A convenient sample preparation protocol for scanning electron microscope examination of xylem-occluding bacterial biofilm on cut flowers and foliage

Microbes and their exopolysaccharides (EPS) can block xylem vessels, thereby increasing the hydraulic resistance and decreasing the vase life of cut flowers and foliage. Scanning electron microscopy (SEM) provides a powerful tool for investigation of bacteria-induced xylem occlusion. However, conventional preparation protocols for SEM involving chemicals can cause loss of hydrated EPS material, and thereby damage the bacterial biofilms during dehydration. A modified chemical fixation protocol involving pre-fixation with 75 mM lysine plus 2.5% glutaraldehyde followed by the normal fixation in 3% glutaraldehyde was, therefore, tested for improved preservation of bacterial biofilm at the stem-ends of cut Acacia holosericea foliage stems. Stem-end segments with different stages of bacterial growth were obtained from stems stood into water. The lysine-based protocol was compared with four other processing protocols of critical point drying (CPD) without fixation (control), freeze-drying (FD), conventional chemical fixation followed by drying with hexamethyldisilazane (HMDS), and conventional chemical fixation with CPD. The non-fixed control. FD and the glutaraldehyde fixation with HMDS drying gave poor preservation of hydrated material, including bacterial EPS. Conventional glutaraldehyde fixation followed by CPD was superior to these three methods in terms of better preserving the EPS. However, this fourth method gave condensation of biofilms during dehydration. In contrast, the modified lysine-based protocol resulted in superior preservation of EPS and biofilm structure. Thus, this fifth method was the most appropriate for examination of bacterial stem-end blockage in cut ornamentals. (C) 2012 Elsevier B.V. All rights reserved.

Interacting effects of vegetation, soils and management on the sensitivity of Australian savanna rangelands to climate change

There is an increasing need to understand what makes vegetation at some locations more sensitive to climate change than others. For savanna rangelands, this requires building knowledge of how forage production in different land types will respond to climate change, and identifying how location-specific land type characteristics, climate and land management control the magnitude and direction of its responses to change. Here, a simulation analysis is used to explore how forage production in 14 land types of the north-eastern Australian rangelands responds to three climate change scenarios of +3A degrees C, +17% rainfall; +2A degrees C, -7% rainfall; and +3A degrees C, -46% rainfall. Our results demonstrate that the controls on forage production responses are complex, with functional characteristics of land types interacting to determine the magnitude and direction of change. Forage production may increase by up to 60% or decrease by up to 90% in response to the extreme scenarios of change. The magnitude of these responses is dependent on whether forage production is water or nitrogen (N) limited, and how climate changes influence these limiting conditions. Forage production responds most to changes in temperature and moisture availability in land types that are water-limited, and shows the least amount of change when growth is restricted by N availability. The fertilisation effects of doubled atmospheric CO2 were found to offset declines in forage production under 2A degrees C warming and a 7% reduction in rainfall. However, rising tree densities and declining land condition are shown to reduce potential opportunities from increases in forage production and raise the sensitivity of pastures to climate-induced water stress. Knowledge of these interactions can be applied in engaging with stakeholders to identify adaptation options.

Cu2+ inhibition of gel secretion in the xylem and its potential implications for water uptake of cut Acacia holosericea stems

Maintaining a high rate of water uptake is crucial for maximum longevity of cut stems. Physiological gel/tylosis formation decreases water transport efficiency in the xylem. The primary mechanism of action for post-harvest Cu2+ treatments in improving cut flower and foliage longevity has been elusive. The effect of Cu2+ on wound-induced xylem vessel occlusion was investigated for Acacia holosericea A. Cunn. ex G. Don. Experiments were conducted using a Cu2+ pulse (5 h, 2.2 mM) and a Cu2+ vase solution (0.5 mM) vs a deionized water (DIW) control. Development of xylem blockage in the stem-end region 10 mm proximal to the wounded stem surface was examined over 21 days by light and transmission electron microscopy. Xylem vessels of stems stood into DIW were occluded with gels secreted into vessel lumens via pits from surrounding axial parenchyma cells. Gel secretion was initiated within 1-2 days post-wounding and gels were detected in the xylem from day 3. In contrast, Cu2+ treatments disrupted the surrounding parenchyma cells, thereby inhibiting gel secretion and maintaining the vessel lumens devoid of occlusions. The Cu2+ treatments significantly improved water uptake by the cut stems as compared to the control. © 2013 Scandinavian Plant Physiology Society.

