PACIFIC NORTHWEST TREE IMPROVEMENT RESEARCH COOPERATIVE

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
123	0612	1080	100%

Progress 07/01/83 to 09/30/14

Outputs
Target Audience:Douglas-fir tree breeders in public agencies and private industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This project has supported the education of ten graduate students and the professional development of three faculty research assistants, two assistant directors, and one project director. We have delivered many workshops or short-courses on a variety of topics to forest genetics practitioners and researchers (6).We have also published a suite of online learning modules using the eXtension online platform. How have the results been disseminated to communities of interest?Results were presented through journals, theses, posters, presentations at conferences, and collaborations with industry and research cooperatives. Results presented annually at the PNWTIRC meeting, in the PNWTIRC Annual Report, and in department seminars. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Highlights in 1985 - 1989 Measurement Methods Study (MMS), three 13-year-old Douglas-fir progeny tests measured for 7 growth and stem form traits to assess genotypic variation and optimal approaches of measurement. Early results of the MMS are 1) measuring DBH is quicker and nearly as effective using calipers as using a D-tape, 2) stem sinuosity can be measured efficiently by visually estimating stem displacement at the second interwhorl below the leader, 3) large variation and moderate heritability for sinuosity indicates that stem straightness could be readily improved, 4) percent genetic gains for branching traits appear to be similar to those for growth traits. Seed from 140 families were sown in 3 nurseries for the Early Testing Study (ETS). Nursery performance of growth, stem form, and adaptive traits were compared with the same families in 10-15-year-old field tests. Two Wood Quality (WQ) projects were conducted. One looked at genetic control of wood density components (i.e., earlywood density, latewood density) in young DF (age 15). The second examined potential for early testing of juvenile wood density (JWD). 1990 - 1994 Analysis of ETS data indicated first-year performance in nursery tests can be effective for low-level culling of families (30-40%) in a testing program for poor height growth, undesirable branch angle, and stem sinuosity. Thus, we can reduce the size and cost of progeny tests without substantially reducing genetic gain. Results from WQ study showed that phenotypic variation in JWD was only about 1/3 - 1/2 that for growth traits in DF. However, narrow-sense heritability was high, suggesting that JWD can readily be improved in DF breeding programs. We studied early selection of wood density (WD) using nursery trees and field trees (age 15). WD assessed at age 1 had a moderate genetic correlation with WD at age 15. Thus, selection in nursery trees will be nearly 50% as effective as delaying selection until age 15. Field Cold Hardiness Study (FCHS) was designed to 1) estimate genetic variation for cold hardiness traits, 2) determine the relationships between different cold hardiness traits, 3) quantify the relationships among cold hardiness, shoot phenology, and growth traits. We measured cold hardiness in the fall, winter, spring; timing of budburst and budset; and duration of shoot elongation in two breeding programs. 1995 - 1999 In a second FCHS, phenology, growth, and cold hardiness were measured repeatedly over a 2-year period in saplings from 80 DF families. Heritability for cold hardiness traits varied seasonally, ranging from low in the fall to very high in the spring. Fall and spring cold hardiness are independent traits, whereas bud phenology is highly correlated with spring cold hardiness. Seedling Drought Physiology Study (SDPS) was designed to examine genetic variation in drought physiology in DF seedlings and to assess physiological mechanisms and methods to measure drought hardiness. Shoot damage due to drought ranged widely among 39 full-sib families, suggesting that selection among families for increased seedling drought hardiness would be quite effective. Field Drought Hardiness Study (FDHS), wood density profiles were analyzed to determine whether annual growth ring variables reflect summer moisture stress in older, field-grown trees. The data indicated that latewood width increased during the summers as soil moisture deficits increased. Thus, latewood growth ring components may be useful for assessing sensitivity of families to drought in sapling-aged trees. A new study, "Age-age correlations of stem defects in coastal DF," progeny test trees scored for stem defects 12 years ago were re-assessed to determine whether defects persisted with age. The validity of early selection for improved stem form was tested. 2000 - 2004 Further analysis of the ETS showed unfavorable correlations between growth potential, stem form, and branching traits in seedlings. Selection for growth alone is expected to increase stem defects in older trees. To avoid negative impacts, stem form and branching traits should be included in nursery-stage selection. Analyses of the SDPS data sugguest: 1) genetic correlations between drought hardiness traits (e.g., shoot damage, xylem cavitation, hydraulic conductivity) in the same year are high, but moderate between years; 2) mild drought often enhances cold hardiness under nursery conditions, whereas severe drought reduces the cold hardiness of age 2 seedlings; 3) cold damage tends to increase in families that set bud late in the fall; 4) drought hardiness traits are unrelated to bud set, second flushing, and cold hardiness. Thus, selection and breeding for improved bud phenology and cold hardiness is unlikely to affect drought hardiness. Early Flowering Study (EFS) treatments applied in the previous spring were measured in two seed orchards on 2- 5-year-old grafts in two different years. The combined GA/girdling treatment significantly increased female flowering, numbers of seed cones and pollen cones, and the proportion of trees bearing cones. Thus, GA plus girdling can enhance cone and seed production in young seed orchards. 9 of the 22 SSR markers developed for the Pollen Contamination Study were used to measure pollen contamination and mating patterns in an OP clonal seed orchard of DF. Pollen contamination was consistently high across the three years (35.3%), and primarily resulted from cross-pollination among the orchard blocks. Pollen contamination varied among clones, and was higher in clones with early female receptivity. 2005 - 2009 We began the Pruning Study at the Vaughn Miniaturized Seed Orchard (MSO). Pruning treatments and collection of flowering data occurred between 2007 - 2009. Timing of pruning seems to affect number of flowers per tree. In Wood Quality Study (WQS), we supervised destructive sampling of progeny tests, gathered field measurements, milled boards from our study logs, tested the stiffness of the kiln-dried lumber, and conducted preliminary data analyses. Microfibril angle was analyzed on a subset of our lumber using the SilviScan system, and all lumber was visually graded. Analyses of measurement and selection strategies for improving wood stiffness were completed. We studied the genetics of wood stiffness in a clonal seed orchard of DF. Indirect estimates of MOE were obtained using two tools that measure acoustic velocity in trees. We observed low to moderate heritabilities for wood stiffness traits using the HM200, whereas clonal variation was non-significant using the ST300 tools. 2010 - 2015 For the Site Characterization Study (SCS), data were obtained for 288 test sites and 4010 parent trees in 36-25 first-generation breeding programs. Soils, climate, and topography data were also collected. In the MSO study, we collected data on numbers of males and female flowers, timing of reproductive and vegetative bud flush, and frost damage in 2010. We also measured the 2011 cone crop and completed the first analyses of crown volume and the cone crops in 2010 and 2011. We completed acoustic velocity (stiffness) measurements at two DF progeny test sites and a western hemlock (WH) progeny test for the "Early genetic selection for wood stiffness in DF and WH" study. Goals are to examine tools and measurement approaches and provide recommendations for breeding programs. We began working with Keith Jayawickrama and the Northwest Advanced Renewables Alliance (NARA) to develop a next-generation SNP genotyping chip for DF and test genomic selection. We continued to develop breeding strategies using SNP markers to improve blister rust resistance in western white pine and improve Douglas-fir using genomic selection. We designed a next-gen SNP chip for DF capable of assaying up to 50K SNPs. Early assessment of the 50K SNP chip (Affymetrix Axiom) is promising. We began the Drought Hardiness Study, measuring growth and drought hardiness traits at three sites in Oregon.

