White Pine Species Resources & Blister Rust Resistance – What Resources are Available for Genomics? PNW Region Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Pacific Northwest Region Regional + Inter-regional Scope – Facilities – Dorena GRC – Field sites – Common garden testing – Clone banking/orchards Scope – Large Scale Operational Testing –& ‘phenotyping’ – Seed storage – 1000’s of seedlots – Species Personnel – >>80 years rust resistance experience Contacts – Partners/Cooperators – U.S – Canada – Europe – Asia – – Breeding and genetic resources of five-needle pines Proposed 2014 Meeting Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

1739.jpg Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; Inoculation Chamber

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Whitebark pine -225 families 2 sources of rust (SY2007) Limber pine (SY2007) WWP (SY2007 ) Sugar Pine (SY2009) SWWP (SY2009) Sugar pine (SY2008) 8 WWP Diallels WWP SY2006 Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; Rust testing at Dorena GRC

Resistance Complete Resistance – ‘MGR’ – Some species Other (partial resistance) – All species Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Species P. monticola (western white pine) P. lambertiana (sugar pine) P. albicaulis (whitebark pine) P. strobiformis* (southwestern white pine) P. flexilis** (limber pine) P. aristata** (Rocky Mountain bristlecone pine) P. longaeva** (Great Basin bristlecone pine) Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Sugar Pine Rust Testing – 1000’s of parent trees – Surviving progeny available Seedlots Available – 1000’s of wild OP and CC’s Field trials – 6 long term (1982/1983) progeny tests ( see FHM poster ) – 5 provenance trials ( last assessed by DGRC ) – 1998 Diallel trial at HC (3 six parent half diallels – 41 of 45 crosses) – Other field trials since 1996 Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Sugar Pine Provenance Field Sites, Five Site Rust Summary, , DRAFT Year# TreesLast# Trees % Alive Mean # BoleMean # Branch Planted InspectionAlive with RustCankers a Notes: Cannon < 1%~1 Much charcoal root rot mortality Fitch-Rantz % Thinned after 2009 evaluation Burnt Timber % trees thinned in 2001 Sundown % Branches pruned Harrel % a Excludes trees without stem symptoms 2010 FHM Poster: Long-term Monitoring of White Pine Blister Rust Infection and Survival at 10 Sugar Pine Evaluation Sites J.L. Hill, R. Sniezko, D. Vogler, J.W. Wright FHM Grant: WC-F Long-term Monitoring of White Pine Blister Rust Infection and Survival at 10 Sugar Pine Evaluation SitesLong-term Monitoring of White Pine Blister Rust Infection and Survival at 10 Sugar Pine Evaluation Sites (Jerry Hill, Rich Sniezko, Ellen Goheen1 of 3 $23,000) FHM Proposal & 62 provenances planted in 4 sites 2 in OR, 2 in CA

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; Sugar Pine Diallels (RV3 – Happy Camp) ~41 families, ~72 trees/fam, Planted 1998

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; B B B B #Stem Symptoms/tree for 6 parents in half diallel (based on 7 of 12 reps ~2007 data) DRAFT & 2012 assessment planned B B B B %52.7%83.7%90.9%87.5% 79.0% 86.3%54.4%86.0%73.1%79.2%98.3% 56.7% 94.8%70.9% 94.8% 79.2%73.1%54.4%86.0%98.3%86.3% 83.7% 79.0% 87.5%81.7%52.7%90.9% 77.5% 63.2% 81.3%80.5%77.7% 89.9%

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; Forest Health Monitoring 2010 National Meeting, Albuquerque, NM s.fed.us/posters/i ndex.shtmlhttp:// s/posters/index.sh tml

