A new data source for fisheries resource assessment: genetic estimates of the effective number of spawners. Final Report to the Fisheries Research and Development Corporation.

The development of innovative methods of stock assessment is a priority for State and Commonwealth fisheries agencies. It is driven by the need to facilitate sustainable exploitation of naturally occurring fisheries resources for the current and future economic, social and environmental well being of Australia. This project was initiated in this context and took advantage of considerable recent achievements in genomics that are shaping our comprehension of the DNA of humans and animals. The basic idea behind this project was that genetic estimates of effective population size, which can be made from empirical measurements of genetic drift, were equivalent to estimates of the successful number of spawners that is an important parameter in process of fisheries stock assessment. The broad objectives of this study were to 1. Critically evaluate a variety of mathematical methods of calculating effective spawner numbers (Ne) by a. conducting comprehensive computer simulations, and by b. analysis of empirical data collected from the Moreton Bay population of tiger prawns (P. esculentus). 2. Lay the groundwork for the application of the technology in the northern prawn fishery (NPF). 3. Produce software for the calculation of Ne, and to make it widely available. The project pulled together a range of mathematical models for estimating current effective population size from diverse sources. Some of them had been recently implemented with the latest statistical methods (eg. Bayesian framework Berthier, Beaumont et al. 2002), while others had lower profiles (eg. Pudovkin, Zaykin et al. 1996; Rousset and Raymond 1995). Computer code and later software with a user-friendly interface (NeEstimator) was produced to implement the methods. This was used as a basis for simulation experiments to evaluate the performance of the methods with an individual-based model of a prawn population. Following the guidelines suggested by computer simulations, the tiger prawn population in Moreton Bay (south-east Queensland) was sampled for genetic analysis with eight microsatellite loci in three successive spring spawning seasons in 2001, 2002 and 2003. As predicted by the simulations, the estimates had non-infinite upper confidence limits, which is a major achievement for the application of the method to a naturally-occurring, short generation, highly fecund invertebrate species. The genetic estimate of the number of successful spawners was around 1000 individuals in two consecutive years. This contrasts with about 500,000 prawns participating in spawning. It is not possible to distinguish successful from non-successful spawners so we suggest a high level of protection for the entire spawning population. We interpret the difference in numbers between successful and non-successful spawners as a large variation in the number of offspring per family that survive – a large number of families have no surviving offspring, while a few have a large number. We explored various ways in which Ne can be useful in fisheries management. It can be a surrogate for spawning population size, assuming the ratio between Ne and spawning population size has been previously calculated for that species. Alternatively, it can be a surrogate for recruitment, again assuming that the ratio between Ne and recruitment has been previously determined. The number of species that can be analysed in this way, however, is likely to be small because of species-specific life history requirements that need to be satisfied for accuracy. The most universal approach would be to integrate Ne with spawning stock-recruitment models, so that these models are more accurate when applied to fisheries populations. A pathway to achieve this was established in this project, which we predict will significantly improve fisheries sustainability in the future. Regardless of the success of integrating Ne into spawning stock-recruitment models, Ne could be used as a fisheries monitoring tool. Declines in spawning stock size or increases in natural or harvest mortality would be reflected by a decline in Ne. This would be good for data-poor fisheries and provides fishery independent information, however, we suggest a species-by-species approach. Some species may be too numerous or experiencing too much migration for the method to work. During the project two important theoretical studies of the simultaneous estimation of effective population size and migration were published (Vitalis and Couvet 2001b; Wang and Whitlock 2003). These methods, combined with collection of preliminary genetic data from the tiger prawn population in southern Gulf of Carpentaria population and a computer simulation study that evaluated the effect of differing reproductive strategies on genetic estimates, suggest that this technology could make an important contribution to the stock assessment process in the northern prawn fishery (NPF). Advances in the genomics world are rapid and already a cheaper, more reliable substitute for microsatellite loci in this technology is available. Digital data from single nucleotide polymorphisms (SNPs) are likely to super cede ‘analogue’ microsatellite data, making it cheaper and easier to apply the method to species with large population sizes.

DNA bar-coding reveals source and patterns of Thaumastocoris peregrinus invasions in South Africa and South America.

