A high-density consensus map of barley linking DArT markers to SSR, RFLP and STS loci and agricultural traits

Background: Molecular marker technologies are undergoing a transition from largely serial assays measuring DNA fragment sizes to hybridization-based technologies with high multiplexing levels. Diversity Arrays Technology (DArT) is a hybridization-based technology that is increasingly being adopted by barley researchers. There is a need to integrate the information generated by DArT with previous data produced with gel-based marker technologies. The goal of this study was to build a high-density consensus linkage map from the combined datasets of ten populations, most of which were simultaneously typed with DArT and Simple Sequence Repeat (SSR), Restriction Enzyme Fragment Polymorphism (RFLP) and/or Sequence Tagged Site (STS) markers. Results: The consensus map, built using a combination of JoinMap 3.0 software and several purpose-built perl scripts, comprised 2,935 loci (2,085 DArT, 850 other loci) and spanned 1,161 cM. It contained a total of 1,629 'bins' (unique loci), with an average inter-bin distance of 0.7 ± 1.0 cM (median = 0.3 cM). More than 98% of the map could be covered with a single DArT assay. The arrangement of loci was very similar to, and almost as optimal as, the arrangement of loci in component maps built for individual populations. The locus order of a synthetic map derived from merging the component maps without considering the segregation data was only slightly inferior. The distribution of loci along chromosomes indicated centromeric suppression of recombination in all chromosomes except 5H. DArT markers appeared to have a moderate tendency toward hypomethylated, gene-rich regions in distal chromosome areas. On the average, 14 ± 9 DArT loci were identified within 5 cM on either side of SSR, RFLP or STS loci previously identified as linked to agricultural traits. Conclusion: Our barley consensus map provides a framework for transferring genetic information between different marker systems and for deploying DArT markers in molecular breeding schemes. The study also highlights the need for improved software for building consensus maps from high-density segregation data of multiple populations.

Increasing the productivity of avocado orchards using high-density plantings: A review

There is strong interest in the use of high-density plantings to increase the productivity of avocado (Persea americana) orchards. Close plantings have the potential for higher yields and returns than standard or traditional plantings, especially in the early years of production. The success of this technology is dependent on the use of methods to control shoot growth and maximise light interception as the trees begin to bear fruit. We reviewed the performance of high-density orchards in different environments, and the success of efforts to control the growth of the trees through the use of dwarfing material, canopy management and growth regulators. Close plantings generally produce higher yields in the first few years of bearing compared with the yields of standard plantings. However, in most growing areas, the trees in the close plantings soon begin to crowd each other and yields decline. This usually occurs despite efforts to control shoot growth by pruning the trees or by applying growth regulators. Efforts to breed dwarfing rootstocks that can control the growth of mature trees have been largely unsuccessful. In the absence of dwarfing material, effective canopy management appears to be the largest barrier to success of high-density orchards. Further research on the use of different pruning strategies and growth regulators to control the growth of the trees and maximise light interception is required. There are potential problems with some of the growth regulators persisting in the harvested fruit and soil under certain circumstances.

Evaluation of exclusion netting for insect pest control and fruit quality enhancement in tree crops

This work evaluated the following aspects of the use of exclusion netting in low chill stone fruit: the efficacy of protection from fruit fly for this highly susceptible crop; the effects on environmental factors; and the effects on crop development. Concurrently, an economic viability study on the use of exclusion netting was undertaken. The trial site was a 0.6-ha block of low chill stone fruit at Nambour, south-east Queensland, Australia. In this area, populations of Queensland fruit fly (Bactrocera tryoni) are known to be substantial, particularly in spring and summer. The trial block contained healthy 4-year-old trees as follows: 96 peach trees (Prunus persica cv. Flordaprince) and 80 nectarine trees (40 P. persica var. nucipersica cv. White Satin and 40 P. persica var. nucipersica cv. Sunwright). Exclusion netting was installed over approximately half of the block in february 2001. The net was a UV-stabilized structural knitted fabric made from high-density polyethylene yarn with a 10-year prorated UV degradation warranty. The results demonstrated the efficacy of exclusion netting in the control of fruit flies. Exclusion netting increased maximum temperatures by 4.4 deg C and decreased minimum temperatures by 0.5 deg C. Although exclusion netting reduced irradiance by approximately 20%, it enhanced fruit development by 7-10 days and improved fruit quality by increasing sugar concentration by 20-30% and colour intensity by 20%.