Long-term Monitoring of White Pine Blister Rust Infection and Survival at 10 Sugar Pine Evaluation Sites R.A. Sniezko 1, R. Danchok 1, S. Long 1, D.P. Savin 1, A. Kegley 1, J. Mayo 1, J. Hill 2 1 Dorena Genetic Resource Center, Shoreview Road Cottage Grove, OR 97424, 2 National Park Service, Southeast Utah Group, 2282 S. West Resource Blvd. Moab, UT MATERIALS AND METHODS Sugar pine seedlings from 53 families were planted in randomized complete block designs at six sites in southwest Oregon in 1982 and 1983 (Table 1; Figure 1). The families were distributed between two Sets and the Sets planted in separate adjacent trials at each site at 2.4 x 2.4 m spacing. Up to 40 seedlings per family were planted at each site in 10 tree row plots (except at Poker where single tree non-contiguous plots were used). The parent trees for all progenies originated from southern OR (Figure 1). 31 families were common to all sites. Based on seedling screening, the families were expected to represent a range in resistance from highly susceptible to partial resistant to major gene resistance (Cr1). The sites were rated for blister rust hazard in the mid-1980’s – and represented a range from low to high rust. Rust hazard was based on infection levels of 50 sampled trees from natural regeneration and the number of Ribes plants per 1/100 th acre plot within the sample area. The sites were assessed for height, survival, blister rust, and other damage at approximately 5, 10, 15 and 25 years after planting. DBH was recorded at the latest assessment (2006 to 2010, depending on the site, so tree age varied from 24 to 29). Figure 1. Location of test sites and parents represented in six progeny tests SITE Elevation (m) Slope (%) Aspect Rust Hazard Year Sown Year Planted Year of Last Inspection # of Trees in Study a # of Families Represented Anchor114310WestMedium Boulder109720EastMedium Hayes86950SouthHigh Jamison85320SouthLow Poker128035WestMedium Rocky97535NorthHigh a Excludes filler/replaced trees Table 1. Descriptions of progeny test sites. Figure 3. Mean and range of family survival at age ‘25’ at each site and averaged over all sites. ACKNOWLEDGMENTS We gratefully acknowledge BLM (and SIS NF) personnel for their insight and lead in establishing, maintaining and assessing these valuable trials since the early 1980’s. For help with the latest assessments, we thank BLM Medford District personnel, notably Jim Brimble and the silviculture group, and Paul LeBlanc and Dave Russell; and Paula Trudeau and Brian Luis (FS). Partial funding of this project was provided FS’s Forest Health Monitoring grant WC-F This proposal was sponsored by Ellen and Don Goheen. Jennifer Christie produced the map. FUTURE ACTIVITIES Several sites have had brush removal and/or been retagged to greatly facilitate future evaluations of these trials. Begin breeding work with the progeny selections. More detailed analysis and summary including: Examination of family variation in blister rust resistance, including whether there is evidence of a virulent vcr1 strain of blister rust, and whether efficacy of resistance varied by site or rust hazard. Further examination of potential family variation in non-rust related mortality and possible maladaptation. Comparative growth of trees with and without cankers. Examination and mapping of rust hazard estimates for 265 plots from the BLM’s 1980s surveys. Figure5. Height (mean and range) at latest assessment (24-29 years) of surviving trees at each site. Height of cankered (red) and non-cankered (green) shown. Large natural sugar pine near Boulder test site INTRODUCTION Sugar pine (Pinus lambertiana) is an important long-lived conifer in forest ecosystems in California and Oregon. Unfortunately, it is extremely susceptible to the white pine blister rust (WPBR), caused by the non-native fungal pathogen Cronartium ribicola. WPBR has caused high mortality of sugar pine in many areas of Oregon, reducing its incidence in many ecosystems as well as its use in reforestation and restoration. Fortunately, there is some genetic resistance to WPBR. The USDA Forest Service (FS) and USDI Bureau of Land Management (BLM) have been working together since the mid-1960’s