Thaumastocoris peregrinus is a recently introduced invertebrate pest of non-native Eucalyptus plantations in the Southern Hemisphere. It was first reported from South Africa in 2003 and in Argentina in 2005. Since then, populations have grown explosively and it has attained an almost ubiquitous distribution over several regions in South Africa on 26 Eucalyptus species. Here we address three key questions regarding this invasion, namely whether only one species has been introduced, whether there were single or multiple introductions into South Africa and South America and what the source of the introduction might have been. To answer these questions, bar-coding using mitochondrial DNA (COI) sequence diversity was used to characterise the populations of this insect from Australia, Argentina, Brazil, South Africa and Uruguay. Analyses revealed three cryptic species in Australia, of which only T. peregrinus is represented in South Africa and South America. Thaumastocoris peregrinus populations contained eight haplotypes, with a pairwise nucleotide distance of 0.2-0.9% from seventeen locations in Australia. Three of these haplotypes are shared with populations in South America and South Africa, but the latter regions do not share haplotypes. These data, together with the current distribution of the haplotypes and the known direction of original spread in these regions, suggest that at least three distinct introductions of the insect occurred in South Africa and South America before 2005. The two most common haplotypes in Sydney, one of which was also found in Brisbane, are shared with the non-native regions. Sydney populations of T. peregrinus, which have regularly reached outbreak levels in recent years, might thus have served as source of these three distinct introductions into other regions of the Southern Hemisphere.

Current and emerging screening methods to identify post-head-emergence frost adaptation in wheat and barley

Cereal crops can suffer substantial damage if frosts occur at heading. Identification of post-head-emergence frost (PHEF) resistance in cereals poses a number of unique and difficult challenges. Many decades of research have failed to identify genotypes with PHEF resistance that could offer economically significant benefit to growers. Research and breeding gains have been limited by the available screening systems. Using traditional frost screening systems, genotypes that escape frost injury in trials due to spatial temperature differences and/or small differences in phenology can be misidentified as resistant. We believe that by improving techniques to minimize frost escapes, such ofalse-positive' results can be confidently identified and eliminated. Artificial freezing chambers or manipulated natural frost treatments offer many potential advantages but are not yet at the stage where they can be reliably used for frost screening in breeding programmes. Here we describe the development of a novel photoperiod gradient method (PGM) that facilitates screening of genotypes of different phenology under natural field frosts at matched developmental stages. By identifying frost escapes and increasing the efficiency of field screening, the PGM ensures that research effort can be focused on finding genotypes with improved PHEF resistance. To maximize the likelihood of identifying PHEF resistance, we propose that the PGM form part of an integrated strategy to (i) source germplasm;(ii) facilitate high throughput screening; and (iii) permit detailed validation. PGM may also be useful in other studies where either a range of developmental stages and/or synchronized development are desired.

Methane in Australian agriculture: current emissions, sources and sinks, and potential mitigation strategies

Methane is a potent greenhouse gas with a global warming potential ∼28 times that of carbon dioxide. Consequently, sources and sinks that influence the concentration of methane in the atmosphere are of great interest. In Australia, agriculture is the primary source of anthropogenic methane emissions (60.4% of national emissions, or 3260kt-1methaneyear-1, between 1990 and 2011), and cropping and grazing soils represent Australia's largest potential terrestrial methane sink. As of 2011, the expansion of agricultural soils, which are ∼70% less efficient at consuming methane than undisturbed soils, to 59% of Australia's land mass (456Mha) and increasing livestock densities in northern Australia suggest negative implications for national methane flux. Plant biomass burning does not appear to have long-term negative effects on methane flux unless soils are converted for agricultural purposes. Rice cultivation contributes marginally to national methane emissions and this fluctuates depending on water availability. Significant available research into biological, geochemical and agronomic factors has been pertinent for developing effective methane mitigation strategies. We discuss methane-flux feedback mechanisms in relation to climate change drivers such as temperature, atmospheric carbon dioxide and methane concentrations, precipitation and extreme weather events. Future research should focus on quantifying the role of Australian cropping and grazing soils as methane sinks in the national methane budget, linking biodiversity and activity of methane-cycling microbes to environmental factors, and quantifying how a combination of climate change drivers will affect total methane flux in these systems.

An overview of land application of pig effluent-P using soil P chemistry and mass balance calculations.

