Gliotoxin effects on fungal growth: Mechanisms and exploitation

Although initially investigated for its antifungal properties, little is actually known about the effect of gliotoxin on Aspergillus fumigatus and other fungi. We have observed that exposure of A. fumigatus to exogenous gliotoxin (14 lg/ml), under gliotoxin-limited growth conditions, results in significant alteration of the expression of 27 proteins (up- and down-regulated >1.9-fold; p < 0.05) including de novo expression of Cu, Zn superoxide dismutase, up-regulated allergen Asp f3 expression and down-regulated catalase and a peroxiredoxin levels. Significantly elevated glutathione GSH levels (p < 0.05), along with concomitant resistance to diamide, were evident in A. fumigatus ∆gliT, lacking gliotoxin oxidoreductase, a gliotoxin self-protection gene. Saccharomyces cerevisiae deletents (∆sod1 and ∆yap1) were hypersensitive to exogenous gliotoxin, while ∆gsh1 was resistant. Significant gliotoxin-mediated (5 µg/ml) growth inhibition (p < 0.001) of Aspergillus nidulans, Aspergillus terreus, Aspergillus niger, Cochliobolus heterostrophus and Neurospora crassa was also observed. Growth of Aspergillus flavus, Fusarium graminearum and Aspergillus oryzae was significantly inhibited (p < 0.001) at gliotoxin (10 lg/ml), indicating differential gliotoxin sensitivity amongst fungi. Re-introduction of gliT into A. fumigatus DgliT, at a different locus (ctsD; AFUA_4G07040, an aspartic protease), with selection on gliotoxin, facilitated deletion of ctsD without use of additional antibiotic selection markers. Absence of ctsD expression was accompanied by restoration of gliT expression, and resistance to gliotoxin. Thus, we propose gliT/gliotoxin as a useful selection marker system for fungal transformation. Finally, we suggest incorporation of gliotoxin sensitivity assays into all future fungal functional genomic studies

The Role of Glutathione S-Transferase GliG in Gliotoxin Biosynthesis in Aspergillus fumigatus

Gliotoxin, a redox-active metabolite, is produced by the opportunistic fungal pathogen Aspergillus fumigatus, and its biosynthesis is directed by the gli gene cluster. Knowledge of the biosynthetic pathway to gliotoxin, which contains a disulfide bridge of unknown origin, is limited, although L-Phe and L-Ser are known biosynthetic precursors. Deletion of gliG from the gli cluster, herein functionally confirmed as a glutathione S-transferase, results in abrogation of gliotoxin biosynthesis and accumulation of 6-benzyl-6-hydroxy-1-methoxy-3-methylenepiperazine- 2,5-dione. This putative shunt metabolite from the gliotoxin biosynthetic pathway contains an intriguing hydroxyl group at C-6, consistent with a gliotoxin biosynthetic pathway involving thiolation via addition of the glutathione thiol group to a reactive acyl imine intermediate. Complementation of gliG restored gliotoxin production and, unlike gliT, gliG was found not to be involved in fungal self-protection against gliotoxin

Malignant mixed tumor in the salivary gland of a cat

The presence of a malignant mixed tumor, also known as a carcinosarcoma, in the salivary gland is very rare. Such tumors, which are typically aggressive, are characterized by the presence of carcinomatous and sarcomatous components. A 9-year-old neutered female domestic short-haired cat presented with swelling in the right mandibular lesion that had rapidly enlarged over the previous 3 weeks. Physical examination revealed a large, fluctuated and painless subcutaneous swelling that was associated with a firm mass. Radiographs of the head revealed a soft-tissue density that involved faint circular calcific opacity. Contrast-enhanced computed tomography revealed that the peripheral capsulated cystic area had a contrast enhanced region without bone lysis. The cat received a total excision of the mass and postoperative radiotherapy. Histopathological analysis of the mass revealed that it was a malignant mixed tumor. Metastasis to the lung was discovered 7 weeks later, at which time treatment was stopped

