Transcriptome Analysis of Capsicum Chlorosis Virus-Induced Hypersensitive Resistance Response in Bell Capsicum

Background Capsicum chlorosis virus (CaCV) is an emerging pathogen of capsicum, tomato and peanut crops in Australia and South-East Asia. Commercial capsicum cultivars with CaCV resistance are not yet available, but CaCV resistance identified in Capsicum chinense is being introgressed into commercial Bell capsicum. However, our knowledge of the molecular mechanisms leading to the resistance response to CaCV infection is limited. Therefore, transcriptome and expression profiling data provide an important resource to better understand CaCV resistance mechanisms. Methodology/Principal Findings We assembled capsicum transcriptomes and analysed gene expression using Illumina HiSeq platform combined with a tag-based digital gene expression system. Total RNA extracted from CaCV/mock inoculated CaCV resistant (R) and susceptible (S) capsicum at the time point when R line showed a strong hypersensitive response to CaCV infection was used in transcriptome assembly. Gene expression profiles of R and S capsicum in CaCV- and buffer-inoculated conditions were compared. None of the genes were differentially expressed (DE) between R and S cultivars when mock-inoculated, while 2484 genes were DE when inoculated with CaCV. Functional classification revealed that the most highly up-regulated DE genes in R capsicum included pathogenesis-related genes, cell death-associated genes, genes associated with hormone-mediated signalling pathways and genes encoding enzymes involved in synthesis of defense-related secondary metabolites. We selected 15 genes to confirm DE expression levels by real-time quantitative PCR. Conclusion/Significance DE transcript profiling data provided comprehensive gene expression information to gain an understanding of the underlying CaCV resistance mechanisms. Further, we identified candidate CaCV resistance genes in the CaCV-resistant C. annuum x C. chinense breeding line. This knowledge will be useful in future for fine mapping of the CaCV resistance locus and potential genetic engineering of resistance into CaCV-susceptible crops.

Establishment of an Australian mild onion industry - the sensory component

In recent years, there has been increasing interest from growers, merchants, supermarkets and consumers in the establishment of a national mild onion industry. Imperative to the success of the emergent industry is the application of the National Mild Onion Certification Scheme that will establish standards and recommendations to be met by growers to allow them to declare their product as certified mild onions. The use of sensory evaluation techniques has played an imperative role throughout the project timeline that has also included varietal evaluation, evaluation of current agronomic practices and correlation of chemical analysis data. Raw onion consumer acceptance testing on five different onion varieties established preferences amongst the varieties for odour, appearance, flavour, texture and overall and differences in the level of pungency and aftertaste perceived. Demographic information was obtained regarding raw and cooked onion use, frequency of consumption and responses to the idea of a mild, less pungent onion. Additionally, focus groups were conducted to further investigate consumer attitudes to onions. Currently, a trained onion panel is being established to evaluate several odour, flavour and aftertaste attributes. Sample assessments will be conducted in January 2004 and correlated with chemical analyses that will hopefully provide the corner-stone for the anticipated Certification Scheme.

Getting value out of your sensory testing - the difference from control test

Non-parametric difference tests such as triangle and duo-trio tests traditionally are used to establish differences or similarities between products. However they only supply the researcher with partial answers and often further testing is required to establish the nature, size and direction of differences. This paper looks at the advantages of the difference from control (DFC) test (also known as degree of difference test) and discusses appropriate applications of the test. The scope and principle of the test, panel composition and analysis of results are presented with the aid of suitable examples. Two of the major uses of the DFC test are in quality control and shelf-life testing. The role DFC takes in these areas and the use of other tests to complement the testing is discussed. Controls or standards are important in both these areas and the use of standard products, mental and written standards and blind controls are highlighted. The DFC test has applications in products where the duo-trio and triangle tests cannot be used because of the normal heterogeneity of the product. While the DFC test is a simple difference test it can be structured to give the researcher more valuable data and scope to make informed decisions about their product.

