Assessment of prey overlap between a native (Polistes humilis) and an introduced (Vespula germanica) social wasp using morphology and phylogenetic analyses of 16S rDNA

In newly invaded communities, interspecific competition is thought to play an important role in determining the success of the invader and its impact on the native community. In southern Australia, the native Polistes humilis was the predominant social wasp prior to the arrival of the exotic Vespula germanica (Hymenoptera: Vespidae). Both species forage for similar resources (water, pulp, carbohydrate and protein prey), and concerns have arisen about potential competition between them. The aim of this study was to identify the protein foods that these wasps feed on. As many prey items are masticated by these wasps to the degree that they cannot be identified using conventional means, morphological identification was complemented by sequencing fragments of the mitochondrial 16S rRNA gene. GenBank searches using blast and phylogenetic analyses were used to identify prey items to at least order level. The results were used to construct complete prey inventories for the two species. These indicate that while P. humilis is restricted to feeding on lepidopteran larvae, V. germanica collects a variety of prey of invertebrate and vertebrate origin. Calculated values of prey overlap between the two species are used to discuss the implications of V. germanica impacting on P. humilis. Results obtained are compared to those gained by solely 'conventional' methods, and the advantages of using DNA-based taxonomy in ecological studies are emphasized.

In Vitro Conditions for a Successful Biolistics transformation System of Pineapples (ananas comosus merr.), in 'Contributing to a Sustainable Future'

In order to develop an efficient and reliable biolistics transformation system for pineapples parameters need to be optimised for growth, survival and development of explants pre- and post transformation. We have optimised in vitro conditions for culture media for the various stages of plant and callus initiation and development, and for effective selection of putative transgenic material. Shoot multiplication and proliferation is best on medium containing MS basic nutrients and vitamins with the addition of 0.1 mg/L myo-inositol, 20 g/L sucrose, 2.5 mg/L BAP and 3 g/L Phytagel, followed by transfer to basic MS medium for further development. Callus production on leaf base explants is best on MS nutrients and vitamins, to which 10 mg/L of BAP and NAA each was added. Optimum explant age for bombardment is 17-35 week old callus, while a pre-bombardment osmoticum treatment in the medium is not required. By comparing several antibiotics as selective agent, it has been established that a two-step selection of 2 fortnightly sub-cultures on 50 μg/mL of geneticin in the culture medium, followed by monthly sub-cultures on 100 μg/mL geneticin is optimal for survival of transgenic callus. Shoot regeneration from callus cultures is optimal on medium containing MS nutrients and vitamins, 5% coconut water and 400 mg/L casein hydrolysate. Plants can be readily regenerated and multiplied from transgenic callus through organogenesis. Rooting of shoots does not require any additional plant hormones to the medium. A transformation efficiency of 1 – 3.5% can be achieved, depending on the gene construct applied.

Transgenic gene silencing strategies for virus control.

Co-suppression of transgenes and their homologous viral sequences by RNA silencing is a powerful strategy for achieving high-level virus resistance in plants. This review provides a brief overview of RNA silencing mechanisms in plants and discusses important transgene construct design features underpinning successful RNA silencing-mediated transgenic virus control. Application of those strategies to protect horticultural and field crops from virus infection and results of field tests are also provided. The effectiveness and stability of RNA-mediated transgenic resistance are assessed taking into account effects of viral, plant and environmental factors.

The essential and non-essential genes of Bovine herpesvirus 1

Bovine herpesvirus 1 (BoHV-1) is an economically important pathogen of cattle associated with respiratory and reproductive disease. To further develop BoHV-1 as a vaccine vector, a study was conducted to identify the essential and non-essential genes required for in vitro viability. Randominsertion mutagenesis utilizing a Tn5 transposition system and targeted gene deletion were employed to construct gene disruption and gene deletion libraries, respectively, of an infectious clone of BoHV-1. Transposon insertion position and confirmation of gene deletion were determined by direct sequencing. The essential or non-essential requirement of either transposed or deleted open reading frames (ORFs) was assessed by transfection of respective BoHV-1 DNA into host cells. Of the 73 recognized ORFs encoded by the BoHV-1 genome, 33 were determined to be essential and 36 to be non-essential for virus viability in cell culture; determining the requirement of the two dual copy ORFs was inconclusive. The majority of ORFs were shown to conform to the in vitro requirements of BoHV-1 homologues encoded by human herpesvirus 1 (HHV-1). However, ORFs encoding glycoprotein K (UL53), regulatory, membrane, tegument and capsid proteins (UL54, UL49.5, UL49, UL35, UL20, UL16 and UL7) were shown to differ in requirement when compared to HHV-1-encoded homologues.