Cotton bunchy top virus and other relatives

Cotton bunchy top virus (CBTV) and the related Cotton leafroll dwarf virus (CLRDV) have caused sporadic disease outbreaks in most cotton regions of the world. Until recently, little was known about the diversity of CBTV or its natural host range. Seven natural field hosts and one experimental host of CBTV have now been identified. These include cotton, Malva parviflora (Marshmallow weed), Abutilon theophrasti (Velvetleaf), Anoda cristata (Spurred anoda), Hibiscus sabdariffa (Rosella), Sida rhombifolia (Paddy’s lucerne), Chamaesyce hirta (Asthma plant) and Gossypium australe. These are currently the only eight known hosts of CBTV. However the virus may have a wider host range than originally thought and include further non-Malvaceae species like asthma plant (family Euphorbiaceae). There are two distinct strains of CBTV in Australia, -A and -B, which have been detected in cotton from numerous locations across almost all growing regions. From 105 samples of cotton that have been positive for CBTV, 6 were infections of strain A only, 60 were strain B only and 64 were a mixed infection of strains A and B. These results indicate the symptoms of cotton bunchy top disease are closely associated with the presence of strain CBTV-B. A diagnostic assay for Cotton leafroll dwarf virus (CLRDV - cotton blue disease) is being developed and applied successfully for the detection of CLRDV samples from Brazil and Thailand. This is the first confirmation of CLRDV from SE-Asia, which may pose an increased biosecurity threat to the Australian industry.

Grazing impacts on the susceptibility of rangelands to wind erosion: The effects of stocking rate, stocking strategy and land condition

An estimated 110 Mt of dust is eroded by wind from the Australian land surface each year, most of which originates from the arid and semi-arid rangelands. Livestock production is thought to increase the susceptibility of the rangelands to wind erosion by reducing vegetation cover and modifying surface soil stability. However, research is yet to quantify the impacts of grazing land management on the erodibility of the Australian rangelands, or determine how these impacts vary among land types and over time. We present a simulation analysis that links a pasture growth and animal production model (GRASP) to the Australian Land Erodibility Model (AUSLEM) to evaluate the impacts of stocking rate, stocking strategy and land condition on the erodibility of four land types in western Queensland, Australia. Our results show that declining land condition, over stocking, and using inflexible stocking strategies have potential to increase land erodibility and amplify accelerated soil erosion. However, land erodibility responses to grazing are complex and influenced by land type sensitivities to different grazing strategies and local climate characteristics. Our simulations show that land types which are more resilient to livestock grazing tend to be least susceptible to accelerated wind erosion. Increases in land erodibility are found to occur most often during climatic transitions when vegetation cover is most sensitive to grazing pressure. However, grazing effects are limited during extreme wet and dry periods when the influence of climate on vegetation cover is strongest. Our research provides the opportunity to estimate the effects of different land management practices across a range of land types, and provides a better understanding of the mechanisms of accelerated erosion resulting from pastoral activities. The approach could help further assessment of land erodibility at a broader scale notably if combined with wind erosion models.

Priority list of endemic diseases for the red meat industries

This report provides a systematic review of the most economically damaging endemic diseases and conditions for the Australian red meat industry (cattle, sheep and goats). A number of diseases for cattle, sheep and goats have been identified and were prioritised according to their prevalence, distribution, risk factors and mitigation. The economic cost of each disease as a result of production losses, preventive costs and treatment costs is estimated at the herd and flock level, then extrapolated to a national basis using herd/flock demographics from the 2010-11 Agricultural Census by the Australian Bureau of Statistics. Information shortfalls and recommendations for further research are also specified. A total of 17 cattle, 23 sheep and nine goat diseases were prioritised based on feedback received from producer, government and industry surveys, followed by discussions between the consultants and MLA. Assumptions of disease distribution, in-herd/flock prevalence, impacts on mortality/production and costs for prevention and treatment were obtained from the literature where available. Where these data were not available, the consultants used their own expertise to estimate the relevant measures for each disease. Levels of confidence in the assumptions for each disease were estimated, and gaps in knowledge identified. The assumptions were analysed using a specialised Excel model that estimated the per animal, herd/flock and national costs of each important disease. The report was peer reviewed and workshopped by the consultants and experts selected by MLA before being finalised. Consequently, this report is an important resource that will guide and prioritise future research, development and extension activities by a variety of stakeholders in the red meat industry. This report completes Phase I and Phase II of an overall four-Phase project initiative by MLA, with identified data gaps in this report potentially being addressed within the later phases. Modelling the economic costs using a consistent approach for each disease ensures that the derived estimates are transparent and can be refined if improved data on prevalence becomes available. This means that the report will be an enduring resource for developing policies and strategies for the management of endemic diseases within the Australian red meat industry.

