Chemical synthesis and structure elucidation of bovine K-casein (1-44)

The caseins (αs1, αs2, β, and κ) are phosphoproteins present in bovine milk that have been studied for over a century and whose structures remain obscure. Here we describe the chemical synthesis and structure elucidation of the N-terminal segment (1–44) of bovine κ-casein, the protein which maintains the micellar structure of the caseins. κ-Casein (1–44) was synthesised by highly optimised Boc solid-phase peptide chemistry and characterised by mass spectrometry. Structure elucidation was carried out by circular dichroism and nuclear magnetic resonance spectroscopy. CD analysis demonstrated that the segment was ill defined in aqueous medium but in 30% trifluoroethanol it exhibited considerable helical structure. Further, NMR analysis showed the presence of a helical segment containing 26 residues which extends from Pro8 to Arg34. This is the first report which demonstrates extensive secondary structure within the casein class of proteins.

How effective are broad-scale nutrient mass balances for determining the sustainability of lot-feed manure application?

Nutrient mass balances have been used to assess a variety of land resource scenarios, at various scales. They are widely used as a simple basis for policy, planning, and regulatory decisions but it is not clear how accurately they reflect reality. This study provides a critique of broad-scale nutrient mass balances, with particular application to the fertiliser use of beef lot-feeding manure in Queensland. Mass balances completed at the district and farm scale were found to misrepresent actual manure management behaviour and potentially the risk of nutrient contamination of water resources. The difficulties of handling stockpile manure and concerns about soil compaction mean that manure is spread thickly over a few paddocks at a time and not evenly across a whole farm. Consequently, higher nutrient loads were applied to a single paddock less frequently than annually. This resulted in years with excess nitrogen, phosphorus, and potassium remaining in the soil profile. This conclusion was supported by evidence of significant nutrient movement in several of the soil profiles studied. Spreading manure is profitable, but maximum returns can be associated with increased risk of nutrient leaching relative to conventional inorganic fertiliser practices. Bio-economic simulations found this increased risk where manure was applied to supply crop nitrogen requirements (the practice of the case study farms, 200-5000 head lot-feeders). Thus, the use of broad-scale mass balances can be misleading because paddock management is spatially heterogeneous and this leads to increased local potential for nutrient loss. In response to the effect of spatial heterogeneity policy makers who intend to use mass balance techniques to estimate potential for nutrient contamination should apply these techniques conservatively.

Is the distribution of Fiji leaf gall in Australian sugarcane explained by variation in the vector Perkinsiella saccharicida?

Fiji leaf gall (FLG) is an important virally induced disease in Australian sugarcane. It is confined to southern canegrowing areas, despite its vector, the delphacid planthopper Perkinsiella saccharicida, occurring in all canegrowing areas of Queensland and New South Wales. This disparity between distributions could be a result of successful containment of the disease through quarantine and/or geographical barriers, or because northern Queensland populations of Perkinsiella may be poorer vectors of the disease. These hypotheses were first tested by investigating variation in the ITS2 region of the rDNA fragment among eastern Australian and overseas populations of Perkinsiella. The ITS2 sequences of the Western Australian P. thompsoni and the Fijian P. vitiensis were distinguishable from those of P. saccharicida and there was no significant variation among the 26 P. saccharicida populations. Reciprocal crosses of a northern Queensland and a southern Queensland population of P. saccharicida were fertile, so they may well be conspecific. Single vector transmission experiments showed that a population of P. saccharicida from northern Queensland had a higher vector competency than either of two southern Queensland populations. The frequency of virus acquisition in the vector populations was demonstrated to be important in the vector competency of the planthopper. The proportion of infected vectors that transmitted the virus to plants was not significantly different among the populations tested. This study shows that the absence of FLG from northern Queensland is not due to a lack of vector competency of the northern population of P. saccharicida.

