In planta localization and interactions of impatiens necrotic spot tospovirus proteins

Impatiens necrotic spot tospovirus (INSV) is a significant pathogen of ornamentals. The tripartite negative- and ambi-sense RNA genome encodes six proteins that are involved in cytoplasmic replication, movement, assembly, insect transmission and defence. To gain insight into the associations of these viral proteins, we determined their intracellular localization and interactions in living plant cells. Nucleotide sequences encoding the nucleoprotein N, non-structural proteins NSs and NSm, and glycoproteins Gn and Gc of a Kentucky isolate of INSV were amplified by RTPCR, cloned, sequenced and transiently expressed as fusions with autofluorescent proteins in leaf epidermal cells of Nicotiana benthamiana. All proteins accumulated at the cell periphery and co-localized with an endoplasmic reticulum marker. The Gc protein fusion also localized to the nucleus. N and NSm protein self-interactions and an NSm-N interaction were observed by using bimolecular fluorescence complementation. A tospovirus NSm homotypic interaction had not been reported previously.

Demonstration reaches: Looking back whilst moving forward with river rehabilitation under the Native Fish Strategy

‘Demonstration reaches’ are sections of river where multiple threats to native fish are addressed through river rehabilitation and strong community participation. They are an important way of promoting the key driving actions of the Murray-Darling Basin Authority's Native Fish Strategy (NFS) by using on-ground community-driven rehabilitation. Measuring rehabilitation success against well-defined targets and using this information to adaptively mange activities is fundamental to the demonstration reach philosophy. Seven years on from the establishment of the first demonstration reach, there are now seven throughout the Murray-Darling Basin (MDB), all in differing states of maturation and but all applying a standardised framework for monitoring native fish outcomes. In this study, we reflect on the role that demonstration reaches have played within the NFS, synthesise some key findings from 32 monitoring and evaluation outputs, and highlight some of the successes and barriers to success. We make recommendations as to how to strengthen the demonstration reach model to ensure it remains a relevant approach for fish habitat rehabilitation beyond the NFS and MDB.

Localization of feeding of Anomalococcus indicus (Hemiptera: Lecanodiaspididae) and supplementary biological notes: Towards the biological management of the invasive tree Vachellia nilotica indica (Fabales: Mimosoideae) in North-Eastern Australia

. Management of the invasive Vachellia nilotica indica infesting tropical grasslands of Northern Australia has remained unsuccessful to date. Presently Anomalococcus indicus is considered a potential agent in the biological management of V. n. indica. Whereas generic biological details of A. indicus have been known, their feeding activity and details of their mouthparts and the sensory structures that are associated with their feeding action are not known. This paper provides details of those gaps. Nymphal instars I and II feed on cortical-parenchyma cells of young stems of V. n. indica, whereas nymphal instars III and adult females feed on phloem elements of older shoots. Nymphal instars and adults (females) trigger stress symptoms in the feeding tissue with cells bearing enlarged and disfigured nuclei, cytoplasmic shrinkage, cytoplasmic trabeculae, abnormal protuberances and uneven cell wall thickening, unusual cell membrane proliferation, and exhausted and necrosed cells. Continuous nutrient extraction by A. indicus can cause stem death. We provide evidence that A. indicus, by virtue of its continuous feeding activity and intense population build up, can be an effective biological-management agent to regulate populations of V. n. indica in infested areas. © 2014 © 2014 Société entomologique de France.

ICPH 2671 – the world's first commercial food legume hybrid

ICRISAT scientists, working with Indian programme counterparts, developed the world's first cytoplasmic-nuclear male sterility (CMS)-based commercial hybrid in a food legume, the pigeonpea [Cajanus cajan (L.) Millsp.]. The CMS, in combination with natural outcrossing of the crop, was used to develop viable hybrid breeding technology. Hybrid ICPH 2671 recorded 47% superiority for grain yield over the control variety ‘Maruti’ in multilocation on-station testing for 4 years. In the on-farm trials conducted in five Indian states, mean yield of this hybrid (1396 kg/ha) was 46.5% greater than that of the popular cv. ‘Maruti’ (953 kg/ha). Hybrid ICPH 2671 also exhibited high levels of resistance to Fusarium wilt and sterility mosaic diseases. The outstanding performance of this hybrid has led to its release for cultivation in India by both a private seed company (as ‘Pushkal’) and a public sector university (as ‘RV ICPH 2671’). Recent developments in hybrid breeding technology and high yield advantages realized in farmers' fields have given hope for a breakthrough in pigeonpea productivity.