Attention is directed at land application of piggery effluent (containing urine, faeces, water, and wasted feed) as a potential source of water resource contamination with phosphorus (P). This paper summarises P-related properties of soil from 0-0.05 m depth at 11 piggery effluent application sites, in order to explore the impact that effluent application has had on the potential for run-off transport of P. The sites investigated were situated on Alfisol, Mollisol, Vertisol, and Spodosol soils in areas that received effluent for 1.5-30 years (estimated effluent-P applications of 100-310000 kg P/ha in total). Total (PT), bicarbonate extractable (PB), and soluble P forms were determined for the soil (0-0.05 m) at paired effluent and no-effluent sites, as well as texture, oxalate-extractable Fe and Al, organic carbon, and pH. All forms of soil P at 0-0.05 m depth increased with effluent application (PB at effluent sites was 1.7-15 times that at no-effluent sites) at 10 of the 11 sites. Increases in PB were strongly related to net P applications (regression analysis of log values for 7 sites with complete data sets: 82.6 % of variance accounted for, p <0.01). Effluent irrigation tended to increase the proportion of soil PT in dilute CaCl2-extractable forms (PTC: effluent average 2.0 %; no-effluent average 0.6%). The proportion of PTC in non-molybdate reactive forms (centrifuged supernatant) decreased (no-effluent average, 46.4 %; effluent average, 13.7 %). Anaerobic lagoon effluent did not reliably acidify soil, since no consistent relationship was observed for pH with effluent application. Soil organic carbon was increased in most of the effluent areas relative to the no-effluent areas. The four effluent areas where organic carbon was reduced had undergone intensive cultivation and cropping. Current effluent management at many of the piggeries failed to maximise the potential for waste P recapture. Ten of the case-study effluent application areas have received effluent-P in excess of crop uptake. While this may not represent a significant risk of leaching where sorption retains P, it has increased the risk of transport of P by run-off. Where such sites are close to surface water, run-off P loads should be managed.

Piggery pond sludge as a nitrogen source for crops. 1. Mineral N supply estimated from laboratory incubations and field application of stockpiled and wet sludge

Piggery pond sludge (PPS) was applied, as-collected (Wet PPS) and following stockpiling for 12 months (Stockpiled PPS), to a sandy Sodosol and clay Vertosol at sites on the Darling Downs of Queensland. Laboratory measures of N availability were carried out on unamended and PPS-amended soils to investigate their value in estimating supplementary N needs of crops in Australia's northern grains region. Cumulative net N mineralised from the long-term (30 weeks) leached aerobic incubation was described by a first-order single exponential model. The mineralisation rate constant (0.057/week) was not significantly different between Control and PPS treatments or across soil types, when the amounts of initial mineral N applied in PPS treatments were excluded. Potentially mineralisable N (No) was significantly increased by the application of Wet PPS, and increased with increasing rate of application. Application of Wet PPS significantly increased the total amount of inorganic N leached compared with the Control treatments. Mineral N applied in Wet PPS contributed as much to the total mineral N status of the soil as did that which mineralised over time from organic N. Rates of C02 evolution during 30 weeks of aerobic leached incubation indicated that the Stockpiled PPS was more stabilised (19-28% of applied organic C mineralised) than the WetPPS (35-58% of applied organic C mineralised), due to higher lignin content in the former. Net nitrate-N produced following 12 weeks of aerobic non-leached incubation was highly correlated with net nitrate-N leached during 12 weeks of aerobic incubation (R^2 = 0.96), although it was <60% of the latter in both sandy and clayey soils. Anaerobically mineralisable N determined by waterlogged incubation of laboratory PPS-amended soil samples increased with increasing application rate of Wet PPS. Anaerobically minemlisable N from field-moist soil was well correlated with net N mineralised during 30 weeks of aerobic leached incubation (R^2 =0.90 sandy soil; R^2=0.93 clay soil). In the clay soil, the amount of mineral N produced from all the laboratory incubations was significantly correlated with field-measured nitrate-N in the soil profile (0-1.5 m depth) after 9 months of weed-free fallow following PPS application. In contrast, only anaerobic mineralisable N was significantly correlated with field nitrate-N in the sandy soil. Anaerobic incubation would, therefore, be suitable as a rapid practical test to estimate potentially mineralisable N following applications of different PPS materials in the field.

