Identification of yeast genes that confer resistance to chitosan oligosaccharide (COS) using chemogenomics

Background: Chitosan oligosaccharide (COS), a deacetylated derivative of chitin, is an abundant, and renewable natural polymer. COS has higher antimicrobial properties than chitosan and is presumed to act by disrupting/permeabilizing the cell membranes of bacteria, yeast and fungi. COS is relatively non-toxic to mammals. By identifying the molecular and genetic targets of COS, we hope to gain a better understanding of the antifungal mode of action of COS. Results: Three different chemogenomic fitness assays, haploinsufficiency (HIP), homozygous deletion (HOP), and multicopy suppression (MSP) profiling were combined with a transcriptomic analysis to gain insight in to the mode of action and mechanisms of resistance to chitosan oligosaccharides. The fitness assays identified 39 yeast deletion strains sensitive to COS and 21 suppressors of COS sensitivity. The genes identified are involved in processes such as RNA biology (transcription, translation and regulatory mechanisms), membrane functions (e.g. signalling, transport and targeting), membrane structural components, cell division, and proteasome processes. The transcriptomes of control wild type and 5 suppressor strains overexpressing ARL1, BCK2, ERG24, MSG5, or RBA50, were analyzed in the presence and absence of COS. Some of the up-regulated transcripts in the suppressor overexpressing strains exposed to COS included genes involved in transcription, cell cycle, stress response and the Ras signal transduction pathway. Down-regulated transcripts included those encoding protein folding components and respiratory chain proteins. The COS-induced transcriptional response is distinct from previously described environmental stress responses (i.e. thermal, salt, osmotic and oxidative stress) and pre-treatment with these well characterized environmental stressors provided little or any resistance to COS. Conclusions: Overexpression of the ARL1 gene, a member of the Ras superfamily that regulates membrane trafficking, provides protection against COS-induced cell membrane permeability and damage. We found that the ARL1 COS-resistant over-expression strain was as sensitive to Amphotericin B, Fluconazole and Terbinafine as the wild type cells and that when COS and Fluconazole are used in combination they act in a synergistic fashion. The gene targets of COS identified in this study indicate that COS’s mechanism of action is different from other commonly studied fungicides that target membranes, suggesting that COS may be an effective fungicide for drug-resistant fungal pathogens.

Federal Insecticide, Fungicide, and Rodenticide Act [microform] : hearing before the Subcommittee on Department Operations, Research, and Foreign Agriculture of the Committee on Agriculture, House of Representatives, Ninety-seventh Congress, second session, on H.R. 5203.

"Serial no. 97-R."

Les maladies des plantes. Leur traitement raisonné et efficace en agriculture et en horticulture,

Mode of access: Internet.

Handbook on pesticides and their uses in Florida agriculture.

Mode of access: Internet.

Handbook of toxicology.

Includes bibliographical references.

Crop protection digest bulletin series.

Reprinted, in part, from various journals.

Metallo-chemotherapy of plant diseases /

Mode of access: Internet.

List of publications on Fermate fungicide, from September 11, 1934 to July 1, 1944,

Mimeographed.

Du Pont products for crop use; a summary of recommended uses, dosages, timing, tolerances and limitations.

Mode of access: Internet.

Insects and fungi injurious to plants, with remedial treatment (pt. II.)

For pt. I see his Standard fungicides and insecticides in agriculture.

Insect and disease control on vegetables and truck crops,

Mode of access: Internet.

Pesticide background statements.

"August 1984"--Vol. 1; "October 1986"--Vol. 1:Supp.; "October 1986"--Vol. 2.

Elicitors of induced disease resistance in postharvest horticultural crops: a brief review

Increasing loss of conventional fungicides due to pathogen resistance and general unacceptability in terms of public and environmental risk have favoured the introduction of integrated pest management (IPM) programmes. Induction of natural disease resistance (NDR) in harvested horticultural crops using physical, biological and/or chemical elicitors has received increasing attention over recent years, it being considered a preferred strategy for disease management. This article reviews the enhancement of constitutive and inducible antifungal compounds and suppression of postharvest diseases through using elicitors. The effect of timing of pre- and/or postharvest elicitor treatment and environment on the degree of elicitation and the potential for inducing local acquired resistance, systemic acquired resistance and/or induced systemic resistance to reduce postharvest disease is discussed. The review highlights that more applied and basic research is required to understand the role that induced NDR can play in achieving practical suppression of postharvest diseases as part of an IPM approach. (C) 2003 Elsevier B.V. All rights reserved.

Postharvest characteristics and handling of litchi fruit - an overview

Litchi ( Litchi chinensis Sonn.) is a tropical to subtropical crop that originated in South-East Asia. Litchi fruit are prized on the world market for their flavour, semi-translucent white aril and attractive red skin. Litchi is now grown commercially in many countries and production in Australia, China, Israel, South Africa and Thailand has expanded markedly in recent years. Increased production has made significant contributions to economic development in these countries, especially those in South-East Asia. Non-climacteric litchi fruit are harvested at their visual and organoleptic optimum. They are highly perishable and, consequently, have a short life that limits marketability and potential expansion of demand. Pericarp browning and pathological decay are common and important defects of harvested litchi fruit. Postharvest technologies have been developed to reduce these defects. These technologies involve cooling and heating the fruit, use of various packages and packaging materials and the application of fungicides and other chemicals. Through the use of fungicides and refrigeration, litchi fruit have a storage life of about 30 days. However, when they are removed from storage, their shelf life at ambient temperature is very short due to pericarp browning and fruit rotting. Low temperature acclimation or use of chitsoan as a coating can extend the shelf life. Sulfur dioxide fumigation effectively reduces pericarp browning, but approval from Europe, Australia and Japan for this chemical is likely to be withdrawn due to concerns over sulfur residues in fumigated fruit. Thus, sulfur-free postharvest treatments that maintain fruit skin colour are increasingly important. Alternatives to SO2 fumigation for control of pericarp browning and fruit rotting are pre-storage pathogen management, anoxia treatment, and dipping in 2% hydrogen chloride solution for 6-8 min following storage at 0 degrees C. Insect disinfestation has become increasingly important for the expansion of export markets because of quarantine issues associated with some fruit fly species. Thus, effective disinfestation protocols need to be developed. Heat treatment has shown promise as a quarantine technology, but it injures pericarp tissue and results in skin browning. However, heat treatment can be combined with an acid dip treatment that inhibits browning. Therefore, the primary aim of postharvest litchi research remains the achievement of highly coloured fruit which is free of pests and disease. Future research should focus on disease control before harvest, combined acid and heat treatments after harvest and careful temperature management during storage and transport.

Methods to evaluate pesticide damage to the biomass of the soil microflora

Respiratory methods to estimate the amount of C in the soil microbial biomass and the relative contributions of prokaryotes and eukaryotes in the biomass were used to evaluate the influence of pesticides on the soil microflora. Experiments were conducted with 5 and 50 micrograms per gram of three fungicides, captan, thiram and verdesan. At 5 micrograms per gram they caused significant decreases (40%) in the biomass; the organomercury fungicide verdesan also caused a shift from fungal to bacterial dominance. Within 8 days, biomass in captan- and thiram-amended soils had recovered to that of controls. Although the fungal to bacterial balance was restored in verdesan-amended soils, biomass recovery was not complete. At 50 micrograms per gram the fungicides caused long-term decreases in the biomass and altered the relative proportions of the bacterial and fungal populations. Verdesan had the greatest effect on soil microbial biomass and competition.