Cucumber mosaic virus infection of kava (Piper methysticum) and implications for cultural control of kava dieback disease

Cucumber mosaic virus (CMV) was found by reverse transcription polymerase chain reaction (RT-PCR) to be not fully systemic in naturally infected kava (Piper methysticum) plants in Fiji. Twenty-six of 48 samples (54%) from various tissues of three recently infected plants were CMV-positive compared with 7/51 samples (14%) from three long-term infections (plants affected by dieback for more than 1 year). The virus was also found to have a limited ability to move into newly formed stems. CMV was detected in only 2/23 samples taken from re-growth stems arising from known CMV infected/dieback affected plants. Mechanical inoculation experiments conducted in Fiji indicate that the known kava intercrop plants banana (Musa spp.), pineapple (Ananas comosus), peanut (Arachis hypogaea) and the common weed Mikania micrantha are potential hosts for a dieback-causing strain of CMV It was not possible to transmit the virus mechanically to the common kava intercrop plants taro (Colocasia esculenta), Xanthosoma sp., sweet potato (Ipomoea batatas), yam (Dioscorea alata), papaya (Carica papaya) or the weed Momordica charantia. Implications of the results of this research on a possible integrated disease management strategy are discussed.

Capsicum chlorosis virus infecting Capsicum annuum in the East Kimberley region of Western Australia

Capsicum chlorosis virus (CaCV) was detected in field grown Capsicum annuum from Kununurra in northeast Western Australia. Identification of the Kununurra isolate (WA-99) was confirmed using sap transmission to indicator hosts, positive reactions with tospovirus serogroup IV-specific antibodies and CaCV-specific primers, and amino acid sequence comparisons that showed >97% identity with published CaCV nucleocapsid gene sequences. The reactions of indicator hosts to infection with WA-99 often differed from those of the type isolate from Queensland. The virus multiplied best when test plants were grown at warm temperatures. CaCV was not detected in samples collected in a survey of C. annuum crops planted in the Perth Metropolitan area.

Carrot as a natural host of Watermelon mosaic virus

Carrot was confirmed as a new natural and experimental host of Watermelon mosaic virus by serology, host reactions and sequence comparisons of the coat protein.

Kinetics of interaction of Cotton Leaf Curl Kokhran Virus-Dabawali (CLCuKV-Dab) coat protein and its mutants with ssDNA

Gemini viral assembly and transport of viral DNA into nucleus for replication, ssentially involve DNA-coat protein interactions. The kinetics of interaction of Cotton LeafCtirl Kokhran Virus-Dabawali recombinant coat protein (rCP) with DNA was studied by electrophoretic mobility shift assay (EMSA) and Surface plasmon resonance (SPR). The rCP interacted with ssDNA with a K-A, of 2.6 +/- 0.29 x 10(8) M-1 in a sequence non-specific manner. The CP has a conserved C2H2 type zinc finger motif composed of residues C68, C72, H81 and H85. Mutation of these residues to alanine resulted in reduced binding to DNA probes. The H85A mutant rCP showed the least binding with approximately 756 fold loss in the association rate and a three order magnitude decrease in the binding affinity as compared to rCP. The CP-DNA interactions via the zinc finger motif could play a Crucial role ill Virus assembly and in nuclear transport. (C) 2009 Elsevier Inc.

Herpesviral haematopoietic necrosis virus (CyHV-2) infection: case studies from commercial goldfish farms