The genetic effective and adult census size of an Australian population of tiger prawns (Penaeus esculentus)

This study compares estimates of the census size of the spawning population with genetic estimates of effective current and long-term population size for an abundant and commercially important marine invertebrate, the brown tiger prawn (Penaeus esculentus). Our aim was to focus on the relationship between genetic effective and census size that may provide a source of information for viability analyses of naturally occurring populations. Samples were taken in 2001, 2002 and 2003 from a population on the east coast of Australia and temporal allelic variation was measured at eight polymorphic microsatellite loci. Moments-based and maximum-likelihood estimates of current genetic effective population size ranged from 797 to 1304. The mean long-term genetic effective population size was 9968. Although small for a large population, the effective population size estimates were above the threshold where genetic diversity is lost at neutral alleles through drift or inbreeding. Simulation studies correctly predicted that under these experimental conditions the genetic estimates would have non-infinite upper confidence limits and revealed they might be overestimates of the true size. We also show that estimates of mortality and variance in family size may be derived from data on average fecundity, current genetic effective and census spawning population size, assuming effective population size is equivalent to the number of breeders. This work confirms that it is feasible to obtain accurate estimates of current genetic effective population size for abundant Type III species using existing genetic marker technology.

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.

Achievements in forest tree improvement in Australia and New Zealand 6: Genetic improvement and conservation of Araucaria cunninghamii in Queensland

Araucaria cunninghamii (hoop pine) typically occurs as an emergent tree over subtropical and tropical rainforests, in a discontinuous distribution that extends from West Irian Jaya at about 0°30'S, through the highlands of Indonesian New Guinea and Papua New Guinea, along the east coast of Australia from 11°39'S in Queensland to 30°35'S in northern New South Wales. Plantations established in Queensland since the 1920s now total about 44000 ha, and constitute the primary source for the continuing supply of hoop pine quality timber and pulpwood, with a sustainable harvest exceeding 440 000 m3 y-1. Establishment of these managed plantations allowed logging of all native forests of Araucaria species (hoop pine and bunya pine, A. bidwillii) on state-owned lands to cease in the late 1980s, and the preservation of large areas of araucarian forest types within a system of state-owned and managed reserves. The successful plantation program with this species has been strongly supported by genetic improvement activities since the late 1940s - through knowledge of provenance variation and reproductive biology, the provision of reliable sources of improved seed, and the capture of substantial genetic gains in traits of economic importance (for example growth, stem straightness, internode length and spiral grain). As such, hoop pine is one of the few tropical tree species that, for more than half a century, has been the subject of continuous genetic improvement. The history of commercialisation and genetic improvement of hoop pine provides an excellent example of the dual economic and conservation benefits that may be obtained in tropical tree species through the integration of gene conservation and genetic improvement with commercial plantation development. This paper outlines the natural distribution and reproductive biology of hoop pine, describes the major achievements of the genetic improvement program in Queensland over the past 50+ y, summarises current understanding of the genetic variation and control of key selection traits, and outlines the means by which genetic diversity in the species is being conserved.

Leptospira weilii serovar Topaz, a new member of the Tarassovi serogroup isolated from a bovine source in Queensland, Australia

This paper reports on a Leptospira isolate of bovine origin and its identification as belonging to a previously unknown serovar, for which the name Topaz is proposed. The isolate (94-79970/3) was cultured from bovine urine from a north Queensland dairy farm in Australia. Strain 94-79970/3 grew at 30 °C in Ellinghausen McCullough Johnson Harris (EMJH) medium but failed to grow at 13 °C in EMJH medium or in the presence of 8-azaguanine. Serologically, strain 94-79970/3 produced titres against the Leptospira borgpetersenii serovar Tarassovi, the reference strain for the Tarassovi serogroup; however, no significant titres to any other serovars within the serogroup were obtained. Using 16S rRNA and DNA gyrase subunit B gene analysis, strain 94-79970/3 was identified as a member of the species Leptospira weilii. We propose that the serovar be named Topaz, after the location where the original isolate was obtained. The reference strain for this serovar is 94-79970/ 3 (=KIT 94-79970/35LT722).

Odour sampling 1: Physical chemistry considerations

The selection of an odour sampling device may influence the composition of the resulting odour sample. Limited comparison of emission rates derived from turbulent and essentially quiescent sampling devices confirms that the emission rates derived from these devices are quite different. There is therefore compelling evidence that current odour sampling practice should have greater regard for fundamental physical and chemical principles, the nature of the odour source and the conditions created by the sampling device. Such consideration may identify the most appropriate situations under which the use of these devices may or may not be correct.