Publications

Type: Journal Articles Status: Published Year Published: 2011 Citation: Chmura, D.J., Anderson, P.D., Howe, G.T., Harrington, C.A., Halofsky, J.E., Peterson, D.L., Shaw, D.C., and St.Clair, J.B. 2011. Forest responses to climate change in the northwestern United States: Ecophysiological foundations for adaptive management. Forest Ecology and Management 261:1121-1142.
Type: Journal Articles Status: Published Year Published: 2011 Citation: Jayawickrama, K.J.S., Ye, T.Z., and Howe, G.T. 2011. Heritabilities, intertrait genetic correlations, GxE interaction and predicted genetic gains for acoustic velocity in mid-rotation coastal Douglas-fir. Silvae Genetica 60:8-18.
Type: Journal Articles Status: Published Year Published: 2010 Citation: Chmura, D.J., Howe, G.T., Anderson, P.D., and St.Clair, J.B. 2010. Adaptation of trees, forests and forestry to climate change. Sylwan 154(9):587-602.
Type: Journal Articles Status: Published Year Published: 2010 Citation: St.Clair, J.B. and Howe, G.T. 2010. Ensuring that forests are adapted to future climates. Northwest Woodlands 26(2):20-21, 31.
Type: Journal Articles Status: Published Year Published: 2009 Citation: St.Clair, J.B. and Howe, G.T. 2009. Genetic options for adapting forests to climate change. Western Forester 54(1): 9-11.
Type: Journal Articles Status: Published Year Published: 2007 Citation: Cherry, M.L., Anekonda, T.S., Albrecht, M.J., and Howe, G.T. 2007. Flower stimulation in young miniaturized seed orchards of Douglas-fir (Pseudotsuga menziesii). Canadian Journal of Forest Research 37(1):1-10.
Type: Book Chapters Status: Published Year Published: 2006 Citation: Howe, G.T., Jayawickrama, K.J., Cherry, M.L., Johnson, G.R, and Wheeler, N.C. 2006. Breeding Douglas-fir. In: Plant breeding reviews, J. Janick (ed). John Wiley and Sons, Inc. 27:245-353.
Type: Other Status: Published Year Published: 2006 Citation: Howe, G.T. 2006. Bibliography of Douglas-fir ecological genetics, evolutionary genetics, physiological genetics, and tree breeding. Pacific Northwest Research Cooperative Report #24, Oregon State University. 20 pp.
Type: Conference Papers and Presentations Status: Published Year Published: 2005 Citation: Howe, D.K., Brunner, A.M., Cherry, M., Krutovsky, K.V., Neale, D.B., and Howe, G.T. 2005. Identifying candidate genes associated with cold hardiness in coastal Douglas-fir using DNA microarrays. In: Proceedings Western Forest Genetics Association 50th Anniversary Meeting: Looking BackLooking Ahead, July 19-21, 2005, Corvallis, Oregon, USA
Type: Other Status: Published Year Published: 2003 Citation: Slavov, G.T. 2003. CTAB protocol for extracting DNA from Douglas-fir seed embryos and megagametophytes. 2p.
Type: Conference Papers and Presentations Status: Published Year Published: 2002 Citation: Jayawickrama, K.J., Bracher, G. and Howe, G.T. 2002. Genetic Improvement of Wood Quality in Coastal Douglas-fir and Western Hemlock. Proceedings joint PNWTIRC/NWTIC workshop held in Corvallis, OR, June 27, 2002.
Type: Journal Articles Status: Published Year Published: 1998 Citation: Krutovskii, K.V., Vollmer, S.S., Sorenson, F.C., Adams, W.T., Knapp, S.J. and Strauss, S.H. 1998. RAPD genome maps of Douglas-fir. Journal of Heredity 89:197-205.
Type: Journal Articles Status: Published Year Published: 1997 Citation: Schermann, N., Adams, W.T., Aitken, S.N. and Bastien, J.-C. 1997. Genetic parameters of stem form traits in a 9-year-old coastal Douglas-fir progeny test in Washington. Silvae Genetica 46:166-170.
Type: Conference Papers and Presentations Status: Published Year Published: 1996 Citation: Aitken, S.N. and Adams, W.T. 1996. Genetics of spring cold hardiness in coastal Douglas-fir. In:Proceedings Western Forest Genetics Association Meeting, Newport, Oregon, July 29-August 1, 1996.
Type: Conference Papers and Presentations Status: Published Year Published: 1996 Citation: Krutovskii, K.V., Vollmer, S.S., Sorensen, F.C., Adams, W.T. and Strauss, S.H. 1996. Genome mapping in inter-varietal hybrids of Douglas-fir. In: Proceedings Western Forest Genetics Association Meeting, Newport, Oregon, July 29-August 1, 1996, p. 23.
Type: Conference Papers and Presentations Status: Published Year Published: 1996 Citation: Krutovskii, K.V., Vollmer, S.S., Sorenson, F.C., Adams, W.T. and Strauss, S.H. 1996. Effects of megagametrophyte removal on DNA yield and early seedling growth in coastal Douglas-fir. In:Proceedings Western Forest Genetics Association Meeting, Newport, Oregon, July 29-August 1, 1996.
Type: Other Status: Published Year Published: 1995 Citation: Adams, W.T. and Aitken, S.N. 1995. Pacific Northwest Tree Improvement Research Cooperative Five-Year Plan (July 1995-June 2000). PNWTIRC Report, Oregon State University, Corvallis, Oregon, 11p.
Type: Conference Papers and Presentations Status: Published Year Published: 1995 Citation: Aitken, S.N. 1995. Effects of selection for increased growth rates on the annual developmental cycle of coastal Douglas-fir. In: Proceedings Canadian Tree Improvement Association and Western Forest Genetics Association Joint Meeting, Victoria, B.C., August 28-September 1, 1995, p. 63.
Type: Journal Articles Status: Published Year Published: 1995 Citation: Aitken, S.N., Kavanagh, K.L. and Yoder, B.J. 1995. Genetic variation in seedling water-use efficiency as estimated by carbon isotope ratios and its relationship to sapling growth in Douglas-fir. Forest Genetics 2:199-206.
Type: Journal Articles Status: Published Year Published: 1994 Citation: Adams, W.T. and Bastien, J.-C. 1994. Genetics of second flushing in a French plantation of coastal Douglas-fir. Silvae Genetica 43:345-352.
Type: Conference Papers and Presentations Status: Published Year Published: 1994 Citation: Adams, W.T. and Bastien, J.-C. 1994. Genetics of second flushing in coastal Douglas-fir. In:Proceedings Joint Conference Western Mensurationists Association and Western Forest Genetics Association, Vancouver, WA, June 27-30, 1994, p. 83.
Type: Journal Articles Status: Published Year Published: 1994 Citation: Vargas-Hernandez, J., Adams, W.T. and Krahmer, R.L. 1994. Family variation in age trends of wood density traits in young coastal Douglas-fir. Wood and Fiber Science 26:229-236.
Type: Journal Articles Status: Published Year Published: 1994 Citation: Kaya, Z., Adams, W.T. and Campbell, R.K. 1994. Adaptive significance of intermittent shoot growth in Douglas-fir seedlings. Tree Physiology 14:1277-1289.
Type: Journal Articles Status: Published Year Published: 1994 Citation: Li, P. and Adams, W.T. 1994. Genetic variation in cambial phenology of coastal Douglas-fir. Canadian Journal of Forest Research 24:1864-1870.
Type: Journal Articles Status: Published Year Published: 1994 Citation: Vargas-Hernandez, J. and Adams, W.T. 1994. Genetic relationships between wood density components and cambial growth rhythm in young coastal Douglas-fir. Canadian Journal of Forest Research 24:1871-1876.
Type: Journal Articles Status: Published Year Published: 1993 Citation: Li, P. and Adams, W.T. 1993. Genetic control of bud phenology in pole-size trees and seedlings of coastal Douglas-fir. Canadian Journal of Forest Research 23:1043-1051.
Type: Journal Articles Status: Published Year Published: 1993 Citation: St. Clair, J.B. and Adams, W.T. 1993. Family composition of Douglas-fir nursery stock as influenced by seed characters, mortality and culling practices. New Forests 7:319-329.
Type: Journal Articles Status: Published Year Published: 1992 Citation: Vargas-Hernandez, J. and Adams, W.T. 1992. Age-age correlations and early selection for wood density in young Douglas-fir. Forest Science 38:467-478.
Type: Journal Articles Status: Published Year Published: 1991 Citation: St. Clair, J.B. and Adams, W.T. 1991. Effects of seed weight and rate of emergence on early growth of open-pollinated Douglas-fir families. Forest Science 37:987-997.
Type: Journal Articles Status: Published Year Published: 1991 Citation: St. Clair, J.B. and Adams, W.T. 1991. Relative family performance and variance structure of open-pollinated Douglas-fir seedlings grown in three competitive environments. Theoretical and Applied Genetics 81:541-550.
Type: Journal Articles Status: Published Year Published: 1991 Citation: Vargas-Hernandez, J. and Adams, W.T. 1991. Genetic variation of wood density components in young coastal Douglas-fir: implications for tree breeding. Canadian Journal of Forest Research 21:1801-1807.
Type: Journal Articles Status: Published Year Published: 1988 Citation: Adams, W.T., Neale, D.B. and Loopstra, C.A. 1988. Verifying controlled crosses in conifer tree-improvement programs. Silvae Genetica 37:147-152.
Type: Other Status: Published Year Published: 1990 Citation: Adams, W.T., Johnson, G., Copes, D.L., Daniels, J., Quam, R.G., Heaman, J.C. and Weber, J. 1990. Information needs of coastal Douglas-fir tree improvement programs. PNWTIRC Report, Oregon State University, Corvallis, Oregon, 18p.
Type: Journal Articles Status: Published Year Published: 1990 Citation: Adams, W.T., Johnson, G., Copes, D.L., Daniels, J., Quam, R.G., Heaman, J.C. and Weber, J. 1990. Is research keeping up with the needs of Douglas-fir tree improvement programs? Western Journal of Applied Forestry 5:135-137.
Type: Journal Articles Status: Published Year Published: 1990 Citation: Adams, W.T. and Joyce, D.G. 1990. Comparison of selection methods for improving volume growth in young coastal Douglas-fir. Silvae Genetica 39:219-226.
Type: Conference Papers and Presentations Status: Published Year Published: 1990 Citation: Adams, W.T., Vargas-Hernandez, J. and Joyce, D. 1990. Selecting for wood density in young coastal Douglas-fir. In: Proceedings Joint Meeting Western Forest Genetics Association and IUFRO Working Parties S2.02-05, -06, -12 and -14 on Douglas-fir, Contorta Pine, Sitka Spruce and Abies Breeding and Genetic Resources, Olympia, Washington, August 20-24, 1990, p. 4.1-4.8.
Type: Theses/Dissertations Status: Published Year Published: 1990 Citation: Li, P. 1990. Genetic variation in phenology of bud and cambial activity in coastal Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco). Ph.D. Dissertation, Oregon State University, Corvallis, Oregon. 147p.
Type: Theses/Dissertations Status: Published Year Published: 1990 Citation: Vargas-Hernandez, J. 1990. Genetic variation of wood density components in coastal Douglas-fir and their relationships to growth rhythm. Ph.D. Dissertation, Oregon State University, Corvallis, Oregon. 123p.
Type: Journal Articles Status: Published Year Published: 1989 Citation: Li, P. and Adams, W.T. 1989. Range-wide patterns of allozyme variation in Douglas-fir (Pseudotsuga menziesii). Canadian Journal of Forest Research 19:149-161.
Type: Theses/Dissertations Status: Published Year Published: 1989 Citation: St. Clair, J.B. 1989. Effects of seed characters and competitive environment on two-year performance of open-pollinated Douglas-fir families. Ph.D. Dissertation, Oregon State University, Corvallis, Oregon. 146p.
Type: Journal Articles Status: Published Year Published: 1988 Citation: Howe, G.T. and Adams, W.T. 1988. Clinometer versus pole measurement of tree heights in young Douglas-fir progeny tests. Western Journal of Applied Forestry 3:86-88.
Type: Conference Papers and Presentations Status: Published Year Published: 1985 Citation: Adams, W.T. and Howe, G.T. 1985. Stem sinuosity measurement in young Douglas-fir progeny tests. In: Proceedings of the IUFRO Working Party on Breeding Strategy for Douglas-fir as an Introduced Species. Working Party: S. 2.02.05. Vienna, Austria, June 1985, pp. 147-159.

Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Douglas-fir tree breeders in public agencies and private industry. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? This project has supported work performed by graduate student Oguz Urhan, and has provided the opportunity for an undergraduate student to participate in data collection and research. How have the results been disseminated to communities of interest? Results were published in refereed journals, one thesis, and one Annual Report. We presented results at the annual meeting of the Pacific Northwest Tree Improvement Research Cooperative, annual meeting of the Center for Advanced Forestry Systems, and in Forest Genetics 2013, a joint meeting of the Canadian Forest Genetics Association, Western Forest Genetics Association, IUFRO Population, Ecological and Conservation Genetics Working Group, and the IUFRO Breeding and Genetic Resources of Pacific Northwest Conifers Working Group. PRESENTATIONS Howe, G.T., Kolpak, S., Urhan. O., Cress, D., Jayawickrama, K., and Ye, T. 2013. Early genetic selection for wood stiffness in Douglas-fir and western hemlock. Presentation In: Center for Advanced Forestry Systems Annual Meeting, April 9-11, 2013, St. Simons Island, Georgia. Howe, G.T., Magalska, L., Jayawickrama, K, Ye, T., Fox, T., Burkhart, H., and Maguire, D. Effects of site and genetics on Douglas-fir growth, stem quality, and adaptability. Presentation In: Center for Advanced Forestry Systems Annual Meeting, April 9-11, 2013, St. Simons Island, Georgia. Howe, G.T., Maguire, D.A, Colin, A.-L., St.Clair, J.B. 2013. Mechanistic growth models: Decomposing phenotypic models into their genetic and environmental components. Presentation In: Center for Advanced Forestry Systems Annual Meeting, April 9-11, 2013, St. Simons Island, Georgia. Magalska, L., Howe, G.T., and Maguire, D.A., 2013. Genetic and environmental control of Douglas-fir stem form. Abstract In: Proceedings of Forest Genetics 2013, July 22-25, 2013, Whistler, British Columbia, Canada. Rust, M.L. and Howe, G.T. 2012. Development of genetic markers for western white pine and Douglas-fir. Presentation In: Center for Advanced Forestry Systems Annual Meeting, April 9-11, 2013, St. Simons Island, Georgia. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? GENETCS OF STEM FORM We studied the genetic and environmental control of Douglas-fir (Pseudotsuga menziesii var. menziesii) stem defects across 22 breeding programs in western Oregon and Washington. Our goal was to understand the relationships between stem defects (forks and ramicorn branches) and growth. We tested the hypotheses that stem defects are associated with progeny test productivity and distance to the coast, and then obtained robust estimates of genetic variances, heritabilities, and genetic correlations that can be used to design effective breeding programs. Stem defects were more frequent on high productivity sites and near the coast. Compared to the least productive site within each breeding program, the proportion of trees with stem defects was about twice as great on the most productive site. For example, the proportion of trees with ramicorn branches increased from 11% to 24%, and the proportion of trees with forks increased from 5% to 14%, between the shortest and tallest plantations. However, the relationships between stem defects and growth varied substantially within and among breeding programs (R2 <= 27%). Stem defects were also more frequent near the coast, but even harder to predict based on the locations of the plantations (R2 <= 18%). Although stem defects are genetically variable, heritable, and have positive genetic correlations with growth, genetic variation and heritabilities for stem defects were low and highly variable. Nonetheless, stem defects can be reduced using direct backward selection, and are expected to increase only a small amount when genotypes are selected based on volume growth alone. Focused breeding could be used to develop low-defect varieties, and these could be deployed to problematic sites. This approach might increase the value of reforestation programs overall, but it will be difficult to deploy low-defect varieties optimally because site productivity and distance to the coast are only weakly associated with stem defects. Although current multi-trait breeding approaches that consider growth and stem defects seem appropriate for most sites, controlled crosses between low-defect parents could be made in seed orchards, and the resulting seedlots could be deployed to sites that are known to be prone to defects (e.g., based on past data). Our results also suggest that (1) it might be possible to improve protocols and training for measuring stem defects, (2) breeders should monitor the among-site relationships between growth and stem defects in advanced generation breeding programs, and (3) low-defect genotypes should be identified and archived so they are available for future breeding. GENETICS OF WOOD STIFFNESS Wood stiffness, or modulus of elasticity (MOE), is an important property of structural wood products. The ability to genetically improve wood stiffness has increased because of the availability of tools that can measure acoustic velocity (AV, an indirect measure of MOE) on standing trees. Our goals were to optimize methods for measuring AV on young (6- to 12-year-old) Douglas-fir and western hemlock, and then use these methods to understand the genetics of wood stiffness. In Phase I, we evaluated four standing-tree tools (TreeSonic-TS, TreeSonic-SD02, Microsecond Timer, and Ultrasonic Timer) by comparing AV-squared measured on standing trees to AV-squared measured on logs cut from these trees (i.e., using the HM200). We found that measuring across a whorl of branches has little adverse effect on AV measurements, and that placing the acoustic sensors on the same-face of the tree was slightly better than placing them on opposite faces. In Phase II, we used full-sib heritabilities (h(fs)^2) to compare the TreeSonic-SD02 and Microsecond Timer, sensor orientations (same face versus opposite face), and flight path adjustments. Because the effect of standing-tree tool and sensor orientation was non-significant, we recommend that breeders use the TreeSonic SD02 and same-face orientation for practical reasons. Using this approach, h(fs)^2 for AV-squared was 0.90 to 0.56 in Douglas-fir and 0.51 in western hemlock, and we concluded that genetic gains of 7.0% to 17.2% can be obtained by measuring 10 or more trees per full-sib family, depending on species and seed production strategy. Because we found no strong statistical support for substantial non-additive genetic variation, the current approach of breeding for additive genetic value, and then collecting open-pollinated seed from orchards seems appropriate for improving wood stiffness. AV-squared had weak genetic correlations with growth traits (height, diameter at breast height, and volume) in Douglas fir (-0.21 to 0.33), but moderately positive correlations in western hemlock (0.36 to 0.54). Therefore, there is ample opportunity to improve the stiffness of young trees without adversely affecting growth. Although mean stiffness was higher for Douglas fir, about 10% of the western hemlock families (7 out of 80) had higher AV-squared, and about 28% of the families (22 out of 80) had higher dynamic MOE than the mean for Douglas fir. This suggests that intensive selection and breeding in western hemlock could be used to create varieties of trees that compare favorably with unselected Douglas-fir. This may improve the ability of western hemlock to compete with Douglas fir where stiffness is important. MINIATURIZED SEED ORCHARDS We studied miniaturized seed orchards of coastal Douglas-fir (Pseudotsuga menziesii var. menziesii) in Oregon and Washington. In Experiments 1 and 2, flowering and cone yields were greater using stem girdles plus stem injections of GA4/7 compared to girdling alone, GA alone, root pruning, and girdling plus fertilization with CaNO3. In Experiment 3, topping and pruning in the summer following flower stimulation minimized crown volume and maximized female and male flower densities. In Experiment 4, the widest spaced orchard (4x6 m) generally produced more flowers and cones per tree, and greater flower and cone densities compared to the higher density orchards (1x3 m and 2x4 m). Initially, (i.e., in 2010), per-hectare cone yields were greatest in the 1x3 orchard, but were greatest in the 4x6 orchard by 2012. In general, per-hectare cone yields were similar or greater in the MSOs compared to nearby conventionally spaced orchards. Trees grafted using scions collected from juvenile trees (age 7 to 8) generally had larger crowns, more female flowers and cones per tree, and greater female flower and cone densities compared to scions collected from middle-aged (age 30 to 31) or mature (56- to 101-year-old) trees. We found no evidence that clonal rows resulted in reduced seed quality relative to nearby conventional orchards.

