and monitoring a consumer‐focused grading system that is able to predict Japanese consumer satisfaction of individual beef cuts prepared by different cooking methods.     

Phylogeography,genetic stocks,and conservation implications for an Australian endemic marine turtle

1. Identification of the geographic extent of population boundaries, the distribution of genetic lineages, and the amount of genetic exchange among breeding groups is needed for effective conservation of vulnerable marine migratory species. This is particularly true of the flatback turtle (Natator depressus), which only breeds in Australia but has extensive migrations that can include international waters.
2. This study investigated the phylogeography and genetic structure among 17 flatback turtle rookeries across their range by sequencing an 810 bp portion of the mitochondrial DNA in 889 samples and genotyping 10 microsatellite loci in 598 samples.
3. There was low phylogenetic divergence among haplotypes and evidence of recent population expansion, likely in the late Pleistocene. A predominant haplotype was found across all rookeries, but other haplotype groups were regionally specific.
4. In general, there was agreement in patterns of genetic differentiation in the mitochondrial DNA and microsatellite data, and in some pairwise comparisons a higher mutation rate of microsatellites provided stronger evidence of differentiation.
5. These results suggest natal philopatry operates in the choice of breeding locations for males as well as females.
6. Evidence of genetic connectivity among neighbouring rookeries led to the identification of seven genetic stocks. Geographic boundaries of rookeries used by genetic stocks varied widely (160–1,300 km), highlighting a need for field studies to better understand movement patterns.
7. Hierarchical analysis of molecular variance identified significant genetic differentiation based upon genetic stock, nesting phenology (summer vs. winter nesters), and a west–east discontinuity across Torres Strait. A pattern of isolation by distance was identified, which was most strongly observed in the microsatellite data.
8. In combination with tagging and telemetry studies, these results will allow better quantification of stock‐specific threats along migratory routes and in foraging habitats. Implications of climate change will be stock specific and may depend upon the extent of genetic connectivity between neighbouring stocks.

     

Zooplankton and epibenthic fauna in shrimp ponds: factors influencing assemblage dynamics

The assemblage composition, biomass and dynamics of zooplankton and epibenthos were examined in a commercial shrimp (penaeid prawn) pond in subtropical Australia. Physicochemical characteristics of the pond water were measured concurrently. Numbers and biomass of zooplankton in the surface tows (140 μm mesh) varied from 111.7 ind. L^?1 (324 μg L^?1) to 8.3 ind. L^?1 (44.2 μg L^?1). Immediately after the ponds were stocked with shrimp postlarvae there was a rapid decline in zooplankton numbers, particularly the dominant larger copepods. We attributed this to predation by the shrimp postlarvae. Subsequent peaks in zooplankton numbers were principally due to barnacle nauplii. Changes in abundance and biomass of the zooplankton assemblage were not correlated with physicochemical characteristics. Epibenthic faunal abundance in the beam trawls (1 mm mesh) peaked at 14 ind. m^?2 and the biomass at 0.8 g m^?2. Unlike zooplankton, the peaks in abundance of epibenthos did not correspond to the peaks in biomass. This was due to the large differences in the size of the dominant taxa across the season. Sergestids (Acetes sibogae) and amphipods were the most abundant taxa in beam trawl samples, with amphipods abundance increasing towards the end of the growout. Negative correlations were found between epibenthos abundance and pH and temperature. These relationships were strongly influenced by the high abundances of amphipods and may reflect an effect on the growth of macroalgae in the pond rather than a direct effect on the epibenthos. No correlations were found between epibenthic fauna biomass and physicochemical parameters. Abundances of epibenthic fauna were not related to zooplankton densities, indicating that this source of food was not likely to be a limiting factor. Neither the pond water exchange regime nor moon phase could explain changes in abundances of zooplankton or epibenthos assemblages. Zooplankton clearly contribute to the nutrition of shrimp postlarvae immediately after stocking. The establishment of an abundant assemblage of zooplankton before stocking shrimp postlarvae would appear to be beneficial, if not essential. Later in the season, zooplankton and epibenthos apparently contribute little to shrimp biomass. Owing to their relatively low biomass, the consumption of shrimp feeds by epibenthos is likely to be insignificant compared with that of the shrimp.     