Predicting the response of wheat (Triticum aestivum L.) to liquid and granular phosphorus fertilisers in Australian soils

Liquid forms of phosphorus (P) have been shown to be more effective than granular P for promoting cereal growth in alkaline soils with high levels of free calcium carbonate on Eyre Peninsula, South Australia. However, the advantage of liquid over granular P forms of fertiliser has not been fully investigated across the wide range of soils used for grain production in Australia. A glasshouse pot experiment tested if liquid P fertilisers were more effective for growing spring wheat (Triticum aestivum L.) than granular P (monoammonium phosphate) in 28 soils from all over Australia with soil pH (H2O) ranging from 5.2 to 8.9. Application of liquid P resulted in greater shoot biomass, as measured after 4 weeks' growth (mid to late tillering, Feeks growth stage 2-3), than granular P in 3 of the acidic to neutral soils and in 3 alkaline soils. Shoot dry matter responses of spring wheat to applied liquid or granular P were related to soil properties to determine if any of the properties predicted superior yield responses to liquid P. The calcium carbonate content of soil was the only soil property that significantly contributed to predicting when liquid P was more effective than granular P. Five soil P test procedures (Bray, Colwell, resin, isotopically exchangeable P, and diffusive gradients in thin films (DGT)) were assessed to determine their ability to measure soil test P on subsamples of soil collected before the experiment started. These soil test values were then related to the dry matter shoot yields to assess their ability to predict wheat yield responses to P applied as liquid or granular P. All 5 soil test procedures provided a reasonable prediction of dry matter responses to applied P as either liquid or granular P, with the resin P test having a slightly greater predictive capacity on the range of soils tested. The findings of this investigation suggest that liquid P fertilisers do have some potential applications in non-calcareous soils and confirm current recommendations for use of liquid P fertiliser to grow cereal crops in highly calcareous soils. Soil P testing procedures require local calibration for response to the P source that is going to be used to amend P deficiency.

Worldwide phylogeography of the globally invasive plant: Jatropha gossypiifolia

Dhileepan, Raghu and colleagues recently published their paper 'Worldwide phylogeography of the globally invasive plant: Jatropha gossypiifolia' in Proceedings of the 16th Australian Weeds Conference. They used chloroplast microsatellites to establish patterns of phylogeographic structure in the native and introduced range of Jatropha gossypiifolia, and to determine the origin(s) of introductions and the level of genetic diversity present in native and introduced populations. J. gossypiifolia exhibited limited phylogeographic structure in its native range which was best explained by contemporary movement associated with the ornamental plant trade. Multiple introductions from diverse source locations and no reduction in genetic diversity was found in the introduced range which includes Australia, Africa and Asia. These results have implications for our current biocontrol project.

Weed management in wide-row cropping systems: a review of current practices and risks for Australian farming systems.

Growing agricultural crops in wide row spacings has been widely adopted to conserve water, to control pests and diseases, and to minimise problems associated with sowing into stubble. The development of herbicide resistance combined with the advent of precision agriculture has resulted in a further reason for wide row spacings to be adopted: weed control. Increased row spacing enables two different methods of weed control to be implemented with non-selective chemical and physical control methods utilised in the wide inter-row zone, with or without selective chemicals used on the on-row only. However, continual application of herbicides and tillage on the inter-row zone brings risks of herbicide resistance, species shifts and/or changes in species dominance, crop damage, increased costs, yield losses, and more expensive weed management technology.

Susceptibility of source plants to sugarcane Fiji disease virus influences the acquisition and transmission of the virus by the planthopper vector Perkinsiella saccharicida

Fiji leaf gall (FLG) caused by Sugarcane Fiji disease virus (SCFDV) is transmitted by the planthopper Perkinsiella saccharicida. FLG is managed through the identification and exploitation of plant resistance. The glasshouse-based resistance screening produced inconsistent transmission results and the factors responsible for that are not known. A series of glasshouse trials conducted over a 2-year period was compared to identify the factors responsible for the erratic transmission results. SCFDV transmission was greater when the virus was acquired by the vector from a cultivar that was susceptible to the virus than when the virus was acquired from a resistant cultivar. Virus acquisition by the vector was also greater when the vector was exposed to the susceptible cultivars than when exposed to the resistant cultivar. Results suggest that the variation in transmission levels is due to variation in susceptibility of sugarcane cultivars to SCFDV used for virus acquisition by the vector.