to select candidate trees and test their progeny for resistance. The resistance screening is done at Dorena Genetic Resource Center (DGRC). Using wind-pollinated progeny of some of the earliest selections from the Pacific Northwest Region’s resistance program, BLM and FS established six field tests (‘progeny’ tests) in the early 1980’s in southern Oregon to evaluate long-term growth, survival and field resistance to WPBR. The FS also established five provenance trials of sugar pine to examine range- wide genetic variation in this species. These trials represent ‘permanent’ plots with known genetic composition and planting dates. These are likely the oldest well documented multi-site test series for disease resistance in sugar pine to WPBR (and perhaps in any conifer to any non- native pathogen in western North America). For this FHM poster, we present an overview of growth, survival, and impacts of WPBR at the six progeny test sites through age 25 (for results through age 15 see f). Future reports will provide additional summary of this trial series (including examination of family variation in rust resistance) and results of the provenance trials. f SUMMARY As anticipated, many families from these field selections are highly susceptible or have only low levels of partial resistance. Breeding in seed orchards or clones banks will be needed to increase resistance levels. The seven families with major gene (Cr1) resistance had the lowest infection and mortality levels. Information on rust hazard can provide land managers with an additional tool for planning sugar pine reforestation or restoration efforts. Because of the multiple sites, known genetic composition and planting dates, and assessments over ~25 years, these plantings provide the most detailed look at the impact of WPBR on sugar pine to date. Figure 4. Canker with little or no swelling. Aecia observed in cracks in bark. RESULTS Although the sites varied in their rust hazard (low to high), by age 25, all 6 sites had high to very high levels of blister rust infection and mortality (Fig. 2). Averaged over all six sites (excludes non-rust mortality): 8.9% of the trees are canker-free (742/8340) 13.5% are alive with cankers 77.6% are dead from rust The sites with lower hazard ratings generally had lower levels of cankering in the early years, but the difference is much less by age 15 or 25. However, sites getting later infections (such as Boulder and Anchor) tended to have much higher survival through age 25 (Fig. 3). Few new cankers were evident since the 15 year assessment. Some cankers produced little or no stem swelling or were only visible due to presence of aecia at the time of assessment (Fig. 4). Thus, many field assessments of blister rust may somewhat underestimate the level and impact of rust infection. Most cankers were low (<1.5 m) at age 15, suggesting that timely branch pruning could complement genetic resistance to increase survival. Mortality from other biotic and abiotic agents was low after age 5 ( %) except for at Jamison Gulch (23.1%). Averaged over the 6 sites, families varied from 40% to 96% mortality. Mean height of trees alive at age 25 varied from 5.9 m at Boulder to 10.7 m at Poker (Fig. 5). Vertical canker, possible expression of partial resistance II. Growth and Survival of Sugar Pine through age 25 in Six Progeny Tests of Low to High Blister Rust Hazard in Southwestern Oregon Living and dead trees at Boulder YearYear Figure 2. Temporal dynamics of blister rust infection (on individual trees) over 25 years in 3 of 6 sugar pine progeny tests. Low Hazard Jamison Medium Hazard Anchor High Hazard Hayes Alive and free of rust Alive with rust Dead with rustDead and free of rustFiller/replaced tree Forest Health Monitoring National Meeting, Tucson, AZ April 2012 Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Western White Pine Rust Testing – 1000’s of parent trees – 8 six-parent half diallels (vcr2 & AVCr2 inoc) – Surviving progeny available Seedlots Available – 1000’s of wild OP and CC’s Field trials – Provenance trial (range-wide) – Diallels and other plantings since 1996 Orchards, clone banks Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

