Cellular and molecular components of plant defence against pathogens

This project investigated how plants respond to invading pathogens using microscopic, biochemical and genetic approaches. The development of transgenic plants containing the green fluorescent protein cloned from jellyfish enabled a new approach to studying plant defence genes. In particular, the role and involvement of the plant gene PAL1 was analysed.

A soil-free plant growth system to facilitate analysis of plant pathogen interactions in roots

Analysis of the interaction of pathogens with plant roots is often complicated by the growth of plants in a soil substrate. A soil-free plant growth system (SPS) was developed that removes the need for a substrate while supporting the growth of seedlings in a nutrient rich, oxygenated environment. The model legume Lupinus angustifolius was used to compare the growth of seedlings within soil and the SPS. Seedlings grown under both conditions were similar in morphology, anatomy and health (measured by leaf chlorophyll abundance) and importantly there was little difference in root growth and development although straighter and fuller root systems were achieved in the SPS. The ease of access to the root system proved efficient for the analysis of root and pathogen interactions with no interference from soil or adhering particulate matter. Following inoculation of L. angustifolius roots with Phytophthora cinnamomi the host/pathogen interaction was easily observed and tissues sampled undamaged.

Soil erosion in developing countries: a socio-economic appraisal

Soil erosion is the single most important environmental degradation problem in the developing world. Despite the plethora of literature that exists on the incidence, causes and impacts of soil erosion, a concrete understanding of this complex problem is lacking. This paper examines the soil erosion problem in developing countries in order to understand the complex inter-relationships between population pressure, poverty and environmental-institutional dynamics. Two recent theoretical developments, namely Boserup's theory on population pressure, poverty and soil erosion and Lopez's theory on environmental and institutional dynamics have been reviewed. The analysis reveals that negative impacts of technical change, inappropriate government policies and poor institutions are largely responsible for the continued soil erosion in developing countries. On the other hand, potential for market-based approaches to mitigate the problem is also low due to the negative externalities involved. A deeper appreciation of institutional and environmental dynamics and policy reforms to strengthen weak institutions may help mitigate the problem.

Surviving suppression : no detectable impacts of Class A foam on soil invertebrates and some Australian native plants

Firefighting foams (Class A foams) are an effective and widespread firefighting tool which are frequently used in environmentally sensitive areas. Firefighting foams are known to be ecologically damaging in aquatic environments, however their impacts at the plant species or ecosystem level are relatively unknown. Reports of shoot damage to plants, suppressed flowering, and changes in plant community composition suggested that the ecological damage caused by their use may be unacceptable. However, applications of foam to seedlings of some Australian plant species from representative and widespread families, showed no detectable impacts on a range of vegetative growth characteristics. Application of 1.0% foam to heathland soils showed no detectable impacts on soil invertebrate Orders sampled over several months. The results are encouraging for the continued use of Class A foam as a fire suppression technique.

Defining plant resistance against Phytophthora Cinnamomi and application of resistance to revegetation

Phytophthora cinnamomi is a soil borne plant pathogen that causes devastating disease in many Australian ecosystems and threatens the survival of native flora. Compared with the number of plant species that are susceptible to P. cinnamomi, only a few species are known to be resistant and control of this pathogen by chemicals is difficult and undesirable in natural systems. The major aim of our research is therefore to characterise natural resistance and determine which signalling pathways and defence responses are involved. Our examination of resistance is being approached at several levels, one of which is through the use of the model plant, Arabidopsis. Previously, Arabidopsis had been shown to display ecotypic variation in responses to P. cinnamomi and we are exploring this further in conjunction with the analysis of a bank of Arabidopsis defence pathway mutants for their responses to the pathogen. These experiments will provide a fundamental basis for further analysis of the defence responses of native plants. Native species (susceptible and resistant) are being assessed for their responses to P. cinnamomi at morphological, biochemical and molecular levels. This research also involves field-based studies of plants under challenge at various sites throughout Victoria, Australia. The focus of this field-based research is to assess the responses of individual species to P. cinnamomi in the natural environment with the goal of identifying individuals within susceptible species that display 'resistance'. Understanding how plants are able to resist this pathogen will enable strategies to be developed to enhance species survival and to restore structure and biodiversity to the ecosystems under threat.

