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 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting1

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 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting2

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 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting3

4. Integrated mapping necessary for finding candidate resistance genes Genetic mapping QTL mapping Physical mapping BAC contigs Start 0 CmSI0050 26984 CmSI0660 1585392 CmSI0532 2341908 CmSI0531 2776209 R04_0775 3077960 CmSI0551 4712728 CmSI0437 6041717 Cmp0028 7163814 End 7403370 Genome Sequence contigs/scaffolds 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting4 CD Nelson, USFS; M. Staton, Clemson; J Carlson, Penn State

5. Chestnut crosses at age 2 Field tests of genetic materials are important for advancing program Inoculations at age 6 SG2-3 EP155 Measure canker sizes, late summer, age 6 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting5 The American Chestnut Foundation

6. Gene Transformation – Transgenic Plant Production Selection of transgenic (TG) colonies with Geneticin TG events grown in flasks TG events in somatic embryogenesis (SE) production Agrobacterium (AGL1) infection of chestnut PEMS TG somatic seedlings in pots TG events grown on plates TG somatic shoots in vitro somatic embryos harvested Air-lift bioreactors make cells for transformation every 2 wk 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting6 S. Merkle, Univ. Georgia

7. Early Leaf Assay Necrotic length (mm) American Chinese American Chinese Error bars = 1 SEM T-test: P<.0001 Chinese chestnut leaves consistently show less necrosis than American; same pattern seen in field inoculations -Combined data from 5 experiments (48-52 inocs per leaf type) 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting7 W. Powell, SUNY- ESF

8. Field Tests, evaluating improvements (e.g., 640 transgenic trees & 534 controls) 85/21/2013Kentucky/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 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting9

10. Thousand Cankers Disease basic 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 5/21/2013Kentucky/Tennessee/SRS Stateline Meeting10