Development and Accreditation of an Applied Climate Education Unit for Sustainable Land Use in Australia

In recent years, there have been significant developments in climate science relevant to agriculture and natural resource management. Assessing impacts of climate variability and use of seasonal climate forecasts have become increasingly important elements in the management "toolkit" for many Australian farmers. Consideration of climate change further increases the need for improved management strategies. While climate risk extension activities have kept pace with advances in climate science, a national review of the Vocational Education and Training system in Australia in relation to "weather and climate" showed that these topics were "poorly represented" at the management level in the Australian Qualifications Framework, and needed increased emphasis. Consequently, a new Unit of Competency concerning management of climatic risk was developed and accredited to address this deficiency. The objective of the unit was to build knowledge and skills for better management of climate variability via the elements of surveying climatic and enterprise data; analysing climatic risks and opportunities; and developing climatic risk management strategies. This paper describes establishment of a new unit for vocational education that is designed to harness recent developments in applied climate science for better management of Australia's highly variable climate. The main benefits of the new unit of competency, "Developing climatic risk management strategies,"were seen as improving decisions in climate and agriculture, and reducing climate risk exposure to enhance sustainable agriculture. The educational unit is now within the scope of agricultural colleges, universities, and registered training organisations as an accredited unit.

(Z)-11-Hexadecenal and (3Z,6Z,9Z)-Tricosatriene: Sex Pheromone Components of the Red Banded Mango Caterpillar Deanolis sublimbalis

The sex pheromone of the red banded mango caterpillar, Deanolis sublimbalis (Lepidoptera: Crambidae), a serious pest of the mango Mangifera indica (Anacardiaceae) in India and Southeast Asia and a recent invader into northern Australia, has been identified. Three candidate compounds were identified from pheromone gland extracts of female moths, using gas chromatography (GC), GC-electroantennographic detection and GC-mass spectrometric analyses, in conjunction with dimethyldisulfide derivatization. Field bioassays established that both (Z)-11-hexadecenal (Z11-16:Ald) and (3Z,6Z,9Z)-tricosatriene (3Z,6Z,9Z-23:Hy) were required for attraction of male D. sublimbalis moths, and 1,000 μg of a 1:1 mix of Z11-16:Ald and 3Z,6Z,9Z-23:Hy was more attractive to male moths than caged virgin females. However, the binary blend was only attractive when the isomeric purity of the monounsaturated aldehyde was >99%, suggesting that the (E)-isomer was inhibitory. Although (Z)-11-hexadecen-1-ol (Z11-16:OH) was tentatively identified in gland extracts, the addition of this compound to the binary blend did not increase the numbers of moths captured. The pheromone can now be used in integrated pest management strategies.

Assessing the relationship between patch type and soil mites: A molecular approach

An urgent need exists for indicators of soil health and patch functionality in extensive rangelands that can be measured efficiently and at low cost. Soil mites are candidate indicators, but their identification and handling is so specialised and time-consuming that their inclusion in routine monitoring is unlikely. The aim of this study was to measure the relationship between patch type and mite assemblages using a conventional approach. An additional aim was to determine if a molecular approach traditionally used for soil microbes could be adapted for soil mites to overcome some of the bottlenecks associated with soil fauna diversity assessment. Soil mite species abundance and diversity were measured using conventional ecological methods in soil from patches with perennial grass and litter cover (PGL), and compared to soil from bare patches with annual grasses and/or litter cover (BAL). Soil mite assemblages were also assessed using a molecular method called terminal-restriction fragment length polymorphism (T-RFLP) analysis. The conventional data showed a relationship between patch type and mite assemblage. The Prostigmata and Oribatida were well represented in the PGL sites, particularly the Aphelacaridae (Oribatida). For T-RFLP analysis, the mite community was represented by a series of DNA fragment lengths that reflected mite sequence diversity. The T-RFLP data showed a distinct difference in the mite assemblage between the patch types. Where possible, T-RFLP peaks were matched to mite families using a reference 18S rDNA database, and the Aphelacaridae prevalent in the conventional samples at PGL sites were identified, as were prostigmatids and oribatids. We identified limits to the T-RFLP approach and this included an inability to distinguish some species whose DNA sequences were similar. Despite these limitations, the data still showed a clear difference between sites, and the molecular taxonomic inferences also compared well with the conventional ecological data. The results from this study indicated that the T-RFLP approach was effective in measuring mite assemblages in this system. The power of this technique lies in the fact that species diversity and abundance data can be obtained quickly because of the time taken to process hundreds of samples, from soil DNA extraction to data output on the gene analyser, can be as little as 4 days.