Climate information needs of Gascoyne-Murchison pastoralists: a representative study of the Western Australian grazing industry

The Gascoyne-Murchison region of Western Australia experiences an arid to semi-arid climate with a highly variable temporal and spatial rainfall distribution. The region has around 39.2 million hectares available for pastoral lease and supports predominantly catle and sheep grazing leases. In recent years a number of climate forecasting systems have been available offering rainfall probabilities with different lead times and a forecast period; however, the extent to which these systems are capable of fulfilling the requirements of the local pastoralists is still ambiguous. Issues can range from ensuring forecasts are issued with sufficient lead time to enable key planning or decisions to be revoked or altered, to ensuring forecast language is simple and clear, to negate possible misunderstandings in interpretation. A climate research project sought to provide an objective method to determine which available forecasting systems had the greatest forecasting skill at times of the year relevant to local property management. To aid this climate research project, the study reported here was undertaken with an overall objective of exploring local pastoralists' climate information needs. We also explored how well they understand common climate forecast terms such as 'mean', median' and 'probability', and how they interpret and apply forecast information to decisions. A stratified, proportional random sampling was used for the purpose of deriving the representative sample based on rainfall-enterprise combinations. In order to provide more time for decision-making than existing operational forecasts that are issued with zero lead time, pastoralists requested that forecasts be issued for May-July and January-March with lead times counting down from 4 to 0 months. We found forecasts of between 20 and 50 mm break-of-season or follow-up rainfall were likely to influence decisions. Eighty percent of pastoralists demonstrated in a test question that they had a poor technical understanding of how to interpret the standard wording of a probabilistic median rainfall forecast. this is worthy of further research to investigate whether inappropriate management decisions are being made because the forecasts are being misunderstood. We found more than half the respondents regularly access and use weather and climate forecasts or outlook information from a range of sources and almost three-quarters considered climate information or tools useful, with preferred methods for accessing this information by email, faxback service, internet and the Department of Agriculture Western Australia's Pastoral Memo. Despite differences in enterprise types and rainfall seasonality across the region we found seasonal climate forecasting needs were relatively consistent. It became clear that providing basic training and working with pastoralists to help them understand regional climatic drivers, climate terminology and jargon, and the best ways to apply the forecasts to enhance decision-making are important to improve their use of information. Consideration could also be given to engaging a range of producers to write the climate forecasts themselves in the language they use and understand, in consultation with the scientists who prepare the forecasts.

Formative evaluation to benchmark and improve climate-based decision support for graziers in Western Queelsland

Researchers developing climate-based forecasts, workshops, software tools and information to aid grazier decisions undertook an evaluation study to enhance planning and benchmark impact. One hundred graziers in Western Queensland were randomly selected from 7 shires and surveyed by mail and telephone (43 respondents) to explore levels of knowledge and use of climate information, practices and information needs. We found 36% of respondents apply the Southern Oscillation Index to property decisions but 92% were unaware El Niño Southern Oscillation’s predictive signal in the region is greater for pasture growth than rainfall, suggesting they may not recognise the potential of pasture growth forecasts. Almost 75% of graziers consider they are conservative or risk averse in their attitude to managing their enterprise. Mail respondents (n= 20) if given a 68%, on average, probability of exceeding median rainfall forecast may change a decision; almost two-thirds vary stocking rate based on forage available, last year’s pasture growth or the Southern Oscillation Index; the balance maintain a constant stocking rate strategy; 90% have access to a computer; 75% to the internet and 95% have a fax. This paper presents findings of the study and draws comparisons with a similar study of 174 irrigators in the Northern Murray-Darling Basin (Aust. J. Exp. Ag. 44, 247-257). New insights and information gained are helping the team better understand client needs and plan, design and extend tools and information tailored to grazier knowledge, practice, information needs and preferences. Results have also provided a benchmark against which to measure project impact and have influenced the team to make important changes to their project planning, activities and methods for transferring technology tailored to grazier preferences.

