1/26/2015. Overview. Types of soil-borne plant pathogens
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1 /6/5 PLP 644 Epidemiology of Plant Diseases Spring 5 Lecture 6: Influence of Pathogen on Disease Development Soil-borne Prof. Dr. Ariena van Bruggen Emerging Pathogens Institute and Plant Pathology Department, IFAS University of Florida at Gainesville Overview Types of soilborne plant Examples of soilborne Factors affecting root-pathogen interactions Pathogen characteristics that influence epidemic development Competitive saprotrophic ability and inoculum potential Quantification of soilborne inoculum DI:ID relationships Conclusions Types of soil-borne plant soil inhabitants competitive saprotrophic ability (on dead organic matter) wide host range secondary or of young plants root inhabitants Little or no competitive saprotrophic ability specialized parasites, primary small or wide host range Examples of ecological types of soilborne Soil inhabitant, good CSA: Pythium, Rhizoctonia, Fusarium; Rhizorhapis and Ralstonia (bacteria) Soil inhabitant, limited CSA: Phytophthora, Verticillium Examples of ecological types of soilborne Soil inhabitant, no CSA: Pratylenchus penetrans Root inhabitant, no CSA: Plasmodiophora, Meloidogyne Infection by soil-borne Some important terms: Rhizoplane- on the root surface Rhizosphere- near the root but within the influence of the root (from exudates, volatiles etc.) Spermoplane- on the surface of seed (in the soil) Spermosphere- near the seed in the soil Factors affecting infection: Saprophytic growth in soil, spermo- and rhizosphere Competitive saprophytic ability Inoculum potential Ability to colonize the spermo- or rhizoplane Ability to infect the roots
2 /6/5 Pathogen characteristics that influence epidemic development of soilborne diseases Pathogen population size (number of propagules) Level of aggressiveness and virulence Inoculum potential Reproduction (sexual, asexual, rate) Ecology Pesticide resistance (fungicides, antibiotics) Dispersal mechanisms Strategies determining the success of infection by soilborne The number of spores and their vitality = inoculum potential Inoculum potential = inoculum density x aggressivity inoculum density = # spores / g soil or meters of hyphae /g soil Aggressivity = number of infection units - can be determined only in bioassays! so: inoculum potential also can only be determined in bioassays quantification interpretation inoculum density easy difficult inoculum potential difficult easy Strategies determining the success of colonisation by soilborne Characteristics ( weapons ) of the colonist = competitive saprotrophic ability Competitive saprotrophic ability =all physiological characteristics that makes an organism suited to colonize dead organic matter Method to determine CSA Fusarium culmorum Cochliobolus 75 sativus Gaeumannomyces 5 graminis 5 5 inoculum added (vol. %) colonised pieces of straw Factors affecting the competitive saprotrophic ability fast germination of survival structures fast growth of mycelium fast formation of enzymes needed to break down the substrate production of antibiotics, toxins or bacteriocins that inhibit competitors tolerance for antibiotics etc. formed by other organisms able to survive unfavorable periods Quantification of soilborne inoculum Extraction and quantification of propagules from soil Soil dilution plating for fungi and bacteria Enrichment with organic matter before dilution plating Warcup plate method (spread soil on plate cover with agar) Anderson sampler with appropriate sieves on layers of petriplates Wet-sieving / flotation possibly in sucrose (sclerotia, nematodes) Disease tests with naturally infested soil Plant seeds / seedlings in soil in a dilution series with sterile soil Baiting techniques Place autoclaved leaves etc in soil, plate after incubation and count positive samples (not really quantitative; why not?) Quantification of soilborne inoculum Dilution end point and most probable number methods Mix field soil with sterilized soil at different ratios Place baits or plants in individual cups or wells Determine dilution at which no more infection occurs use MPN table or computer program to calculate MPN/ g dry soil Immunological techniques Soil extracts in ELISA tests with mono-/ polyclonal antibodies Immunomagnetic capture Test kits available, but quantification is still problematic DNA techniques Extraction of DNA; amplify by PCR with spec. primers (qualitative) Same but use quantitative or qpcr (also called real-time or rtpcr); this is semiquantitative