Publications

Type: Journal Articles Status: Published Year Published: 2013 Citation: Howe, G.T., Yu, J., Knaus, B., Cronn, R., Kolpak, S., Dolan, P., Lorenz. W.W., Dean, J.F.D. 2013. A SNP resource for Douglas-fir: de novo transcriptome assembly and SNP detection and validation. BMC Genomics 14:137.
Type: Theses/Dissertations Status: Published Year Published: 2013 Citation: Urhan, Oguz S. 2013. Early genetic selection for wood stiffness in juvenile Douglas-fir and western hemlock. M.S. Thesis, Oregon State University, Corvallis, OR. 92 pp.

Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: The Pacific Northwest Tree Improvement Research Cooperative has four areas of focus: (1) genetics of wood stiffness in Douglas-fir and western hemlock; (2) miniaturized seed orchards (MSOs); (3) Douglas-fir site characterization; and (4) molecular markers for tree breeding. In 2012, we completed the bulk of the analyses on the genetics of wood stiffness in young trees of Douglas-fir and western hemlock. Specifically, we: (1) completed the measurements of acoustic velocity (stiffness) at the remaining test sites of Fir Grove (Douglas-fir) and Toledo (western hemlock); (2) analyzed the relative merits of tools and measurement approaches; (3) compared estimated stiffness between Douglas-fir and western hemlock; and (4) evaluated sampling strategies. For the MSO project, we: (1) recorded flower, cone, and crown measurements at Plum Creek's MSO; (2) completed the outline for the MSO publication; (3) conducted statistical analysis of MSO data; and (4) acquired and summarized MSO data from the Lebanon Forest Regeneration Center and Meridian seed orchards. For the site characterization project, we: (1) completed the stem form genetic analyses and new site analyses; and (2) updated the site characterization dataset with newly available soils data from the Natural Resources Conservation Service (NRCS). For the molecular marker work, we initiated a new project aimed at developing low-cost SNP genotyping assays that can be used for parentage analysis in Douglas-fir and western white pine. This latter project is being done in collaboration with the Inland Empire Tree Improvement Cooperative. Our work has been disseminated in refereed publications, at the annual meeting at the Pacific Northwest Tree Improvement Research Cooperative (PNWTIRC), at other forestry research cooperative meetings. PARTICIPANTS: Glenn T. Howe, Principal Investigator; Scott Kolpak, Research Coordinator; Keith Jayawickrama, Director of the Northwest Tree Improvement Cooperative; Terrance Ye, Quantitative Geneticist for the Northwest Tree Improvement Cooperative; David W. Cress, Olympic Resource Management; Kori Ault, Program Manager; Lauren Magalska, Graduate Research Assistant; Oguz Urhan, Graduate Research Assistant; and Jim Smith, Sara Lipow, Jeff DeBell, Marc Rust, and Anthony Davis, collaborators. TARGET AUDIENCES: Douglas-fir and western hemlock tree breeders in public agencies and private industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. Our wood quality research will help ensure Douglas-fir's niche in domestic and international timber markets by providing tree breeders with the tools to maintain wood stiffness in the face of shorter rotations. We developed approaches for measuring and selecting for wood stiffness in genetic test plantations that are now being used to improve Douglas-fir wood stiffness. Our miniaturized seed orchard research will help seed orchard managers produce seed with greater genetic gains at an earlier age. In 2013, we will summarize the results from all of our seed orchard studies and provide recommendations to seed orchard managers. Our site characterization research will give forest mangers an enhanced ability to predict the growth, stem quality, and adaptability of Douglas-fir plantations; manage their land base; refine breeding and deployment zones; and understand the potential effects of climate change. Our research on molecular genetic markers (SNPs) will allow breeders to adopt new approaches to tree breeding including an approach to marker-assisted selection called genomic selection.

Publications

Lorenz, W.W., D.B. Neale, K.D. Jermstad, G.T. Howe, D.L. Rogers, J.M. Bordeaux, S. Ayyampalayam and J.F.D. Dean. 2012. Conifer DBMagic: A database housing multiple de novo transcriptome assemblies for twelve diverse conifer species. Tree Genet. Genomes:1477-1485.

Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: The Pacific Northwest Tree Improvement Research Cooperative has four areas of focus: (1) genetics of wood stiffness in Douglas-fir and western hemlock; (2) miniaturized seed orchards (MSOs); (3) Douglas-fir site characterization; and (4) molecular markers for tree breeding. The wood stiffness and molecular marker research overlaps with another project that ended in 2011 entitled "Genetics of Douglas-fir Wood Stiffness and Strength." In 2011, we continued our research on wood stiffness in young trees (6 to 12 years old) of Douglas-fir and western hemlock. We completed a phenotypic study in operational plantations designed to determine which measurement tools and approaches work best for estimating wood stiffness of standing trees. We tested 4 tools available from Fakopp Enterprise: (1) TreeSonic with standard sensors; (2) TreeSonic with smaller SD-02 sensors; (3) Microsecond Timer with SD-02 sensors; and (4) Ultrasonic Timer with SD-02 sensors. In addition, we also tested the effect of placing the sensors on the same side versus the opposite side of the tree, and the effect of spanning versus not spanning a whorl of branches. After analyzing these results, we began testing the best approaches in three young genetic test plantations of Douglas-fir and western hemlock. Our accomplishments at the Plum Creek's MSO include: (1) flower stimulation of 640 trees using CaNO3 and girdling; and (2) seed orchard measurements of the number of seed cones and rate of operation cone collecting. We completed the data analysis and interpretation of results from our Douglas-fir site characterization research. We developed new molecular genetic markers (SNPs, or single nucleotide polymorphic markers) by first assembling a consensus transcriptome from next-generation sequence data that consists of 25,002 unigenes and 102,623 singletons. We then identified 281,192 SNP markers and constructed and tested an Illumina Infinium SNP genotyping array that can assay 8,769 SNPs. Our work has been disseminated in refereed publications, at the annual meeting at the Pacific Northwest Tree Improvement Research Cooperative (PNWTIRC), at other forestry research cooperative meetings, and through the Conifer Translational Genomics Network newsletter at the Dendrome webpage hosted by one of our collaborators at the University of California at Davis. PARTICIPANTS: Glenn T. Howe, Principal Investigator; Scott Kolpak, Research Coordinator; Keith Jayawickrama, Director of the Northwest Tree Improvement Cooperative; Terrance Ye, Quantitative Geneticist for the Northwest Tree Improvement Cooperative; David Briggs, Professor, University of Washington; and David W. Cress, Olympic Resource Management; Elizabeth Etherington, Program Manager; Kori Ault, Program Manager; Lauren Magalska, Graduate Research Assistant; Oguz Urhan, Graduate Research Assistant. TARGET AUDIENCES: Douglas-fir and western hemlock tree breeders in public agencies and private industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. Our wood quality research will help ensure Douglas-fir's niche in domestic and international timber markets by providing tree breeders with the tools to maintain wood stiffness in the face of shorter rotations. We developed approaches for measuring and selecting for wood stiffness in genetic test plantations that are now being used to improve Douglas-fir wood stiffness. Our miniaturized seed orchard research will help seed orchard managers produce seed with greater genetic gains at an earlier age. In 2012, we will summarize the results from all of our seed orchard studies and provide recommendations to seed orchard managers. Our site characterization research will give forest mangers an enhanced ability to predict the growth, stem quality, and adaptability of Douglas-fir plantations; manage their land base; refine breeding and deployment zones; and understand the potential effects of climate change. Our research on molecular genetic markers (SNPs) will allow breeders to adopt new approaches to tree breeding including an approach to marker-assisted selection called genomic selection.