The distribution and spread of citrus canker in Emerald, Australia

Citrus canker is a disease of citrus and closely related species, caused by the bacterium Xanthomonas citri subsp. citri. This disease, previously exotic to Australia, was detected on a single farm [infested premise-1, (IP1). IP is the terminology used in official biosecurity protocols to describe a locality at which an exotic plant pest has been confirmed or is presumed to exist. IP are numbered sequentially as they are detected] in Emerald, Queensland in July 2004. During the following 10 months the disease was subsequently detected on two other farms (IP2 and IP3) within the same area and studies indicated the disease first occurred on IP1 and spread to IP2 and IP3. The oldest, naturally infected plant tissue observed on any of these farms indicated the disease was present on IP1 for several months before detection and established on IP2 and IP3 during the second quarter (i.e. autumn) 2004. Transect studies on some IP1 blocks showed disease incidences ranged between 52 and 100% (trees infected). This contrasted to very low disease incidence, less than 4% of trees within a block, on IP2 and IP3. The mechanisms proposed for disease spread within blocks include weather-assisted dispersal of the bacterium (e.g. wind-driven rain) and movement of contaminated farm equipment, in particular by pivot irrigator towers via mechanical damage in combination with abundant water. Spread between blocks on IP2 was attributed to movement of contaminated farm equipment and/or people. Epidemiology results suggest: (i) successive surveillance rounds increase the likelihood of disease detection; (ii) surveillance sensitivity is affected by tree size; and (iii) individual destruction zones (for the purpose of eradication) could be determined using disease incidence and severity data rather than a predefined set area.

Thermal tolerance and potential distribution of Carvalhotingis visenda (Hemiptera: Tingidae), a biological control agent for cat's claw creeper, Macfadyena unguis-cati (Bignoniaceae)

The specialist tingid, Carvalhotingis visenda, is a biological control agent for cat's claw creeper, Macfadyena unguis-cati (Bignoniaceae). Cat's claw creeper is an invasive liana with a wide climatic tolerance, and for biological control to be effective the tingid must survive and develop over a range of temperatures. We evaluated the effect of constant temperatures (0-45°C) on the survival and development of C. visenda. Adults showed tolerance for wider temperature ranges (0-45°C), but oviposition, egg hatching and nymphal development were all affected by both high (>30°C) and low (<20°C) temperatures. Temperatures between 20°C and 30°C are the most favourable for adult survival, oviposition, egg hatching and nymphal development. The ability of adults and nymphs to survive for a few days at high (40°C and 45°C) and low (0°C and 5°C) temperatures suggest that extreme temperature events, which usually occur for short durations (hours) in cat's claw creeper infested regions in Queensland and New South Wales states are not likely to affect the tingid population. The potential number of generations (egg to adult) the tingid can complete in a year in Australia ranged from three to eight, with more generations in Queensland than in New South Wales.

Taxonomy, distribution and ecology of Zoysia macrantha desv., an Australian native species with turf breeding potential

Only three of the 11 species in the genus Zoysia Willd. have thus far contributed to commercially available turfgrass varieties. One of the neglected taxa is Z. macrantha Desv., an Australian native species further divided into two subspecies. The coarser Z. macrantha subsp. macrantha occurs on sand dunes, headlands and tidal areas along eastern and southeastern coasts from about 23 to 38°S latitude. The shorter, denser-growing Z. macrantha subsp. walshii M.E. Nightingale is found on the southern mainland (South Australia and Victoria from longitude 137° to 148°E and at latitudes higher than 36°S), adjacent offshore islands, and northern, eastern and central Tasmania to 43°S growing on the edges of coastal, sub-coastal and even inland salt lakes, in riverine environments, and from moist grassy depressions (both coastal and inland) to rocky headlands. The latter subspecies has the more discontinuous and specialised distribution, largely determined by the need for an appropriate level of peat, clay or silt in the soil to maintain adequate moisture during the dry summers in southern Australia while at the same time avoiding anything more than temporary waterlogging. It grows on low fertility soils ranging from strongly acid to neutral or mildly alkaline, and is often very closely grazed by marsupials. Both subspecies are salt and drought tolerant, but not notably shade tolerant. Their potential to add greater drought tolerance in particular to the Asian Zoysia material in current use through future breeding programs is discussed.