Plant pathogens causing vegetation dieback: a serious threat to the conservation of small mammals in Australia

The soil-borne plant pathogen Phytophthora cinnamomi occurs in most Australian states. It is pathogenic to many Australian species, particularly the Proteaceae, Fabaceae, Dillineaceae and Epacridaceae. In Western Australia, c. 2000 of the 9000 endemic plant species are directly affected by the disease. The epidemic of plant deaths caused by P. cinnamomi is recognised as one of 11 Key Threatening Processes to the Australian Environment, and is now also acknowledged as a potential threat fauna in a range of communities. The implications of landscape modification due to the effects of P. cinnamomi dieback prompted our research, designed to measure the distribution and abundance of small mammals in disease-affected ecosystems. This study was in the Jarrah (Eucalyptus marginata) forests in the Darling Range, Western Australia and measured the distribution and abundance of one small mammal species, the Mardo (Antechinus flavipes) by Elliott trapping in forests with (1) high, (2) mixed and (3) no evidence of Phytophthora dieback. Trap success was highest in sites with no effect of Phytophthora (7.3 animals per 100 trap nights), whereas the lowest trap success was recorded at the high impact sites (0.67 animals per 100 trap night). There was a significant difference in trap success of Mardos in Elliott trapping over 1800 trap nights (x²= 23.19, d.f = 5, p < 0.001). An examination of the distribution of individuals and sexes suggests that Phytophthora-affected sites act as sinks for Mardos, while source areas are healthy, unaffected Jarrah forest.

Impact of saltmarsh disturbance on seed quality of Sarcocornia (Sarcocornia quinqueflora), a food plant of an endangered Australian parrot

The article presents a study which evaluates the impact of saltmarsh perturbation on seed quality of Sarcocornia which is a food for the endangered Orange-bellied parrot in Australia. It notes the significant impact of grazing on the energy and availability of Sarcocornia seeds in saltmarshes and indicates that graze-free saltmarshes with regular inundation provide the highest potential for Sarcocornia seed availability. Further, the substantial factor of soil characteristics and salinity of flood water and inundation on the seed production of Sarcocornia is also discussed.

Effect of Phytophthora cinnamomi on the habitat utilization of Antechinus stuartii in a Victorian forest

Phytophthora cinnamomi (Cinnamon fungus) is a pathogenic soil fungus which infects plant communities along the south-eastern coast of Australia, and the south-western corner of Western Australia. The symptoms of this disease include chlorosis, death of branches (ie. ‘dieback’), retarded growth and the eventual death of infected plants. This leads to devastating effects upon plant communities by altering both the structural and floristic characteristics of these communities. Small mammal species are dependent on specific features of their habitat such as vegetation structure and floristics. This thesis investigated alterations to the habitat of the insectivorous marsupial mouse, Antechinus stuartii, due to the presence of P. cinnamomi. The study was undertaken in an area of an open forest in the Brisbane Ranges, Victoria. Significant changes were found in both the floristic composition and structure of the vegetation at study sites infected with P, cinnamomi, compared to uninfected sites. The habitat utilization by A. stuartii of uninfected and infected vegetation was investigated using live trapping and radio-telemetric techniques. Capture rates were higher at sites uninfected by P. cinnamomi, and both male and females selected areas free from infection. Home range areas of males were significantly larger than those of females as assessed by telemetry. Both sexes spent a high proportion of time in areas dominated by Xanthorrhoea australis (Austral grass tree). There were significant relationships between the abundance of A. stuartii and the denseness of vegetation above 1 metre in height, and in particular, the proportion of cover afforded by X. australis. There were no significant differences in the cover of Eucalyptus spp. between uninfected and infected sites, but there were significantly more nest hollows in infected areas. The abundance of invertebrates was examined using pitfall traps. There were no significant differences in the abundance of the larger invertebrate taxa at infected and uninfected sites, but higher abundances of some micro-invertebrate groups in infected areas were recorded. The most likely factors considered to be influential in the habitat selection of A. stuartii were vegetation structure, and the presence of X. australis. To assess whether these factors were important the leaves of X. australis were removed with a brushcutter, to mimic the early effects of infection with P. cinnamomi. Animals did not respond to the alteration of vegetation structure in the short term (3-4 days). Longer-term experiments are required to assess the habitat utilization of A. stuartii at different periods following habitat manipulation. The implications of the presence of P. cinnamomi on the conservation of fauna are discussed. The destructive nature of the pathogen, and the slow rate of recovery from the disease, means that P. cinnamomi can be considered a threatening process to plant communities and the fauna that reside within that habitat. Future management of this disease within natural areas must therefore be cognisant of the potential of P. cinnamomi to significantly affect faunal as well as vegetative communities.