Herpesviral haematopoietic necrosis is a disease of goldfish, Carassius auratus, caused by Cyprinid herpesvirus-2 (CyHV-2) infection. Quantitative PCR was carried out on tissue homogenates from healthy goldfish fingerlings, broodfish, eggs and fry directly sampled from commercial farms, from moribund fish submitted to our laboratory for disease diagnosis, and on naturally-infected CyHV-2 carriers subjected to experimental stress treatments. Healthy fish from 14 of 18 farms were positive with copy numbers ranging from tens to 10(7) copies mu g(-1) DNA extracted from infected fish. Of 118 pools of broodfish tested, 42 were positive. The CyHV-2 was detected in one lot of fry produced from disinfected eggs. Testing of moribund goldfish, in which we could not detect any other pathogens, produced 12 of 30 cases with 10(6)-10(8) copies of CyHV-2 mu g(-1) DNA extracted. Subjecting healthy CyHV-2 carriers to cold shock (22-10 degrees C) but not heat, ammonia or high pH, increased viral copy numbers from mean copy number (+/- SE) of 7.3 +/- 11 to 394 +/- 55 mu g(-1) DNA extracted after 24 h. CyHV-2 is widespread on commercial goldfish farms and outbreaks apparently occur when healthy carriers are subjected to a sharp temperature drop followed by holding at the permissive temperature for the disease.

A one-tube fluorescent assay for the quarantine detection and identification of Tilletia indica and other grass bunts in wheat

A molecular assay with enhanced specificity and sensitivity has been developed to assist in the surveillance of Karnal bunt, a quarantineable disease with a significant impact on international trade. The protocol involves the release of DNA from spores, PCR amplification to enrich Tilletia-specific templates from released DNA and a five-plex, real-time PCR assay to detect, identify and distinguish T. indica and other Tilletia species (T. walkeri, T. ehrhartae, T. horrida and a group comprising T. caries, T. laevis, T. contraversa, T. bromi and T. fusca) in wheat grains. This fluorescent molecular tool has a detection sensitivity of one spore and thus bypasses the germination step, which in the current protocol is required for confirmation when only a few spores have been found in grain samples. The assay contains five dual-labelled, species-specific probes and associated species-specific primer pairs in a PCR mix in one tube. The different amplification products are detected simultaneously by five different fluorescence spectra. This specific and sensitive assay with reduced labour and reagent requirements makes it an effective and economically sustainable tool to be used in a Karnal bunt surveillance program. This protocol will also be valuable for the identification of some contaminant Tilletia sp. in wheat grains.

Development of near infrared analysis of faeces to estimate non-grass proportions in diets selected by cattle grazing tropical pastures

Grass (monocots) and non-grass (dicots) proportions in ruminant diets are important nutritionally because the non-grasses are usually higher in nutritive value, particularly protein, than the grasses, especially in tropical pastures. For ruminants grazing tropical pastures where the grasses are C-4 species and most non-grasses are C-3 species, the ratio of C-13/C-12 in diet and faeces, measured as delta C-13 parts per thousand, is proportional to dietary non-grass%. This paper describes the development of a faecal near infrared (NIR) spectroscopy calibration equation for predicting faecal delta C-13 from which dietary grass and non-grass proportions can be calculated. Calibration development used cattle faeces derived from diets containing only C-3 non-grass and C-4 grass components, and a series of expansion and validation steps was employed to develop robustness and predictive reliability. The final calibration equation contained 1637 samples and faecal delta C-13 range (parts per thousand) of [12.27]-[27.65]. Calibration statistics were: standard error of calibration (SEC) of 0.78, standard error of cross-validation (SECV) of 0.80, standard deviation (SD) of reference values of 3.11 and R-2 of 0.94. Validation statistics for the final calibration equation applied to 60 samples were: standard error of prediction (SEP) of 0.87, bias of -0.15, R-2 of 0.92 and RPD of 3.16. The calibration equation was also tested on faeces from diets containing C-4 non-grass species or temperate C-3 grass species. Faecal delta C-13 predictions indicated that the spectral basis of the calibration was not related to C-13/C-12 ratios per se but to consistent differences between grasses and non-grasses in chemical composition and that the differences were modified by photosynthetic pathway. Thus, although the calibration equation could not be used to make valid faecal delta C-13 predictions when the diet contained either C-3 grass or C-4 non-grass, it could be used to make useful estimates of dietary non-grass proportions. It could also be ut :sed to make useful estimates of non-grass in mixed C-3 grass/non-grass diets by applying a modified formula to calculate non-grass from predicted faecal delta C-13. The development of a robust faecal-NIR calibration equation for estimating non-grass proportions in the diets of grazing cattle demonstrated a novel and useful application of NIR spectroscopy in agriculture.