Effects of added shelter and stocking density on growth of sleepy cod Oxyeleotris lineolatus in ponds

Sleepy cod Oxyeleotris lineolatus is a species of freshwater goby in demand in Australian markets by consumers of Asian origin. It is related to marble goby Oxyeleotris marmoratus, the most expensive freshwater food fish in Asia, which is cultured throughout southeast Asia in ponds and cages. The performance of sleepy cod in culture conditions was investigated to assess the viability of farming them in northern Australia. Sleepy cod fingerlings (62.8 +/- 0.8 mm total length and 2.56 +/- 0.095 g) were stocked into experimental ponds at 32,857 fish/ha, and grown out for 8 mo. Shelter was provided in each of three replicate ponds and was absent in three control ponds. The provision of shelter in juvenile growout was found to be of no benefit, although fish in ponds provided with shelter weighed slightly more per unit length than fish in ponds without shelter. Cannibalism was not a problem in growout, and survival was close to 100%. After the shelter trial was completed, fish were graded into large and small classes (three replicates of each), and grown out without shelter at the same density for 158 d. Following that, fish were again graded, and the largest 30% retained from growout at a density of 8,857 fish/ha (large, 198 +/-6.44 g) or 10,000 fish/ha (small, 48.9 +/-1.27 g). These were grown out for 188 d. Growth of selected stock at low densities was slower than earlier growth rates, although smaller fish gained weight more rapidly than larger fish. Growth rates were better than the only published data for marble goby. Further investigation into high density culture and different genotypes of sleepy cod needs to be undertaken to determine the viability of pond culture.

Row spacing and planting density effects on the growth and yield of sugarcane. 1. Responses in fumigated and non-fumigated soil.

It has been reported that high-density planting of sugarcane can improve cane and sugar yield through promoting rapid canopy closure and increasing radiation interception earlier in crop growth. It is widely known that the control of adverse soil biota through fumigation (removes soil biological constraints and improves soil health) can improve cane and sugar yield. Whether the responses to high-density planting and improved soil health are additive or interactive has important implications for the sugarcane production system. Field experiments established at Bundaberg and Mackay, Queensland, Australia, involved all combinations of 2-row spacings (0.5 and 1.5 m), two planting densities (27 000 and 81 000 two-eyed setts/ha), and two soil fumigation treatments (fumigated and non-fumigated). The Bundaberg experiment had two cultivars (Q124, Q155), was fully irrigated, and harvested 15 months after planting. The Mackay experiment had one cultivar (Q117), was grown under rainfed conditions, and harvested 10 months after planting. High-density planting (81 000 setts/ha in 0.5-m rows) did not produce any more cane or sugar yield at harvest than low-density planting (27 000 setts/ha in 1.5-m rows) regardless of location, crop duration (15 v. 10 months), water supply (irrigated v. rainfed), or soil health (fumigated v. non-fumigated). Conversely, soil fumigation generally increased cane and sugar yields regardless of site, row spacing, and planting density. In the Bundaberg experiment there was a large fumigation x cultivar x density interaction (P<0.01). Cultivar Q155 responded positively to higher planting density in non-fumigated soil but not in fumigated soil, while Q124 showed a negative response to higher planting density in non-fumigated soil but no response in fumigated soil. In the Mackay experiment, Q117 showed a non-significant trend of increasing yield in response to increasing planting density in non-fumigated soil, similar to the Q155 response in non-fumigated soil at Bundaberg. The similarity in yield across the range of row spacings and planting densities within experiments was largely due to compensation between stalk number and stalk weight, particularly when fumigation was used to address soil health. Further, the different cultivars (Q124 and Q155 at Bundaberg and Q117 at Mackay) exhibited differing physiological responses to the fumigation, row spacing, and planting density treatments. These included the rate of tiller initiation and subsequent loss, changes in stalk weight, and propensity to lodging. These responses suggest that there may be potential for selecting cultivars suited to different planting configurations.