Sorption and Fractionation of Copper in soil at a sewage irrigation farm in Australia

Copper (Cu) is an important heavy metal to be considered in soil contamination, because high concentrations of copper in soil produce toxic effects and may accumulate in plant tissues. In Australia's oldest sewage irrigation farm, located in Werribee, Victoria, soil in the land filtration area is contaminated by Cu. However, Cu content in herbage tissues is in the normal range and has been trending downward since 1979. Therefore, studies on the sorption capacity and sequential extraction of Cu in soil at the Werribee Farm is of significance, not only for better understanding the mechanism of transport, chemical processes, and plant uptake of Cu, but also in providing information for the practical management of sewage farm soils. Methods of combining sorption isotherms with sequential extraction procedures were adopted, and the results showed that the soil in the land filtration area at Werribee Farm has a high sorption capacity for Cu, and distribution coefficients, K_f of Cu, were 629 L kg^-1 in surface soils (0-20 cm) and 335 L kg^-1 in subsurface soils (20-40 cm). The sequential extraction fractions demonstrate that exchangeable and carbonate fractions are very low, only comprising 3.49 to 5.49% of total copper. The other fractions are also discussed. This characteristic of Cu in soil is related to the low concentration of Cu in plant tissues.

Profiling secondary metabolites of plant roots inoculated with Phytophthora cinnamomi

The soil-borne pathogen, Phytophthora cinnamomi, continues to cause severe dieback in Australian native forest species and is of great international significance due to its global distribution. This research established a protocol to successfully identify phyto-chemicals associated with the defense response of plants challenged by the disease caused by Phytophthora cinnamomi.

Defining plant resistance to Phytophthora cinnamomi : a standardized approach to assessment

Phytophthora cinnamomi is a soil-borne plant pathogen that causes devastating disease in agricultural and natural systems worldwide. While a small number of species survive infection by the pathogen without producing disease symptoms, the nature of resistance, especially under controlled conditions, remains poorly understood. At present, there are no standardized criteria by which resistance or susceptibility to P. cinnamomi can be assessed, and we have used five parameters consisting of plant fresh weight, root growth, lesion length, relative chlorophyll content of leaves and pathogen colonization of roots to analyse responses to the pathogen. The parameters were tested using two plant species, Zea mays and Lupinus angustifolius, through a time course study of the interactions and resistance and susceptibility defined 7days after inoculation. A scoring system was devised to enable differentiation of these responses. In the resistant interaction with Z. mays, there was no significant difference in fresh weight, root length and relative chlorophyll content in inoculated compared with control plants. Both lesion size and pathogen colonization of root tissues were limited to the site of inoculation. Following inoculation L. angustifolius showed a significant reduction in plant fresh weight and relative leaf chlorophyll content, cessation of root growth and increased lesion lengths and pathogen colonization. We propose that this technique provides a standardized method for plant-P. cinnamomi interactions that could be widely used to differentiate resistant from susceptible species.

