Inconsistent Transmission of Banana Bunchy Top Virus in Micropropagated Bananas and its Implication for Germplasm Screening

Banana bunchy top virus (BBTV) was readily transmitted through tissue culture in banana (Mum sp.) cv. Lady finger (AAB) and Cavendish cv. Williams (AAA). Lines derived from infected and healthy field plants had similar in vitro multiplication rates. BBTV infected in vitro cultures displayed symptoms of stunting, leaf curling, chlorotic and green flecks, and poor root growth. Symptoms became milder with time, and were often difficult to discern in older, rapidly multiplying cultures. A triple antibody sandwich ELISA using polyclonal and monoclonal antibodies was very efficient for detecting BBTV in vitro. Symptomless, ELISA-negative plants arose in 10 out of 11 lines derived from BBTV-infected field plants and first appeared after 9 months continuous in vitro culture at a constant 28OC. Meristem tip culture or heat therapy was not used. These plants remained symptomless and ELISA-negative after planting out in the glasshouse (individual plants checked for up to 16 months). The implications of this inconsistent transmission of BBTV for germplasm indexing and exchange are discussed.

Subsoil constraints to grain production in the cropping soils of the north-eastern region of Australia: an overview

In dryland agricultural systems of the subtropical, semi-arid region of north-eastern Australia, water is the most limiting resource. Crop productivity depends on the efficient use of rainfall and available water stored in the soil during fallow. Agronomic management practices including a period of fallow, stubble retention, and reduced tillage enhance reserves of soil water. However, access to stored water in these soils may be restricted by the presence of growth-limiting conditions in the rooting zone of the crop. These have been termed as subsoil constraints. Subsoil constraints may include compacted or gravel layers (physical), sodicity, salinity, acidity, nutrient deficiencies, presence of toxic elements (chemical) and low microbial activity (biological). Several of these constraints may occur together in some soils. Farmers have often not been able to obtain the potential yield determined by their prevailing climatic conditions in the marginal rainfall areas of the northern grains region. In the past, the adoption of soil management practices had been largely restricted to the top 100 mm soil layer. Exploitation of the subsoil as a source of water and nutrients has largely been overlooked. The key towards realising potential yields would be to gain better understanding of subsoils and their limitations, then develop options to manage them practically and economically. Due to the complex nature of the causal factors of these constraints, efforts are required for a combination of management approaches rather than individual options, with the aim to combat these constraints for sustainable crop production, managing natural resources and avoiding environmental damage.

Subsoil constraints in Vertosols: Crop water use nutrient concentration, and grain yields of bread wheat, durum wheat, barley, chickpea and canola

Single or multiple factors implicated in subsoil constraints including salinity, sodicity, and phytotoxic concentrations of chloride (Cl) are present in many Vertosols including those occurring in Queensland, Australia. The variable distribution and the complex interactions that exist between these constraints limit the agronomic or management options available to manage the soil with these subsoil constraints. The identification of crops and cultivars adapted to these adverse subsoil conditions and/or able to exploit subsoil water may be an option to maintain productivity of these soils. We evaluated relative performance of 5 winter crop species, in terms of grain yields, nutrient concentration, and ability to extract soil water, grown on soils with various levels and combinations of subsoil constraints in 19 field experiments over 2 years. Subsoil constraints were measured by levels of soil Cl, electrical conductivity of the saturation extract (ECse), and exchangeable sodium percentage (ESP). Increasing levels of subsoil constraints significantly decreased maximum depth of water extraction, grain yield, and plant-available water capacity for all the 5 crops and more so for chickpea and durum wheat than bread wheat, barley, or canola. Increasing soil Cl levels had a greater restricting effect on water availability than did ECse and ESP. We developed empirical relationships between soil Cl, ECse, and ESP and crop lower limit (CLL) for estimating subsoil water extraction by 5 winter crops. However, the presence of gypsum influenced the ability to predict CLL based on the levels of ECse. Stronger relationships between apparent unused plant-available water (CLL - LL15; LL15 is lower limit at -1.5 MPa) and soil Cl concentrations than ESP or ECse suggested that the presence of high Cl in these soils most likely inhibited the subsoil water extraction by the crops. This was supported by increased sodium (Na) and Cl concentration with a corresponding decrease in calcium (Ca) and potassium (K) in young mature leaf of bread wheat, durum wheat, and chickpea with increasing levels of subsoil constraints. Of the 2 ions, Na and Cl, the latter appears to be more damaging than the former, resulting in plant dieback and reduced grain yields.