Publications

Magalska, L.E. 2011. Identifying site characteristics that explain variation in Douglas-fir site productivity and stem form. Master's Thesis, Oregon State University, Corvallis, OR. 161pp.
Jayawickrama, K.J.S., Ye, T.Z., and Howe, G.T. 2010. Heritabilities, intertrait genetic correlations, GxE interaction and predicted genetic gains for acoustic velocity in mid-rotation coastal Douglas-fir. Silvae Genet. 60(1):8-18.
Vikram, V., Cherry, M.L., Briggs, D., Cress, D.W., Evans, R., and Howe, G.T. 2011. Stiffness of Douglas-fir lumber: Effects of wood properties and genetics. Can. J. For. Res. 41(6): 1160-1173.

Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: The Pacific Northwest Tree Improvement Research Cooperative has three areas of focus: (1) genetics of Douglas-fir wood stiffness; (2) miniaturized seed orchards (MSOs); and (3) Douglas-fir site characterization. The work on wood stiffness partially overlaps with a second project entitled "Genetics of Douglas-fir Stiffness and Strength." In 2010, we initiated new research on wood stiffness in trees aged 6 to 12 years. We previously studied stiffness in 25 year-old trees (see Vikram et al., in press), but information on younger trees is also needed because selections in breeding programs are often made at much younger ages. Our first step is to determine which measurement tool and approach works best for estimating wood stiffness of standing trees. We are doing this by examining phenotypic correlations between indirect measures of wood stiffness on standing trees versus more accurate and precise measurements made on logs using the Hitman 200 acoustic measuring tool. We have already estimated standing tree stiffness in two plantations of Douglas-fir and western hemlock using 4 tools available from Fakopp Enterprise: (1) TreeSonic with standard sensors; (2) TreeSonic with smaller SD-02 sensors; (3) Microsecond Timer with SD-02 sensors; and (4) Ultrasonic Timer with SD-02 sensors. In addition to testing these 4 tools, we are also testing the effect of placing the sensors on the same side versus the opposite side of the tree, and the effect of spanning versus not spanning a whorl of branches. Once we determine the best measurement approach, this approach will be used to estimate genetic parameters, heritabilities, and genetic gains for wood stiffness in young progeny test trees. Accomplishments at Plum Creek's MSO include: (1) flower stimulation of 640 trees using GA4/7 and girdling; and (2) seed orchard measurements, including crown size, timing of vegetative bud flush, timing of floral bud flush, number of female flowers, number of female flowers damaged by frost, and number of seed cones. The site characterization project focused on data analysis. We developed models to predict progeny test survival, growth, and stem form from climatic, topographic, and soils information. Growth traits included height, diameter, and basal area at ages 5, 10, and/or 15. Stem form traits included sinuosity, forking, and ramicorn branching. We analyzed 205 progeny test sites belonging to 32 testing programs managed by the Northwest Tree Improvement Cooperative. The elevations of the progeny test sites ranged from 15 to 1090 meters, with an average of 3500 trees per site (400 to 9400). We used Random Forests to rank the ability of independent variables to explain variation in survival, growth, and stem form, and multiple regression to develop predictive models. Our work has been disseminated to the academic and industrial communities through the Conifer Translational Genomics Network newsletter at the Dendrome webpage hosted by our collaborator at the University of California at Davis. Updated information is also provided at the Pacific Northwest Tree Improvement Research Cooperative annual meeting and newsletter that is distributed to PNWTIRC members. PARTICIPANTS: Glenn T. Howe, Principal Investigator; Scott Kolpak, Research Coordinator; Elizabeth Etherington, Program Manager; Jianbin Yu, Post-Doc; Lauren Magalska, Graduate Student. TARGET AUDIENCES: Douglas-fir tree breeders in public agencies and private industry. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. Our wood quality research will help ensure Douglas-fir's niche in domestic and international timber markets by providing tree breeders with the tools to maintain wood stiffness in the face of shorter rotations. Our miniaturized seed orchard research will help seed orchard managers produce seed with greater genetic gains at an earlier age. Our site characterization research will give forest mangers an enhanced ability to predict the growth, stem quality, and adaptability of Douglas-fir plantations; manage their land base; refine breeding and deployment zones; and understand the potential effects of climate change.

Publications

Vikram, V., M.L. Cherry, D. Briggs, D.W. Cress, R. Evans and G.T. Howe. 2011. Stiffness of Douglas-fir lumber: effects of wood properties and genetics. Can. J. For. Res. (In Press).

Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: We have four areas of continuing emphasis: (1) genetics of Douglas-fir wood stiffness; (2) genetics of cold adaptation; (3) miniaturized seed orchards (MSOs); and (4) Douglas-fir site characterization. Wood stiffness (acoustic velocity) measurements were taken on standing trees using the Fakopp TreeSonic tool, and on logs using the Fibre-gen Director Hitman (HM200). TreeSonic measurements were recorded on 167 parents (528 trees) in two BLM seed orchards prior to harvesting. HM200 measurements were taken on logs from 407 trees of the same clones after the trees were harvested. In addition, HM200 measurements were taken during the thinning of two first-generation progeny test sites representing BLM's Lorane breeding program. These tests include progeny (families) of 167 parents that were measured in the seed orchards: 437 progeny from 114 parents were measured at the Carpenter Bypass site, and 427 progeny from 119 parents were measured at Hawley Creek. Finally, stiffness measurements began on a second set of clones and progeny from BLM's breeding unit 33 (BU-33). We began taking TreeSonic measurements on 160 parents at BLM's Horning Seed Orchard, and measurements were completed on the progeny of 131 parents at two first-generation progeny test sites. Vegetative bud phenology was measured on 592 clonally-replicated parents in 8 grafted seed orchards, and flowering phenology was measured on 284 of these parents. Artificial cold-hardiness testing was done on 160 families at two progeny test sites (low and high elevation) in the Washington Cascades. Accomplishments at Plum Creek's MSO in 2009 include: (1) flower stimulation using GA4/7 and girdling (1000 of 2000 trees in the orchard) and (2) seed cone counts on all trees. The Douglas-fir site characterization project focused on acquiring location and other information for progeny tests sites and parent-trees for 25 breeding programs. The spatial information was gathered from Northwest Tree Improvement Cooperative records, directly from industrial collaborators, or from digital aerial photos. The parent and progeny test site locations were verified using aerial photos. Data for climate, soils, and topography were downloaded and processed to create GIS layers covering the range of coastal Douglas-fir. The gathering and processing of spatial information for other breeding programs continues as well as preliminary data analyses. Our work has been disseminated to the academic and industrial communities through the Conifer Translational Genomics Network newsletter at the Dendrome webpage hosted by our collaborator at the University of California at Davis. Updated information is also provided at the Pacific Northwest Tree Improvement Research Cooperative (PNWTIRC) annual meeting and newsletter that is distributed to PNWTIRC members. PARTICIPANTS: Glenn T. Howe, Principal Investigator; Scott Kolpak, Research Coordinator; Elizabeth Etherington, Program Manager; Jianbin Yu, Post-Doc; Lauren Magalska, Graduate Student. TARGET AUDIENCES: Douglas-fir tree breeders in public agencies and private industry. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. Our wood quality research will help ensure Douglas-fir's niche in domestic and international timber markets by providing tree breeders with the tools to maintain wood stiffness in the face of shorter rotations. Our miniaturized seed orchard research will help seed orchard managers produce seed with greater genetic gains at an earlier age. Our site characterization research will give forest mangers an enhanced ability to predict the growth, stem quality, and adaptability of Douglas-fir plantations; manage their land base; refine breeding and deployment zones; and understand the potential effects of climate change.

Publications

Vikram, V. 2008. Stiffness of Douglas-fir lumber: Effects of wood properties and genetics. M.S. Thesis. Oregon State University, Corvallis. 78 p. Available from the Scholars Archive database at Oregon State University.

Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: We currently have three major research projects: genetics of Douglas-fir wood stiffness, miniaturized seed orchards (MSOs), and Douglas-fir site characterization. Our wood quality research combines a number of novel elements, including the evaluation of new acoustic tools that can be used to obtain indirect estimates of wood stiffness on standing trees and logs, comparisons of wood stiffness estimated indirectly with stiffness measured directly on lumber harvested from the same trees, evaluation of wood stiffness of seed orchard parents and their progeny growing in genetic test plantations, and discovery of genes associated with wood properties using genomic approaches. To accomplish these goals, we are collaborating with the Stand Management Cooperative at the University of Washington, scientists at the University of California at Davis, the Genetics Team at the USFS Pacific Northwest Research Station, and Olympic Resource Management. We have demonstrated that gains can be made in wood stiffness. Acoustic tools are valuable for assessing wood quality, but basal wood density is not a good measure of bending stiffness. Clonal seed orchards, which are easily measured, can be used to assess wood quality in breeding programs. Selection for growth will not have adverse effects on bending stiffness or acoustic velocity, but will adversely affect wood density. The objectives of the Douglas-fir site characterization project are to understand how stand growth, stem quality, and adaptability are influenced by site characteristics such as climate, weather, topography, and soils; the mechanistic basis of genotype by site interactions; and the effects of seed source and genotype transfer among sites. To understand genotype by environmental interactions, we will characterize the sites from which the progeny-tested parents originated using map-based data. Phenotypic analyses will explore which site variables best explain stand-level variation in plantation growth, stem quality, and adaptability. Genetic analyses will investigate how parental site characteristics are related to progeny performance at each test location, and the extent to which seed sources and families can be successfully moved among sites. Miniaturized seed orchards are orchards in which the trees are planted at close spacings in clonal rows, and then maintained at a height of only 2 to 4 m. Our largest MSO experiment is a long-term test of alternative MSO designs that was established at the Plum Creek Seed Orchard Complex in western Oregon. We completed the grafting for this experiment in 2004, and plan to begin flower stimulation in the spring of 2009. We also carried out a pruning study at the Roseburg Products Vaughn seed orchard between 2005 and 2008. This experiment was designed to determine the best time to prune the crowns of MSO trees. Pruning is needed to keep the trees small, but reduces the number of cones, thereby adversely affecting seed production. Preliminary analyses suggest that cone and seed production are directly related to crown size, and that mortality may be greater when the trees are pruned shortly before bud flush in the year of flower stimulation. PARTICIPANTS: Glenn T. Howe, Principal Investigator; Marilyn L. Cherry, Co-Principal Investigator; Vikas Vikram, Graduate Student; Lauren Magalska, Graduate Student TARGET AUDIENCES: Douglas-fir tree breeders in public agencies and private industry. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. Our wood quality research will help ensure Douglas-fir's niche in domestic and international timber markets by providing tree breeders with the tools to maintain wood stiffness in the face of shorter rotations. Our miniaturized seed orchard research will help seed orchard managers produce seed with greater genetic gains at an earlier age. Our site characterization research will give forest mangers an enhanced ability to predict the growth, stem quality, and adaptability of Douglas-fir plantations; manage their land base; refine breeding and deployment zones; and understand the potential effects of climate change.

Publications

Cherry, M.L., V. Vikram, D. Briggs, D.W. Cress and G.T. Howe. 2008. Genetic variation in direct and indirect measures of wood stiffness in coastal Douglas-fir. Canadian Journal of Forest Research 38(9):2476-2486.

Progress 01/01/07 to 12/31/07

Outputs
OUTPUTS: Our work involves field and laboratory data collection, data analysis, and interpretation and writeup of results. The seed orchard containing maternal parents of the study's progeny test material was rogued. During harvesting, we collected wood disks from the base and top of every 17' butt log, and measured wood density of each disk. The acoustic modulus of elasticity (MOEa) of the 17' butt logs were measured using the Fibre-gen Director ST300 (ST300) and Fibre-gen Director HM200 (HM200). Foliage was collected from all orchard trees for DNA isolation and candidate gene analysis. Prior to thinning the three progeny tests, we measured diameter at breast height on the 25-year-old trees from all 130 families. MOEa was measured on standing trees on a subset of 50 families (8 trees per family) at two of the test sites (Shine and Opsata) using the ST300. The Shine and Opsata progeny tests had been thinned during the fall, but data were collected at the time of harvesting only at the Shine site. Half of the trees at the Shine progeny test were felled by chainsaw, and the logs were skidded to a landing where they were processed. After the logs were delimbed at the landing, acoustic velocities were measured with the HM200. Wood disks were collected from the base of every log, and green wood densitywas measured in the field. Wood disks were later shipped to Corvallis, kiln-dried, and weighed again. A subset of harvested trees from the Shine site (the same trees from which the ST300 measurements were taken) were cut to 9' butt logs and shipped to Corvallis for milling into lumber. A portable WoodMizer sawmill was used to mill each log into 1.5" x 3.5" x 7' boards. These boards were then kiln-dried at OSU, and were used to obtain direct measurements of wood stiffness through bending tests. The Watershed progeny test was thinned in the spring using a Timberjack harvester. Trees were delimbed at the stump, laid on the ground, and HM200 measurements were taken. During the past year, we isolated DNA from approximately 180 trees in the Hood Canal Seed Orchard, and began developing molecular genetic markers for 19 wood property candidate genes. We are targeting these 19 genes because they play key roles in the formation of wood based on studies in loblolly pine. We are developing genetic markers called single nucleotide polymorphisms (SNPs), which are single-letter changes in the DNA code that occur between different alleles (copies) of the same gene. To date, we have identified 11 of our target genes in Douglas-fir, and have developed SNP markers for these genes and then genotyped the parents. Data analysis and reporting of results was carried out during the past year. To date, we have determined that gains can be made in wood stiffness. Acoustic tools are valuable for assessing wood quality, but basal wood density is not a good measure of bending stiffness. Clonal seed orchards, which are easily measured, can be used to assess wood quality in breeding programs. Selection for growth will not have adverse effects on bending stiffness or acoustic velocity, but will adversely affect wood density. PARTICIPANTS: Glenn T. Howe, Principal Investigator; Marilyn L. Cherry, Co-Principal Investigator; Vikas Vikram, Graduate Research Assistant; David Briggs, Stand Management Cooperative, University of Washington; Daniel W. Cress, Olympic Resource Management; Brad St. Clair, USFS Pacific Northwest Research Station; and Dave Neale, University of California at Davis.

Impacts
Wood stiffness is the most important property of structural lumber. Because juvenile wood is less stiff than mature wood, the quality of Douglas-fir lumber may decline as rotation lengths decrease and proportionally more of the wood is derived from the juvenile core of the tree. Furthermore, because wood traits of coniferous species are often highly heritable, and can be improved through selection and breeding, it may be valuable to incorporate wood stiffness into Douglas-fir breeding programs. Nonetheless, direct measures of wood stiffness are costly and require destructive sampling. Therefore, alternatives to destructive testing are needed to measure wood stiffness prior to harvest. For these reasons, we are carrying out extensive research on the measurement, quantitative genetics, and molecular genetics of Douglas-fir wood stiffness and strength. This study has determined that indirect and/or nondestructive methods of evaluating wood stiffness and strength are useful in genetic tests. Furthermore, we will develop protocols and recommendations for using nondestructive test procedures in tree improvement programs.

Publications

No publications reported this period

Progress 01/01/06 to 12/31/06

Outputs
We currently have two major research projects: (1) genetics of Douglas-fir wood stiffness and strength; and (2) miniaturized seed orchards (MSOs). Our wood quality research combines a number of novel elements, including the evaluation of new acoustic tools that can be used to obtain indirect estimates of wood stiffness on standing trees and logs, comparisons of wood stiffness estimated indirectly from the acoustic tools with stiffness measured directly on lumber harvested from the same trees, evaluation of wood stiffness of seed orchard parents and their progeny growing in genetic test plantations, and discovery of genes associated with wood properties using genomic approaches. To accomplish these goals, we are collaborating with the Stand Management Cooperative at the University of Washington, scientists at the University of California at Davis, the Genetics Team at the USFS Pacific Northwest Research Station, and Olympic Resource Management. Some of the key questions we hope to answer are: "What are the potential genetic gains for wood stiffness?," "What are the most efficient approaches for improving wood stiffness in operational breeding programs?," "Can acoustic tools such as the Fibre-gen HM200 and ST300 be used to measure and select for wood stiffness in operational programs?," "Can we improve wood stiffness by simply measuring and selecting for increased wood specific gravity?," and "Will wood stiffness and specific gravity decline if we select and breed for volume growth alone?" Miniaturized seed orchards are orchards in which the trees are planted at close spacings in clonal rows, and then maintained at a height of only 2 to 4 m. We are undertaking experiments that are designed to help us develop methods for establishing and managing miniaturized seed orchards of Douglas-fir. Previously, we tested flower stimulation techniques for very young grafts of Douglas-fir. This research demonstrated that male and female flowering can be stimulated on very young grafts using a combination of girdling and gibberellic acid. Because the treated trees had higher mortality than the untreated trees, the best approach would be to delay flower stimulation until the grafts are 5 years old. Our largest MSO experiment is a long-term test of alternative MSO designs that was established at the Plum Creek Seed Orchard Complex in western Oregon. We completed the grafting for this experiment in 2004, and the trees should be large enough to begin testing crown management treatments as early as the summer of 2007. We also initiated a pruning study at the Roseburg Products Vaughn seed orchard in the spring of 2005. This experiment is designed to determine the best time to prune the crowns of MSO trees. Pruning is needed to keep the trees small, but is also expected to reduce the number of cones, thereby adversely affecting seed production. We hypothesize that the timing (season and frequency) of crown pruning can be physiologically optimized to maximize seed production. Eventually, we expect to test one or more of the best pruning treatments from this experiment at the Plum Creek MSO, in addition to other crown management treatments.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. Our wood quality research will lead to ensuring Douglas-fir's niche in domestic and international timber markets by maintaining wood stiffness and strength. Our results will also assist the forest sector in cost-saving and increasing product value. We will identify wood properties that can be incorporated into breeding programs to improve wood stiffness and strength.

