Phytophthora cinnamomi and Australia’s biodiversity : impacts, predictions and progress towards control

Phytophthora cinnamomi continues to cause devastating disease in Australian native vegetation and consequently the disease is listed by the Federal Government as a process that is threatening Australia’s biodiversity. Although several advances have been made in our understanding of how this soil-borne pathogen interacts with plants and of how we may tackle it in natural systems, our ability to control the disease is limited. The pathogen occurs widely across Australia but the severity of its impact is most evident within ecological communities of the south-west and south-east of the country. A regional impact summary for all states and territories shows the pathogen to be the cause of serious disease in numerous species, a significant number of which are rare and threatened. Many genera of endemic taxa have a high proportion of susceptible species including the iconic genera Banksia, Epacris and Xanthorrhoea. Long-term studies in Victoria have shown limited but probably unsustainable recovery of susceptible vegetation, given current management practices. Management of the disease in conservation reserves is reliant on hygiene, the use of chemicals and restriction of access, and has had only limited effectiveness and not provided complete control. The deleterious impacts of the disease on faunal habitat are reasonably well documented and demonstrate loss of individual animal species and changes in population structure and species abundance. Few plant species are known to be resistant to P. cinnamomi; however, investigations over several years have discovered the mechanisms by which some plants are able to survive infection, including the activation of defence-related genes and signalling pathways, the reinforcement of cell walls and accumulation of toxic metabolites. Manipulation of resistance and resistance-related mechanisms may provide avenues for protection against disease in otherwise susceptible species. Despite the advances made in Phytophthora research in Australia during the past 40 years, there is still much to be done to give land managers the resources to combat this disease. Recent State and Federal initiatives offer the prospect of a growing and broader awareness of the disease and its associated impacts. However, awareness must be translated into action as time is running out for the large number of susceptible, and potentially susceptible, species within vulnerable Australian ecological communities.

Androgenic activity of effluent from forty-five municipal waste water treatment plants in Victoria, Australia

It is well known that waste water treatment plant (WWTP) effluents are estrogenic. There has been much less consideration of the androgenic activity of WWTP effluents. To partly address the shortage of information on androgens in Australian WWTP effluents, in August 2006, and again in 2007, we collected discharges from up to 45 Victorian WWTPs (~25% of all WWTPs in Victoria), grouped by treatment process, i.e. activated sludge, extended aeration, and lagoon based treatment, and measured the total estrogenic, androgenic, retinoic acid, and aromatic hydrocarbon hydrogenase activity of the effluents using a hybrid yeast bioassay. This paper will concentrate on the androgenic activity and male hormone concentrations.

Incorporation of oxygen into the succinate co-product of iron(II) and 2-oxoglutarate dependent oxygenases from bacteria, plants and humans.

The ferrous iron and 2-oxoglutarate (2OG) dependent oxygenases catalyse two electron oxidation reactions by coupling the oxidation of substrate to the oxidative decarboxylation of 2OG, giving succinate and carbon dioxide coproducts. The evidence available on the level of incorporation of one atom from dioxygen into succinate is inconclusive. Here, we demonstrate that five members of the 2OG oxygenase family, AlkB from Escherichia coli, anthocyanidin synthase and flavonol synthase from Arabidopsis thaliana, and prolyl hydroxylase domain enzyme 2 and factor inhibiting hypoxia-inducible factor-1 from Homo sapiens all incorporate a single oxygen atom, almost exclusively derived from dioxygen, into the succinate co-product.

Endemic freshwater finfish of Asia : distribution and conservation status

Freshwater finfish species richness and level of endemism in East, and South and South-East Asia that included 17 nations were studied using available databases, and included nation-wise distribution, habitat types, and conservation status. The number of endemic finfish species in the region was 559, belonging to 47 families. Families Cyprinidae and Balitoridae accounted for 43.5% and 16.2% of the total number of endemic species in the region, respectively, followed by Sisoridae (25), Gobiidae (20), Melanotaeniidae (19), and Bagridae (16), and the other 41 families had at least one endemic species. Nation-wise the most number of endemic freshwater finfish species occur in India (191), followed by China (88), Indonesia (84), and Myanmar (60). In India, the endemic species accounted for 26.4% of the native freshwater fish fauna, followed by South Korea (16.9%), the Philippines, (16.3%) and Myanmar (15.7%).