Effect of Packaging Materials and Storage on Major Volatile Compounds in Three Australian Native Herbs

Lemon myrtle, anise myrtle, and Tasmanian pepper leaf are commercial Australian native herbs with a high volatile or essential oil content. Packaging of the herbs in high- or low-density polyethylene (HDPE and LDPE) has proven to be ineffective in preventing a significant loss of volatile components on storage. This study investigates and compares the effectiveness of alternate high-barrier property packaging materials, namely, polyvinylidene chloride coated polyethylene terephthalate/casted polypropylene (PVDC coated PET/CPP) and polyethylene terephthalate/polyethylene terephthalate/aluminum foil/linear low-density polyethylene (PET/PET/Foil/LLDPE), in prevention of volatile compound loss from the three native herbs stored at ambient temperature for 6 months. Concentrations of major volatiles were monitored using gas chromatography?mass spectrometry (GC-MS) techniques. After 6 months of storage, the greatest loss of volatiles from lemon myrtle was observed in traditional LDPE packaging (87% loss) followed by storage in PVDC coated PET/CPP (58% loss) and PET/PET/Foil/LLDPE (loss of 23%). The volatile loss from anise myrtle and Tasmanian pepper leaf stored in PVDC coated PET/CPP and PET/PET/Foil/LLDPE packaging was <30%. This study clearly indicates the importance of selecting the correct packaging material to retain the quality of herbs with high volatile content.

Modified atmosphere packaging effects on the postharvest quality of papaya fruit

Export of Fijian papaya (Carica papaya) fruit to destinations such as New Zealand has increased significantly over the last several years. Shipment by sea rather than air is the preferred method, given the capacity for larger volumes and reductions in cost. Long shipping times, however, can compromise fruit quality, although the use of modified atmosphere packaging (MAP) may provide a viable solution for extending fruit storage life. In a collaborative ACIAR project, Australian and Fijian researchers investigated the potential of using MAP to extend storage life of a Fijian papaya ('Fiji Red') fruit based on simulated sea transport conditions. Fruit were packed in one of three MAP environments within cartons, consisting of either a (1) Low Density Polyethylene (LDPE) bag with 10 g of KMnO4, (2) Polyamide Film (PF) bag with macro-perforations or (3) without a bag (control fruit). Fruit were held for 1, 2 or 3 weeks at 10°C before being unpacked, ripened and assessed for quality. On day 6 after outturn, fruit with the highest overall quality were those held in LDPE bags. LDPE fruit generally coloured up faster at outturn than PF or control fruit, had less overall moisture loss and scored high in flavour. Headspace carbon dioxide and oxygen concentrations within the LDPE bags were also near recommended levels for maintaining optimum storage-life quality. The LDPE bag provided the most suitable conditions for long term storage of fresh papaya fruit and is therefore the recommended MAP type for use with sea freight export out of Fiji.

New tools for management of phosphine resistance

Phosphine is the primary fumigant used to protect the majority of the world' s grain and a variety of other stored commodities from insect pests. Phosphine is playing an increasingly important role in the protection of commodities for two primary reasons. Firstly, use of the alternative fumigant, methyl bromide, has been sharply curtailed and is tightly regulated due to its role in ozone depletion, and secondly, consumers are becoming increasingly intolerant of contact pesticides. Niche alternatives to phosphine exist, but they suffer from a range of factors that limit their use, including: 1) Limited commercial adoption due to expense or slow mode of action; 2) Poor efficacy due to low toxicity, rapid sorption, limited volatility or high density; 3) Public health concerns due to toxicity to handlers or nearby residents, as well as risk of explosion; 4) Poor consumer acceptance due to toxic residues or smell. These same factors limit the prospects of quickly identifying and deploying a new fumigant. Given that resistance toward phosphine is increasing among insect pests, improved monitoring and management of resistance is a priority. Knowledge of the mode of action of phosphine as well as the mechanisms of resistance may also greatly reduce the effort and expense of identifying synergists or novel replacement compounds.