Publications

Howe, G.T., K.J. Jayawickrama, M.L. Cherry, G.R. Johnson and N.C. Wheeler. 2006. Breeding Douglas-fir. p. 245-353 In: Plant Breeding Reviews. 27. J. Janick, ed. John Wiley and Sons, Inc.

Progress 01/01/05 to 12/31/05

Outputs
The PNWTIRC, in conjunction with other organizations, hosted the Western Forest Genetics Association meeting in 2005. A new long-term study on the genetics of Douglas-fir wood quality was initiated in 2005 after being approved by PNWTIRC members at the 2005 Annual Meeting. Wood quality has a high impact on timber value, but current methods of evaluating wood quality prior to harvest are inadequate. The use of wood quality traits in tree improvement programs requires the use of rapid measurement techniques that are preferably non-destructive and applicable to small trees. The Wood Quality Study will explore whether nondestructive or post-harvest assessment methods can be developed for use in tree improvement programs. The study will also estimate potential genetic gains for wood quality strength and stiffness traits. By measuring parents in seed orchards, we may be able to efficiently and cheaply estimate the wood quality of their progeny. We will compare wood stiffness and strength of clonal orchard parents with that of their progeny. Because of its high economic importance, wood quality is one of the key areas of interest for gene association mapping. The associations between wood quality phenotypes and candidate genes will be investigated in Douglas-fir. The goal of this work is to identify molecular genetic markers that are associated with desirable wood properties in Douglas-fir. Ultimately these markers may be used by breeders to improve wood stiffness and strength. Progeny tests associated with one tree improvement program series in Washington are being thinned, and will provide the study materials. During the summer of 2005, trees will be measured for diameter and standing tree acoustic velocity. Harvesting will commence in the fall of 2005. Nine foot butt logs will be measured for log acoustic velocity and wood disks will be measured for specific gravity and ring count traits. A subsample of about 8 logs from each of 50 families per site will be milled into 2 x 4 lumber, and tested for wood stiffness and strength. A new PNWTIRC-supported graduate student will be working on a portion of the study for his thesis.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. Our wood quality research will lead to ensuring Douglas-firs niche in domestic and international timber markets by maintaining wood stiffness and strength. Our results will also assist the forest sector in cost-saving and increasing product value. We will identify wood properties that can be incorporated into breeding programs to improve wood stiffness and strength.

Publications

Bower, A.D., W.T. Adams, D. Birkes and D. Nalle. 2005. Response of annual growth ring components to soil moisture deficit in young, plantation-grown Douglas-fir in coastal British Columbia. Canadian Journal of Forest Research 35:2491-2499.

Progress 01/01/04 to 12/31/04

Outputs
The PNWTIRC, in conjunction with other organizations, held a workshop on "Cold Hardiness Testing in Advanced-generation Cooperative Genetic Improvement Programs." The Pollen Contamination Study, which characterized DNA-based SSR markers, was completed with the Ph.D. thesis defense of a PNWTIRC-supported graduate student and final publication of results. The Early Flowering Study, which demonstrated that seed production is possible on young grafted trees with gibberellic acid and girdling treatments, was also completed, and a manuscript of the results is in preparation. A new study on the genetics of wood quality is now being designed, and work will commence in the spring of 2005.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. The Pollen Contamination Study demonstrated that SSR markers are a valuable tool for Douglas-fir tree improvement programs. The Early Flowering Study indicates the potential for obtaining commercially harvestable crops in just a few years after orchard establishment. Benefits also include a shortened time period between generations in breeding programs and improved capture of genetic gains from seed orchards.

Publications

Cherry, M.L. and G.T. Howe, eds. 2004. Proceedings, Genetics and Growth Modeling Workshop. November 4-6, 2003, Vancouver, WA. Pacific Northwest Research Cooperative Report #21. Oregon State Univ., Corvallis. 171 p.
Slavov, G.T., G.T. Howe, A.V. Gyaourova, D.S. Birkes and W.T. Adams. 2004. Estimating pollen flow using SSR markers: The effect of mistyping on paternity exclusion. Pacific Northwest Tree Improvement Cooperative Report, PNWTIRC #20. 33 p.
Slavov, G.T., G.T. Howe, A.V. Gyaourova, D.S. Birkes and W.T. Adams. 2004. Pollen Flow (PFL): A computer program for estimating pollen flow using highly variable markers and paternity exclusion.
Slavov, G.T., G.T. Howe, I. Yakovlev, K.J. Edwards, K.V. Krutovskii, G.A. Tuskan, J.E. Carlson, S.H. Strauss and W.T. Adams. 2004. Highly variable SSR markers in Douglas-fir: Mendelian inheritance and map locations. Theoretical and Applied Genetics 108:873-880.
Slavov, G.T. 2004. Development and Application of SSR Markers for Measuring Gene Flow in Douglas-fir. Ph.D. Dissertation. Oregon State Univ., Corvallis.

Progress 01/01/03 to 12/31/03

Outputs
PNWTIRC staff published three refereed journal articles. The PNWTIRC, in conjunction with other agencies, held a workshop entitled "Genetics and Growth Modeling Workshop." In the Pollen Contamination Study, characterization of DNA-based SSR markers was completed by mapping 20 of our 22 highly variable SSRs to 10 linkage groups in Douglas-fir. A subset of these SSRs was used to measure seed orchard seed and pollen contamination and to determine proportional parental contributions to orchard seedlots. SSRs were also used to measure the success of supplemental mass pollination. Final measurements were taken in 2003 in the Early Flowering Study following gibberellic acid (GA) and girdling treatments applied during 2001 and 2002 to 2- and 4-year-old grafts. The greatest number of cones was produced using a combined GA and girdling treatment. Flower stimulation treatments did not impede tree growth. Graduate Student = 1.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of regional tree improvement programs. The Pollen Contamination Study is demonstrating that SSR markers are a valuable tool for Douglas-fir tree improvement programs. The Early Flowering Study indicates the potential for obtaining commercially harvestable crops in just a few years after orchard establishment. Benefits also include a shortened time period between generations in breeding programs and improved capture of genetic gains from seed orchards.

Publications

Slavov, G.T., S.P. DiFazio and S.H. Strauss. 2003. Gene flow in forest trees: gene migration patterns and landscape modeling of transgene dispersion in hybrid poplar. In: Proceedings of the International Conference on Introgression from Genetically Modified Plants into Wild Relatives, January 21-24, Amsterdam, The Netherlands. (In Press).
Slavov, G.T., I. Yakovlev, G.T. Howe and S.H. Strauss. 2003. Identification and characterization of microsatellite markers in Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco). Abstract. In: Dynamics and conservation of genetic diversity in forest ecosystems. International Conference, December 2-5, 2002, Strasbourg, France. http://www.pierroton.inra.fr/genetics/Dygen/abstracts.pdf.
Vargas-Hernandez, J.J., W.T. Adams and D.G. Joyce. 2003. Quantitative genetic structure of stem form and branching traits in Douglas-fir seedlings and implications for early selection. Silvae Genetica 52(1): 36-44.
Howe, G.T., S.N. Aitken, D.B. Neale, K.D. Jermstad, N.C. Wheeler and T.H.H. Chen. 2003. From genotype to phenotype: Unraveling the complexities of cold adaptation in forest trees. Can. J. Botany 81(12): 1247-1266.

Progress 01/01/02 to 12/31/02

Outputs
PNWTIRC staff published two refereed journal articles, one abstract in conference proceedings, and a book chapter was accepted for publication. In addition, PNWTIRC conducted a workshop entitled "Genetic Improvement of Wood Quality in Coastal Douglas-fir and Western Hemlock." In the Pollen Contamination Study we developed 7 new SSR genetic markers, adding to the 15 SSR markers we developed last year. We subsequently conducted detailed analyses of our full set of 22 SSRs. Based on their ease of scoring and repeatability, at least 15 of these markers are suitable for measuring pollen contamination. In the spring of 2002, we measured the results of the early flowering treatments that were applied in the spring of 2001 for the Early Flowering Study. Gibberellic acid (GA) and girdling treatments were applied to 2- and 4-year-old grafts in two young seed orchards. The combined GA/girdling treatment significantly increased female flowering on both the 2- and 4-year-old grafts, but only increased male flowering on the older trees. The same treatments were applied in the spring of 2002, and new treatments were applied to test different levels of GA.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative conducts genetics and breeding research in support of tree improvement programs in the region. The Pollen Contamination Study will help increase genetic gains by using the identified SSRs to measure and manage pollen contamination in seed orchards. Our SSR markers will also be valuable for identifying genotypes in tree improvement programs, gene conservation, population genetic studies, and gene mapping. The predicted benefits of our Early Flowering Study will be to reduce the time lag between seed orchard establishment and seed production. This will shorten the generation time in breeding programs and improve the capture of genetic gains from seed orchards.