Limitations of soil microbial biomass carbon as an indicator of soil pollution in the field

The size of the soil microbial biomass carbon (SMBC) has been proposed as a sensitive indicator for measuring the adverse effects of contaminants on the soil microbial community. In this study of Australian agricultural systems, we demonstrated that field variability of SMBC measured using the fumigation-extraction procedure limited its use as a robust ecotoxicological endpoint. The SMBC varied up to 4-fold across control samples collected from a single field site, due to small-scale spatial heterogeneity in the soil physicochemical environment. Power analysis revealed that large numbers of replicates (3-93) were required to identify 20% or 50% decreases in the size of the SMBC of contaminated soil samples relative to their uncontaminated control samples at the 0.05% level of statistical significance. We question the value of the routine measurement of SMBC as an ecotoxicological endpoint at the field scale, and suggest more robust and predictive microbiological indicators.

Soil sorption-desorption of phosphorus from piggery effluent compared with inorganic sources

The leaching of phosphorus (P) within soils can be a limiting consideration for the sustainable operation of intensive livestock enterprises. Sorption curves are widely used to assist estimation of P retention, though the effect of effluent constituents on their accuracy is not well understood. We conducted a series of P-sorption-desorption batch experiments with an Oxic Haplustalf (soil 1), Haplusterts (soils 2 and 3), and a Natrustalf (soil 4). Phosphorus sources included effluent, orthophosphate-P in a matrix replicating the effluent's salt constituents (the reference solution), and an orthophosphate-P solution. Treated soils were incubated for up to 193 days before sequential desorption extraction. Effluent constituents, probably the organic or particulate components, temporarily increased the vulnerability of sorbed-P to desorption. The increase in vulnerability was removed by 2-113 days of incubation (25 degrees C). Despite vigorous extraction for 20 consecutive days, some P sorbed as part of the treatments of soils 1 and 2 was not desorbed. The increased vulnerability due to effluent constituents lasted a maximum of about one cropping season and, for all other treatments, adsorption curves overestimated vulnerability to desorption. Therefore, adsorption curves provide a conservative estimate of vulnerability to desorption where effluent is used in continued crop production in these soils.

The environmental fate of diuron under a conventional production regime in a sugarcane farm during the plant cane phase

BACKGROUND: Field studies of diuron and its metabolites 3-(3,4-dichlorophenyl)-1-methylurea (DCPMU), 3,4-dichlorophenylurea (DCPU) and 3,4-dichloroaniline (DCA) were conducted in a farm soil and in stream sediments in coastal Queensland, Australia. RESULTS: During a 38 week period after a 1.6 kg ha^-1 diuron application, 70-100% of detected compounds were within 0-15 cm of the farm soil, and 3-10% reached the 30-45 cm depth. First-order t1/2 degradation averaged 49 ± 0.9 days for the 0-15, 0-30 and 0-45 cm soil depths. Farm runoff was collected in the first 13-50 min of episodes lasting 55-90 min. Average concentrations of diuron, DCPU and DCPMU in runoff were 93, 30 and 83-825 µg L^-1 respectively. Their total loading in all runoff was >0.6% of applied diuron. Diuron and DCPMU concentrations in stream sediments were between 3-22 and 4-31 µg kg^-1 soil respectively. The DCPMU/diuron sediment ratio was >1. CONCLUSION: Retention of diuron and its metabolites in farm topsoil indicated their negligible potential for groundwater contamination. Minimal amounts of diuron and DCMPU escaped in farm runoff. This may entail a significant loading into the wider environment at annual amounts of application. The concentrations and ratio of diuron and DCPMU in stream sediments indicated that they had prolonged residence times and potential for accumulation in sediments. The higher ecotoxicity of DCPMU compared with diuron and the combined presence of both compounds in stream sediments suggest that together they would have a greater impact on sensitive aquatic species than as currently apportioned by assessments that are based upon diuron alone.