Seminal plasma proteome of eletroejaculated Bos indicus bulls

The present study describes the seminal plasma proteome of Bos indicus bulls. Fifty-six, 24-month old Australian Brahman sires were evaluated and subjected to electroejaculation. Seminal plasma proteins were separated by 2-D SDS-PAGE and identified by mass spectrometry. The percentage of progressively motile and morphologically normal sperm of the bulls were 70.4±2.3 and 64±3.2%, respectively. A total of 108 spots were identified in the 2-D maps, corresponding to 46 proteins. Binder of sperm proteins accounted for 55.8% of all spots detected in the maps and spermadhesins comprised the second most abundant constituents. Other proteins of the Bos indicus seminal plasma include clusterin, albumin, transferrin, metalloproteinase inhibitor 2, osteopontin, epididymal secretory protein E1, apolipoprotein A-1, heat shock 70kDa protein, glutathione peroxidase 3, cathelicidins, alpha-enolase, tripeptidyl-peptidase 1, zinc-alpha-2-glycoprotein, plasma serine protease inhibitor, beta 2-microglobulin, proteasome subunit beta type-4, actin, cathepsins, nucleobinding-1, protein S100-A9, hemoglobin subunit alpha, cadherin-1, angiogenin-1, fibrinogen alpha and beta chain, ephirin-A1, protein DJ-1, serpin A3-7, alpha-2-macroglobulin, annexin A1, complement factor B, polymeric immunoglobulin receptor, seminal ribonuclease, ribonuclease-4, prostaglandin-H2 D-isomarase, platelet-activating factor acetylhydrolase, and phosphoglycerate kinase In conclusion, this work uniquely portrays the Bos indicus seminal fluid proteome, based on samples from a large set of animals representing the Brahman cattle of the tropical Northern Australia. Based on putative biochemical attributes, seminal proteins act during sperm maturation, protection, capacitation and fertilization.

Australian bat lyssavirus infection in two horses

In May 2013, the first cases of Australian bat lyssavirus infections in domestic animals were identified in Australia. Two horses (filly-H1 and gelding-H2) were infected with the Yellow-bellied sheathtail bat (YBST) variant of Australian bat lyssavirus (ABLV). The horses presented with neurological signs, pyrexia and progressing ataxia. Intra-cytoplasmic inclusion bodies (Negri bodies) were detected in some Purkinje neurons in haematoxylin and eosin (H&E) stained sections from the brain of one of the two infected horses (H2) by histological examination. A morphological diagnosis of sub-acute moderate non-suppurative, predominantly angiocentric, meningo-encephalomyelitis of viral aetiology was made. The presumptive diagnosis of ABLV infection was confirmed by the positive testing of the affected brain tissue from (H2) in a range of laboratory tests including fluorescent antibody test (FAT) and real-time PCR targeting the nucleocapsid (N) gene. Retrospective testing of the oral swab from (H1) in the real-time PCR also returned a positive result. The FAT and immunohistochemistry (IHC) revealed an abundance of ABLV antigen throughout the examined brain sections. ABLV was isolated from the brain (H2) and oral swab/saliva (H1) in the neuroblastoma cell line (MNA). Alignment of the genome sequence revealed a 97.7% identity with the YBST ABLV strain.

Polybridge Technical Report

This study examined the physical and chemical properties of a novel, fully-recirculated prawn and polychaete production system that incorporated polychaete-assisted sand filters (PASF). The aims were to assess and demonstrate the potential of this system for industrialisation, and to provide optimisations for wastewater treatment by PASF. Two successive seasons were studied at commercially-relevant scales in a prototype system constructed at the Bribie Island Research Centre in Southeast Queensland. The project produced over 5.4 tonnes of high quality black tiger prawns at rates up to 9.9 tonnes per hectare, with feed conversion of up to 1.1. Additionally, the project produced about 930 kg of high value polychaete biomass at rates up to 1.5 kg per square metre of PASF, with the worms feeding predominantly on waste nutrients. Importantly, this closed production system demonstrated rapid growth of healthy prawns at commercially relevant production levels, using methods that appear feasible for application at large scale. Deeper (23 cm) PASF beds provided similar but more reliable wastewater treatment efficacies compared with shallower (13 cm) beds, but did not demonstrate significantly greater polychaete productivity than (easier to harvest) shallow beds. The nutrient dynamics associated with seasonal and tidal operations of the system were studied in detail, providing technical and practical insights into how PASF could be optimised for the mitigation of nutrient discharge. The study also highlighted some of the other important advantages of this integrated system, including low sludge production, no water discharge during the culture phase, high ecosystem health, good prospects for biosecurity controls, and the sustainable production of a fishery-limited resource (polychaetes) that may be essential for the expansion of prawn farming industries throughout the world. Regarding nutrient discharge from this prototype mariculture system, when PASF was operating correctly it proved feasible to have no water (or nutrient) discharge during the entire prawn growing season. However, the final drain harvest and emptying of ponds that is necessary at the end of the prawn farming season released 58.4 kg ha-1 of nitrogen and 6 kg ha-1 of phosphorus (in Season 2). Whilst this is well below (i.e., one-third to one-half of) the current load-based licencing conditions for many prawn farms in Australia, the levels of nitrogen and chlorophyll a in the ponds remained higher than the more-stringent maximum limits at the Bribie Island study site. Zero-net-nutrient discharge was not achieved, but waste nutrients were low where 5.91 kg of nitrogen and 0.61 kg of phosphorus was discharged per tonne of prawns produced. This was from a system that deployed PASF at 14.4% of total ponded farm area which treated an average of 5.8% of pond water daily and did not use settlement ponds or other natural or artificial water remediation systems. Four supplemental appendices complement this research by studying several additional aspects that are central to the industrialisation of PASF. The first details an economic model and decision tool which allows potential users to interactively assess construction and operational variables of PASF at different scales. The second provides the qualitative results of a prawn maturation trial conducted collaboratively with the Commonwealth Scientific and Industrial Research Organisation (CSIRO) to assess dietary inclusions of PASF-produced worms. The third provides the reproductive results from industry-based assessments of prawn broodstock produced using PASF. And the fourth appendix provides detailed elemental and nutritional analyses of bacterial biofilm produced by PASF and assesses its potential to improve the growth of prawns in recirculated culture systems.