Mile-a-minute (Mikania micrantha): its distribution, growth and physical and socio-economic impacts in Papua New Guinea

Mikania micrantha, Kunth. H.B.K (Asteraceae) or mile-a-minute is a weed of Neotropical origin in 17 Pacific Island countries. It is becoming increasingly regarded as an invasive weed in Papua New Guinea and is now the focus of an Australian Government-funded biological control program. As part of the program, growth rates, distribution and physical and socia-economic impacts were studied to obtain baseline data and to assist with the field release of biological control agents. Through public awareness campaigns and dedicated surveys, mikania has been reported in most lowland provinces. It is particularly widespread in East New Britain and West New Britain Province. In field trials, mikania grew more than 1 metre per month in open sunny areas but slightly slower when growing under cocoa. The weed invades a wide range of land types, impacting on plantations and food gardens, smothering pawpaw, young cocoa, banana, taro, young oil palms and ornamental plants. In socia-economic surveys, mikania was found to have severe impacts on crop production and income generated through reduced yields and high weeding costs. These studies suggest that there would be substantial benefits to the community if biological control of mikania is successful.

Mikania micrantha Kunth (Asteraceae) (Mile-a-Minute): Its Distribution and Physical and Socioeconomic Impacts in Papua New Guinea

Mikania micrantha or mile-a-minute is regarded as a major invasive weed in Papua New Guinea (PNG) and is now the target of a biological control program. As part of the program, distribution and physical and socioeconomic impacts of M. micrantha were studied to obtain baseline data and to assist with field release of biological control agents. Through public awareness campaigns and dedicated surveys, M. micrantha has been reported in all 15 lowland provinces. It is particularly widespread in East New Britain, as well as in West New Britain and New Ireland. A CLIMEX model suggests that M. micrantha has the potential to continue to spread throughout all lowland areas in PNG. The weed was found in a wide range of land uses, impacting on plantations and food gardens and smothering papaya, young cocoa, banana, taro, young oil palms, and ornamental plants. In socioeconomic surveys, M. micrantha was found to have severe impacts on crop production and income generated through reduced yields and high weeding costs, particularly in subsistence mixed cropping systems. About 89% of all respondents had M. micrantha on their land, and 71% of respondents had to weed monthly. Approximately 96% of respondents in subsistence mixed cropping systems used only physical means of control compared with 68% of respondents in other farming systems. About 45% of all respondents estimated that M. micrantha causes yield losses in excess of 30%. These studies suggest that there would be substantial benefits to landholders if biological control of M. micrantha were to be successful.

The Distribution of Henipaviruses in Southeast Asia and Australasia: Is Wallace's Line a Barrier to Nipah Virus?

Nipah virus (NiV) (Genus Henipavirus) is a recently emerged zoonotic virus that causes severe disease in humans and has been found in bats of the genus Pteropus. Whilst NiV has not been detected in Australia, evidence for NiV-infection has been found in pteropid bats in some of Australia's closest neighbours. The aim of this study was to determine the occurrence of henipaviruses in fruit bat (Family Pteropodidae) populations to the north of Australia. In particular we tested the hypothesis that Nipah virus is restricted to west of Wallace's Line. Fruit bats from Australia, Papua New Guinea, East Timor and Indonesia were tested for the presence of antibodies to Hendra virus (HeV) and Nipah virus, and tested for the presence of HeV, NiV or henipavirus RNA by PCR. Evidence was found for the presence of Nipah virus in both Pteropus vampyrus and Rousettus amplexicaudatus populations from East Timor. Serology and PCR also suggested the presence of a henipavirus that was neither HeV nor NiV in Pteropus alecto and Acerodon celebensis. The results demonstrate the presence of NiV in the fruit bat populations on the eastern side of Wallace's Line and within 500 km of Australia. They indicate the presence of non-NiV, non-HeV henipaviruses in fruit bat populations of Sulawesi and Sumba and possibly in Papua New Guinea. It appears that NiV is present where P. vampyrus occurs, such as in the fruit bat populations of Timor, but where this bat species is absent other henipaviruses may be present, as on Sulawesi and Sumba. Evidence was obtained for the presence henipaviruses in the non-Pteropid species R. amplexicaudatus and in A. celebensis. The findings of this work fill some gaps in knowledge in geographical and species distribution of henipaviruses in Australasia which will contribute to planning of risk management and surveillance activities.