Sampling and management of Bemisia tabaci (Genn.) biotype B in Australian cotton

Data on seasonal population abundance of Bemisia tabaci biotype B (silverleaf whitefly (SLW)) in Australian cotton fields collected over four consecutive growing seasons (2002/2003-2005/2006) were used to develop and validate a multiple-threshold-based management and sampling plan. Non-linear growth trajectories estimated from the field sampling data were used as benchmarks to classify adult SLW field populations into six density-based management zones with associated control recommendations in the context of peak flowering and open boll crop growth stages. Control options based on application of insect growth regulators (IGRs) are recommended for high-density populations (>2 adults/leaf) whereas conventional (non-IGR) products are recommended for the control of low to moderate population densities. A computerised re-sampling program was used to develop and test a binomial sampling plan. Binomial models with thresholds of T=1, 2 and 3 adults/leaf were tested using the field abundance data. A binomial plan based on a tally threshold of T=2 adults/leaf and a minimum sample of 20 leaves at nodes 3, 4 or 5 below the terminal is recommended as the most parsimonious and practical sampling protocol for Australian cotton fields. A decision support guide with management zone boundaries expressed as binomial counts and control options appropriate for various SLW density situations is presented. Appropriate use of chemical insecticides and tactics for successful field control of whiteflies are discussed.

Control of rabbits in arid Australia: destroying the drought refuge

Rabbits continued to infest Bulloo Downs in southwest Queensland even after rabbit haemorrhagic disease virus (RHDV) had effectively reduced rabbit populations to very low levels in most other arid parts of Australia. Control efforts for over 100 years have all appeared unable to stop rabbits causing damage to cattle production and native plants and animals in the area. In 2001 an experiment established to measure the benefit of rabbit control to biodiversity and cattle production showed warren ripping to cause an immediate reduction in rabbit activity. Three months after ripping there were still 98% fewer rabbits in ripped plots despite these plots being exposed to invasion from surrounding populations. The cost of ripping was high because of the high density of warrens and is prohibitive for a full-scale programme. Nevertheless, ripping warrens just in the rabbit’s drought refuge (2002 -2004) appears to have effectively controlled rabbits over the entire property. Following one good season rabbits still have not recovered where the drought refuge was effectively ripped. Destroying warrens in the areas where rabbits survived droughts achieved a reduction in rabbits of over 99% ompared to a similar area near Coongie Lakes in South Australia. Low rabbit numbers allowed cattle to continue to be run on the property even though the area experienced seven consecutive years with below average rainfall. It still remains to be seen whether rabbits can recover from this low population-base during a run of good seasons. If rabbit numbers remain suppressed after a run of good seasons then rabbit control by destruction of drought refuge could be repeated at Coongie Lakes and other drought refuge areas in the arid zone. Identification and treatment of areas similar to Bulloo Downs where rabbits survive drought may relieve a very large area of arid Australia from the damage caused by rabbits.

The value of having no wild rabbits in south-east Queensland

Rabbits released in Australia in 1859 spread to most areas of suitable habitat by 1910 causing great damage to the environment and primary industries. Measurement of damage is essential to justify spending money and utilising resources to remove rabbits. Damage to pasture and biodiversity may be irreversible and therefore difficult to measure without comparison with an area that has never suffered such damage. A rabbit proof fence completed in 1906 protected a large part of south east Queensland from rabbits. The Darling Downs Moreton Rabbit Board (DDMRB) continues to maintain the fence and keep the area relatively free of rabbits. This area is unique because it is highly suitable for rabbits and yet it has never ‘experienced’ the damage caused by plagues of uncontrolled rabbits. A study site was established where the DDMRB fence separates an area heavily used by rabbits (‘dirty side’) from an area that has never been infested by rabbits (‘clean side’). The number and location of all rabbit warrens and log piles were recorded. The absence of warrens from the ‘clean side’ shows clearly that the rabbit proof fence has prevented rabbits from establishing warren systems. The ‘dirty side’ is characterised by a high number of warrens, a high density of rabbits, fewer pasture species and low macropod activity. Future work will determine whether the rabbit populations are viable in the absence of rabbit warrens. We plan to radio collar rabbits on both sides of the fence to measure their survival rate. In selected warrens and log piles of varying degrees of complexity and size, rabbits will be trapped and information on reproduction and age structure will be collected. This will allow better targeting of the source of rabbits during control operations. Once the initial comparative analysis of the site has been completed, all rabbit warrens will be destroyed on the dirty side of the fence. After rabbits are removed from this area, monitoring will continue to determine if pasture and biodiversity on opposite sides of the fence begin to mirror each other.