Simulation of growth and development of diverse legume species in APSIM

This paper describes the physiological basis and validation of a generic legume model as it applies to 4 species: chickpea (Cicer arietinum L.), mungbean (Vigna radiata (L.) Wilczek), peanut (Arachis hypogaeaL.), and lucerne (Medicago sativa L.). For each species, the key physiological parameters were derived from the literature and our own experimentation. The model was tested on an independent set of experiments, predominantly from the tropics and subtropics of Australia, varying in cultivar, sowing date, water regime (irrigated or dryland), row spacing, and plant population density. The model is an attempt to simulate crop growth and development with satisfactory comprehensiveness, without the necessity of defining a large number of parameters. A generic approach was adopted in recognition of the common underlying physiology and simulation approaches for many legume species. Simulation of grain yield explained 77, 81, and 70% of the variance (RMSD = 31, 98, and 46 g/m2) for mungbean (n = 40, observed mean = 123 g/m2), peanut (n = 30, 421 g/m2), and chickpea (n = 31, 196 g/m2), respectively. Biomass at maturity was simulated less accurately, explaining 64, 76, and 71% of the variance (RMSD = 134, 236, and 125 g/m2) for mungbean, peanut, and chickpea, respectively. RMSD for biomass in lucerne (n = 24) was 85 g/m2 with an R2 of 0.55. Simulation accuracy is similar to that achieved by single-crop models and suggests that the generic approach offers promise for simulating diverse legume species without loss of accuracy or physiological rigour

Scope for improved eco-efficiency varies among diverse cropping systems

Global food security requires eco-efficient agriculture to produce the required food and fiber products concomitant with ecologically efficient use of resources. This eco-efficiency concept is used to diagnose the state of agricultural production in China (irrigated wheat–maize double-cropping systems), Zimbabwe (rainfed maize systems), and Australia (rainfed wheat systems). More than 3,000 surveyed crop yields in these three countries were compared against simulated grain yields at farmer-specified levels of nitrogen (N) input. Many Australian commercial wheat farmers are both close to existing production frontiers and gain little prospective return from increasing their N input. Significant losses of N from their systems, either as nitrous oxide emissions or as nitrate leached from the soil profile, are infrequent and at low intensities relative to their level of grain production. These Australian farmers operate close to eco-efficient frontiers in regard to N, and so innovations in technologies and practices are essential to increasing their production without added economic or environmental risks. In contrast, many Chinese farmers can reduce N input without sacrificing production through more efficient use of their fertilizer input. In fact, there are real prospects for the double-cropping systems on the North China Plain to achieve both production increases and reduced environmental risks. Zimbabwean farmers have the opportunity for significant production increases by both improving their technical efficiency and increasing their level of input; however, doing so will require improved management expertise and greater access to institutional support for addressing the higher risks. This paper shows that pathways for achieving improved eco-efficiency will differ among diverse cropping systems

Linocin and OmpW Are Involved In Attachment Of The Cystic Fibrosis-Associated Pathogen Burkholderia Cepacia Complex To Lung Epithelial Cells and Protect Mice Against Infection

Burkholderia cepacia complex (Bcc) causes chronic opportunistic lung infections in people with cystic fibrosis (CF) resulting in a gradual lung function decline and, ultimately, patient death. Bcc is a complex of eighteen species and is rarely eradicated once a patient is colonised, therefore vaccination may represent a better therapeutic option. We developed a new proteomics approach to identify bacterial proteins that are involved in attachment of Bcc to lung epithelial cells. Fourteen proteins were reproducibly identified by 2-DE from four Bcc strains, representative of two Bcc species: B. cenocepacia, the most virulent and B. multivorans, the most frequently acquired. Seven proteins were identified in both species, but only two were common to all four strains, Linocin and OmpW. Both proteins were selected based previously published data on these proteins in other species. The E. coli strains expressing recombinant Linocin and OmpW showed enhanced attachment (4.2- and 3.9-fold) to lung cells, compared to control, confirming that both proteins are involved in host cell attachment. Immunoproteomic analysis using serum from Bcc colonised CF patients confirmed that both proteins elicit potent humoral responses in vivo. Mice immunised with either recombinant Linocin or OmpW were protected from B. cenocepacia and B. multivorans challenge. Both antigens induced potent antigen-specific antibody responses and stimulated strong cytokine responses. In conclusion, our approach identified adhesins that induced excellent protection against two Bcc species and are promising vaccine candidates for a multi-subunit vaccine. Furthermore, it highlights the potential of our proteomics approach to identify potent antigens against other difficult pathogens

A proteomic approach to investigating gene cluster expression and secondary metabolite functionality in Aspergillus fumigatus.