Impact of subsoil constraints on wheat yield and gross margin on fine-textured soils of the southern Victorian Mallee

The APSIM-Wheat module was used to investigate our present capacity to simulate wheat yields in a semi-arid region of eastern Australia (the Victorian Mallee), where hostile subsoils associated with salinity, sodicity, and boron toxicity are known to limit grain yield. In this study we tested whether the effects of subsoil constraints on wheat growth and production could be modelled with APSIM-Wheat by assuming that either: (a) root exploration within a particular soil layer was reduced by the presence of toxic concentrations of salts, or (b) soil water uptake from a particular soil layer was reduced by high concentration of salts through osmotic effects. After evaluating the improved predictive capacity of the model we applied it to study the interactions between subsoil constraints and seasonal conditions, and to estimate the economic effect that subsoil constraints have on wheat farming in the Victorian Mallee under different climatic scenarios. Although the soils had high levels of salinity, sodicity, and boron, the observed variability in root abundance at different soil layers was mainly related to soil salinity. We concluded that: (i) whether the effect of subsoil limitations on growth and yield of wheat in the Victorian Mallee is driven by toxic, osmotic, or both effects acting simultaneously still requires further research, (ii) at present, the performance of APSIM-Wheat in the region can be improved either by assuming increased values of lower limit for soil water extraction, or by modifying the pattern of root exploration in the soil pro. le, both as a function of soil salinity. The effect of subsoil constraints on wheat yield and gross margin can be expected to be higher during drier than wetter seasons. In this region the interaction between climate and soil properties makes rainfall information alone, of little use for risk management and farm planning when not integrated with cropping systems models.

Quantifying Transmission of Campylobacter jejuni in Commercial Broiler Flocks

Since meat from poultry colonized with Campylobacter spp. is a major cause of bacterial gastroenteritis, human exposure should be reduced by, among other things, prevention of colonization of broiler flocks. To obtain more insight into possible sources of introduction of Campylobacter into broiler flocks, it is essential to estimate the moment that the first bird in a flock is colonized. If the rate of transmission within a flock were known, such an estimate could be determined from the change in the prevalence of colonized birds in a flock over time. The aim of this study was to determine the rate of transmission of Campylobacter using field data gathered for 5 years for Australian broiler flocks. We used unique sampling data for 42 Campylobacter jejuni-colonized flocks and estimated the transmission rate, which is defined as the number of secondary infections caused by one colonized bird per day. The estimate was 2.37 +/- 0.295 infections per infectious bird per day, which implies that in our study population colonized flocks consisting of 20,000 broilers would have an increase in within-flock prevalence to 95% within 4.4 to 7.2 days after colonization of the first broiler. Using Bayesian analysis, the moment of colonization of the first bird in a flock was estimated to be from 21 days of age onward in all flocks in the study. This study provides an important quantitative estimate of the rate of transmission of Campylobacter in broiler flocks, which could be helpful in future studies on the epidemiology of Campylobacter in the field.

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.

Transmission of Japanese Encephalitis Virus from the Black Flying Fox, Pteropus alecto, to Culex annulirostris Mosquitoes, Despite the Absence of Detectable Viremia.

To determine the potential role of flying foxes in transmission cycles of Japanese encephalitis virus (JEV) in Australia, we exposed Pteropus alecto (Megachiroptera: Pteropididae) to JEV via infected Culex annulirostris mosquitoes or inoculation. No flying foxes developed symptoms consistent with JEV infection. Anti-JEV IgG antibodies developed in 6/10 flying foxes exposed to infected Cx. annulirostris and in 5/5 inoculated flying foxes. Low-level viremia was detected by real-time reverse transcriptase polymerase chain reaction in 1/5 inoculated flying foxes and this animal was able to infect recipient mosquitoes. Although viremia was not detected in any of the 10 flying foxes that were exposed to JEV by mosquito bite, two animals infected recipient mosquitoes. Likewise, an inoculated flying fox without detectable viremia infected recipient mosquitoes. Although infection rates in recipient mosquitoes were low, the high population densities in roosting camps, coupled with migratory behavior indicate that flying foxes could play a role in the dispersal of JEV.