Root-lesion nematodes (Pratylenchus thornei and P. neglectus): A review of recent progress in managing a significant pest of grain crops in northern Australia

Two species of root-lesion nematode (predominantly Pratylenchus thornei but also P. neglectus) are widespread pathogens of wheat and other crops in Australia's northern grain belt, a subtropical region with deep, fertile clay soils and a summer-dominant rainfall pattern. Losses in grain yield from P. thornei can be as high as 70% for intolerant wheat cultivars. This review focuses on research which has led to the development of effective integrated management programs for these nematodes. It highlights the importance of correct identification in managing Pratylenchus species, reviews the plant breeding work done in developing tolerant and resistant cultivars, outlines the methods used to screen for tolerance and resistance, and discusses how planned crop sequencing with tolerant and partially resistant wheat cultivars, together with crops such as sorghum, sunflower, millets and canaryseed, can be used to reduce nematode populations and limit crop damage. The declining levels of soil organic matter in cropped soils are also discussed with reference to their effect on soil health and biological suppression of root-lesion nematodes.

Fate of applied biosolids nitrogen in a cut and remove forage system on an alluvial clay loam soil

The fate of nitrogen (N) applied in biosolids was investigated in a forage production system on an alluvial clay loam soil in south-eastern Queensland, Australia. Biosolids were applied in October 2002 at rates of 6, 12, 36, and 54dryt/ha for aerobically digested biosolids (AE) and 8, 16, 48, and 72dryt/ha for anaerobically digested biosolids (AN). Rates were based on multiples of the Nitrogen Limited Biosolids Application rate (0.5, 1, 3, and 4.5NLBAR) for each type of biosolid. The experiment included an unfertilised control and a fertilised control that received multiple applications of synthetic fertiliser. Forage sorghum was planted 1 week after biosolids application and harvested 4 times between December 2002 and May 2003. Dry matter production was significantly greater from the biosolids-treated plots (21-27t/ha) than from the unfertilised (16t/ha) and fertilised (18t/ha) controls. The harvested plant material removed an extra 148-488kg N from the biosolids-treated plots. Partial N budgets were calculated for the 1NLBAR and 4.5NLBAR treatments for each biosolids type at the end of the crop season. Crop removal only accounted for 25-33% of the applied N in the 1NLBAR treatments and as low as 8-15% with 4.5NLBAR. Residual biosolids N was predominantly in the form of organic N (38-51% of applied biosolids N), although there was also a significant proportion (10-23%) as NO3-N, predominantly in the top 0.90m of the soil profile. From 12 to 29% of applied N was unaccounted for, and presumed to be lost as gaseous nitrogen and/or ammonia, as a consequence of volatilisation or denitrification, respectively. In-season mineralisation of organic N in biosolids was 43-59% of the applied organic N, which was much greater than the 15% (AN)-25% (AE) expected, based on current NLBAR calculation methods. Excessive biosolids application produced little additional biomass but led to high soil mineral N concentrations that were vulnerable to multiple loss pathways. Queensland Guidelines need to account for higher rates of mineralisation and losses via denitrification and volatilisation and should therefore encourage lower application rates to achieve optimal plant growth and minimise the potential for detrimental impacts on the environment.