DArT markers: Diversity analyses and mapping in Sorghum bicolor

The sequential nature of gel-based marker systems entails low throughput and high costs per assay. Commonly used marker systems such as SSR and SNP are also dependent on sequence information. These limitations result in high cost per data point and significantly limit the capacity of breeding programs to obtain sufficient return on investment to justify the routine use of marker-assisted breeding for many traits and particularly quantitative traits. Diversity Arrays Technology (DArT™) is a cost effective hybridisation-based marker technology that offers a high multiplexing level while being independent of sequence information. This technology offers sorghum breeding programs an alternative approach to whole-genome profiling. We report on the development, application, mapping and utility of DArT™ markers for sorghum germplasm. Results: A genotyping array was developed representing approximately 12,000 genomic clones using PstI+BanII complexity with a subset of clones obtained through the suppression subtractive hybridisation (SSH) method. The genotyping array was used to analyse a diverse set of sorghum genotypes and screening a Recombinant Inbred Lines (RIL) mapping population. Over 500 markers detected variation among 90 accessions used in a diversity analysis. Cluster analysis discriminated well between all 90 genotypes. To confirm that the sorghum DArT markers behave in a Mendelian manner, we constructed a genetic linkage map for a cross between R931945-2-2 and IS 8525 integrating DArT and other marker types. In total, 596 markers could be placed on the integrated linkage map, which spanned 1431.6 cM. The genetic linkage map had an average marker density of 1/2.39 cM, with an average DArT marker density of 1/3.9 cM. Conclusion: We have successfully developed DArT markers for Sorghum bicolor and have demonstrated that DArT provides high quality markers that can be used for diversity analyses and to construct medium-density genetic linkage maps. The high number of DArT markers generated in a single assay not only provides a precise estimate of genetic relationships among genotypes, but also their even distribution over the genome offers real advantages for a range of molecular breeding and genomics applications.

Identification of quantitative trait loci for resistance to two species of root-lesion nematode (Pratylenchus thornei and P. neglectus) in wheat

Pratylenchus thornei and P. neglectus are two species of root-lesion nematode that cause substantial yield losses in wheat. No commercially available wheat variety has resistance to both species. A doubled-haploid population developed from a cross between the synthetic hexaploid wheat line CPI133872 and the bread wheat Janz was used to locate and tag quantitative trait loci (QTLs) associated with resistance to both P. thornei and P. neglectus. Wheat plants were inoculated with both species of nematode in independent replicated glasshouse trials repeated over 2 years. Known locations of wheat microsatellite markers were used to construct a framework map. After an initial single-marker analysis to detect marker-trait linkages, chromosome regions associated with putative QTLs were targetted with microsatellite markers to increase map density in the chromosome regions of interest. In total, 148 wheat microsatellite markers and 21 amplified fragment length polymorphism markers were mapped. The codominant microsatellite marker Xbarc183 on the distal end of chromosome 6DS was allelic for resistance to both P. thornei and P. neglectus. The QTL were designated QRlnt.lrc-6D.1 and QRlnn.lrc-6D.1, for the 2 traits, respectively. The allele inherited from CPI133872 explained 22.0-24.2% of the phenotypic variation for P. thornei resistance, and the allele inherited from Janz accounted for 11.3-14.0% of the phenotypic variation for P. neglectus resistance. Composite interval mapping identified markers that flank a second major QTL on chromosome 6DL (QRlnt.lrc-6D.2) that explained 8.3-13.4% of the phenotypic variation for P. thornei resistance. An additional major QTL associated with P. neglectus resistance was detected on chromosome 4DS (QRlnn.lrc-4D.1) and explained a further 10.3-15.4% of the phenotypic variation. The identification and tagging of nematode resistance genes with molecular markers will allow appropriate allele combinations to be selected, which will aid the successful breeding of wheat with dual nematode resistance.

