Single gene resistance to Fusarium oxysporum f. sp. cubense Race 4 in the wild banana Musa acuminata subsp. malaccensis

Fusarium wilt of banana, caused by the fungal pathogen Fusarium oxysporum f. sp. cubense (Foc), is one of the most destructive diseases of banana. A particularly virulent strain of the pathogen, tropical race 4 (TR4), presents an emerging threat to banana producing regions throughout the world. No commercially acceptable banana cultivar is resistant to TR4 and, as with all strains of the Fusarium wilt pathogen, there is no effective chemical control. Genetic resistance to TR4 has been observed in the diploid wild banana Musa acuminata subsp. malaccensis, which has consequently received attention as a potential source of Fusarium resistance genes. The aim of this research was to determine the pattern of inheritance of the resistance trait by screening plants for resistance to Foc subtropical race 4 (SR4) and TR4. Our results showed that the F1 progeny of self-fertilized malaccensis plants challenged in pot trials against SR4 (VCGs 0120, 0129, 01211) and TR4 (VCG 01213/16) segregated for resistance according to a Mendelian ratio of 3:1 which is consistent with a single dominant gene hypothesis.

Morphological Characteristics of Genetically-Modified Pineapple Fruit

Genetic engineering is an attractive method for changing a single characteristic of ‘Smooth Cayenne’ pineapple, without altering its other desirable attributes. Techniques used in pineapple transformation, however, such as tissue culture and biolistic-mediated or Agrobacterium-mediated gene insertion are prone to somaclonal variation, resulting in the production of several morphological mutations (Smith et al., 2002). Fruit mutations can include distortion in fruit shape (round ball, conical, fan-shaped), reduced fruit size, multiple crowns, crownless fruit, fruitless crowns, and spiny crown leaves (Dalldorf, 1975; Sanewski et al., 1992). The present paper describes the variability in fruit-shape mutations between transgenic and non-transgenic fruit, and its subsequent impact on organoleptic characteristics.

Sigastus weevil - an emerging pest of macadamias in north Queensland

A large weevil was found infesting macadamia nuts on the Atherton Tableland during the 1994/95 season. It was unrepresented in various Australian insect collections but thought to belong to the genus Sigastus. This paper reports some preliminary studies on its biology, pest status and control. From 4-6 weeks after first nut-set adult females commence laying single eggs through the husk, after first scarifying an oviposition site. The nut stalk is then cleaved leading to rapid abscission. Nuts were generally attacked up until hard shell formation. Weevil larvae consumed whole kernels, with % survival higher and larval duration shorter in larger nuts. Infestation rates increased with increasing nut diameter, reaching 72.8% of fallen nuts by mid-October. A crop loss of 30% could be attributed to weevils in an unsprayed orchard. However, adult weevils are very susceptible to both carbaryl and methidathion sprays. In addition, exposure of infested nuts to full sunlight over several weeks kills 100% of larvae. Crops should be surveyed for weevil damage from the 5-10 mm diameter stage until mid-December. Methidathion used as an initial spray for fruitspotting bugs should provide control. Organic growers are advised to sweep infested nuts into mown interrows where solarisation will kill larvae.

The effect of row configuration on yield reliability in grain sorghum: II. Modelling the effects of row configuration

In recent years many sorghum producers in the more marginal (<600 mm annual rainfall) cropping areas of Qld and northern NSW have utilised skip row configurations in an attempt to improve yield reliability and reduce sorghum production risk. But will this work in the long run? What are the trade-offs between productivity and risk of crop failure? This paper describes a modelling and simulation approach to study the long-term effects of skip row configurations. Detailed measurements of light interception and water extraction from sorghum crops grown in solid, single and double skip row configurations were collected from three on-farm participatory research trials established in southern Qld and northern NSW. These measurements resulted in changes to the model that accounted for the elliptical water uptake pattern below the crop row and reduced total light interception associated with the leaf area reduction of the skip configuration. Following validation of the model, long-term simulation runs using historical weather data were used to determine the value of skip row sorghum production as a means of maintaining yield reliability in the dryland cropping regions of southern Qld and northern NSW.