Summative evaluation of climate application activities with pastoralists in western Queensland

Survey methods were engaged to measure the change in use and knowledge of climate information by pastoralists in western Queensland. The initial mail survey was undertaken in 2000-01 (n=43) and provided a useful benchmark of pastoralists climate knowledge. Two years of climate applications activities were completed and clients were re-surveyed in 2003 (n=49) to measure the change in knowledge and assess the effectiveness of the climate applications activities. Two methods were used to assess changes in client knowledge, viz., self-assessment and test questions. We found that the use of seasonal climate forecasts in decision making increased from 36% in 2001 (n=42) to 51% in 2003 (n=49) (P=0.07). The self-assessment technique was unsatisfactory as a measure of changing knowledge over short periods (1-3 years), but the test question technique was successful and indicated an improvement in climate knowledge among respondents. The increased levels of use of seasonal climate forecasts in management and improved knowledge was partly attributed to the climate applications activities of the project. Further, those who used seasonal forecasting (n=25) didn't understand key components of forecasts (e.g. probability, median) better than those who didn't use seasonal forecasts (n=24) (P>0.05). This identifies the potential for misunderstanding and misinterpretation of forecasts among users and highlights the need for providers of forecasts to understand the difficulties and prepare simply written descriptions of forecasts and disseminate these with the maps showing probabilities. The most preferred means of accessing climate information were internet, email, 'The Season Ahead' newsletter and newspaper. The least preferred were direct contact with extension officers and attending field days and group meetings. Eighty-six percent of respondents used the internet and 67% used ADSL broadband internet (April 2003). Despite these findings, extension officers play a key role in preparing and publishing the information on the web, in emails and newsletters. We also believe that direct contact with extension officers trained in climate applications is desirable in workshop-like events to improve knowledge of the difficult concepts underpinning climate forecasts, which may then stimulate further adoption.

Effect of changes in hook pattern and size on catch rate, hooking location, injury and bleeding for a number of tropical reef fish species

The Queensland Great Barrier Reef line fishery in Australia is regulated via a range of input and output controls including minimum size limits, daily catch limits and commercial catch quotas. As a result of these measures a substantial proportion of the catch is released or discarded. The fate of these released fish is uncertain, but hook-related mortality can potentially be decreased by using hooks that reduce the rates of injury, bleeding and deep hooking. There is also the potential to reduce the capture of non-target species though gear selectivity. A total of 1053 individual fish representing five target species and three non-target species were caught using six hook types including three hook patterns (non-offset circle, J and offset circle), each in two sizes (small 4/0 or 5/0 and large 8/0). Catch rates for each of the hook patterns and sizes varied between species with no consistent results for target or non-target species. When data for all of the fish species were aggregated there was a trend for larger hooks, J hooks and offset circle hooks to cause a greater number of injuries. Using larger hooks was more likely to result in bleeding, although this trend was not statistically significant. Larger hooks were also more likely to foul-hook fish or hook fish in the eye. There was a reduction in the rates of injuries and bleeding for both target and non-target species when using the smaller hook sizes. For a number of species included in our study the incidence of deep hooking decreased when using non-offset circle hooks, however, these results were not consistent for all species. Our results highlight the variability in hook performance across a range of tropical demersal finfish species. The most obvious conservation benefits for both target and non-target species arise from using smaller sized hooks and non-offset circle hooks. Fishers should be encouraged to use these hook configurations to reduce the potential for post-release mortality of released fish.

A field investigation of solubility and food chain accumulation of biosolid-cadmium across diverse soil types

One of the pathways for transfer of cadmium (Cd) through the food chain is addition of urban wastewater solids (biosolids) to soil, and many countries have restrictions on biosolid use to minimize crop Cd contamination. The basis of these restrictions often lies in laboratory or glasshouse experimentation of soil-plant transfer of Cd, but these studies are confounded by artefacts from growing crops in controlled laboratory conditions. This study examined soil to plant (wheat grain) transfer of Cd under a wide range of field environments under typical agronomic conditions, and compared the solubility and bioavailability of Cd in biosolids to soluble Cd salts. Solubility of biosolid Cd (measured by examining Cd partitioning between soil and soil solution) was found to be equal to or greater than that of soluble Cd salts, possibly due to competing ions added with the biosolids. Conversely, bioavailability of Cd to wheat and transfer to grain was less than that of soluble Cd salts, possibly due to addition of Zn with the biosolids, causing reduced plant uptake or grain loading, or due to complexation of soluble Cd2+ by dissolved organic matter.