Variation in acquisition of Fiji disease virus by Perkinsiella saccharicida (Hemiptera: Delphacidae).

Fiji leaf gall, caused the Fiji disease virus (genus Fijivirus, family Reoviridae, FDV), is a serious disease of sugarcane, Saccharum officinarum L., in Australia and several other Asia-Pacific countries. In Australia FDV is transmitted only by the planthopper Perkinsiella saccharicida Kirkaldy (Hemiptera: Delphacidae), in a propagative manner. Successful transmission of FDV by single planthoppers confined to individual virus free plants is highly variable, even under controlled conditions. The research reported here addresses two possible sources of this variation: 1) gender, wing form, and life stage of the planthopper; and 2) genotype of the source plant. The acquisition of FDV by macropterous males, macropterous females, brachypterous females, and nymphs of P. saccharicida from infected plants was investigated using reverse transcription-polymerase chain reaction to diagnose FDV infection in the vector. The proportion of individuals infected with FDV was not statistically related to life stage, gender, or adult wing form of the vector. The acquisition of FDV by P. saccharicida from four cultivars of sugarcane was compared to assess the influence of plant genotype on acquisition. Those planthopper populations reared on diseased 'NCo310' plants had twice as many infected planthoppers as those reared on 'Q110', 'WD1', and 'WD2'. Therefore, variation in FDV acquisition in this system is not the result of variation in the gender, wing form and life stage of the P. saccharicida vectors. The cultivar used as the source plant to rear vector populations does affect the proportion of infected planthoppers in a population.

Grader grass (Themeda quadrivalvis): changing savannah ecosystems.

Wayne Vogler and Nikki Owen recently published their paper 'Grader grass (Themeda quadrivalvis): changing savannah ecosystems' in Proceedings of the 16th Australian Weeds Conference. Grader grass is an invasive exotic 'high biomass' grass from India that is increasing its distribution in northern Australia. It is unpalatable and can dominate ecosystems, thereby decreasing grazing animal production, degrading conservation areas and increasing fire intensity and hazard. They studied aspects of its biology at a field site in north Queensland where the initial biomass of the grass layer was found to be 70% grader grass. Grader grass also produced 80% of the seed input into this ecosystem during the first growing season. These factors, in combination with a large viable seed bank and rapid germination at the start of the wet season, demonstrate the potential of grader grass to dominate and degrade the savannah ecosystems of northern Australia.

Surveillance protocols for management of invasive plants: modelling Chilean needle grass (Nassella neesiana) in Australia

Aim: To develop a surveillance support model that enables prediction of areas susceptible to invasion, comparative analysis of surveillance methods and intensity and assessment of eradication feasibility. To apply the model to identify surveillance protocols for generalized invasion scenarios and for evaluating surveillance and control for a context-specific plant invasion. Location: Australia. Methods: We integrate a spatially explicit simulation model, including plant demography and dispersal vectors, within a Geographical Information System. We use the model to identify effective surveillance protocols using simulations of generalized plant life-forms spreading via different dispersal mechanisms in real landscapes. We then parameterize the surveillance support model for Chilean needle grass [CNG; Nassella neesiana (Trin. & Rupr.) Barkworth], a highly invasive tussock grass, which is an eradication target in south-eastern Queensland, Australia. Results: General surveillance protocols that can guide rapid response surveillance were identified; suitable habitat that is susceptible to invasion through particular dispersal syndromes should be targeted for surveillance using an adaptive seek-and-destroy method. The search radius of the adaptive method should be based on maximum expected dispersal distances. Protocols were used to define a surveillance strategy for CNG, but simulations indicated that despite effective and targeted surveillance, eradication is implausible at current intensities. Main conclusions: Several important surveillance protocols emerged and simulations indicated that effectiveness can be increased if they are followed in rapid response surveillance. If sufficient data are available, the surveillance support model should be parameterized to target areas susceptible to invasion and determine whether surveillance is effective and eradication is feasible. We discovered that for CNG, regardless of a carefully designed surveillance strategy, eradication is implausible at current intensities of surveillance and control and these efforts should be doubled if they are to be successful. This is crucial information in the face of environmentally and economically damaging invasive species and large, expensive and potentially ineffective control programmes.