Genetic variability of Tomato spotted wilt virus in Australia and validation of real time RT-PCR for its detection in single and bulked leaf samples

The potential for large-scale use of a sensitive real time reverse transcription polymerase chain reaction (RT-PCR) assay was evaluated for the detection of Tomato spotted wilt virus (TSWV) in single and bulked leaf samples by comparing its sensitivity with that of DAS-ELISA. Using total RNA extracted with RNeasy® or leaf soak methods, real time RT-PCR detected TSWV in all infected samples collected from 16 horticultural crop species (including flowers, herbs and vegetables), two arable crop species, and four weed species by both assays. In samples in which DAS-ELISA had previously detected TSWV, real time RT-PCR was effective at detecting it in leaf tissues of all 22 plant species tested at a wide range of concentrations. Bulk samples required more robust and extensive extraction methods with real time RT-PCR, but it generally detected one infected sample in 1000 uninfected ones. By contrast, ELISA was less sensitive when used to test bulked samples, once detecting up to 1 infected in 800 samples with pepper but never detecting more than 1 infected in 200 samples in tomato and lettuce. It was also less reliable than real time RT-PCR when used to test samples from parts of the leaf where the virus concentration was low. The genetic variability among Australian isolates of TSWV was small. Direct sequencing of a 587 bp region of the nucleoprotein gene (S RNA) of 29 isolates from diverse crops and geographical locations yielded a maximum of only 4.3% nucleotide sequence difference. Phylogenetic analysis revealed no obvious groupings of isolates according to geographic origin or host species. TSWV isolates, that break TSWV resistance genes in tomato or pepper did not differ significantly in the N gene region studied, indicating that a different region of the virus genome is responsible for this trait.

Pasteurella multocida Expresses Two Lipopolysaccharide Glycoforms Simultaneously, but Only a Single Form Is Required for Virulence: Identification of Two Acceptor-Specific Heptosyl I Transferases

Lipopolysaccharide (LPS) is a critical virulence determinant in Pasteurella multocida and a major antigen responsible for host protective immunity. In other mucosal pathogens, variation in LPS or lipooligosaccharide structure typically occurs in the outer core oligosaccharide regions due to phase variation. P. multocida elaborates a conserved oligosaccharide extension attached to two different, simultaneously expressed inner core structures, one containing a single phosphorylated 3-deoxy-D-manno-octulosonic acid (Kdo) residue and the other containing two Kdo residues. We demonstrate that two heptosyltransferases, HptA and HptB, add the first heptose molecule to the Kdo1 residue and that each exclusively recognizes different acceptor molecules. HptA is specific for the glycoform containing a single, phosphorylated Kdo residue (glycoform A), while HptB is specific for the glycoform containing two Kdo residues (glycoform B). In addition, KdkA was identified as a Kdo kinase, required for phosphorylation of the first Kdo molecule. Importantly, virulence data obtained from infected chickens showed that while wild-type P. multocida expresses both LPS glycoforms in vivo, bacterial mutants that produced only glycoform B were fully virulent, demonstrating for the first time that expression of a single LPS form is sufficient for P. multocida survival in vivo. We conclude that the ability of P. multocida to elaborate alternative inner core LPS structures is due to the simultaneous expression of two different heptosyltransferases that add the first heptose residue to the nascent LPS molecule and to the expression of both a bifunctional Kdo transferase and a Kdo kinase, which results in the initial assembly of two inner core structures.

Genetic control of propagation traits in a single Corymbia torelliana x Corymbia variegata family

Genetic control of vegetative propagation traits was described for a second-generation, outbred, intersectional hybrid family (N = 208) derived from two species, Corymbia torelliana (F. Muell.) K.D. Hill & L.A.S. Johnson and Corymbia variegata (F. Muell.) K.D. Hill & L.A.S. Johnson, which contrast for propagation characteristics and in their capacity to develop lignotubers. Large phenotypic variances were evident for rooting and most other propagation traits, with significant proportions attributable to differences between clones (broad-sense heritabilities 0.2-0.5). Bare root assessment of rooting rate and root quality parameters tended to have the highest heritabilities, whereas rooting percentage based on root emergence from pots and shoot production were intermediate. Root biomass and root initiation had the lowest heritabilities. Strong favourable genetic correlations were found between rooting percentage and root quality traits such as root biomass, volume, and length. Lignotuber development on a seedling was associated with low rooting and a tendency to poor root quality in cuttings and was in accord with the persistence of species parent types due to gametic phase disequilibrium. On average, nodal cuttings rooted more frequently and with higher quality root systems, but significant cutting type x genotype interaction indicated that for some clones, higher rooting rates were obtained from tips. Low germination, survival of seedlings, and rooting rates suggested strong hybrid breakdown in this family.