Dynamic Oryza Genomes: Repetitive DNA Sequences as Genome Modeling Agents

Repetitive sequences, primarily transposable elements form an indispensable part of eukaryotic genomes. However, little is known about how these sequences originate, evolve and function in context of a genome. In an attempt to address this question, we performed a comparative analysis of repetitive DNA sequences in the genus Oryza, representing ~15 million years of evolution. Both Class I and Class II transposable elements, through their expansion, loss and movement in the genome, were found to influence genome size variation in this genus. We identified 38 LTRretrotransposon families that are present in 1,500 or more copies throughout Oryza, and many are preferentially amplified in specific lineages. The data presented here, besides furthering our understanding of genome organization in the genus Oryza, will aid in the assembly, annotation and analysis of genomic data, as part of the future genome sequencing projects of O. sativa wild relatives.     

Seroepidemiology of leptospirosis in livestock in Trinidad

A study was conducted to determine the seroprevalence of leptospirosis and infecting serovars across livestock (cattle, sheep, goats, and pigs) in Trinidad using the microscopic agglutination test with an international panel of 23 serovars. Of a total of 590 cattle tested, 21.5% were seropositive with agglutinations to 13 of the 23 antigens used in the panel. Icterohaemorrhagiae (9.3%), Sejroe (4.1%), Ballum (4.1%), and Autumnalis (1.9%) were the predominant serogroups detected in the cattle sampled (n = 590). Of 222 sheep tested, 5.0% were seropositive with agglutinations to five serovars belonging to two serogroups. These serogroups were Autumnalis at 2.7%, and Icterohaemorrhagiae at 2.3% of all sheep tested (n = 222). Of a total of 180 goats tested, 3.3% were seropositive, all agglutinating to the Icterohaemorrhagiae serogroup, 1.7% to serovar Copenhageni, 1.1% to serovar Mankarso, and 0.6% to serovar Icterohaemorrhagiae. Among pigs (n = 200), 5.0% were seropositive for five serovars belonging to three serogroups. These serogroups were Icterohaemorrhagiae at 2.5%, Australis at 2%, and Ballum at 0.5%. Overall, age and sex of animals were not significantly associated with leptospirosis with the exception of cattle where age was a significant factor for seropositivity. It was concluded that for livestock, leptospirosis may be an important zoonotic and economic disease, particularly in the case of cattle. It is imperative that the impact of leptospirosis on abortion, stillbirths, and decreased milk production in livestock in the country be assessed.     

Genotyping by sequencing of rice interspecific backcross inbred lines identifies QTLs for grain weight and grain length

Grain weight and grain length are the most stable components of rice yield and important indicators of consumer preference. Considering the potentials of wild rice and to enhance the rice yields to meet the increasing demands, 185 Backcross Inbred Lines (BILs) in the background of O. sativa ssp. indica cv. PR114, including 63 rufi-BILs derived from O. rufipogon IRGC104433 and 122 glumae-BILs from O. glumaepatula IRGC104387 were evaluated for mapping QTLs for yield and yield component traits using Genotyping by Sequencing (GBS). Phenotypic evaluation of BILs in three seasons spanning two locations revealed significant differences compared with recurrent parent. BILs which did not show significant differences for any trait under investigation, or similar based on pedigree, were excluded from GBS. Some glumae-BILs had to be excluded from mapping QTLs due to less sequence information. A custom designed approach for GBS data analysis identified 3322 informative SNPs in 55 rufi-BILs and 3437 informative SNPs in 79 glumae-BILs. QTL mapping identified one QTL for thousand grain weight (qtgw5.1), two for grain width (qgw5.1, qgw5.2) and one for grain length (qgl7.1) in rufi-BILs. In the glumae-BILs, three QTL for thousand grain weight (qtgw2.1, qtgw3.1, qtgw6.1) and two for grain length (qgl3.1, qgl7.1) were identified. Most of the grain weight and width QTL showed positive additive effect contributed by wild species allele, whereas the grain length QTL showed positive additive effect contributed by recurrent parent allele. Based on their physical position, none of the QTLs were found similar to previously cloned QTLs. QTLs for grain traits identified from low yielding wild relatives of rice reveals their significance in improving further the rice yields and widen the genetic base of cultivated rice.