Forest Health Initiative: A Model for Rapid Response to Emerging Pests

1. Forest Health Initiative:A Model for Rapid Response to Emerging Pests C. Dana Nelson, Project Leader/Research Geneticist dananelson@fs.fed.us Southern Research Station Southern Institute of Forest Genetics Harrison Experimental Forest Saucier, MS Kentucky/Tennessee/SRS Stateline Meeting

2. Forest Health Crisis • Invasive pests, climate change • New approaches and tools required to slow decline and recover health • Forest Health Initiative developed to test a hypothesis: • that a coordinated effort in biotechnology research will lead to resistant trees capable of restoring a species in a relevant time frame • American chestnut selected as test case • www.foresthealthinitiative.org Kentucky/Tennessee/SRS Stateline Meeting

3. Forest Health Initiative (FHI) • Key components of FHI hypothesis: • Coordinated effort • Steering Committee, Social/Environmental Group, Science Advisory Committee, PIs, Cooperating Scientists and their research teams • Biotechnology research • Biological Sciences effort– genetics, molecular biology, pathology • Resistant trees in Relevant time • ensure blight resistance through seeds from adapted and diverse American chestnuts • 3 years to a ‘plantable tree’ • tangible progress w/array of promising trees going to field tests • infrastructure/know-how-- to plant and grow trees in reintroduction/restoration Kentucky/Tennessee/SRS Stateline Meeting

4. Start 0 CmSI0050 26984 CmSI0660 1585392 CmSI0532 2341908 CmSI0531 2776209 R04_0775 3077960 CmSI0551 4712728 CmSI0437 6041717 Cmp0028 7163814 End 7403370 Integrated mapping necessary for finding candidate resistance genes Genetic mapping QTL mapping Physical mapping BAC contigs Genome Sequence contigs/scaffolds Kentucky/Tennessee/SRS Stateline Meeting CD Nelson, USFS; M. Staton, Clemson; J Carlson, Penn State

5. Field tests of genetic materials are important for advancing program SG2-3 Chestnut crosses at age 2 EP155 Inoculations at age 6 Measure canker sizes, late summer, age 6 Kentucky/Tennessee/SRS Stateline Meeting The American Chestnut Foundation

6. Gene Transformation – Transgenic Plant Production Agrobacterium (AGL1) infection of chestnut PEMS TG somatic seedlings in pots Selection of transgenic (TG) colonies with Geneticin TG somatic shoots in vitro Air-lift bioreactors make cells for transformation every 2 wk TG events in somatic embryogenesis (SE) production TG events grown in flasks TG events grown on plates somatic embryos harvested Kentucky/Tennessee/SRS Stateline Meeting S. Merkle, Univ. Georgia

7. Early Leaf Assay American Necrotic length (mm) Chinese American Chinese Error bars = 1 SEM T-test: P<.0001 • Combined data from 5 experiments (48-52 inocs per leaf type) Chinese chestnut leaves consistently show less necrosis than American; same pattern seen in field inoculations Kentucky/Tennessee/SRS Stateline Meeting W. Powell, SUNY- ESF

8. Field Tests, evaluating improvements(e.g., 640 transgenic trees & 534 controls) Kentucky/Tennessee/SRS Stateline Meeting W. Powell, SUNY- ESF

9. FHI Conclusions • FHI hypothesis is tentatively accepted • ‘plantable trees’ have been achieved in short period • know-how & infrastructure developed • Continuing work will • evaluate the new materials under field conditions • advance winners to additional field tests • apply mapping resources to breeding programs • Adapt FHI to new case • e.g., thousand cankers disease Kentucky/Tennessee/SRS Stateline Meeting

10. Thousand Cankers Diseasebasic biological sciences needs • Screening methods for assaying resistance • insect vector and/or fungus pathogen • Genetics of host resistance/susceptibility • Markers and genes for resistance • Biotech tools for engineering resistance • Breeding populations for deploying resistance • Cooperators with trees of interest and land for field testing Kentucky/Tennessee/SRS Stateline Meeting