Plant extract assisted extracellular tannase production by Aspergillus sp MIK 23

An extracellular tannase (E.C. 3.1.1.20) producing fungal strain was isolated from soil and identified as Aspergillus sp MIK23. Out of various plant extracts, Terminalia chebula powder (TCP) in the optimized medium enhanced enzyme production. Maximum yield of tannase (3 IU ml-1) was obtained with glucose (10 g/L), urea (2 g/L), and yeast extract (2.5 g/L) when inoculated with 10% inoculum in 48 h. An initial medium at pH 6.0 and a cultivation temperature of 37 0C was found to be optimum for enzyme production. Metal ions Mg2+, Zn2+, Ca2+, Cu2+ and Cd2+ did not improve enzyme activity, whereas, Ca2+, Fe2+ and Hg2+ repressed enzyme activity. The enzyme was purified using ammonium sulfate precipitation followed by Q-sepharose ion-exchange chromatography. The enzyme was purified to 42-fold with an overall recovery of 20. The pH and temperature optima of the purified tannase were found to be 7.0 and 37°C, respectively.

Is there more soil carbon under nitrogen-fixing trees than under non-nitrogen-fixing trees in mixed-species restoration plantings?

Afforestation of agricultural land provides an important opportunity to mitigate climate change by storing carbon (C) in both plant biomass and the soil. Here we present results of a study in which we sought to determine whether soil under nitrogen(N)-fixing trees contained more C than soil under non-N-fixing trees in mixed-species plantings, and thus if inclusion of N-fixers is beneficial in terms of increasing soil C sequestration. Soils were sampled directly beneath N-fixing and non-N-fixing tree species in riparian and upland mixed-species plantings in southeastern Australia. Soil C and N contents were assessed at both the landscape and individual planting scales. At the landscape scale, there were higher levels of soil C and N under N-fixing trees compared with non-N-fixing trees. At the individual planting scale, the patterns were less clear with both large increases and decreases occurring across the range of sites. The results presented here indicate that the inclusion of N-fixers may help to increase soil C, and N, but that the response may be site- and species-specific. © 2014 Elsevier B.V.

Nurse plant effects of a dominant shrub (Duma florulenta) on understorey vegetation in a large, semi-arid wetland in relation to flood frequency and drying

 Questions: Do nurse plant interactions significantly influence understorey vegetation diversity in a large, semi-arid, shrub-dominated wetland? How do the modes and net effects of nurse plant interactions vary spatially along a flood frequency gradient, and temporally in response to drying? Location: Narran Lakes Ramsar site, New South Wales, Australia. Methods: Microhabitat characteristics, understorey vegetation and germinable soil seed banks were investigated in shrub and open habitats across a flood frequency gradient in a large, semi-arid wetland dominated by open shrubland under productive conditions following floodwater recession and again following 6 mo of drought. Split-plot ANOVA and multivariate analyses were used to determine the effects of shrubs on microhabitat character, understorey vegetation cover, species diversity, richness and composition and germinable soil seed banks. Results: Microhabitat characteristics, including canopy cover, litter cover and soil character, all differed between shrub and open habitats, especially in the most frequently flooded sites. Under productive conditions following flooding, lignum shrubs suppressed understorey vegetation cover but increased species richness at the site scale across the flood frequency gradient and, in the most frequently flooded sites, supported higher species density at a microhabitat scale. Under dry conditions, lignum shrubs had a positive effect on understorey vegetation cover, species richness and species density across the flood frequency gradient, but particularly in frequently flooded sites. A significant difference in soil seed bank composition between shrub and open habitats was only observed in frequently flooded sites. Conclusions: Nurse plant interactions appear to play an important role in determining understorey vegetation diversity in the lignum shrubland of the Narran Lakes wetland system. The modes and net effects of these nurse plant interactions vary in space and time in relation to flood history and drying. Positive interactions, probably involving microhabitat amelioration, appear to be particularly important to plant diversity and abundance under dry conditions. Under more favourable wetter conditions, lignum shrubs also contribute to understorey vegetation diversity by facilitating the establishment of different species than those dominating open habitats. Our findings have implications for the management of perennial shrubs and hydrological regimes in such wetlands.