Infection of mungbean seed by Macrophomina phaseolina is more likely to result from localized pod infection than from systemic plant infection

The ubiquitous fungal pathogen Macrophomina phaseolina is best known as causing charcoal rot and premature death when host plants are subject to post-flowering stress. Overseas reports of M.phaseolina causing a rapid rot during the sprouting of Australian mungbean seed resulted in an investigation of the possible modes of infection of seed. Isolations from serial portions of 10 mungbean plants naturally infected with the pathogen revealed that on most plants there were discrete portions of infected tissue separated by apparently healthy tissue. The results from these studies, together with molecular analysis of isolates collected from infected tissue on two of the plants, suggested that aerial infection of aboveground parts by different isolates is common. Inoculations of roots and aboveground parts of mungbean plants at nine temperaturexsoil moisture incubation combinations and of detached green pods strongly supported the concept that seed infection results from infection of pods by microsclerotia, rather than from hyphae growing systemically through the plant after root or stem infection. This proposal is reinforced by anecdotal evidence that high levels of seed infection are common when rainfall occurs during pod fill, and by the isolation of M.phaseolina from soil peds collected on pods of mungbean plants in the field. However, other experiments showed that when inoculum was placed within 130mm of a green developing pod and a herbicide containing paraquat and diquat was sprayed on the inoculated plants, M.phaseolina was capable of some systemic growth from vegetative tissue into the pods and seeds.

Networks of stored grain diseases and pests: Strategies for sampling and mitigation

Pathogens and pests of stored grains move through complex dynamic networks linking fields, farms, and bulk storage facilities. Human transport and other forms of dispersal link the components of this network. A network model for pathogen and pest movement through stored grain systems is a first step toward new sampling and mitigation strategies that utilize information about the network structure. An understanding of network structure can be applied to identifying the key network components for pathogen or pest movement through the system. For example, it may be useful to identify a network node, such as a local grain storage facility, through which grain from a large number of fields will be accumulated and move through the network. This node may be particularly important for sampling and mitigation. In some cases more detailed information about network structure can identify key nodes that link two large sections of the network, such that management at the key nodes will greatly reduce the risk of spread between the two sections. In addition to the spread of particular species of pathogens and pests, we also evaluate the spread of problematic subpopulations, such as subpopulations with pesticide resistance. We present an analysis of stored grain pathogen and pest networks for Australia and the United States.

Cuelure but not zingerone make the sex pheromone of male Bactrocera tryoni (Tephritidae: Diptera) more attractive to females

In tephritid fruit flies of the genus Bactrocera Macquart, a group of plant derived compounds (sensu amplo ‘male lures’) enhance the mating success of males that have consumed them. For flies responding to the male lure methyl eugenol, this is due to the accumulation of chemicals derived from the male lure in the male rectal gland (site of pheromone synthesis) and the subsequent release of an attractive pheromone. Cuelure, raspberry ketone and zingerone are a second, related group of male lures to which many Bactrocera species respond. Raspberry ketone and cuelure are both known to accumulate in the rectal gland of males as raspberry ketone, but it is not known if the emitted male pheromone is subsequently altered in complexity or is more attractive to females. Using Bactrocera tryoni as our test insect, and cuelure and zingerone as our test chemicals, we assess: (i) lure accumulation in the rectal gland; (ii) if the lures are released exclusively in association with the male pheromone; and (iii) if the pheromone of lure-fed males is more attractive to females than the pheromone of lure-unfed males. As previously documented, we found cuelure was stored in its hydroxyl form of raspberry ketone, while zingerone was stored largely in an unaltered state. Small but consistent amounts of raspberry ketone and β-(4-hydroxy-3-methoxyphenyl)-propionic acid were also detected in zingerone-fed flies. Males released the ingested lures or their analogues, along with endogenous pheromone chemicals, only during the dusk courtship period. More females responded to squashed rectal glands extracted from flies fed on cuelure than to glands from control flies, while more females responded to the pheromone of calling cuelure-fed males than to control males. The response to zingerone treatments in both cases was not different from the control. The results show that male B. tryoni release ingested lures as part of their pheromone blend and, at least for cuelure, this attracts more females.