Three-dimensional reconstruction of black tiger prawn (Penaeus monodon) spermatozoa using serial block-face scanning electron microscopy

Serial Block-Face Scanning Electron Microscopy (SBF-SEM) was used in this study to examine the ultrastructural morphology of Penaeus monodon spermatozoa. SBF-SEM provided a large dataset of sequential electron-microscopic-level images that facilitated comprehensive ultrastructural observations and three-dimensional reconstructions of the sperm cell. Reconstruction divulged a nuclear region of the spermatophoral spermatozoon filled with decondensed chromatin but with two apparent levels of packaging density. In addition, the nuclear region contained, not only numerous filamentous chromatin elements with dense microregions, but also large centrally gathered granular masses. Analysis of the sperm cytoplasm revealed the presence of degenerated mitochondria and membrane-less dense granules. A large electron-lucent vesicle and "arch-like" structures were apparent in the subacrosomal area, and an acrosomal core was found in the acrosomal vesicle. The spermatozoal spike arose from the inner membrane of the acrosomal vesicle, which was slightly bulbous in the middle region of the acrosomal vesicle, but then extended distally into a broad dense plate and to a sharp point proximally. This study has demonstrated that SBF-SEM is a powerful technique for the 3D ultrastructural reconstruction of prawn spermatozoa, that will no doubt be informative for further studies of sperm assessment, reproductive pathology and the spermiocladistics of penaeid prawns, and other decapod crustaceans. J. Morphol., 2016. (c) 2016 Wiley Periodicals, Inc.

Understanding urban rail travel for improved patronage forecasting

Improved forecasting of urban rail patronage is essential for effective policy development and efficient planning for new rail infrastructure. Past modelling and forecasting of urban rail patronage has been based on legacy modelling approaches and often conducted at the general level of public transport demand, rather than being specific to urban rail. This project canvassed current Australian practice and international best practice to develop and estimate time series and cross-sectional models of rail patronage for Australian mainland state capital cities. This involved the implementation of a large online survey of rail riders and non-riders for each of the state capital cities, thereby resulting in a comprehensive database of respondent socio-economic profiles, travel experience, attitudes to rail and other modes of travel, together with stated preference responses to a wide range of urban travel scenarios. Estimation of the models provided a demonstration of their ability to provide information on the major influences on the urban rail travel decision. Rail fares, congestion and rail service supply all have a strong influence on rail patronage, while a number of less significant factors such as fuel price and access to a motor vehicle are also influential. Of note, too, is the relative homogeneity of rail user profiles across the state capitals. Rail users tended to have higher incomes and education levels. They are also younger and more likely to be in full-time employment than non-rail users. The project analysis reported here represents only a small proportion of what could be accomplished utilising the survey database. More comprehensive investigation was beyond the scope of the project and has been left for future work.

Fugacity coefficients. Enthalpy, and second virial coefficients of boron trifluoride

Fugacity coefficients and isothermal changes of enthalpy have been calculated and reported. The calculations cover a temperature range of 0° to 75°C. up to gas densities of 1.0 gram per cc. The generalized Benedict-Webb-Rubin constants evaluated from generalized PVT relations is found to predict the experimental data with an over-all absolute deviation of 3.1%. Second virial coefficients and potential energy parameters for Lennard-Jones (12-6) potential energy function are reported also.