Availability of soil potassium and diagnostic soil tests

Thirty-seven surface (0-0.10 or 0-0.20 m) soils covering a wide range of soil types (16 Vertosols, 6 Ferrosols, 6 Dermosols, 4 Hydrosols, 2 Kandosols, 1 Sodosol, 1 Rudosol, and 1 Chromosol) were exhaustively cropped in 2 glasshouse experiments. The test species were Panicum maximum cv. Green Panic in Experiment A and Avena sativa cv. Barcoo in Experiment B. Successive forage harvests were taken until the plants could no longer grow in most soils because of severe potassium (K) deficiency. Soil samples were taken prior to cropping and after the final harvest in both experiments, and also after the initial harvest in Experiment B. Samples were analysed for solution K, exchangeable K (Exch K), tetraphenyl borate extractable K for extraction periods of 15 min (TBK15) and 60 min (TBK60), and boiling nitric acid extractable K (Nitric K). Inter-correlations between the initial levels of the various soil K parameters indicated that the following pools were in sequential equilibrium: solution K, Exch K, fast release fixed K [estimated as (TBK15-Exch K)], and slow release fixed K [estimated as (TBK60-TBK15)]. Structural K [estimated as (Nitric K-TBK60)] was not correlated with any of the other pools. However, following exhaustive drawdown of soil K by cropping, structural K became correlated with solution K, suggesting dissolution of K minerals when solution K was low. The change in the various K pools following cropping was correlated with K uptake at Harvest 1 ( Experiment B only) and cumulative K uptake ( both experiments). The change in Exch K for 30 soils was linearly related to cumulative K uptake (r = 0.98), although on average, K uptake was 35% higher than the change in Exch K. For the remaining 7 soils, K uptake considerably exceeded the change in Exch K. However, the changes in TBK15 and TBK60 were both highly linearly correlated with K uptake across all soils (r = 0.95 and 0.98, respectively). The slopes of the regression lines were not significantly different from unity, and the y-axis intercepts were very small. These results indicate that the plant is removing K from the TBK pool. Although the change in Exch K did not consistently equate with K uptake across all soils, initial Exch K was highly correlated with K uptake (r = 0.99) if one Vertosol was omitted. Exchangeable K is therefore a satisfactory diagnostic indicator of soil K status for the current crop. However, the change in Exch K following K uptake is soil-dependent, and many soils with large amounts of TBK relative to Exch K were able to buffer changes in Exch K. These soils tended to be Vertosols occurring on floodplains. In contrast, 5 soils (a Dermosol, a Rudosol, a Kandosol, and 2 Hydrosols) with large amounts of TBK did not buffer decreases in Exch K caused by K uptake, indicating that the TBK pool in these soils was unavailable to plants under the conditions of these experiments. It is likely that K fertiliser recommendations will need to take account of whether the soil has TBK reserves, and the availability of these reserves, when deciding rates required to raise exchangeable K status to adequate levels.

Comparison of odour emission rates measured from various sources using two sampling devices

Two commonly used sampling devices (a wind tunnel and the US EPA dynamic emission chamber), were used to collect paired samples of odorous air from a number of agricultural odour sources. The odour samples were assessed using triangular, forced-choice dynamic olfactometry. The odour concentration data was combined with the flushing rate data to calculate odour emission rates for both devices on all sources. Odour concentrations were consistently higher in samples collected with a flux chamber (ratio ranging from 10:7 to 5:1, relative to wind tunnel samples), whereas odour emission rates were consistently larger when derived from wind tunnels (ratio ranging from 60:1 to 240:1, relative to flux chamber values). A complex relationship existed between emission rate estimates derived from each device, apparently influenced by the nature of the emitting surface. These results have great significance for users of odour dispersion models, for which an odour emission rate is a key input parameter.