Statistically significant relationships discerned between the number of indigenous and endemic species richness to land area (X_la in 10³ km²) of the nations in the region were, Y_in = 218.961 Ln(X_la) – 843.1 (R² = 0.735; P < 0.001) and Y_e = 28.445 Ln X_la−134.47 (R² = 0.534; P < 0.01), respectively, and between indigenous and endemic species richness was Y_e = 0.079X_n− 1.558 (R² = 0.235; P < 0.05).

The overall conservation status of endemic finfish in Asia was satisfactory in that only 92 species were in some state of vulnerability, of which 37 species (6.6%) are endangered or critically endangered. However, the bulk of these species (83.7%) were cave- and or lake-dwelling fish. However, nation-wise, the endemic freshwater finfish fauna of the Philippines and Sri Lanka, based on the imperilment index, were found to be in a highly vulnerable state. Among river basins, the Mekong Basin had the highest number of endemic species (31.3%). The discrepancies between databases are highlighted and the need to consolidate information among databases is discussed. It is suggested that the Mekong Basin be considered as a biodiversity hotspot, and appropriate management strategies be introduced in this regard.

Solar energy use for energy savings in dairy processing plants

New Zealand is one of the world’s largest producers of dairy products and has a climate with high levels of solar radiation; however, the use of solar energy in the dairy processing industry has received limited attention. An examination of historical records found that the annual peak in New Zealand milk production and processing occurs at a time when solar radiation levels are increasing markedly. An F-Chart analysis was used to simulate the performance of large-area arrays of solar collectors and to determine their suitability for heating and cooling in a dairy processing environment. For the study four types of solar collectors were analysed: glazed flat plates, evacuated tubes, evacuated tubes with CPC reflectors and a building-integrated solar collector under development at the University of Waikato (UoW). It was found that of these echnologies, both flat plate and evacuated tubes with CPC reflectors could make useful heating and cooling contributions. Furthermore, the solar fraction was determined mainly by the collector area to storage volume ratio. Finally, it was found that the UoW building-integrated solar collector could make a significant contribution to energy use in dairies and may be an attractive future technology for the industry.