Bats and Emerging Zoonoses: Henipaviruses and SARS.

Nearly 75% of all emerging infectious diseases (EIDs) that impact or threaten human health are zoonotic. The majority have spilled from wildlife reservoirs, either directly to humans or via domestic animals. The emergence of many can be attributed to predisposing factors such as global travel, trade, agricultural expansion, deforestation habitat fragmentation, and urbanization; such factors increase the interface and or the rate of contact between human, domestic animal, and wildlife populations, thereby creating increased opportunities for spillover events to occur. Infectious disease emergence can be regarded as primarily an ecological process. The epidemiological investigation of EIDs associated with wildlife requires a trans-disciplinary approach that includes an understanding of the ecology of the wildlife species, and an understanding of human behaviours that increase risk of exposure. Investigations of the emergence of Nipah virus in Malaysia in 1999 and severe acute respiratory syndrome (SARS) in China in 2003 provide useful case studies. The emergence of Nipah virus was associated with the increased size and density of commercial pig farms and their encroachment into forested areas. The movement of pigs for sale and slaughter in turn led to the rapid spread of infection to southern peninsular Malaysia, where the high-density, largely urban pig populations facilitated transmission to humans. Identifying the factors associated with the emergence of SARS in southern China requires an understanding of the ecology of infection both in the natural reservoir and in secondary market reservoir species. A necessary extension of understanding the ecology of the reservoir is an understanding of the trade, and of the social and cultural context of wildlife consumption. Emerging infectious diseases originating from wildlife populations will continue to threaten public health. Mitigating and managing the risk requires an appreciation of the connectedness between human, livestock and wildlife health, and of the factors and processes that disrupt the balance.

The physiological and behavioral responses of steers to gaseous ammonia in simulated long distance transport by ship.

Ammonia (NH3) can accumulate in high density cattle accommodation during live export shipments and could potentially threaten the animals' health and welfare. The effects of 4 NH3 concentrations, control (<8), 15, 30, and 45 ppm, on the physiology and behavior of steers were recorded. The animals were held for 12 d under a micro-climate and stocking density similar to shipboard conditions experienced on voyages from Australia to the Middle East during the northern hemispheric summer. In bronchoalveolar lavage samples, ammonia increased (P < 0.05) macrophage activity in proportion to NH3 concentration and it increased (P < 0.05) neutrophil percentage at 30 and 45 ppm, indicating active pulmonary inflammation. It also increased (P < 0.05) lacrimation, nasal secretions and coughing, particularly at 45 ppm, indicating that the NH3 was irritating the mucous membranes of the eyes, nasal cavity and respiratory tract. Ammonia had no effect (P > 0.05) on hematological parameters or body weight. Twenty-eight days after exposure to NH3, the steers' pulmonary macrophage activity and neutrophil levels had returned to normal. It was concluded that ammonia concentrations of 30 and 45 ppm induced temporary inflammatory responses which indicate an adverse effect on the welfare of steers.

