1. Andrew David, Associate Professor Department of Forest Resources University of Minnesota and Mike Reichenbach, Extension Educator University of Minnesota February, 2010

2. Overview  Emerald ash borer  Genetic conservation  How to get involved -- seed collection Courtesy of Mark Abrahamson, MDA

3. Emerald Ash Borer  Exotic, invasive insect native to Asia  Found 2002 -- Detroit, MI & Windsor, ONT Courtesy of Mark Abrahamson, MDA

4. Where is EAB?

5.  Houston County April 2009 found in Victory, WI – just across border  Ramsey and Hennepin Counties May 14 th, 2009 discovered in St. Paul – South St. Anthony neighborhood by Rainbow Tree Care EAB quarantines in MN Courtesy of Mark Abrahamson, MDA

6. EAB Lifecycle JFMAMJJASOND Immature (under bark) Adult (free living) David Cappaert, MSU Courtesy of Mark Abrahamson, MDA

7. How Does EAB Kill Trees? Courtesy of Mark Abrahamson, MDA

8. Why is EAB a threat?  All ash species -- seedlings or mature, healthy or not.  Abundant host Courtesy of Mark Abrahamson, MDA

9. EAB – Lack of Control Options  No long range pheromones  Chemical options  Biological controls released 2007  No known resistance in ash populations  Quarantine & eradication S. Perez / The Detroit News

10. Ash are  Culturally important  Ecologically important 51% of the lowland hardwood cover type 149 wildlife species  Economically important Annual ash harvest – 71,000 cords Value -- $15 million dollars per year Courtesy of Michael Benedict

11. Based on these facts it is prudent and proactive to prepare for an invasion of EAB in Minnesota.

12. Project Goals 1. Protect the genetic variation of Minnesota’s ash resource by collecting seed and creating an ex situ seed bank. 2. Evaluate collection strategies using molecular tools to determine the most efficient method to collect ash seed.

13. Three Sampling Strategies  Population Collections  Ecoregion Collections  Volunteer Collections

14. Population Collections  15-20 populations per species  At least 20 trees per population  Minimum of 50 viable seeds per tree  Trees separated by 150 feet or more  Seed sent to NRCS Plant Introduction Station in Ames, IA

16. Ecoregion Collections  Omernik Level III ecoregions  10 – 15 individuals per ecoregion  Seed sent to National Seed Laboratory in Dry Branch, GA

17. Volunteer Collections  Variety of sources  Primarily single tree collections  Seed sent to USDA ARS National Center for Genetic Resources Preservation in Fort Collins, CO

18. Collections Online  Total Collections Green ash – 299 trees Black ash – 205 trees http://forest.nrri.umn.edu/ash/ash-seed-conservation

19. Collection strategies aim to  Capture more than 80% of the genetic variation in the population  Capture traits adapted to local growing conditions

20. Molecular Analysis M B B G G B B B B B G G G G LeavesSeed M B B G G B B B B B G G G G LeavesSeed FM4 FM11

21. Molecular Marker Update  Working with 17 markers 6 optimized in black & green ash 2 others promising for black & green ash 4 additional promising for black ash only 5 show no amplification in either species  Goal is 6 – 8 markers in each species

22. Seed collection -- 2010  Seed from any area is needed  Native wild trees Black ash Green ash White ash

23. USDA Forest Service www.na.fs.fed.us/fhp/eab/maps/maps.shtm Wild trees – Native Variability

24. Seed clusters Seed from native wild trees  Identify the species of ash  Find trees with an abundant seed  Obtain permission to collect Courtesy of R. P. Karrfalt, USDA Forest Service

25. The brown seed color indicates the seed is ripe. Courtesy of R. P. Karrfalt, USDA Forest Service When are seeds ready to collect?

26. The seed coat is brown The seed fills the fruit Whole fruit Fruit torn open exposing seed A firm full sized embryo Radicle (seed root) Cotyledons (seed leaves) Courtesy of R. P. Karrfalt, USDA Forest Service The seed is ready.

27. This seed is not ready. Whole uncut white ash fruit Fruit cut longitudinally Immature seed Placental tissue Courtesy of R. P. Karrfalt, USDA Forest Service

28. A weevil larva has destroyed this seed. Larva Head of larva Courtesy of R. P. Karrfalt, USDA Forest Service

29. These fruits apparently have been attacked by a seed bug. Possible seed bug wounds Courtesy of R. P. Karrfalt, USDA Forest Service

30. Use one data collection form for each tree. Fill out the data sheet as the first step. Number the forms and the trees

31. Write the tree number on the bag. Use paper bags. Courtesy of R. P. Karrfalt, USDA Forest Service

32. Quantity of Seed Try to get 1” to 2” of seed covering the bottom of a large paper bag from each tree. Courtesy of R. P. Karrfalt, USDA Forest Service

33. Bags should be marked with the tree number. Use one bag per tree. Put the collection form in the bag. Courtesy of R. P. Karrfalt, USDA Forest Service

34. Insert at twig sample with the seed. Courtesy of R. P. Karrfalt, USDA Forest Service

35. Insert the seed collection form in the bag, then staple the bag shut. Courtesy of R. P. Karrfalt, USDA Forest Service

36. Send forms and seed to Andrew David University of Minnesota North Central Research and Outreach Center 1861 E Hwy 169 Grand Rapids, MN 55744 Questions: Call or email Mike Reichenbach 218-726-6470 or reich027@umn.edu

37. Seed storage  Cleaned at U of M Research and Outreach Center, Grand Rapids, MN

38. Seed storage  Cold room, 3 0 C between 2 to 4 weeks Weevil larvae will leave the seeds  Weevil invested seeds are separated  Good seed dried to 8% moisture  Stored at -8 0 C (17 0 F) to -20 0 C (-4 0 F)  Viability retained 15-20 years

39.  Andrew David – University of Minnesota  Shanelle Dube – Itasca Community College/ University of Minnesota  Mike Benedict – Bureau of Indian Affairs  David Heinzen, MN Dept of Natural Resources, Resource Assessment  Julie Hendrickson, University of Minnesota  Egon Humenberger, University of Minnesota  Rick Klevorn, MN Dept of Natural Resources, Forestry  Mike Reichenbach, University of Minnesota Extension  Keith Webb – Itasca Community College/ University of Minnesota

40. Thank you  If you have any questions contact Mike Reichenbach reich027@umn.edureich027@umn.edu  Visit http://forest.nrri.umn.edu/ashhttp://forest.nrri.umn.edu/ash  Please complete the evaluation forms and leave them at the back of the room.