A combined proteomics and metabolomics approach was utilised to advance the identification and characterisation of secondary metabolites in Aspergillus fumigatus. Here, implementation of a shotgun proteomic strategy led to the identification of non-redundant mycelial proteins (n = 414) from A. fumigatus including proteins typically under-represented in 2-D proteome maps: proteins with multiple transmembrane regions, hydrophobic proteins and proteins with extremes of molecular mass and pI. Indirect identification of secondary metabolite cluster expression was also achieved, with proteins (n = 18) from LaeA-regulated clusters detected, including GliT encoded within the gliotoxin biosynthetic cluster. Biochemical analysis then revealed that gliotoxin significantly attenuates H2O2-induced oxidative stress in A. fumigatus (p>0.0001), confirming observations from proteomics data. A complementary 2-D/LC-MS/MS approach further elucidated significantly increased abundance (p<0.05) of proliferating cell nuclear antigen (PCNA), NADH-quinone oxidoreductase and the gliotoxin oxidoreductase GliT, along with significantly attenuated abundance (p<0.05) of a heat shock protein, an oxidative stress protein and an autolysis-associated chitinase, when gliotoxin and H2O2 were present, compared to H2O2 alone. Moreover, gliotoxin exposure significantly reduced the abundance of selected proteins (p<0.05) involved in de novo purine biosynthesis. Significantly elevated abundance (p<0.05) of a key enzyme, xanthine-guanine phosphoribosyl transferase Xpt1, utilised in purine salvage, was observed in the presence of H2O2 and gliotoxin. This work provides new insights into the A. fumigatus proteome and experimental strategies, plus mechanistic data pertaining to gliotoxin functionality in the organism