Characterisation of chicken Campylobacter jejuni isolates using resolution optimised single nucleotide polymorphisms and binary gene markers

The principal objective of this study was to determine if Campylobacter jejuni genotyping methods based upon resolution optimised sets of single nucleotide polymorphisms (SNPs) and binary genetic markers were capable of identifying epidemiologically linked clusters of chicken-derived isolates. Eighty-eight C. jejuni isolates of known flaA RFLP type were included in the study. They encompassed three groups of ten isolates that were obtained at the same time and place and possessed the same flaA type. These were regarded as being epidemiologically linked. Twenty-six unlinked C. jejuni flaA type I isolates were included to test the ability of SNP and binary typing to resolve isolates that were not resolved by flaA RFLP. The remaining isolates were of different flaA types. All isolates were typed by real-time PCR interrogation of the resolution optimised sets of SNPs and binary markers. According to each typing method, the three epidemiologically linked clusters were three different clones that were well resolved from the other isolates. The 26 unlinked C. jejuni flaA type I isolates were resolved into 14 SNP-binary types, indicating that flaA typing can be unreliable for revealing epidemiological linkage. Comparison of the data with data from a fully typed set of isolates associated with human infection revealed that abundant lineages in the chicken isolates that were also found in the human isolates belonged to clonal complex (CC) -21 and CC-353, with the usually rare C-353 member ST-524 being especially abundant in the chicken collection. The chicken isolates selected to be diverse according to flaA were also diverse according to SNP and binary typing. It was observed that CC-48 was absent in the chicken isolates, despite being very common in Australian human infection isolates, indicating that this may be a major cause of human disease that is not chicken associated.

The use of high resolution melting (HRM) to map single nucleotide polymorphism markers linked to a covered smut resistance gene in barley

Using an established genetic map, a single gene conditioning covered smut resistance, Ruh.7H, was mapped to the telomere region of chromosome 7HS in an Alexis/Sloop doubled haploid barley population. The closest marker to Ruh.7H, abg704 was 7.5 cM away. Thirteen loci on the distal end of 7HS with potential to contain single nucleotide polymorphisms (SNPs) were identified by applying a comparative genomics approach using rice sequence data. Of these, one locus produced polymorphic co-dominant bands of different size while two further loci contained SNPs that were identified using the recently developed high resolution melting (HRM) technique. Two of these markers flanked Ruh.7H with the proximal marker located 3.8 cM and the distal marker 2.7 cM away. This is the first report on the application of the HRM technique to SNP detection and to rapid scoring of known cleaved amplified polymorphic sequence (CAPS) markers in plants. This simple, precise post-PCR technique should find widespread use in the fine-mapping of genetic regions of interest in complex cereal and other plant genomes.

Post capture necrosis of the feet caused by soft-catch traps – incidents, cause and prevention.

When recapturing satellite collared wild dogs that had been trapped one month previous in padded foothold traps, we noticed varying degrees of pitting on the pads of their trapped paw. Veterinary advice, based on images taken of the injuries, suggests that the necrosis was caused by vascular compromise. Five of six dingoes we recaptured had varying degrees of necrosis restricted only to the trapped foot and ranging from single 5 mm holes to 25% sections of the toe pads missing or deformed, including loss of nails. The traps used were rubber-padded, two–coiled, Victor Soft Catch #3 traps. The springs are not standard Victor springs but were Beefer springs; these modifications slightly increase trap speed and the jaw pressure on the trapped foot. Despite this modification the spring pressure is still relatively mild in comparison to conventional long spring or four-coiled wild dog traps. The five wild dogs developing necrosis were trapped in November 2006 at 5-6 months of age. Traps were checked each morning so the dogs were unlikely to have been restrained in the trap for more than 12 hours. All dogs exhibited a small degree of paw damage at capture which presented itself as a swollen paw and compression at the capture point. In contrast, eight wild dogs, 7-8 month-old, were captured two months later in February. Upon their release, on advice from a veterinarian, we massaged the trapped foot to get blood flow back in to the foot and applied a bruise treatment (Heparinoid 8.33 mg/ml) to assist restoring blood flow. These animals were subsequently recaptured several months later and showed no signs of necrosis. While post-capture foot injuries are unlikely to be an issue in conventional control programs where the animal is immediately destroyed, caution needs to be used when releasing accidentally captured domestic dogs or research animals captured in rubber-padded traps. We have demonstrated that 7-8 month old dogs can be trapped and released without any evidence of subsequent necrosis following minimal veterinary treatment. We suspect that the rubber padding on traps may increase the tourniquet effect by wrapping around the paw and recommend the evaluation of offset laminated steel jaw traps as an alternative. Offset laminated steel jaw traps have been shown to be relatively humane producing as few foot injuries as rubber-jawed traps.

