Southern Illinois University

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Presentation transcript:

Southern Illinois University New Approaches to Study the Cyst Nematode/Sudden Death Syndrome Interaction in Soybean Michael Schmidt Jason P. Bond Ahmad Fakhoury Southern Illinois University

Distribution of SCN and SDS SCN SCN & SDS

Soybean Cyst Nematode Heterodera glycines Many HG types classified Resistance is qualitative/quantitative

Root Knot Nematode Two species of concern in Illinois Meloidogyne incognita (southern root-knot) Has over 500 different hosts including corn Meloidogyne hapla (northern root-knot) More limited host range, corn is a non host Both species are important in soybean, crops, orchards, vineyards, etc.

Sudden Death Syndrome Fusarium virguliforme (Fv). Soilborne, root rotting fungus that colonizes tap root and crown. Produces a toxin causing foliar symptoms (scorch).

Components of SDS Resistance Foliar symptoms - resistance to toxin effects, measured as DX. DX=DS*DI/9. DS= disease severity valued on a 1-9 scale. DI=disease incidence as a percentage of plants in plot having disease. Root infection/colonization - resistance to the fungal pathogen

SCN Interactions with SDS Synergistic – Roy et al., 1989- greenhouse McLean and Lawrence, 1993 - greenhouse Rupe et al., 1991; field trials Hershman et al., 1990, field trials Xing and Westphal, 2006 - microplots Additive Gao et al., 2006 - greenhouse

Limitations - SCN/SDS Research Field studies were/are conducted in fields with SCN SCN densities – generally the resolution at field level. Providing an environment suitable for expression of SDS. Difficult y in quantifying Fv in soil and roots. Need for the right compliment of soybean isolines contrasting for SCN and SDS resistance.

New Tools to Study the Nematode/Fv/Soybean Interaction Develop a transformation system for Fv Generate fluorescing strains of the fungus – GFP = Green fluorescing protein. Identify GFP phenotypes differing in aggressiveness toward soybean. To study the infection & colonization To screen for resistance Producing tools Transformation system Tagged Production of tagged mutants

Restriction Enzyme Mediated Integration (REMI) Linearized Plasmid Fungal DNA Integration of the Linearized Plasmid New Phenotypes Developed Phenotypic Analysis Isolation & Characterization of Regions Flanking the Integration Characterizing the disrupted gene(s)

GFP Strain in Soybean Roots

REMI Results An aggressive Fv isolate known as Mont1 (Monticello, IL) was chosen as source population for transformation. Several GFP strains were produced. An aggressive strain of Fv was identified. A non-aggressive strain was identified.

OBJECTIVE: To use the aggressive and non-aggressive strains to: Determine the role of SCN and RKN in the ability of Fv to penetrate and colonize soybean roots. Determine the role of SCN and RKN in the ability of Fv to cause SDS foliar symptoms. OBJECTIVE: To use the aggressive and non-aggressive strains to:

Experimental Design Greenhouse experiment. Greenhouse experiment. Five soybean cultivars different in their resistance to SCN, RKN, and SDS. Cultivar s were challenged with the GFP-expressing aggressive strain and the GFP-expressing non-aggressive strain. Cultivars were co-challenged with SCN and RKN. The experiment consisted of 30 treatments replicated 5 times. Duration of the experiment was 15 days. Fv root colonization quantified via QPCR. SDS leaf scorch evaluated on a 1-9 scale.

Cultivars Cultivar H. glycines M. incognita F. virguliforme Forrest R P94M50 S Spencer GH3983 LS97-1610

Inoculum Mock Inoculation Aggressive strain Non-aggressive strain SCN + aggressive strain SCN + non-aggressive strain RKN + aggressive strain Five cultivars x Six inoculum levels = 30 treatments. Inoculum: SCN=2300egs/plant, RKN=2400 eggs/plant, Fv=7,700,000spores.

Greenhouse Experiment Waterbath system maintains constant temperature across all experimental units. Five replications, sample size of 3 plants. ~0.5 liter soil/container.

Fungus Fungus + SCN Fungus +RKN Cultivar H. glycines M. incognita F. virguliforme Forrest R P94M50 S Spencer GH3983 LS97-1610

Cultivar H. glycines M. incognita F. virguliforme Forrest R P94M50 S Spencer GH3983 LS97-1610

Cultivar H. glycines M. incognita F. virguliforme Forrest R P94M50 S Spencer GH3983 LS97-1610

Cultivar H. glycines M. incognita F. virguliforme Forrest R P94M50 S Spencer GH3983 LS97-1610

Results Co-inoculation with Fv and either SCN or RKN lead to a significant increase in Fv root colonization and SDS foliar symptoms for all cultivars. Co-inoculating with the fungus and the nematode can overwhelm plant resistance to SDS under greenhouse conditions.