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.

Cell wall degrading enzymes and interaction between Trichoderma Aggressivum and Agaricus Bisporus

Agaricus bisporus is the most commonly cultivated mushroom in North America and has a great economic value. Green mould is a serious disease of A. bisporus and causes major reductions in mushroom crop production. The causative agent of green mould disease in North America was identified as Trichoderma aggressivum f. aggressivum. Variations in the disease resistance have been shown in the different commercial mushroom strains. The purpose of this study is to continue investigations of the interactions between T. aggressivum and A. bisporus during the development of green mould disease. The main focus of the research was to study the roles of cell wall degrading enzymes in green mould disease resistance and pathogenesis. First, we tried to isolate and sequence the N-acetylglucosaminidase from A. bisporus to understand the defensive mechanism of mushroom against the disease. However, the lack of genomic and proteomic information of A. bisporus limited our efforts. Next, T. aggressivum cell wall degrading enzymes that are thought to attack Agaricus and mediate the disease development were examined. The three cell wall degrading enzymes genes, encoding endochitinase (ech42), glucanase (fJ-1,3 glucanase) and protease (prb 1), were isolated and sequenced from T. aggressivum f. aggressivum. The sequence data showed significant homology with the corresponding genes from other fungi including Trichoderma species. The transcription levels of the three T. aggressivum cell wall degrading enzymes were studied during the in vitro co-cultivation with A. bisporus using R T -qPCR. The transcription levels of the three genes were significantly upregulated compared to the solitary culture levels but were upregulated to a lesser extent in co-cultivation with a resistant strain of A. bisporus than with a sensitive strain. An Agrobacterium tumefaciens transformation system was developed for T. aggressivum and was used to transform three silencing plasmids to construct three new T. aggressivum phenotypes, each with a silenced cell wall degrading enzyme. The silencing efficiency was determined by RT-qPCR during the individual in vitro cocultivation of each of the new phenotypes with A. bisporus. The results showed that the expression of the three enzymes was significantly decreased during the in vitro cocultivation when compared with the wild type. The phenotypes were co-cultivated with A. bisporus on compost with monitoring the green mould disease progression. The data indicated that prbi and ech42 genes is more important in disease progression than the p- 1,3 glucanase gene. Finally, the present study emphasises the role of the three cell wall degrading enzymes in green mould disease infection and may provide a promising tool for disease management.

Diseases and pests of ornamental plants, a manual for growers,

Mimeographed.

Protein subcellular targeting inTrichoderma aggressivum f. aggressivum

Trichoderma aggressivum f. aggressivum is a filamentous soil fungus. Green mold disease of commercial mushrooms caused by this species in North America has resulted in millions of dollars in lost revenue within the mushroom growing industry. Research on the molecular level of T aggressivum have jus t begun with the goal of understanding the functions of each gene and protein, and their expression control. Protein targeting has not been well studied in this species yet. Therefore, the intent of this study was to test the protein localization and production levels in T aggressivum with green fluorescent protein (GFP) with an intron and tagged with either nuclear localization signal (NLS) or an endoplasmic reticulum retention signal (KDEL). Two GFP constructs (with and without the intron) were used as controls in this study. All four constructs were successfully transferred into T aggressivum and all modified strains showed similar growth characteristics as the wild type non-transformed isolate. GFP expression was detected from all modified T aggressivum with confocal microscopy and the expression was similar in all four strains. The intron tested in this study had no or very minor effects as GFP expression was similar with or without it. The GFP signal increased over a 5 day period for all transformants, while the GFP to total protein ratio decreased over the same period for all transformants. The GFP-KDEL transformant showed similar protein expression level and localization as did the control transformant lacking the KDEL retention signal. The GFP-NLS transformant similarly failed to localize GFP into nucleus as fluorescence with this strain was virtually identical to the GFP transformant lacking the NLS. Thus, future research is required to find effective localization signals for T aggressivum.

Healthy hardwoods. A field guide to pests, diseases and nutritional disorders in subtropical hardwoods.

Healthy hardwoods: A field guide to pests, diseases and nutritional disorders in subtropical hardwoods can be used to help identify the common damaging insects, fungi and nutritional disorders in young eucalypt (Eucalyptus and Corymbia species) plantations in subtropical eastern Australia. This guide includes photographs of each insect, fungus and nutritional disorder and the damage they cause, along with a brief description to aid identification. A brief host list for insects and fungi, including susceptibility and occurrence, is provided as a guide only. A hand lens will be useful, especially to identify fungi. Although it is possible to identify insects and fungi from these photographs, laboratory examination will sometimes be necessary. For example, microscopes and culturing media might be used to identify fungi. Information about four exotic pests and diseases has also been included in the Biosecurity threats chapter. Potentially, these would have a severe impact on plantation and natural forests if introduced into Australia. To prevent establishment of these pests, early detection and identification is crucial. If an exotic insect or disease is suspected, then an immediate response is required. Usually, the first response will be to contact the nearest Australian Quarantine and Inspection Service office or forestry agency to seek advice.