Lichen flora of the Mount Donna Buang Scenic Reserve, Victoria

This report investigated the lichen flora of the Mt Donna Buang Scenic Reserve in Victoria, There were several aims: to describe the lichens of the region, to produce a pictorial key enabling field identification and to determine any distribution patterns. A floristic survey covering approximately 50 square km was undertaken to determine lichen diversity of the region generally. Lichens were sampled along roads, tracks, walking trails and in sections of bush, taking into account forest type and, particularly, areas that were lichen rich. Seventy-five lichen species in 43 genera and 27 families were identified and described from the region. An unknown, species H, also was described. Of the 76 lichen species, 22 were crastose and the remainder macrolichens. The best represented families were: Cladoniaceae (8 species), Hypogymniaceae (6), Lobariaceae (7), Lecideaceae (6), Pannariaceae (6) and Parmeliaccae (6). This study described 12 species (17%) which previously were not known for Victoria and which are a first record for the state. These include: Cladonia sarmentosa (J.D. Hook & Taylor) Dodge, Graphis librata Knight, Parmelinopsis neodamaziana (Elix & Johnston) Elix & Hale, Pertusaria novaezelandiae Szatala, Placopsis pardlina f. microphylla Lamb, Porina leptalea AX. Sm., Pseudocyphellaria ardesiaca Galloway, Trapeliopsis congregant (Zahlbr.) Brako, Menegazzia myriotrema (Mull. Arg.) P. James, Bunodophoron scrobiculatum (Church. Bab,) Wedin, Parmelia testacea Stirton and Menegazzia purpurascens S. Louwhoff sp. nov.. The last eight species are new to the mainland and, apart from Menegazzia purpurascens, previously were known only from Tasmania. Five main elements of distribution were identified for the lichen flora of the Mt Donna Buang Scenic Reserve: cosmopolitan, austral/australasian, paleotropical, pantropical and western pacific. The majority of species (68%) had austral/australasian distributions, eleven (16%) were endemic to Australia and nine (13%) occurred only in Tasmania , Victoria and New Zealand. A pictorial, dichotomous key was constructed for the lichen flora of the Mt Donna Buang Scenic Reserve. Previously, keys to the lichen flora of Tasmanian rainforests were suggested as appropriate to similar areas in Victoria, however, the Victorian forests include a significant sclerophyll element The key presented is specific for the study site but is appropriate to similar regions in Victoria and has been tested in a number of these areas. The key was designed to be ‘user-friendly’ so that the experienced and inexperienced alike are able to use it. A more detailed investigation of the lichen flora of the Mt Donna Buang Scenic Reserve was carried out in order to determine distribution. A total of 50 quadrats, each 20m x 20m in size, were sampled. Within each, the dominant vegetation type was determined and individuals were identified and location noted. The cover abundance of each lichen species on each individual tree was estimated using a modified Braun-Blanquet scale. A total of 710 trees, representing 13 different species, were examined. Nothofagus cunninghamii (Hook.) Oerst, Eucalyptus regnans R Mull., Acacia dealbata Link, A. melanoxylon R. Br., Hedycarya angustifolia A. Cunn. and Atherosperma moschatum Labill. were the six most common tree species encountered at the study site. Nothofagus cunninghamii supported the greatest lichen diversity (39 species), although most species occurred on less than 10% of the trees. The majority of lichens occurring on N. cunninghamii A. melanoxylon, A. dealbata and H. angustifolia were foliose or crustose, those on Ł. regnans fruticose and foliose and those on A moschatum crustose. Bunodophoron australe was the only lichen species at the study site to occur on one host, Nothofagus cunninghamiL Many occurred on a number of different hosts, but were most common on one particular tree species. The distribution of lichens at the study site was analysed with a rnultivariate statistical package (PATN) which dealt with ‘pattern analysis’. The program ‘SSH’ in PATN which uses the Bray-Curtis ordination technique, was used to create scatterplots displaying the degree of dissimilarity between quadrats in terms of presence/absence of lichen species. The program ‘TWAY’ in PATN was used to construct a two way table to display which lichen species occurred in each vegetation type. The pattern analysis revealed that the lichens of the Mt Donna Buang Scenic Reserve were not restricted to any particular forest type, but particular lichens, or groups of lichens, tended to predominate in certain vegetation communities. This concurs with work done by others in Tasmanian forests. Quadrats which were situated in cool temperate rainforest were grouped more closely with each other than with quadrats in other vegetation types. These also supported the greatest number of lichen species. This was not surprising since N. cunninghamii the dominant tree species in cool temperate rainforest, supported the greatest lichen diversity.

Study of the natriuretic peptide hormone system in plants

In this study, both physiological and cellular effects are elicited by natriuretic peptides (NPs), a novel type of plant hormone. It was found that rat ANP (rANP) influenced stomatal opening movement in Tradescantia sp., where a significant increase in stomatal opening was observed in the presence of 1 µM rANP. Furthermore, this effect is mediated by cGMP, a (putative) second messenger of NPs. Two inhibitors of guanylyl cyclase, LY 83583 and methylene blue, inhibited rANP-induced stomatal opening. In contrast, stomatal opening is induced in a concentration dependent manner by the cell permeant cGMP analogue 8-Br-cGMP. In addition it was found, that like in animals, the secondary structure of rANP is essential for rANP responses. Linearised rANP is biologically inactive. Since ANP elicit plant responses, an attempt was made to isolate NP analogues from plants. A protocol for partially purifying NP from plants was developed. It was found that two fractions eluted from an immunoaffinity chromatography column (0.5 M KCI eluted fraction and 0.75 M KCI eluted fraction) were biologically active. The level of cGMP in response to NPs was also tested. It is suggested that the receptor of NP is specific since only 0.75 M KCI eluted fractions increased cGMP levels in Zea mays root stele tissue. rANP did not elicit an effect on cGMP levels in this tissue and LY 83583 did not affect this response. It is therefore argued that a plant specific biologically active NP system is present in the stele and it is predicted that NPs modulate solute movement in this tissue. NPs also influence K⁺, Na⁺ and H⁺ fluxes in Zea mays root stele. Increase in both K⁺ and Na+ uptake were observed after 30 min., while H⁺ flux shifted immediately toward influx in the presence of both 0.5 and 0.75 KCI eluted fractions. Finally, a model is proposed for the effect of NPs on solute movement and its signalling system in plants.