The big pond debate.

Over recent decades, Australian piggeries have commonly employed anaerobic ponds to treat effluent to a standard suitable for recycling for shed flushing purposes and for irrigation onto nearby agricultural land. Anaerobic ponds are generally sized according to the Rational Design Standard (RDS) developed by Barth (1985), resulting in large ponds, which can be expensive to construct, occupy large land areas, and are difficult and expensive to desludge, potentially disrupting the whole piggery operation. Limited anecdotal and scientific evidence suggests that anaerobic ponds that are undersized according to the RDS, operate satisfactorily, without excessive odour emission, impaired biological function or high rates of solids accumulation. Based on these observations, this paper questions the validity of rigidly applying the principles of the RDS and presents a number of alternate design approaches resulting in smaller, more highly loaded ponds that are easier and cheaper to construct and manage. Based on limited data of pond odour emission, it is suggested that higher pond loading rates may reduce overall odour emission by decreasing the pond volume and surface area. Other management options that could be implemented to reduce pond volumes include permeable pond covers, various solids separation methods, and bio-digesters with impermeable covers, used in conjunction with biofilters and/or systems designed for biogas recovery. To ensure that new effluent management options are accepted by regulatory authorities, it is important for researchers to address both industry and regulator concerns and uncertainties regarding new technology, and to demonstrate, beyond reasonable doubt, that new technologies do not increase the risk of adverse impacts on the environment or community amenity. Further development of raw research outcomes to produce relatively simple, practical guidelines and implementation tools also increases the potential for acceptance and implementation of new technology by regulators and industry.

A Practitioner-Oriented Decision Support Process for Forestry Pest Management

The article discusses a new decision support process for forestry pest management. Over the past few years, DSS have been introduced for forestry pest management, providing forest growers with advice in areas such as selecting the most suitable pesticide and relevant treatment. Most of the initiatives process knowledge from various domains for providing support for specific decision making problems. However, very few studies have identified the requirements of developing a combined process model in which all relevant practitioners can contribute and share knowledge for effective decision making; such an approach would need to include the decision makers’ perspective along with other relevant attributes such as the problem context and relevant policies. We outline a decision support process for forestry pest management, based on the design science research paradigm, in which a focus group technique has application to acquire both expert and practical knowledge in order to construct the DSS solution.

Simple software processes and tests improve the reliability and usefulness of a model.

Models are abstractions of reality that have predetermined limits (often not consciously thought through) on what problem domains the models can be used to explore. These limits are determined by the range of observed data used to construct and validate the model. However, it is important to remember that operating the model beyond these limits, one of the reasons for building the model in the first place, potentially brings unwanted behaviour and thus reduces the usefulness of the model. Our experience with the Agricultural Production Systems Simulator (APSIM), a farming systems model, has led us to adapt techniques from the disciplines of modelling and software development to create a model development process. This process is simple, easy to follow, and brings a much higher level of stability to the development effort, which then delivers a much more useful model. A major part of the process relies on having a range of detailed model tests (unit, simulation, sensibility, validation) that exercise a model at various levels (sub-model, model and simulation). To underline the usefulness of testing, we examine several case studies where simulated output can be compared with simple relationships. For example, output is compared with crop water use efficiency relationships gleaned from the literature to check that the model reproduces the expected function. Similarly, another case study attempts to reproduce generalised hydrological relationships found in the literature. This paper then describes a simple model development process (using version control, automated testing and differencing tools), that will enhance the reliability and usefulness of a model.

TU10025 Erosion control - turf research and development facility

The aim of this project is to construct a large-scale erosion control education and demonstration facility at Redland Research Station. This will be done in collaboration with the Australian turf industry (as members of the steering committee) and consultant researcher Dr Rob Loch (project partner). The project will employ a part-time industry development officer (IDO) for Turf Australia to increase engagement with the project by the target audience. The project’s main strategy is to extend the research results from HAL funded project Optimising Turf Use to Minimise Soil Erosion on Construction Sites TU08033 so that the maximum return on investment can be derived for the turf levy payers and HAL from that study.