Benefits of Adding Physiological Measurements to Long-term Monitoring Efforts

1. Benefits of Adding Physiological Measurements to Long-term Monitoring Efforts Calcium Depletion and Red Spruce / Sugar Maple Decline as Examples

2. USDA Forest Service Paul Schaberg Kelly Baggett John Bennink Brynne Lazarus Paula Murakami

3. The University of Vermont Rubenstein School, Burlington, VT Donald DeHayes Gary Hawley Cathy Borer Tammy Coe Michelle Turner Brett Huggett Josh Halman and many students…

4. Winter Injury of Red Spruce

5. Cold tolerance of several north temperate conifers (sampled in Wolcott, VT on March 3, 1998) Species Mean cold tolerance (ºC) Red spruce -38.1 White pine -59.5 Eastern hemlock -61.0 White spruce -90.0 Red pine -90.0 DeHayes et al. 2001

6. Mechanisms for acid-induced freezing injury of red spruce H+ displacement of membrane-associated Ca (-20%, P<0.05) Acid deposition (H+) Membrane destabilization (+20%, P<0.05) & reduction in messenger Ca Foliar Ca leaching (2-8X increase, P<0.05) Reduced cold tolerance (-10%, P<0.05) Increased susceptibility to freezing injury DeHayes et al. 1999, Schaberg et al. 2000, Schaberg et al. 2001

7. Unresolved issues… • Relevance to field? • Soil Ca depletion? • Just cold tolerance? • Just red spruce?

8. Providing a Broader Context • Field verification • Soil-based Ca depletion • Stress response systems • Other tree species

9. 2003 – high winter injury year

10. Historic Context of 2003 Winter Injury

11. 2003 sampling: 185 plots at 28 sites in VT, NH, eastern NY, and Northwestern MALazarus et al. 2004

12. 2003 sampling: 185 plots at 28 sites in VT, NH, eastern NY, and Northwestern MA • 96% dominant and co- • dominant trees effected • 65% current-year foliage lost • 56% buds killed Lazarus et al. 2004

13. Statistical spatial analysis -significantly greater injury at: • Higher elevations • Western longitudes • West-facing plots Lazarus et al. 2006

14. Providing a Broader Context • Field verification • Soil-based Ca depletion • Stress response systems • Other tree species

15. HBEF watershed 1 (wollastonite) compared to watershed 6 (reference) WollastoniteTreated Oct. 1999 (CaSiO3) Reference

16. Red Spruce Winter injury Slide 15\ Hawley et al. 2006

17. Providing a Broader Context • Field verification • Soil-based Ca depletion • Stress response systems • Other tree species

18. Environmental Change/Stress Signal Temperature extremes, fungal elicitors, wounding, drought, ozone, etc. Signal transduction: Ca moves from area of high concentration (e.g., extracellular mCa or organelle stores) to the low Ca cytosol. Cytoplasmic Ca binds to specific proteins (e.g. calmodulin). Ca2+ Ca2+ protein Ca2+ Ca2+ vacuole Denzyme activity, genetic transcription, etc. changes in cell physiology cell membrane cell wall Plant Adaptation to Environmental Change/Stress Alterations in cold tolerance, stomatal conductance, antimicrobial defense, carbon allocation, etc.

19. Our Overarching Hypothesis Acid Rain, Calcium Depletion and Immune Dysfunction

20. HBEF watershed 1 (wollastonite) compared to watershed 6 (reference) WollastoniteTreated Oct. 1999 (CaSiO3) Reference

21. Red Spruce Winter injury Slide 15\ Hawley et al. in press

22. Foliar Cold Tolerance * P = 0.0431 Halman et al. in prep.

23. Carbohydrate Metabolism • Cell energy relations • Sugars important to cold tolerance • Osmotic control during freezing • Act as an “antifreeze” agent • Direct cryoprotection of biomolecules • Influenced by Ca deficiency

24. Foliar Carbohydrates * * * * Fig. 2. Total sugar levels in current-year foliage were significantly greater in WS1 than in WS6 in both fall and winter collections; * = (p< 0.05)

25. Individual Sugars Table 1. Individual sugar concentrations (mean ± SE, n = 30) in current-year foliage (mg g-1). Three major sugars tested had significantly greater means between watersheds from the fall collection. Data collected from winter collection showed significant differences between watersheds in sucrose and glucose. Note: * = p<0.1; ** = p<0.05

26. Antioxidant Enzymes • Enzymes responsible for scavenging harmful reactive oxygen species • Inadequate function can result in cellular and membrane dysfunction, and/or cell death • Assaying ascorbate peroxidase (APX) – activity dependent upon Ca availability

27. * * Fig. 4. APX activity (mean ± SE, n = 30) from current-year foliage. Winter APX activity is elevated in Ca-addition watershed yet relatively unchanged in reference. * = p < 0.1

28. Providing a Broader Context • Field verification • Soil-based Ca depletion • Stress response systems • Other tree species

29. Field ExamplesCa Depletion and Tree Decline • Red Spruce Winter Injury • Sugar Maple Decline • Flowering Dogwoods and Anthracnose • Norway spruce freezing injury • Hemlock Woolly Adelgid Infestation

30. Sugar Maple Decline • Nutritional predisposition • (D cations – Ca, Mg) • Disproportionate decline • following secondary • stress

31. Soil Ca depletion Secondary stresses drought insects disease pollutants etc. mCa disruption Maple decline Mechanism of Maple Decline? Impairment of stress response system

32. NAMP and HHS Plots Predicted High Ca (6) Predicted Low Ca (8) 6 sugar maples per site 84 total trees

33. Till Source Model: Source Envelope Glacial Movement -Courtesy of Scott Bailey et al. 32 km Designated site 60° •26 HHS & NAMP sites were selected for TSM predictions. • Latitudinal and longitudinal coordinates were used to generate a source envelope. • 8 sites predicted to contain low levels of Ca, 6 were predicted to have high levels of Ca.

34. Schaberg et al2006 Schaberg et al., 2006

35. Schaberg et al2006 Schaberg et al., 2006

36. Schaberg et al2006 Schaberg et al., 2006

37. Hubbard Brook Experimental Forest • NuPert Plots • 12 (45m x 45m) plots • Control plots – marginal Ca nutrition • Randomized Ca or Al additions – begun 1995 • Sugar Maple Dominant Species

38. Minimum level for healthy sugar maple Kolb and McCormick (1993) Total Foliar Cation Analysis Treatment means with different letters are significantly different (P ≤ 0.05) based on orthogonal contrasts: Ca vs. Al and Control; Al vs. Control

39. increased dieback decreased dieback Crown Health Evaluation

40. Treatments Began Tree Increment Growth

41. Ice Storm Treatments Began Tree Increment Growth

42. Wound Closure Amount Results • Differences associated w/ Ca fertilization were most pronounced in wounds lower of the tree stem (100cm-130cm, P = 0.0016) Huggett et al. submitted to CJFR

43. Sugar Maple Shoot Cold Tolerance Site Soil Ca Status Cold Tolerance (oC) March 8, 2006

44. Providing a Broader Context • Field verification • Soil-based Ca depletion • Stress response systems • Other tree species

45. Synergies: Physiology and Monitoring Gain information about the relevance of experimental evidence to trees/forests in the “real world”

46. Synergies: Physiology and Monitoring Physiological measurements can add a mechanistic understanding to changes in forest health and productivity noted via monitoring

47. Questions?