Phylodynamic evidence of the migration of turnip mosaic potyvirus from Europe to Australia and New Zealand

Turnip mosaic virus (TuMV) is a potyvirus that is transmitted by aphids and infects a wide range of plant species. We investigated the evolution of this pathogen by collecting 32 isolates of TuMV, mostly from Brassicaceae plants, in Australia and New Zealand. We performed a variety of sequence-based phylogenetic and population genetic analyses of the complete genomic sequences and of three non-recombinogenic regions of those sequences. The substitution rates, divergence times and phylogeographical patterns of the virus populations were estimated. Six inter- and seven intralineage recombination-type patterns were found in the genomes of the Australian and New Zealand isolates, and all were novel. Only one recombination-type pattern has been found in both countries. The Australian and New Zealand populations were genetically different, and were different from the European and Asian populations. Our Bayesian coalescent analyses, based on a combination of novel and published sequence data from three nonrecombinogenic protein-encoding regions, showed that TuMV probably started to migrate from Europe to Australia and New Zealand more than 80 years ago, and that distinct populations arose as a result of evolutionary drivers such as recombination. The basal-B2 subpopulation in Australia and New Zealand seems to be older than those of the world-B2 and -B3 populations. To our knowledge, our study presents the first population genetic analysis of TuMV in Australia and New Zealand. We have shown that the time of migration of TuMV correlates well with the establishment of agriculture and migration of Europeans to these countries.

Diagnostic Molecular Markers for Phosphine Resistance in U.S. Populations of Tribolium castaneum and Rhyzopertha dominica

Stored product beetles that are resistant to the fumigant pesticide phosphine (hydrogen phosphide) gas have been reported for more than 40 years in many places worldwide. Traditionally, determination of phosphine resistance in stored product beetles is based on a discriminating dose bioassay that can take up to two weeks to evaluate. We developed a diagnostic cleaved amplified polymorphic sequence method, CAPS, to detect individuals with alleles for strong resistance to phosphine in populations of the red flour beetle, Tribolium castaneum, and the lesser grain borer, Rhyzopertha dominica, according to a single nucleotide mutation in the dihydrolipoamide dehydrogenase (DLD) gene. We initially isolated and sequenced the DLD genes from susceptible and strongly resistant populations of both species. The corresponding amino acid sequences were then deduced. A single amino acid mutation in DLD in populations of T.castaneum and R.dominica with strong resistance was identified as P45S in T.castaneum and P49S in R.dominica, both collected from northern Oklahoma, USA. PCR products containing these mutations were digested by the restriction enzymes MboI and BstNI, which revealed presence or absence, respectively of the resistant (R) allele and allowed inference of genotypes with that allele. Seven populations of T.castaneum from Kansas were subjected to discriminating dose bioassays for the weak and strong resistance phenotypes. Application of CAPS to these seven populations confirmed the R allele was in high frequency in the strongly resistant populations, and was absent or at a lower frequency in populations with weak resistance, which suggests that these populations with a low frequency of the R allele have the potential for selection of the strong resistance phenotype. CAPS markers for strong phosphine resistance will help to detect and confirm resistant beetles and can facilitate resistance management actions against a given pest population.

The global distribution of Banana bunchy top virus reveals little evidence for frequent recent, human-mediated long distance dispersal events

Banana bunchy top virus (BBTV; family Nanoviridae, genus Babuvirus) is a multi-component single-stranded DNA virus, which infects banana plants in many regions of the world, often resulting in large-scale crop losses. Weanalyzed 171 banana leaf samples from fourteen countries and recovered, cloned, and sequenced 855 complete BBTV components including ninety-four full genomes. Importantly, full genomes were determined from eight countries, where previously no full genomes were available (Samoa, Burundi, Republic of Congo, Democratic Republic of Congo, Egypt, Indonesia, the Philippines, and the USA [HI]). Accounting for recombination and genome component reassortment, we examined the geographic structuring of global BBTV populations to reveal that BBTV likely originated in Southeast Asia, that the current global hotspots of BBTV diversity are Southeast Asia/Far East and India, and that BBTV populations circulating elsewhere in the world have all potentially originated from infrequent introductions. Most importantly, we find that rather than the current global BBTV distribution being due to increases in human-mediated movements of bananas over the past few decades, it is more consistent with a pattern of infrequent introductions of the virus to different parts of the world over the past 1,000 years.