Non-target impacts of poison baiting for predator control in Australia

1. Mammalian predators are controlled by poison baiting in many parts of the world, often to alleviate their impacts on agriculture or the environment. Although predator control can have substantial benefits, the poisons used may also be potentially harmful to other wildlife. 2. Impacts on non-target species must be minimized, but can be difficult to predict or quantify. Species and individuals vary in their sensitivity to toxins and their propensity to consume poison baits, while populations vary in their resilience. Wildlife populations can accrue benefits from predator control, which outweigh the occasional deaths of non-target animals. We review recent advances in Australia, providing a framework for assessing non-target effects of poisoning operations and for developing techniques to minimize such effects. We also emphasize that weak or circumstantial evidence of non-target effects can be misleading. 3. Weak evidence that poison baiting presents a potential risk to non-target species comes from measuring the sensitivity of species to the toxin in the laboratory. More convincing evidence may be obtained by quantifying susceptibility in the field. This requires detailed information on the propensity of animals to locate and consume poison baits, as well as the likelihood of mortality if baits are consumed. Still stronger evidence may be obtained if predator baiting causes non-target mortality in the field (with toxin detected by post-mortem examination). Conclusive proof of a negative impact on populations of non-target species can be obtained only if any observed non-target mortality is followed by sustained reductions in population density. 4. Such proof is difficult to obtain and the possibility of a population-level impact cannot be reliably confirmed or dismissed without rigorous trials. In the absence of conclusive evidence, wildlife managers should adopt a precautionary approach which seeks to minimize potential risk to non-target individuals, while clarifying population-level effects through continued research.

Ultraviolet radiation effects on fruit surface respiration and chlorophyll fluorescence

High-value fruit crops are exposed to a range of environmental conditions that can reduce fruit quality. Solar injury (SI) or sunburn is a common disorder in tropical, sub-tropical, and temperate climates and is related to: 1) high fruit surface temperature; 2) high visible light intensity; and, 3) ultraviolet radiation (UV). Positional changes in fruit that are caused by increased weight or abrupt changes that result from summer pruning, limb breakage, or other damage to the canopy can expose fruit to high solar radiation levels, increased fruit surface temperatures, and increased UV exposure that are higher than the conditions to which they are adapted. In our studies, we examined the effects of high fruit surface temperature, saturating photosynthetically-active radiation (PAR), and short-term UV exposure on chlorophyll fluorescence, respiration, and photosynthesis of fruit peel tissues from tropical and temperate fruit in a simulation of these acute environmental changes. All tropical fruits (citrus, macadamia, avocado, pineapple, and custard apple) and the apple cultivars 'Gala', 'Gold Rush', and 'Granny Smith' increased dark respiration (A0) when exposed to UV, suggesting that UV repair mechanisms were induced. The maximum quantum efficiency of photosystem II (Fv/Fm) and the quantum efficiency of photosystem II (ΦII) were unaffected, indicating no adverse effects on photosystem II (PSII). In contrast, 'Braeburn' apple had a reduced Fv/Fm with no increase in A0 on all sampling dates. There was a consistent pattern in all studies. When Fv/Fm was unaffected by UV treatment, A0 increased significantly. Conversely, when Fv/Fm was reduced by UV treatment, then A0 was unaffected. The pattern suggests that when UV repair mechanisms are effective, PSII is adequately protected, and that this protection occurs at the cost of higher respiration. However, when the UV repair mechanisms are ineffective, not only is PSII damaged, but there is additional short-term damage to the repair mechanisms, indicated by a lack of respiration to provide energy.

Rice molecular breeding laboratories in the genomics era: Current status and future considerations

Using DNA markers in plant breeding with marker-assisted selection (MAS) could greatly improve the precision and efficiency of selection, leading to the accelerated development of new crop varieties. The numerous examples of MAS in rice have prompted many breeding institutes to establish molecular breeding labs. The last decade has produced an enormous amount of genomics research in rice, including the identification of thousands of QTLs for agronomically important traits, the generation of large amounts of gene expression data, and cloning and characterization of new genes, including the detection of single nucleotide polymorphisms. The pinnacle of genomics research has been the completion and annotation of genome sequences for indica and japonica rice. This information-coupled with the development of new genotyping methodologies and platforms, and the development of bioinformatics databases and software tools-provides even more exciting opportunities for rice molecular breeding in the 21st century. However, the great challenge for molecular breeders is to apply genomics data in actual breeding programs. Here, we review the current status of MAS in rice, current genomics projects and promising new genotyping methodologies, and evaluate the probable impact of genomics research. We also identify critical research areas to "bridge the application gap" between QTL identification and applied breeding that need to be addressed to realize the full potential of MAS, and propose ideas and guidelines for establishing rice molecular breeding labs in the postgenome sequence era to integrate molecular breeding within the context of overall rice breeding and research programs.