WWP Field Trials

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; WWP diallel (example: 1 of 8) in SY1998 – inoculated with [1] vcr2 source of rust, and [2] AVCr2 source of rust DRAFT vcr2 AVCr2

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; (a)(b) (Maps produced by H.Lintz) (a) Geographic distribution of parent trees of P. albicaulis families in blister rust testing (inoculated as of 2010); (b) Geographic sources of P. albicaulis with seed collected as of 2009 (P. albicaulis range from Little 1971). Whitebark Pine

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; 100’s of Needle spots Cankers, 13 months post -inoculation. ‘Needle Shed ‘- 50 needle spots at 1 st assessment in 2009, but is still canker-free in winter 2010 Bark reaction 23 months after inoculation

Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service; Distribution of canker-free seedlings using family means* ( DRAFT) WHITEBARK PINE – FAMILY WHITE PINE BLISTER RUST RESISTANCE FREQUENCY *early data (13 months after inoculation) for one of the two trials at Dorena GRC involving 225 families (using Avcr2 source of rust).

Southwestern white pine MGR & Partial Resistance identified from trees in same stand (Sniezko et al. 2008) Larger trial underway – 3 NF’s, 40 families Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Some recent publications Hamlin, J.; Kegley, A.; Sniezko, R.A Genetic Variation of Whitebark Pine (Pinus albicaulis) Provenances and Families from Oregon and Washington in Juvenile Height Growth and Needle Color. In: Keane, R.E.; Tomback, D.F.; Murray, M.P.; and Smith, C.M., eds The future of high-elevation, five-needle white pines in Western North America: Proceedings of the High Five Symposium. Proceedings RMRS-P-63. Fort Collins, CO: U.S. Department of Agriculture, Forest Service, Rocky Mountain Research Station: Sniezko, R.A.; Mahalovich, M.F.; Schoettle, A.W.; Vogler, D.R Past and current investigations of the genetic resistance to Cronartium ribicola in high-elevation five-needle pines. In: Keane, R.E.; Tomback, D.F.; Murray, M.P.; and Smith, C.M., eds The future of high-elevation, five-needle white pines in Western North America: Proceedings of the High Five Symposium. Proceedings RMRS-P-63. Fort Collins, CO: U.S. Department of Agriculture, Forest Service, Rocky Mountain Research Station: Zamany, A.; Liu, J.J.; Ekramoddoullah, A.; Sniezko, R Antifungal activity of a Pinus monticola antimicrobial peptide 1 (Pm-AMP1) and its accumulation in western white pine infected with Cronartium ribicola. Can. J. Microbiology. 57: 667– 679 (2011); doi: /W Liu, J.J.; Sniezko, R. A.; Ekramoddoullah, A Association of a novel Pinus monticola chitinase gene (PmCh4B) with quantitative resistance to Cronartium ribicola. Phytopathology 101: Zhang, X.Y.; Lu, Q.; Sniezko, R.A.; Song, R.Q.; Man, G Blister rusts in China: hosts, pathogens, and management. Forest Pathology 40: Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;

Some recent publications (cont’d) Sniezko, R.A.; Schoettle, A.; Dunlap, J.; Vogler, D.; Conklin, D.; Bower, A.; Jensen, C.; Mangold, R.; Daoust, D.; Man, G Ex situ gene conservation in high elevation white pine species in the United States:a beginning. In: Keane, R.E.; Tomback, D.F.; Murray, M.P.; and Smith, C.M., eds The future of high-elevation, five-needle white pines in Western North America: Proceedings of the High Five Symposium. Proceedings RMRS-P-63. Fort Collins, CO: U.S. Department of Agriculture, Forest Service, Rocky Mountain Research Station: situ gene conservation in high elevation white pine species in the United States:a beginning Sniezko, R.A; Hill, J.; Danchok, R.S.; Kegley, A.J.; Long, S.; Mayo, J.B.; Smith, A.J White pine blister rust resistance in a seven year old field trial of 28 western white pine (Pinus monticola) families in the Coast Range of Oregon. APS 2010 Meeting, Charlotte, NC. Phytopathology 100:S120. (abstract). Sniezko, R.A.; Kegley, A.; Danchok, R.; Hamlin, J.; Long, S Resistance to Cronartium ribicola in whitebark pine – family variation and effect of inoculum density. APS 2009 Meeting, August 1-5, Portland, OR. Phytopathology 99:S122 (abstract). Sniezko, R.A.; Kegley, A.J.; Danchok, R., 2008: White pine blister rust resistance in North American, Asian, and European species – results from artificial inoculation trials in Oregon. Annals of Forest Research 51, 53–66. [Online]. Available: Strobusphere Mtg- Happy Camp, May 10, 2012 (R.Sniezko, USDA Forest Service;