Dynamo models of grand minima

Since a universally accepted dynamo model of grand minima does not exist at the present time, we concentrate on the physical processes which may be behind the grand minima. After summarizing the relevant observational data, we make the point that, while the usual sources of irregularities of solar cycles may be sufficient to cause a grand minimum, the solar dynamo has to operate somewhat differently from the normal to bring the Sun out of the grand minimum. We then consider three possible sources of irregularities in the solar dynamo: (i) nonlinear effects; (ii) fluctuations in the poloidal field generation process; (iii) fluctuations in the meridional circulation. We conclude that (i) is unlikely to be the cause behind grand minima, but a combination of (ii) and (iii) may cause them. If fluctuations make the poloidal field fall much below the average or make the meridional circulation significantly weaker, then the Sun may be pushed into a grand minimum.

Is meridional circulation important in modelling irregularities of the solar cycle?

We explore the importance of meridional circulation variations in modelling the irregularities of the solar cycle by using the flux transport dynamo model. We show that a fluctuating meridional circulation can reproduce some features of the solar cycle like the Waldmeier effect and the grand minimum. However, we get all these results only if the value of the turbulent diffusivity in the convection zone is reasonably high.

An estimate of the number of tropical tree species

The high species richness of tropical forests has long been recognized, yet there remains substantial uncertainty regarding the actual number of tropical tree species. Using a pantropical tree inventory database from closed canopy forests, consisting of 657,630 trees belonging to 11,371 species, we use a fitted value of Fisher's alpha and an approximate pantropical stem total to estimate the minimum number of tropical forest tree species to fall between similar to 40,000 and similar to 53,000, i.e., at the high end of previous estimates. Contrary to common assumption, the Indo-Pacific region was found to be as species-rich as the Neotropics, with both regions having a minimum of similar to 19,000-25,000 tree species. Continental Africa is relatively depauperate with a minimum of similar to 4,500-6,000 tree species. Very few species are shared among the African, American, and the Indo-Pacific regions. We provide a methodological framework for estimating species richness in trees that may help refine species richness estimates of tree-dependent taxa.

Correct biological timing in Arabidopsis requires multiple light-signaling pathways.

Circadian oscillators provide rhythmic temporal cues for a range of biological processes in plants and animals, enabling anticipation of the day/night cycle and enhancing fitness-associated traits. We have used engineering models to understand the control principles of a plant's response to seasonal variation. We show that the seasonal changes in the timing of circadian outputs require light regulation via feed-forward loops, combining rapid light-signaling pathways with entrained circadian oscillators. Linear time-invariant models of circadian rhythms were computed for 3,503 circadian-regulated genes and for the concentration of cytosolic-free calcium to quantify the magnitude and timing of regulation by circadian oscillators and light-signaling pathways. Bioinformatic and experimental analysis show that rapid light-induced regulation of circadian outputs is associated with seasonal rephasing of the output rhythm. We identify that external coincidence is required for rephasing of multiple output rhythms, and is therefore important in general phase control in addition to specific photoperiod-dependent processes such as flowering and hypocotyl elongation. Our findings uncover a fundamental design principle of circadian regulation, and identify the importance of rapid light-signaling pathways in temporal control.

The circadian oscillator gene GIGANTEA mediates a long-term response of the Arabidopsis thaliana circadian clock to sucrose.

Circadian clocks are 24-h timing devices that phase cellular responses; coordinate growth, physiology, and metabolism; and anticipate the day-night cycle. Here we report sensitivity of the Arabidopsis thaliana circadian oscillator to sucrose, providing evidence that plant metabolism can regulate circadian function. We found that the Arabidopsis circadian system is particularly sensitive to sucrose in the dark. These data suggest that there is a feedback between the molecular components that comprise the circadian oscillator and plant metabolism, with the circadian clock both regulating and being regulated by metabolism. We used also simulations within a three-loop mathematical model of the Arabidopsis circadian oscillator to identify components of the circadian clock sensitive to sucrose. The mathematical studies identified GIGANTEA (GI) as being associated with sucrose sensing. Experimental validation of this prediction demonstrated that GI is required for the full response of the circadian clock to sucrose. We demonstrate that GI acts as part of the sucrose-signaling network and propose this role permits metabolic input into circadian timing in Arabidopsis.