Distribution and characterization of Poleroviruses affecting food legumes in Central/West Asia and North Africa (CWANA) region and Australia

During the past 15 years, surveys to identify virus diseases affecting cool-season food legume crops in Australia and 11 CWANA countries (Algeria, China, Egypt, Ethiopia, Lebanon, Morocco, Sudan, Syria, Tunisia, Uzbekistan and Yemen) were conducted. More than 20,000 samples were collected and tested for the presence of 14 legume viruses by the tissue-blot immunoassay (TBIA) using a battery of antibodies, including the following Luteovirus monoclonal antibodies (McAbs): a broad-spectrum legume Luteovirus (5G4), BLRV, BWYV, SbDV and CpCSV. A total of 195 Luteovirus samples were selected for further testing by RT-PCR using 7 primers (one is degenerate, and can detect a wide range of Luteoviridae virus species and the other six are species-specific primers) at the Virology Laboratory, QDAF, Australia, during 2014. A total of 145 DNA fragments (represented 105 isolates) were sequenced. The following viruses were characterized based on molecular analysis: BLRV from Lebanon, Morocco, Tunisia and Uzbekistan; SbDV from Australia, Syria and Uzbekistan; BWYV from Algeria, China, Ethiopia, Lebanon, Morocco, Sudan, Tunisia and Uzbekistan; CABYV from Algeria, Lebanon, Syria, Sudan and Uzbekistan; CpCSV from Algeria, Ethiopia, Lebanon, Morocco, Syria and Tunisia, and unknown Luteoviridae species from Algeria, Ethiopia, Morocco, Sudan, Uzbekistan and Yemen. This study has clearly shown that there are a number of Polerovirus species, in addition to BWYV, all can produce yellowing/stunting symptoms in pulses (e.g. CABYV, CpCSV, and other unknown Polerovirus species). Based on our knowledge this is the first report of CABYV affecting food legumes. Moreover, there was about 95% agreement between results obtained from serological analysis (TBIA) and molecular analysis for the detection of BLRV and SbDV. Whereas, TBIA results were not accurate when using CpCSV and BWYV McAbs . It seems that the McAbs for CpCSV and BWYV used in this study and those available worldwide, are not virus species specific. Both antibodies, reacted with other Polerovirus species (e.g. CABYV, and unknown Polerovirus). This highlights the need for more accurate characterization of existing antibodies and where necessary the development of better, virus-specific antibodies to enable their use for accurate diagnosis of Poleroviruses.

Climate savvy grazing (Developing improved grazing and related practices to assist beef production enterprises across northern Australia to adapt to a changing and more variable climate)

There is uncertainty over the potential changes to rainfall across northern Australia under climate change. Since rainfall is a key driver of pasture growth, cattle numbers and the resulting animal productivity and beef business profitability, the ability to anticipate possible management strategies within such uncertainty is crucial. The Climate Savvy Grazing project used existing research, expert knowledge and computer modelling to explore the best-bet management strategies within best, median and worse-case future climate scenarios. All three scenarios indicated changes to the environment and resources upon which the grazing industry of northern Australia depends. Well-adapted management strategies under a changing climate are very similar to best practice within current climatic conditions. Maintaining good land condition builds resource resilience, maximises opportunities under higher rainfall years and reduces the risk of degradation during drought and failed wet seasons. Matching stocking rate to the safe long-term carrying capacity of the land is essential; reducing stock numbers in response to poor seasons and conservatively increasing stock numbers in response to better seasons generally improves profitability and maintains land in good condition. Spelling over the summer growing season will improve land condition under a changing climate as it does under current conditions. Six regions were included within the project. Of these, the Victoria River District in the Northern Territory, Gulf country of Queensland and the Kimberley region of Western Australia had projections of similar or higher than current rainfall and the potential for carrying capacity to increase. The Alice Springs, Maranoa-Balonne and Fitzroy regions had projections of generally drying conditions and the greatest risk of reduced pasture growth and carrying capacity. Encouraging producers to consider and act on the risks, opportunities and management options inherent in climate change was a key goal of the project. More than 60,000 beef producers, advisors and stakeholders are now more aware of the management strategies which build resource resilience, and that resilience helps buffer against the effects of variable and changing climatic conditions. Over 700 producers have stated they have improved confidence, skills and knowledge to attempt new practices to build resilience. During the course of the project, more than 165 beef producers reported they have implemented changes to build resource and business resilience.