Crop Updates 2006 - Cereals

This session covers twenty nine papers from different authors: PLENARY 1. The 2005 wheat streak mosaic virus epidemic in New South Wales and the threat posed to the Western Australian wheat industry, Roger Jones and Nichole Burges, Department of Agriculture SOUTH COAST AGRONOMY 2. South coast wheat variety trial results and best options for 2006, Mohammad Amjad, Ben Curtis and Wal Anderson, Department of Agriculture 3. Dual purpose winter wheats to improve productivity, Mohammad Amjad and Ben Curtis, Department of Agriculture 4. South coast large-scale premium wheat variety trials, Mohammad Amjad and Ben Curtis, Department of Agriculture 5. Optimal input packages for noodle wheat in Dalwallinu – Liebe practice for profit trial, Darren Chitty, Agritech Crop Research and Brianna Peake, Liebe Group 6. In-crop risk management using yield prophet®, Harm van Rees1, Cherie Reilly1, James Hunt1, Dean Holzworth2, Zvi Hochman2; 1Birchip Cropping Group, Victoria; 2CSIRO, Toowoomba, Qld 7. Yield Prophet® 2005 – On-line yield forecasting, James Hunt1, Harm van Rees1, Zvi Hochman2,Allan Peake2, Neal Dalgliesh2, Dean Holzworth2, Stephen van Rees1, Trudy McCann1 and Peter Carberry2; 1Birchip Cropping Group, Victoria; 2CSIRO, Toowoomba, Qld 8. Performance of oaten hay varieties in Western Australian environments, Raj Malik and Kellie Winfield, Department of Agriculture 9. Performance of dwarf potential milling varieties in Western Australian environments, Kellie Winfield and Raj Malik, Department of Agriculture 10. Agronomic responses of new wheat varieties in the Southern agricultural region of WA, Brenda Shackley and Judith Devenish, Department of Agriculture 11. Responses of new wheat varieties to management factors in the central agricultural region of Western Australia, Darshan Sharma, Steve Penny and Wal Anderson,Department of Agriculture 12. Sowing time on wheat yield, quality and $ - Northern agricultural region, Christine Zaicou-Kunesch, Department of Agriculture NUTRITION 13.The most effective method of applying phosphorus, copper and zinc to no-till crops, Mike Bolland and Ross Brennan, Department of Agriculture 14. Uptake of K from the soil profile by wheat, Paul Damon and Zed Rengel, Faculty of Natural and Agricultural Sciences, University of Western Australia 15. Reducing nitrogen fertiliser risks, Jeremy Lemon, Department of Agriculture 16. Yield Prophet® and canopy management, Harm van Rees1, Zvi Hochman2, Perry Poulton2, Nick Poole3, Brooke Thompson4, James Hunt1; 1Birchip Cropping Group, Victoria; 2CSIRO, Toowoomba, Qld; 3Foundation for Arable Research, New Zealand; 4Cropfacts, Victoria 17. Producing profits with phosphorus, Stephen Loss, CSBP Ltd, WA 18. Potassium response in cereal cropping within the medium rainfall central wheatbelt, Jeff Russell1, Angie Roe2 and James Eyres2, Department of Agriculture1, Farm Focus Consultants, Northam2 19. Matching nitrogen supply to wheat demand in the high rainfall cropping zone, Narelle Simpson, Ron McTaggart, Wal Anderson, Lionel Martin and Dave Allen, Department of Agriculture DISEASES 20. Comparative study of commercial wheat cultivars and differential lines (with known Pm resistance genes) to powdery mildew response, Hossein Golzar, Manisha Shankar and Robert Loughman, Department of Agriculture 21. On farm research to investigate fungicide applications to minimise leaf disease impacts in wheat – part II, Jeff Russell1, Angie Roe2and James Eyres2, Department of Agriculture1, and Farm Focus Consultants, Northam2 22. Disease resistance update for wheat varieties in WA, Manisha Shankar, John Majewski, Donna Foster, Hossein Golzar, Jamie Piotrowski, Nicole Harry and Rob Loughman, Department of Agriculture 23. Effect of time of stripe rust inoculum arrival on variety response in wheat, Manisha Shankar, John Majewski and Rob Loughman, Department of Agriculture 24. Fungicide seed dressing management of loose smut in Baudin barley, Geoff Thomas and Kith Jayasena, Department of Agriculture PESTS 25. How to avoid insect contamination in cereal grain at harvest, Svetlana Micic, Paul Matson and Tony Dore, Department of Agriculture ABIOTIC 26. Environment – is it as important as variety in sprouting tolerance? Thomas (Ben) Biddulph1, Dr Daryl Mares1, Dr Julie Plummer1 and Dr Tim Setter2, School of Plant Biology, University of Western Australia1 and Department of Agriculture2 27. Frost or fiction, Garren Knell, Steve Curtin and Wade Longmuir, ConsultAg Pty Ltd, WA 28. High moisture wheat harvesting in Esperance 2005, Nigel Metz, South East Premium Wheat Growers Association (SEPWA) Projects Coordinator, Esperance, WA SOILS 28. Hardpan penetration ability of wheat roots, Tina Botwright Acuña and Len Wade, School of Plant Biology, University of Western Australia MARKETS 29. Crop shaping to meet predicted market demands for wheat in the 21st Century, Cindy Mills and Peter Stone,Australian Wheat Board, Melbourn

Body composition at birth and its relationship with neonatal anthropometric ratios: the newborn body composition study of the INTERGROWTH-21(st) project.

Background We aimed to describe newborn body composition and identify which anthropometric ratio (weight/length; BMI; or ponderal index, PI) best predicts fat mass (FM) and fat-free mass (FFM). Methods Air-displacement plethysmography (PEA POD) was used to estimate FM, FFM, and body fat percentage (BF%). Associations between FFM, FM, and BF% and weight/length, BMI, and PI were evaluated in 1,019 newborns using multivariate regression analysis. Charts for FM, FFM, and BF% were generated using a prescriptive subsample (n=247). Standards for the best-predicting anthropometric ratio were calculated utilizing the same population used for the INTERGROWTH-21(st) Newborn Size Standards (n=20,479). Results FFM and FM increased consistently during late pregnancy. Differential FM, BF%, and FFM patterns were observed for those born preterm (34(+0)-36(+6) weeks' gestation) and with impaired intrauterine growth. Weight/length by gestational age (GA) was a better predictor of FFM and FM (adjusted R(2)=0.92 and 0.71, respectively) than BMI or PI, independent of sex, GA, and timing of measurement. Results were almost identical when only preterm newborns were studied. We present sex-specific centiles for weight/length ratio for GA. Conclusions Weight/length best predicts newborn FFM and FM. There are differential FM, FFM, and BF% patterns by sex, GA, and size at birth