Determining changes in the distribution and abundance of a Rhyzopertha dominica phosphine resistance allele in farm grain storages using a DNA marker

BACKGROUND: The lesser grain borer, Rhyzopertha dominica (F.), is a highly destructive pest of stored grain that is strongly resistant to the fumigant phosphine (PH3). Phosphine resistance is due to genetic variants at the rph2 locus that alter the function of the dihydrolipoamide dehydrogenase (DLD) gene. This discovery now enables direct detection of resistance variants at the rph2 locus in field populations. RESULTS: A genotype assay was developed for direct detection of changes in distribution and frequency of a phosphine resistance allele in field populations of R. dominica. Beetles were collected from ten farms in south-east Queensland in 2006 and resampled in 2011. Resistance allele frequency increased in the period from 2006 to 2011 on organic farms with no history of phosphine use, implying that migration of phosphine-resistant R. dominica had occurred from nearby storages. CONCLUSION: Increasing resistance allele frequencies on organic farms suggest local movement of beetles and dispersal of insects from areas where phosphine has been used. This research also highlighted for the first time the utility of a genetic DNA marker in accurate and rapid determination of the distribution of phosphine-resistant insects in the grain value chain. Extending this research over larger landscapes would help in identifying resistance problems and enable timely pest management decisions. © 2013 Society of Chemical Industry © 2013 Society of Chemical Industry 69 6 June 2013 10.1002/ps.3514 Rapid Report Rapid Report © 2013 Society of Chemical Industry.

Strong resistance to phosphine in the rusty grain beetle, Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae): its characterisation, a rapid assay for diagnosis and its distribution in Australia

BACKGROUND: The recent development of very high resistance to phosphine in rusty grain beetle, Cryptolestes ferrugineus (Stephens), seriously threatens stored-grain biosecurity. The aim was to characterise this resistance, to develop a rapid bioassay for its diagnosis to support pest management and to document the distribution of resistance in Australia in 20072011. RESULTS: Bioassays of purified laboratory reference strains and field-collected samples revealed three phenotypes: susceptible, weakly resistant and strongly resistant. With resistance factors of > 1000 x , resistance to phosphine expressed by the strong resistance phenotype was higher than reported for any stored-product insect species. The new time-to-knockdown assay rapidly and accurately diagnosed each resistance phenotype within 6 h. Although less frequent in western Australia, weak resistance was detected throughout all grain production regions. Strong resistance occurred predominantly in central storages in eastern Australia. CONCLUSION: Resistance to phosphine in the rusty grain beetle is expressed through two identifiable phenotypes: weak and strong. Strong resistance requires urgent changes to current fumigation dosages. The development of a rapid assay for diagnosis of resistance enables the provision of same-day advice to expedite resistance management decisions. (c) 2012 Commonwealth of Australia. Published by John Wiley & Sons, Ltd.