Poor adoption of ley-pastures in south-west Queensland: biophysical, economic and social constraints.

The present review identifies various constraints relating to poor adoption of ley-pastures in south-west Queensland, and suggests changes in research, development and extension efforts for improved adoption. The constraints include biophysical, economic and social constraints. In terms of biophysical constraints, first, shallower soil profiles with subsoil constraints (salt and sodicity), unpredictable rainfall, drier conditions with higher soil temperature and evaporative demand in summer, and frost and subzero temperature in winter, frequently result in a failure of established, or establishing, pastures. Second, there are limited options for legumes in a ley-pasture, with the legumes currently being mostly winter-active legumes such as lucerne and medics. Winter-active legumes are ineffective in improving soil conditions in a region with summer-dominant rainfall. Third, most grain growers are reluctant to include grasses in their ley-pasture mix, which can be uneconomical for various reasons, including nitrogen immobilisation, carryover of cereal diseases and depressed yields of the following cereal crops. Fourth, a severe depletion of soil water following perennial ley-pastures (grass + legumes or lucerne) can reduce the yields of subsequent crops for several seasons, and the practice of longer fallows to increase soil water storage may be uneconomical and damaging to the environment. Economic assessments of integrating medium- to long-term ley-pastures into cropping regions are generally less attractive because of reduced capital flow, increased capital investment, economic loss associated with establishment and termination phases of ley-pastures, and lost opportunities for cropping in a favourable season. Income from livestock on ley-pastures and soil productivity gains to subsequent crops in rotation may not be comparable to cropping when grain prices are high. However, the economic benefits of ley-pastures may be underestimated, because of unaccounted environmental benefits such as enhanced water use, and reduced soil erosion from summer-dominant rainfall, and therefore, this requires further investigation. In terms of social constraints, the risk of poor and unreliable establishment and persistence, uncertainties in economic and environmental benefits, the complicated process of changing from crop to ley-pastures and vice versa, and the additional labour and management requirements of livestock, present growers socially unattractive and complex decision-making processes for considering adoption of an existing medium- to long-term ley-pasture technology. It is essential that research, development and extension efforts should consider that new ley-pasture options, such as incorporation of a short-term summer forage legume, need to be less risky in establishment, productive in a region with prevailing biophysical constraints, economically viable, less complex and highly flexible in the change-over processes, and socially attractive to growers for adoption in south-west Queensland.

Nutritional constraints on displacement of Fusarium pseudograminearum from cereal straw by antagonists.

Displacement of the fungus Fusarium pseudograminearum from stubble by antagonists is a potential means of biocontrol of crown rot in cereals. The role of carbon and nitrogen nutrition in interactions between the pathogen and the antagonists Fusarium equiseti, Fusarium nygamai, Trichoderma harzianum and the non-antagonistic straw fungus Alternaria infectoria was investigated. Sole carbon source utilization patterns on Biolog plates were similar among the three Fusarium species, suggesting a possible role for competition. However, carbon niche overlap was unlikely to be important in antagonism by T. harzianum. Straw medium supplemented with sugars generally reduced the inhibitory effect of antagonists on growth of F. pseudograminearum in dual culture, indicating that availability of simple carbon sources does not limit antagonism. Adding nitrogen as urea, nitrate or ammonium to straw medium had little effect on antagonism by F. equiseti and F. nygamai, but ammonium addition removed the inhibitory effect of T. harzianum on growth of F. pseudograminearum. Displacement of F. pseudograminearum from straw by all fungi in a Petri dish assay was greater when urea or nitrate was used as a nitrogen source than with ammonium. All forms of nitrogen significantly increased displacement of F. pseudograminearum from straw under simulated field conditions when straws were either inoculated with T. harzianum or exposed to resident soil microbes. However, in 2 out of 3 experiments urea and nitrate were more effective than ammonium. The results suggest that availability of nitrogen, but not carbon, is limiting the activities of antagonists of F. pseudograminearum in straw, and the way nitrogen is applied can influence the rate of displacement and mortality of the pathogen in host residues.