Co-evolution of wild rabbits (Oryctolagus cuniculus) and RHDV: resistance and virulence

Rabbit Haemorrhagic Disease Virus (RHDV) was introduced to Australia in 1995 for the control of wild rabbits. Initial outbreaks greatly reduced rabbit numbers and the virus has continued to control rabbits to varying degrees in different parts of Australia. However, recent field evidence suggests that the virus may be becoming less effective in those areas that have previously experienced repeated epizootics causing high mortality. There are also reports of rabbits returning to pre-1995 density levels, Virus and host can be expected to co-evolve. The host will develop resistance to the virus with the virus subsequently changing to overcome that resistance. It has been 12 years since the release of RHDV and it is an opportune time to examine where the dynamic currently stands between RHDV and rabbits. Laboratory challenge tests have indicated that resistance to RHDV has developed to different degrees in populations throughout Australia. In one population a low dose (1:25 dilution) of Czech strain RHDV failed to infect a single susceptible rabbit, yet infected a low to high (up to 73%) percentage across other populations tested. Different selection pressures are present in these populations and will be driving the level of resistance being seen. The mechanisms and genetics behind the development of resistance are also important as the on-going use of RHDV as a control tool in the management of rabbits relies on our understanding of factors influencing the efficacy of the virus. Understanding how resistance has developed may provide clues on how best to use the virus to circumvent these mechanisms. Similarly, it will help in managing populations that have yet to develop high levels of resistance.

Single kernel NIR assessment for QTL identification

Development and evaluation of a single kernel NIR assessment method for improving baley malting quality QTL identification.

African mahogany (Khaya senegalensis) plantations in Australia - status, needs and progress

The Australian African mahogany estate comprises over 12,000 ha of industrial plantations, farm-forestry plots and trials, virtually all derived from Africa-sourced wild seed. However, the better trees have given high-value products such as veneers, high-grade boards and award-winning furniture. Collaborative conservation and improvement by the Northern Territory (NT) and Queensland governments since 2000 realised seed orchards, hedge gardens and genetic tests revealing promising clones and families. Private sector R&D since the mid 2000s includes silvicultural-management and wood studies, participatory testing of government material and establishing over 90 African provenances and many single-tree seedlots in multisite provenance and family trials. Recent, mainly public sector research included a 5-agency project of 2009-12 resulting in advanced propagation technologies and greater knowledge of biology, wood properties and processing. Operational priority in the short term should focus on developing seed production areas and ‘rolling front’ clonal seed orchards. R&D priorities should include: developing and implementing a collaborative improvement strategy based on pooled resources; developing non-destructive evaluation of select-tree wood properties, micropropagation (including field testing of material from this source) to ‘industry ready’ and a select-tree index; optimising seed production in orchards; advancing controlled pollination techniques; and maximising benefits from the progeny, clone and provenance trials. Australia leads the world in improvement and ex situ conservation of African mahogany based on the governments’ 13-year program and more recent industry inputs such that accumulated genetic resources total over 120 provenances and many families from 15 of the 19 African countries of its range. Having built valuable genetic resources, expertise, technologies and knowledge, the species is almost ‘industry ready’. The industry will benefit if it exploits the comparative advantage these assets provide. However the status of much of the diverse germplasm introduced since the mid 2000s is uncertain due to changes in ownership. Further, recent reductions of government investment in forestry R&D will be detrimental unless the industry fills the funding gaps. Expansion and sustainability of the embryonic industry must capitalise on past and current R&D, while initiating and sustaining critical new work through all-stakeholder collaboration.

Allelic variation at a single gene increases food value in a drought-tolerant staple cereal

The production of adequate agricultural outputs to support the growing human population places great demands on agriculture, especially in light of ever-greater restrictions on input resources. Sorghum is a drought-adapted cereal capable of reliable production where other cereals fail, and thus represents a good candidate to address food security as agricultural inputs of water and arable land grow scarce. A long-standing issue with sorghum grain is that it has an inherently lower digestibility. Here we show that a low-frequency allele type in the starch metabolic gene, pullulanase, is associated with increased digestibility, regardless of genotypic background. We also provide evidence that the beneficial allele type is not associated with deleterious pleiotropic effects in the modern field environment. We argue that increasing the digestibility of an adapted crop is a viable way forward towards addressing food security while maximizing water and land-use efficiency.