Studies on the composition and nutritional value of the cultivated mushroom, agaricus bisporus (Lange) Sing

The effects of various cultural conditions on the composition and nutritional quality of Agaricus bisporus (Lange) Sing. were investigated. Variation in composition was found between different classes of sample. Sampling techniques were standardised to allow for major variations in the different developmental stages and culture ages. Fruitbodies were found to be of low calorific value but contained protein of high digestibility and quality, containing all the essential amino acids required by man. Quantitative estimates of the sulphur-containing amino acids indicated that fruitbodies were deficient in methionine and cysteine. The extent of water application and the supplementation of conventional substrates with various nitrogen-containing substances, influenced yield and composition, establishing the importance of these two factors in the physiology of fruitbodies and cultural management. Storage conditions influenced composition, high temperatures being deleterious to the nutritional value of fruitbodies. Submerged culture techniques were used to investigate the effects of various nutrients on growth and composition of mushroom mycelium, with special reference to the sulphur-containing amino acids. Yield and composition were greatly affected by the carbon:nitrogen ratio of the medium and by the nitrogen source. Significant increases in mycelial methionine content were observed on the addition of inorganic sulphate, the methionine derivative N-acetyl-L-methionine, and L-methionine. A greater increase in methionine content was obtained when the biomass of a thermophilic bacterium isolated from compost was used as a nitrogen source.

The importance of cotton disease surveys in Queensland for monitoring endemic diseases and detecting new pathogens and pests

The cotton industry in Australia funds biannual disease surveys conducted by plant pathologists. The objective of these surveys is to monitor the distribution and importance of key endemic pests and record the presence or absence of new or exotic diseases. Surveys have been conducted in Queensland since 2002/03, with surveillance undertaken by experienced plant pathologists. Monitoring of endemic diseases indicates the impact of farming practices on disease incidence and severity. The information collected gives direction to cotton disease research. Routine diagnostics has provided early detection of new disease problems which include 1) the identification of Nematospora coryli, a pathogenic yeast associated with seed and internal boll rot; and 2) Rotylenchulus reniformis, a plant-parasitic nematode. This finding established the need for an intensive survey of the Theodore district revealing that reniform was prevalent across the district at populations causing up to 30% yield loss. Surveys have identified an exotic defoliating strain (VCG 1A) and non-defoliating strains of Verticillium dahliae, which cause Verticillium wilt. An intensive study of the diversity of V. dahliae and the impact these strains have on cotton are underway. Results demonstrate the necessity of general multi-pest surveillance systems in broad acre agriculture in providing (1) an ongoing evaluation of current integrated disease management practices and (2) early detection for a suite of exotic pests and previously unknown pests.

Weekly report on insects, diseases, and crop development to cooperative extension agents.

Mode of access: Internet.

The casing layer in the culture of Agaricus bisporus, the cultivated mushroom

The casing layer is an essential component of the system employed in the culture of Agaricus bisporus. The literature appropriate to the casing layer is fully reviewed, including aspects relating to fructification and morphogenesis in A.bisporus, together with an appraisal of the various media employed, their properties and functions, and the commercial significance of the casing layer. Equipment is described for use in experiments in mushroom culture, based on a scaled-down version of normal growing technique, allowing the analysis of both weights and number of fruitbodies forming, which was useful in assessing the effects of different casing treatments. The basic steps in the production of fruitbodies in A.bisporus.are described, including a photographic study of the colonisation of casing and fructification. Various alterations to the physical structure of peat/chalk casing mixtures were found to have an effect on fructification; those causing an opening-out of the casing structure tended to give better yields, especially in the early stages of production. It was shown that, in order to obtain greater yield through casing amendment, fructification must be stimulated, giving increased numbers of fruitbodies, disproportionate to their total weight and consequently of lower mean weight. A synthetic casing medium based on the light glass-like mineral, perlite, was developed. The best formula obtained was -.1 part perlite: 1 part montmorillonite clay (by weight): 3 parts 0.01% glucose solution. Perlite/montmorillonite casing could be improved by adding compost colonised by mycelium of A.bisporus, or adding a peat-chalk casing extract. Perlite was also found to be suitable for admixture with the standard casing medium and a mixture of equal parts by volume performed as well as the peat/chalk casing normally used.

Soil Health, Soil Biology, Soilborne Diseases and Sustainable Agriculture

Soil Health, Soil Biology, Soilborne Diseases and Sustainable Agriculture provides readily understandable information about the bacteria, fungi, nematodes and other soil organisms that not only harm food crops but also help them take up water and nutrients and protect them from root diseases. Complete with illustrations and practical case studies, it provides growers and their consultants with holistic solutions for building an active and diverse soil biological community capable of improving soil structure, enhancing plant nutrient uptake and suppressing root pests and pathogens.