Repair and tolerance of DNA damage in higher plants

DNA repair mechanisms constitute an essential cellular response to DNA damage arising either from metabolic processes or from environmental sources such as ultraviolet radiation. Repair of these lesions may be via direct reversal, or by processes such as nucleotide excision repair (NER), a coordinated pathway in which lesions and the surrounding nucleotides are excised and replaced via DNA resynthesis. The importance of repair is illustrated by human disease states such as xeroderma pigmentosum and Cockayne's syndrome which result from defects in the NER system arising from mutations in XP- genes or XP- and CS- genes respectively Little detail is known of DNA damage repair processes in plants, despite the economic and ecological importance of these organisms. This study aimed to expand our knowledge of the process of NER in plants, largely via a polymerase chain reaction (PCR)-based approach involving amplification, cloning and characterisation of plant genomic DNA and cDNA. Homologues of the NER components XPF/RAD1 and XPD/RAD3 were isolated as both genomic and complete cDNA sequences from the model dicotyledonous plant Arabidopsis thaliana. The sequence of the 3'-untranslated region of atXPD was also determined. Comparison of genomic and cDNA sequences allowed a detailed analysis of gene structures, including details of intron/exon processing. Variable transcript processing to produce three distinct transcripts was found in the case of atXPF. In an attempt to validate the proposed homologous function of these cDNAs, assays to test complementation of resistance to ultraviolet radiation in the relevant yeast mutants were performed. Despite extensive amino acid sequence conservation, neither plant cDNA was able to restore UV-resistance. As the yeast RAD3 gene product is also involved in vivo in transcription, and so is required for viability, the atXPD cDNA was tested in a complementation assay for this function in an appropriate yeast mutant. The plant cDNA was found to substantially increase the viability of the yeast mutant. The structural and functional significance of these results is discussed comparatively with reference to yeast, human and other known homologues. Other putative NER homologues were identified in A. thaliana database sequences, including those of ERCC1/RAD10 and XPG/ERCC5/RAD2, and are now the subjects of ongoing investigations. This study also describes preliminary investigations of putative REVS and RAD30 translesion synthesis genes from A. thaliana.

In vitro and immunological assessment of the estrogenic activity and concentrations of 17β-estradiol, estrone, and ethinyl estradiol in treated effluent from 45 wastewater treatment plants in Victoria, Australia

The project was conducted between May 2006 and September 2007, and involved the collection of effluent samples from 45 wastewater treatment plants (WWTPs). The 45 WWTPs included 16 lagoon-based plants and 29 with activated sludge-based processes. Permission was obtained from all the relevant water authorities to collect samples of final effluent at point of discharge to the environment, whether that was to a creek, a river, the ocean, or the land. Samples were collected on two occasions, namely, in August 2006 (winter) and late February–early March 2007 (summer), and subjected to a number of biological and chemical analyses, including toxicity tests, measurement of hormonal (estrogenic) activity using yeast-based bioassays, and measurement of specific hormonal concentrations using enzyme-linked immunosorbent assays (ELISAs). Almost all of the effluents examined showed estrogenic activity: in winter, no activity to 73 ng/l 17β-estradiol equivalents (EEQ); and in summer, no activity to 20 ng/l EEQ. On the whole, the levels of estrogenic activity observed were comparable with the range recently reported in Australia and New Zealand using human estrogen receptor-based assays (“not detected” to ~10 ng/l EEQ). The low/no bioassay response was confirmed by the chemical assessment of estradiol, estrone, and ethinyl estradiol concentrations by ELISA, which returned concentrations of these compounds for the most part below 10 ng/l.

Conservation of the endemic dwarf carnivores of Cozumel Island, Mexico.

Cozumel Island, Mexico, harbours two endemic species of dwarf procyonids: the Pygmy Raccoon Procyon pygmaeus and the Dwarf Coati Nasua nelsoni. Both species are Critically Endangered, and are among the world’s most threatened Carnivora. Here we summarise the research we have been conducting on their ecology, evolution, genetics, and conservation. We also summarise the conservation initiatives we have been undertaking and promoting in order to advance the conservation of these unique species and their habitats. This effort illustrates the importance of an interdisciplinary approach in conservation science and action in maximising effectiveness. Nevertheless, the precarious status of the species make it imperative to continue and expand the work we have carried out in Cozumel to prevent two imminent global extinctions.

Antibody-mediated protection of plants against infection from potyviruses

Transgenic plants expressing single-chain antibodies have been produced to investigate the feasibility of antibody-mediated broad-spectrum protection against plant virus infections. This study indicates that protection against a wide range of plant viruses can be achieved in transgenic plants expressing a single antibody construct.