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Microbes and Mulches

Microbes and Mulches. Marianne Powell Masters Student Dr. Debra Inglis, Graduate Advisor Dr. Marion Brodhagen, Committee Member; USDA Lab Host Dr. Carol Miles, Committee Member Dr. Karen Leonas, Materials Lab Host Graduate Student Symposium SCRI Team Meeting Lubbock, TX June 2, 2011.

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Microbes and Mulches

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  1. Microbes and Mulches Marianne Powell Masters Student Dr. Debra Inglis, Graduate Advisor Dr. Marion Brodhagen, Committee Member; USDA Lab Host Dr. Carol Miles, Committee Member Dr. Karen Leonas, Materials Lab Host Graduate Student Symposium SCRI Team Meeting Lubbock, TX June 2, 2011 1

  2. Today’s Outline • Isolation of potential biodegradable mulch utilizing microbes from TN, TX, and WA mesh bag studies A. Colony forming units per gram of mulch B. Number of isolates that degraded BDMs C. Conclusions from the Mesh Bag Study II. 2010 WA disease assessments in the field over time A. Tomato diseases and disorders in the BDM experiment B. Koch’s Postulates of Verticillium wilt on lettuce C. Conclusions from the 2010 growing season 2

  3. Mesh Bag Study: Isolate Microbes Utilizing BDM Material as C Source Visit to Brodhagen laboratory at USDA ARS Forage Seed and Cereal Research Unit, Corvallis, OR 3

  4. BDM Samples Examined 4

  5. Microbe Isolation and BDM Biodegradation Assay 1 3 2 Multipipetter Serial dilutions 0.5 g mulch in PBS 5 6 4 Minimal media with BDM sample After incubation Dilution plates 5

  6. 1: WA/IN/SB Fungi 1st BDM Test 2: TN/IN/BT #9 Bacteria 1000X, ˽ = 3.0 µm 4: TN/IN/BT #9 Bacteria 75X,growing on mulch 3: TX/OUT/BB #2 Fungi 20X, growing on mulch 6

  7. Fifty microbes capable of utilizing BDMs were obtained from TN and TX. 36 fungal and 14 bacterial isolates have been shown to degrade SB, BB, and BT. WAsamples are still in progress. 8

  8. Biodegradation Assay Conclusions • We have developed a novel assay for testing BDM utilization by soil bacteria and fungi. • We have isolated native bacteria and fungi capable of degrading each of the BDMs from TN and TX. • The Brodhagen lab will confirm our initial results in future testsand identify microbes with universal bacteria and fungal primers • Some of these isolates are likely to be redundant (e.g. same species). • In future studies, BDM degradation by these microbes could potentially be tested via composting studies - ASTM D5338 (2003) TN/IN/BT/ #15 Fungi 9

  9. Tomato Disease Ratings in WA BDM Study • Weekly visual assessments of disease incidence and severity Physiological leaf roll Leaf mold suspect (HT) Verticillium wilt suspect (HT) 10

  10. Culturing Fungal Organisms from Symptomatic Tissues ITS-PCR sequencing Morphology ID Morphology ID Ulocladium spp. Botrytis spp. Fulvia fulva 11

  11. AUDPC for Physiological Leaf Roll BT BB SB NM 12

  12. Pathogens in Nearby Plots Lettuce is susceptible to Verticillium wilt, caused by V. dahliae; we isolated and confirmed (ITS) V. tricorpus. 13

  13. Source of Verticillium sp. on Lettuce? Seedborne? Soilborne? Soil dilution plates from plots with diseased lettuce Tested seeds from lettuce seed lot for 2010 planting V. Colonies growing on NP-10 Counting V. spp. 14

  14. Lettuce: Verticillium Assays From the seed? From the soil? 15

  15. Completing Koch’s Postulates on Lettuce Inoculate seedling Grow inoculum Evaluate disease Culture and characterize 16

  16. Conclusions for 2010 • V. tricorpus isolated from lettuce in 2010 is pathogenic on lettuce and this is a first report for WA state (Japan) • The non-mulch and spundbond-PLA treatments in the WA open field plots showed reduced physiological leaf roll severity and this could be an important clue to as what causes the disorder, which is currently unknown. • No BDM treatment differences to leaf mold (F. fulva) noted on tomato cultivar ‘Celebrity’, but Celebrity showed resistance in 2010 (Inglis tomato cultivar study finding). 17

  17. Special Thanks • Dr. Debra Ann Inglis • Babette Gunderson • Jeremy Cowen • Dr. Marion Brodhagen • Ashley Florence • Carl Evans

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