Detection of Chlamydiaceae DNA in veterinary specimens using a family-specific PCR

Aims: The aim of this work was to develop a rapid molecular test for the detection of the Chlamydiaceae family, irrespective of the species or animal host. Methods and Results: The method described herein is a polymerase chain reaction targeting the 16S rRNA gene of the Chlamydiaceae family, and the results demonstrate that the test reacts with five reference Chlamydiaceae but none of the 19 other bacterial species or five uninfected animal tissues tested. The results also indicate the enhanced sensitivity of this test when compared with conventional culture or serology techniques. This is demonstrated through parallel testing of six real clinical veterinary cases and confirmatory DNA sequence analysis. Conclusions, Significance and Impact of the Study: This test can be used by veterinary diagnostic laboratories for rapid detection of Chlamydiaceae in veterinary specimens, with no restriction of chlamydial species or animal host. The test does not differentiate chlamydial species, and if required, speciation must be carried out retrospectively using alternate methods. However, for the purpose of prescribing therapy for chlamydiosis, this test would be an invaluable laboratory tool.

Purification of immunoglobulins from chicken sera by thiophilic gel chromatography

Immunoglobulin Y is different from most of the other immunoglobulins because it does not bind protein A or protein G. Thiophilic gel chromatography has been successfully used to purify IgY from chicken egg yolk, but the technology has not previously been used to purify IgY from serum. In this research note, we describe the optimization of T-gel chromatography for purification of IgY from serum. Data are provided on the recovery and purity of IgY obtained using potassium sulfate buffers of different concentrations. Decreasing the strength of potassium sulfate buffer from 0.5 to 0.3 M did not alter the amount of IgY recovered but increased the purity. Using 0.3 M potassium sulphate, we recovered approximately 63.7% of the serum Ig as almost pure IgY.

Development and validation of an ELISA for detecting antibodies to Eimeria tenella in chickens

The aim of this study was to develop and validate an ELISA for detecting chicken antibodies to Eimeria tenella. An initial comparison of merozoite and sporozoite antigen preparations revealed few differences in their ability to monitor the onset, kinetics and magnitude of the antibody response suggesting that both antigens would be equally useful for development of an ELISA. Furthermore the cross-reactivity of these antigens with sera from birds infected with chicken Eimeria species was similar. The merozoite antigen was selected for further evaluation because it was easier to prepare. Discrimination between sera from birds experimentally infected with E. tenella and birds maintained in an Eimeria-free isolation facility was excellent. In sera collected from free-range layers and commercial broilers there also appeared to be clear discrimination between infected and uninfected birds. The ELISA should prove useful for monitoring infectivity in vaccination programmes in layer and breeder flocks and for assessing the effectiveness of biosecurity measures in broiler flocks.

Antibody response against endogenous stages of an attenuated strain of Eimeria tenella

The application of attenuated vaccines for the prevention of chicken coccidiosis has increased exponentially in recent years. In Eimeria infections, protective immunity is thought to rely on a strong cell mediated response with antibodies supposedly playing a minor role. However, under certain conditions antibodies seem to be significant in protection. Furthermore, antibodies could be useful for monitoring natural exposure of flocks to Eimeria spp. and for monitoring the infectivity of live vaccines. Our objective was to investigate the chicken antibody response to the different parasite lifecycle stages following infection with an attenuated strain of Eimeria tenella. Western blotting analysis of parasite antigens prepared from the lining of caeca infected with the attenuated strain of E. tenella revealed two dominant antigens of 32 and 34 kDa, apparently associated with trophozoites and merozoites that were present at high concentrations between 84 and 132 h post-infection. When cryosections of caeca infected with E. tenella were probed with IgY purified from immune birds the most intense reaction was observed with the asexual stages. Western blotting analysis of proteins of purified sporozoites and third generation merozoites and absorption of stage-specific antibodies from sera suggested that a large proportion of antigens is shared by the two stages. The time-courses of the antibody response to sporozoite and merozoite antigens were similar but varied depending on the inoculation regime and the degree of oocyst recirculation.

Variability in odour reception in the peripheral sensory system of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Mixtures of single odours were used to explore the receptor response profile across individual antennae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Seven odours were tested including floral and green-leaf volatiles: phenyl acetaldehyde, benzaldehyde, b-caryophyllene, limonene, a-pinene, 1-hexanol, 3Z-hexenyl acetate. Electroantennograms of responses to paired mixtures of odours showed that there was considerable variation in receptor tuning across the receptor field between individuals. Data from some moth antennae showed no additivity, which indicated a restricted receptor profile. Results from other moth antennae to the same odour mixtures showed a range of partial additivity. This indicated that a wider array of receptor types was present in these moths, with a greater percentage of the receptors tuned exclusively to each odour. Peripheral receptor fields show variation in the spectrum of response within a population (of moths) when exposed to high doses of plant volatiles. This may be related to recorded variation in host choice within moth populations as reported by other authors.