3 /6/5 A lot of discussion in 97 s, 8 s and 9 s Van der Plank (975 Principles of Plant Infection ): six basic types of responses for the DI:ID relationship ) Disease is directly proportional to inoculum dose (straight line passing through the origin (,); slope is infection efficiency; IE=constant) Data from Schein (964) In this case, the spores 8 neither interact nor 6 IE =.9 compete for sites. 4 Disease intensity Colonies cm Urediniospores Spore density cm - Six basic types of responses for the DI:ID relationship ) DI:ID decreases as inoculum increases; something limits IE (availability of susceptible host sites or inhibitory interactions among propagules) ) Antagonistic response: the DI decreases with increases in ID (accumulation of inhibitory products in the presence of crowded spores) Number of infections Number of spores Six basic types of responses for the DI:ID relationship 4) Synergistic response: DI:ID increases with increases in ID 5) ID has to exceed a certain threshold 4 before the first infection (a large number of spores is needed to attain the probability that one spore will reach a susceptible site 6) Impossible response where disease seemingly occurs without inoculum! (curves like #6 in the literature; be suspicious!) Number of infections 6 5 Number of spores Basic assumption under Van der Plank s curves: All propagules act independently, independent-action model Each organism has a certain probability of causing an infection Total probability of infection from a given dose is the statistical combination of the probabilities for all the individual organisms The model of independent action: y = N [ - exp(-ax)] y = number of infections, x = number of spores, N = total number of susceptible sites, and a = the rate parameter that determines the proportion of effective units of inoculum High values of a mean very efficient inoculum Low values of N mean low numbers of susceptible sites With high a and low N, the monomolecular DI:ID curve has a strong curvature DI:ID relationship: y = N [ - exp(-ax)] high a low a high N high a high N low a Ralph Baker (Colorado State Univ.) published extensively on the ID:DI relationship in the 97s and 98s, mainly of Rhizoctonia solani on radish Baker s concepts were severely criticized and a series of letters to the editor in Phytopathology resulted Four possible kinds of infection courts and their interaction with inoculum (Baker et al., Phytopathology 57:66-666): Fixed court non-motile inoculum e.g. hypocotyl and sclerotia; infection occurs by chance when a sclerotium is next to a suitable infection court based on the density of inoculum and the density of infection courts.
4 /6/5 Four possible kinds of infection courts and their interaction with inoculum (continued): fixed infection court- motile inoculum; e.g., hypocotyl and zoospores or growing hyphae moving infection court- non-motile inoculum; e.g., root tip and sclerotia moving infection court and motile inoculum; e.g., root tip and zoospores or growing hyphae. Moving infection courts and motile inoculum would reduce the distance between propagules and infection court and thus increase the likelihood of infection Pathogen propagules considered to be arranged in soil as points at the vertices of a tetrahedron (simplest geometric form to utilize all space efficiently) To describe the ID:DI relationship mathematically, they plotted log (I) [successful infections] vs. log (D) [distance between propagules]. The slopes for fixed inoculum (with either fixed or moving infection courts) was about.67 The slopes for motile inoculum were about. Baker interpreted the slopes of.67 as a rhizoplane effect and slopes of. as a rhizosphere effect. Rhizosphere rhizoplane hypothesis by Baker Baker et al. (Phytopathology 6:8-9): log [ln (/(-y))] vs. log (ID) to linearize many curves Most slopes had values either close to. or close to.67 Baker interpreted the log-log slopes of. as a rhizosphere effect and the slopes of.67 (/) as a rhizoplane effect Many plant pathologists who worked with soil-borne infections used Baker s analyses, but there were sceptics Grogan (UCD) called this transformation a triple log transformation Baker et al., 98 Rhizosphere-rhizoplane hypothesis - criticism by Grogan Van der Plank rejects Baker s / hypothesis because: root diseases actually do not differ from leaf diseases; / slopes are also found for leaf diseases. Baker s models do not predict ID:DI curves for known root diseases from prior work. Baker s models are based on three erroneous assumptions: a) spores are mathematical points in soil with no dimensions or volume; b) spores are uniformly distributed (vertices of a tetrahedron); c) infection is NOT restricted by the number of susceptible sites. 4
5 /6/5 DI:ID (or Pf:Pi) relationships for nematodes Various models relate final density of nematodes (Pf) after harvest of the crop to the initial density (Pi) before the crop is sown A common structure of these models is: /α is slope of curve in origin /β is horizontal asymptote Valid for cyst and root knot nematodes with one generation per season Van den Berg: also valid for migratory nematodes (Pratylenchus penetrans) DI-ID (or Pf:Pi) relationships for nematodes Van Den Berg used Pf:Pi relations for single crops to predict population dynamics and crop loss for four different crops in different rotations (used complex nonlinear models and Monte Carlo simulations) Van den Berg and Rossing, 5 Conclusion Many pathogen characteristics determine epidemic development by soilborne Which factors? What is competitive saprotrophic ability? How can you measure it? What is inoculum potential? How can you measure it? How can you quantify inoculum density? Which technique is best and why? What is the relationship between inoculum density and disease intensity? What was the DI:ID controversy about? 5
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