Polychaete-assisted sand filters – prawn farm wastewater remediation trial

Medium bedding sand which is commonly available in coastal sedimentary deposits, and a marine polychaete-worm species from Moreton Bay recently classified as Perinereis helleri (Nereididae), were deployed in a simple low-maintenance sand filter design that potentially has application at large scale. Previous work had shown that this physical and biological combination can provide a new option for saline wastewater treatment, since the worms help to prevent sand filter blocking with organic debris and offer a profitable by-product. To test the application of this new concept in a commercial environment, six 1.84 m2 Polychaete-assisted sand filters were experimentally tested for their ability to treat wastewater from a semi-intensive prawn culture pond. Polychaetes produced exclusively on the waste nutrients that collected in these gravity-driven sand filters were assessed for their production levels and nutritional contents. Water parameters studied included temperature, salinity, pH, dissolved oxygen (DO), oxidation/ reduction potential (redox), suspended solids, chlorophyll a, biological oxygen demand (BOD), and common forms of nitrogen and phosphorus. Pond water which had percolated through the sand bed had significantly lower pH, DO and redox levels compared with inflow water. Suspended solids and chlorophyll a levels were consistently more than halved by the process. Reductions in BOD appeared dependant on regular subsurface flows. Only marginal reductions in total nitrogen and phosphorus were documented, but their forms were altered in a potentially useful way: dissolved forms (ammonia and orthophosphate) were generated by the process, and this remineralisation also seemed to be accentuated by intermittent flow patterns. Flow rates of approximately 1,500 L m-2 d-1 were achieved suggesting that a 1 ha polychaete bed of this nature could similarly treat the discharge from a 10 ha semi-intensive prawn farm. Sixteen weeks after stocking sand beds with one-month-old P. helleri, over 3.6 kg of polychaete biomass (wet weight) was recovered from the trial. Production on a sand bed area basis was 328 g m-2. Similar (P>0.05) overall biomass production was found for the two stocking densities tested (2000 and 6000 m-2; n = 3), but survival was lower and more worms were graded as small (<0.6 g) when produced at the higher density (28.2 ± 1.5 % and approx. 88 %, respectively) compared with the lower density (46.8 ± 4.4 % and approx. 76 %, respectively). When considered on a weight for weight basis, about half of the worm biomass produced was generally suitable for use as bait. The nutritional contents of the worms harvested were analysed for different stocking densities and graded sizes. These factors did not significantly affect their percentages of dry matter (DM) (18.23 ± 0.57 %), ash (19.77 ± 0.80 % of DM) or gross energy 19.39 ± 0.29 MJ kg-1 DM) (n = 12). Although stocking density did not affect the worms’ nitrogen and phosphorus contents, small worms had a higher mean proportion of nitrogen and phosphorus (10.57 ± 0.17 % and 0.70 ± 0.01 % of DM, respectively) than large worms (9.99 ± 0.12 % and 0.65 ± 0.01 % of DM, respectively) (n = 6). More lipid was present in large worms grown at the medium density (11.20 ± 0.19 %) compared with the high density (9.50 ± 0.31 %) and less was generally found in small worms (7.1-7.6 % of DM). Mean cholesterol and total phospholipid levels were 5.24 ± 0.15 mg g-1 and 13.66 ± 2.15 mg g-1 DM, respectively (n = 12). Of the specific phospholipids tested, phosphatidyl-serine or sphingomyelin were below detection limits (<0.05 mg g-1), whilst mean levels of phosphatidyl-ethanolamine, phosphatidyl-inositol, phosphatidyl-choline and lysophosphatidyl-choline were 6.89 ± 1.09, 0.89 ± 0.26, 4.04 ± 1.17 and 1.84 ± 0.37 mg g-1, respectively (n = 12). Culture density generally had a more pronounced effect on phospholipid contents than did size of worms. By contrast, worm size had a more pronounced effect on total fatty acid contents, with large worms containing significantly higher (P<0.001) levels on a DM basis (46.88 ± 2.46 mg g-1) than smaller worms (27.76 ± 1.28 mg g-1). A very broad range of fatty acids were detected with palmitic acid being the most heavily represented class (up to 14.23 ± 0.49 mg g-1 DM or 27.28 ± 0.22 % of total fatty acids). Other heavily represented classes included stearic acid (7.4-8.8 %), vaccenic acid (6.8-7.8 %), arachidonic acid (3.5-4.4 %), eicosapentaenoic acid (9.9-13.8 %) and docosenoic acid (5.7-7.0 %). Stocking density did not affect (P>0.05) the levels of amino acids present in polychaete DM, but there was generally less of each amino acid tested on a weight per weight basis in large worms than in small worms. This difference was significant (P<0.05) for the most heavily represented classes being glutamic acid (73-77 mg g-1), aspartic acid (50-54 mg g-1), and glycine (46-53 mg g-1). These results demonstrate how this polychaete species can be planted and sorted at harvest according to various strategies aimed at providing biomass with specific physical and nutritional qualities for different uses.