Puccinia psidii in Queensland, Australia: disease symptoms, distribution and impact

Puccinia psidii has long been considered a significant threat to Australian plant industries and ecosystems. In April 2010, P. psidii was detected for the first time in Australia on the central coast of New South Wales (NSW). The fungus spread rapidly along the east coast and in December 2010 was found in Queensland (Qld) followed by Victoria a year later. Puccinia psidii was initially restricted to the southeastern part of Qld but spread as far north as Mossman. In Qld, 48 species of Myrtaceae are considered highly or extremely susceptible to the disease. The impact of P. psidii on individual trees and shrubs has ranged from minor leaf spots, foliage, stem and branch dieback to reduced fecundity. Tree death, as a result of repeated infection, has been recorded for Rhodomyrtus psidioides. Rust infection has also been recorded on flower buds, flowers and fruits of 28 host species. Morphological and molecular characteristics were used to confirm the identification of P. psidii from a range of Myrtaceae in Qld and compared with isolates from NSW and overseas. A reconstructed phylogeny based on the LSU and SSU regions of rDNA did not resolve the familial placement of P. psidii, but indicated that it does not belong to the Pucciniaceae. Uredo rangelii was found to be con-specific with all isolates of P. psidii in morphology, ITS and LSU sequence data, and host range.

Dietary media for mass rearing of rusty grain beetle, Cryptolestes ferrugineus (Stephens) and flat grain beetle, Cryptolestes pusillus (Schonherr) (Coleoptera: Cucujidae)

We developed a suitable diet for mass rearing of Cryptolestes ferrugineus (Stephens) populations under laboratory conditions. Recently, this pest has developed strong level of resistance to phosphine in Australia, and therefore, a significant amount of research has been directed towards its management. In total, nineteen grain-based diets, containing rolled oats, various combinations of cracked grains and flours of wheat, sorghum, maize and barley were tested. Each diet contained a small proportion of wheat germ (4.5% w/w) and torula yeast (0.5% w/w). Experiments were conducted at fixed temperature and relative humidity regimes of 30 ± 2 °C and 70 ± 2%, respectively, and replicated three times. Adults (n = 40) of a laboratory strain of C. ferrugineus were introduced into each diet, removed after 14 days and total numbers of live adult progeny were recorded. The following diets resulted in highest live progeny production: barley flour (95%) (607.67 ± 11.21) = rolled oats (75%) + cracked sorghum (20%) (597.33 ± 33.79) ≥ wheat flour (47.5%) + barley flour (47.5%) (496.67 ± 52.93) > cracked sorghum (95%) (384.00 ± 60.66). The performance of these four diets was then tested with field-collected populations of C. ferrugineus and Cryptolestes pusillus (Schonherr). The diets based on rolled oats + cracked sorghum, wheat flour + barley flour, and barley flour alone consistently produced highest progeny numbers in field-collected populations of both species, with mean progeny numbers ranging from 359.9 to 478.5. The multiplication of C. pusillus was significantly higher than C. ferrugineus on all four diets. Our findings will help in mass rearing of healthy cultures of C. ferrugineus and C. pusillus that will greatly facilitate laboratory and field research and in particular, in developing management tactics for these species.

Cotton root systems and recovery of applied P and K fertilisers

With potential to accumulate substantial amounts of above-ground biomass, at maturity an irrigated cotton crop can have taken up more than 20 kg/ha phosphorus and often more than 200 kg/ha of potassium. Despite the size of plant accumulation of P and K, recovery of applied P and K fertilisers by the crop in our field experiment program has poor. Processing large amounts of mature cotton plant material to provide a representative sample for chemical analysis has not been without its challenges, but the questions regarding mechanism of where, how and when the plant is acquiring immobile nutrients remain. Dry matter measured early in the growing season (squaring, first white flower) have demonstrated a 50% increase in crop biomass to applied P (in particular), but it represents only 20% of the total P accumulation by the plant. By first open boll (and onwards), no response in dry matter or P concentration could be detected to P application. A glasshouse study indicated P recovery was greater (to FOB) where it was completely mixed through a profile as opposed to a banded application method suggesting cotton prefers a more diffuse distribution. The relative effects of root morphology, mycorrhizal fungi infection, seasonal growth patterns and how irrigation is applied are areas for future investigation on how, when and where cotton acquires immobile nutrients.