Routes of Hendra Virus Excretion in Naturally-Infected Flying-Foxes: Implications for Viral Transmission and Spillover Risk

Pteropid bats or flying-foxes (Chiroptera: Pteropodidae) are the natural host of Hendra virus (HeV) which sporadically causes fatal disease in horses and humans in eastern Australia. While there is strong evidence that urine is an important infectious medium that likely drives bat to bat transmission and bat to horse transmission, there is uncertainty about the relative importance of alternative routes of excretion such as nasal and oral secretions, and faeces. Identifying the potential routes of HeV excretion in flying-foxes is important to effectively mitigate equine exposure risk at the bat-horse interface, and in determining transmission rates in host-pathogen models. The aim of this study was to identify the major routes of HeV excretion in naturally infected flying-foxes, and secondarily, to identify between-species variation in excretion prevalence. A total of 2840 flying-foxes from three of the four Australian mainland species (Pteropus alecto, P. poliocephalus and P. scapulatus) were captured and sampled at multiple roost locations in the eastern states of Queensland and New South Wales between 2012 and 2014. A range of biological samples (urine and serum, and urogenital, nasal, oral and rectal swabs) were collected from anaesthetized bats, and tested for HeV RNA using a qRT-PCR assay targeting the M gene. Forty-two P. alecto (n = 1410) had HeV RNA detected in at least one sample, and yielded a total of 78 positive samples, at an overall detection rate of 1.76% across all samples tested in this species (78/4436). The rate of detection, and the amount of viral RNA, was highest in urine samples (>serum, packed haemocytes >faecal >nasal >oral), identifying urine as the most plausible source of infection for flying-foxes and for horses. Detection in a urine sample was more efficient than detection in urogenital swabs, identifying the former as the preferred diagnostic sample. The detection of HeV RNA in serum is consistent with haematogenous spread, and with hypothesised latency and recrudesence in flying-foxes. There were no detections in P. poliocephalus (n = 1168 animals; n = 2958 samples) or P. scapulatus (n = 262 animals; n = 985 samples), suggesting (consistent with other recent studies) that these species are epidemiologically less important than P. alecto in HeV infection dynamics. The study is unprecedented in terms of the individual animal approach, the large sample size, and the use of a molecular assay to directly determine infection status. These features provide a high level of confidence in the veracity of our findings, and a sound basis from which to more precisely target equine risk mitigation strategies.

Natural host range, thrips and seed transmission of distinct Tobacco streak virus strains in Queensland, Australia

Diseases caused by Tobacco streak virus (TSV) have resulted in significant crop losses in sunflower and mung bean crops in Australia. Two genetically distinct strains from central Queensland, TSV-parthenium and TSV-crownbeard, have been previously described. They share only 81% total-genome nucleotide sequence identity and have distinct major alternative hosts, Parthenium hysterophorus (parthenium) and Verbesina encelioides (crownbeard). We developed and used strain-specific multiplex Polymerase chain reactions (PCRs) for the three RNA segments of TSV-parthenium and TSV-crownbeard to accurately characterise the strains naturally infecting 41 hosts species. Hosts included species from 11 plant families, including 12 species endemic to Australia. Results from field surveys and inoculation tests indicate that parthenium is a poor host of TSV-crownbeard. By contrast, crownbeard was both a natural host of, and experimentally infected by TSV-parthenium but this infection combination resulted in non-viable seed. These differences appear to be an effective biological barrier that largely restricts these two TSV strains to their respective major alternative hosts. TSV-crownbeard was seed transmitted from naturally infected crownbeard at a rate of between 5% and 50% and was closely associated with the geographical distribution of crownbeard in central Queensland. TSV-parthenium and TSV-crownbeard were also seed transmitted in experimentally infected ageratum (Ageratum houstonianum) at rates of up to 40% and 27%, respectively. The related subgroup 1 ilarvirus, Ageratum latent virus, was also seed transmitted at a rate of 18% in ageratum which is its major alternative host. Thrips species Frankliniella schultzei and Microcephalothrips abdominalis were commonly found in flowers of TSV-affected crops and nearby weed hosts. Both species readily transmitted TSV-parthenium and TSV-crownbeard. The results are discussed in terms of how two genetically and biologically distinct TSV strains have similar life cycle strategies in the same environment.