Isolation and characterization of novel microsatellite markers and their application for diversity assessment in cultivated groundnut (Arachis hypogaea)

Background: Cultivated peanut or groundnut (Arachis hypogaea L.) is the fourth most important oilseed crop in the world, grown mainly in tropical, subtropical and warm temperate climates. Due to its origin through a single and recent polyploidization event, followed by successive selection during breeding efforts, cultivated groundnut has a limited genetic background. In such species, microsatellite or simple sequence repeat (SSR) markers are very informative and useful for breeding applications. The low level of polymorphism in cultivated germplasm, however, warrants a need of larger number of polymorphic microsatellite markers for cultivated groundnut. Results: A microsatellite- enriched library was constructed from the genotype TMV2. Sequencing of 720 putative SSR-positive clones from a total of 3,072 provided 490 SSRs. 71.2% of these SSRs were perfect type, 13.1% were imperfect and 15.7% were compound. Among these SSRs, the GT/CA repeat motifs were the most common (37.6%) followed by GA/CT repeat motifs (25.9%). The primer pairs could be designed for a total of 170 SSRs and were optimized initially on two genotypes. 104 (61.2%) primer pairs yielded scorable amplicon and 46 (44.2%) primers showed polymorphism among 32 cultivated groundnut genotypes. The polymorphic SSR markers detected 2 to 5 alleles with an average of 2.44 per locus. The polymorphic information content (PIC) value for these markers varied from 0.12 to 0.75 with an average of 0.46. Based on 112 alleles obtained by 46 markers, a phenogram was constructed to understand the relationships among the 32 genotypes. Majority of the genotypes representing subspecies hypogaea were grouped together in one cluster, while the genotypes belonging to subspecies fastigiata were grouped mainly under two clusters. Conclusion. Newly developed set of 104 markers extends the repertoire of SSR markers for cultivated groundnut. These markers showed a good level of PIC value in cultivated germplasm and therefore would be very useful for germplasm analysis, linkage mapping, diversity studies and phylogenetic relationships in cultivated groundnut as well as related Arachis species.

Evolutionary relationships between bat coronaviruses and their hosts

Recent studies have suggested that bats are the natural reservoir of a range of coronaviruses (CoVs), and that rhinolophid bats harbor viruses closely related to the severe acute respiratory syndrome (SARS) CoV, which caused an outbreak of respiratory illness in humans during 2002-2003. We examined the evolutionary relationships between bat CoVs and their hosts by using sequence data of the virus RNA-dependent RNA polymerase gene and the bat cytochrome b gene. Phylogenetic analyses showed multiple incongruent associations between the phylogenies of rhinolophid bats and their CoVs, which suggested that host shifts have occurred in the recent evolutionary history of this group. These shifts may be due to either virus biologic traits or host behavioral traits. This finding has implications for the emergence of SARS and for the potential future emergence of SARS-CoVs or related viruses.

Phylogenetic considerations for predicting the host range of Ustilago sporoboli-indici, a potential biological control agent for Sporobolus species in Australia

The ribosomal DNA internal transcribed spacer region was amplified and sequenced from a selection of specimens of the Sporobolus smut Ustilago sporoboli-indici. Phylogenetic comparison with other Ustilago and Sporisorium species revealed strong support for an evolutionary radiation of Ustilago species infecting the Chloridoideae and Pooideae, of which U. sporoboli-indici forms a major lineage. Comparisons are made with other groups of plant pathogenic fungi, and it is concluded that phylogenetic analyses of potential biocontrol agents are useful for identifying pathogens that are derived from evolutionary lineages that parasitize a wide range of unrelated plants. Such pathogens are less desirable as biocontrol agents as they may have a greater likelihood of infecting plants outside their normal host ranges.

Australian and Thai isolates of Burkholderia pseudomallei are distinct by multilocus sequence typing: Revision of a case of mistaken identity

A recent study using multilocus sequence typing (MLST) of Burkholderia pseudomallei isolates found a sequence type (ST60) to be common to both Thailand and Australia, contradicting earlier studies showing complete distinction between isolates from these regions. The ST60 isolates reportedly from Australia had been obtained for MLST from United Kingdom and U.S. collections. We have located and characterized the original Australian isolates; they were collected in 1983, and they are neither ST60 nor B. pseudomallei isolates. The B. pseudomallei MLST database has been corrected, and there is no ST common to isolates verified as obtained from Australia or from Thailand.