Should the 40-year-old practice of releasing virulent myxoma virus to control rabbits (Oryctolagus cuniculus) be continued?

Release of virulent myxoma virus has been a key component of rabbit-control operations in Queensland, Australia, since the 1960s but its use rests on anecdotal reports. During a routine operation to release virulent myxoma virus we found no evidence to support the continued regular use of the technique in south-west Queensland. Radio-tagged rabbits inoculated with virulent myxoma virus contracted the disease but failed to pass enough virus to other rabbits to spread the disease. Rabbits with clinical signs of myxomatosis that were shot were infected with field strain derived from the original laboratory strain released in 1950 rather than the virulent strain that has been released annually. There was no change in rabbit survival or abundance caused by the release. Nevertheless, the release of virulent virus may be useful against isolated pockets of rabbits mainly because field strains are less likely to be present. Such pockets are more common now that rabbit haemorrhagic disease virus is established in Queensland.

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.

Buffel grass in Queensland's semi-arid woodlands: response to local and landscape scale variables, and relationship with grass, forb and reptile species.

Buffel grass [Pennisetum ciliare (L.) Link] has been widely introduced in the Australian rangelands as a consequence of its value for productive grazing, but tends to competitively establish in non-target areas such as remnant vegetation. In this study, we examined the influence landscape-scale and local-scale variables had upon the distribution of buffel grass in remnant poplar box (Eucalyptus populnea F. Muell.) dominant woodland fragments in the Brigalow Bioregion, Queensland. Buffel grass and variables thought to influence its distribution in the region were measured at 60 sites, which were selected based on the amount of native woodland retained in the landscape and patch size. An information-theoretic modelling approach and hierarchical partitioning revealed that the most influential variable was the percent of retained vegetation within a 1-km spatial extent. From this, we identified a critical threshold of similar to 30% retained vegetation in the landscape, above which the model predicted buffel grass was not likely to occur in a woodland fragment. Other explanatory variables in the model were site based, and included litter cover and long-term rainfall. Given the paucity of information on the effect of buffel grass upon biodiversity values, we undertook exploratory analyses to determine whether buffel grass cover influenced the distribution of grass, forb and reptile species. We detected some trends; hierarchical partitioning revealed that buffel grass cover was the most important explanatory variable describing habitat preferences of four reptile species. However, establishing causal links - particularly between native grass and forb species and buffel grass - was problematic owing to possible confounding with grazing pressure. We conclude with a set of management recommendations aimed at reducing the spread of buffel grass into remnant woodlands.

A review of current knowledge of the weedy species Themeda quadrivalvis (grader grass)

This review of grader grass (Themeda quadrivalvis) attempts to collate current knowledge and identify knowledge gaps that may require further research. Grader grass is a tropical annual grass native to India that is now spread throughout many of the tropical regions of the world. In Australia, it has spread rapidly since its introduction in the 1930s and is now naturalised in the tropical areas of Queensland, the Northern Territory and Western Australia and extends south along the east coast to northern New South Wales. It is a vigorous grass with limited palatability, that is capable of invading native and improved pastures, cropping land and protected areas such as state and national parks. Grader grass can form dense monocultures that reduce biodiversity, decrease animal productivity and increase the fire hazard in the seasonally dry tropics. Control options are based on herbicides, grazing management and slashing, while overgrazing appears to favour grader grass. The effect of fire on grader grass is inconclusive and needs to be defined. Little is known about the biology and impacts of grader grass in agricultural and protected ecosystems in Australia. In particular, information is needed on soil seed bank longevity, seed production, germination and growth, which would allow the development of management strategies to control this weedy grass.