Publications

Anekonda, T.S., M.C. Lomas, W.T. Adams, K.L. Kavanagh and S.N. Aitken. 2002. Genetic variation in drought hardiness of coastal Douglas-fir seedlings from British Columbia. Canadian Journal of Forest Research. (In press).
DiFazio, S.P., G.T. Slavov, J. Burczyk, S. Leonardi and S.H. Strauss. 2002. Gene flow from tree plantations and implications for transgenic risk assessment. In: Plantation forest biotechnology for the 21st century. Walter, C. and M. Carson, eds. Research Signpost. Trivandrum, India. (Accepted for publication).
Slavov, G.T., S.P. DiFazio and S.H. Strauss. 2002. Gene flow in forest trees: From empirical estimates to transgenic risk assessment. p. 113-133 In: Proceedings Scientific Methods Workshop: Ecological and agronomic consequences of gene flow from transgenic crops to wild relatives, March 5-6, Columbus, OH. http://www.biosci.ohio state.edu/~lspencer/Proceedings.pdf
Adams, W.T., S.N. Aitken, D.G. Joyce, G.T. Howe and J. Vargas-Hernandez. 2001. Evaluating efficacy of early testing for stem growth in coastal Douglas-fir. Silvae Genetica 50:167-175.

Progress 01/01/01 to 12/31/01

Outputs
PNWTIRC staff published four refereed journal articles, three abstracts in conference proceedings, and an annual report in 2001. In the Seedling Drought Physiology Study, 39 full-sib families showed considerable genetic variation in bud set, second flushing and cold hardiness under both well-watered and moisture-stress regimes. Although mild drought enhances cold hardiness under routine nursery management, severe drought applied in this study reduced cold hardiness of the full-sib families. Family rankings for bud set, second flushing and cold hardiness remained similar across the moisture regimes. There was a tendency for fall cold injury to be increased in trees that set bud late in fall, second flushed and grew taller. Drought hardiness traits, such as drought-induced foliage damage, xylem cavitation and hydraulic conductivity, are essentially uncorrelated with bud set, second flushing, and cold hardiness. These observations suggest that selection for improved bud phenology and cold hardiness is unlikely to affect drought hardiness in coastal Douglas-fir. We developed 15 promising SSR markers in the Pollen Contamination Study. These markers were derived from five new genomic libraries that we constructed using recently improved molecular techniques. At least 62 potentially promising SSR markers remain to be tested.

Impacts
A primary goal of the Pacific Northwest Tree Improvement Research Cooperative is to conduct quantitative genetics and breeding research on Douglas-fir growth, adaptability and wood quality traits in support of tree improvement programs in the region. Cold and drought hardiness screening methods and early testing procedures developed by the Cooperative will help in the development of improved Douglas-fir varieties, while also reducing progeny test costs. In addition, genetic markers will be useful for accurately quantifying and reducing pollen contamination levels in seed orchards.

Publications

Adams, W.T., S.N. Aitken, D.G. Joyce, G.T. Howe and J. Vargas-Hernandez. 2002. Evaluating efficiency of early testing for stem growth in coastal Douglas-fir. Silvae Genetica. (In Press).
Anekonda, T.S. 2001. Extreme growth phenotypes of trees are caused by differences in energy metabolism. Thermochimica Acta 373:125-132.
Anekonda, T.S. 2001. Genetics of cold and drought hardiness in coastal Douglas-fir. In: Proceedings of the Western Forest Genetics Association Meeting, July 30-August 2, Davis, CA.
Anekonda, T.S. 2001. A perspective on the application of biological calorimetry to tree physiology and forest genetics studies. XII Conference of the International Society for Biological Calorimetry, September 7-11, Santiago de Compostela, Spain.
Howe, G.T., T.S. Anekonda and G. Slavov. 2001. Pacific Northwest Tree Improvement Research Cooperative Annual Report 2000-01. Forest Research Laboratory, Oregon State University, Corvallis. 33 pp.
Howe, G.T. 2001. Physiology and genetics of dormancy-related traits in Populus. In: Proceedings of the Western Forest Genetics Association Meeting, July 30-August 2, Davis, CA.

Progress 01/01/00 to 12/31/00

Outputs
Five papers resulting from Coop's Cold Hardiness research and one each from Seedling Drought Hardiness and Age-age Genetic Correlations in Stem Defects were published in 2000. In the Seedling Drought Physiology Study, 39 coastal Douglas-fir families varied considerably in their response to summer drought applied in the nursery, in terms of growth increment, xylem cavitation, hydraulic conductivity, and shoot damage. Genetic correlations between drought hardiness traits in the same year were high, and were low to moderate between different years. Seedling growth potential of families under well-watered conditions is uncorrelated with drought hardiness, suggesting that selection for growth traits will not reduce drought hardiness of the selected trees. Moderate drought applied to seedlings during the second growing season greatly reduced their growth increment in the following recovery year. In the Field Drought Study, due to their low heritability and difficulty of measurement, there seems to be little practical utility to using drought sensitivity coefficients to assess drought hardiness in older, field-grown trees. The lack of correlation between diameter at breast-height and drought sensitivity coefficients is encouraging because like in seedlings, it suggests that selection for rapid stem growth in older trees will not inadvertently reduce hardiness to summer drought. The relationship between drought hardiness in seedlings and the ability to grow under summer moisture deficits later in the rotation is unclear. Additional analysis of data from the Cooperative's Early Testing Study showed unfavorable correlations between growth potential and stem form and branching traits in seedlings, such that selection for growth alone is expected to increase the number of whorls with steep-angle branches and stem sinuosity in older trees. To avoid negative impacts, appropriate stem form and branching traits should be included, along with growth traits in nursery-stage selection. Alternatively, growth potential could be emphasized in initial nursery culling, with stem form and branching traits dealt with later in field selections. Pollen Contamination Study of our Coop showed that microsatellite and minisatellite DNA markers are highly polymorphic and very effective for mating studies, and four to six microsatellite marker loci should be sufficient for pollen contamination estimation in Douglas-fir seed-orchards.

Impacts
A primary goal of Pacific Northwest Tree Improvement Research Cooperative is to conduct quantitative genetics and breeding research on Douglas-fir growth and adaptability traits in support of tree improvement programs in the region. Cold and drought hardiness screening methods developed by the Coop will not only help developing improved Douglas-fir varieties, but also provide information useful for predicting stability and productivity of Douglas-fir families in operational field conditions.

Publications

O'Neill, G.A., W.T. Adams and S.N. Aitken. 2000. Quantitative genetics of spring and fall cold hardiness in seedlings from two Oregon populations of coastal Douglas-fir. Forest Ecology and Management. (In Press).
Temel, F. and W.T. Adams. 2000. Persistence and age-age genetic correlations of stem defects in coastal Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco). Forest Genetics 7:145-153.
Adams, T. and T. Anekonda. 2000. Pacific Northwest Tree Improvement Research Coop. Western Forester 45:13.
Adams, W.T. and J. Burczyk. 2000. Magnitude and implications of gene flow in gene conservation reserves. p. 215-224 In: Forest Conservation Genetics: Principles and Practices. A. Young, D. Boshier and T. Boyle, eds. CSIRO Publishing, Collingwood, Victoria, Australia.
Anekonda, T.S. and W.T. Adams. 2000. Genetics of dark respiration and its relationship with drought hardiness in coastal Douglas-fir. Thermochimica Acta 349:69-77.
Anekonda, T.S., W.T. Adams and S.N. Aitken. 2000. Cold hardiness testing for Douglas-fir tree improvement programs: Guidelines for a simple, robust, and inexpensive screening method. Western Journal of Applied Forestry 15:129-136.
Anekonda, T.S., W.T. Adams, S.N. Aitken, D.B. Neale, K.D. Jermstad and N.C. Wheeler. 2000. Genetics of cold hardiness in a cloned full-sib family of coastal Douglas-fir. Canadian Journal of Forest Research 30: 837-840.
Anekonda, T.S., M.C. Lomas and W.T. Adams. 2000. Genetic variation in drought hardiness of coastal Douglas-fir (Pseudotsuga menziesii var. menziesii) seedlings. Joint meeting of the North American Forest Biology Workshop and Western Forest Genetics Association, July 17-20, Merida, Mexico. (Abstract).
Jermstad, K.D., D.L. Bassoni, N.C. Wheeler, T.S. Anekonda, S.N. Aitken, W.T. Adams and D.B. Neale. 2000. Mapping of quantitative trait loci controlling adaptive traits in coastal Douglas-fir: II. Spring and fall cold hardiness. Theoretical and Applied Genetics. (In Press).
O'Neill, G.A., S.N. Aitken and W.T. Adams. 2000. Genetic selection for cold hardiness in coastal Douglas-fir seedlings and saplings. Canadian Journal of Forest Research. (In Press).

Progress 01/01/99 to 12/31/99

Outputs
Work continued analyzing results from three studies on the genetics of drought hardiness in coastal Douglas-fir. Shoot damage varied widely among 39 full-sib families subjected to a severe drought in a nursery trial (3rd growing season), suggesting that selection among families for increased seedling drought hardiness at the seedling stage would be quite effective. There appears to be little genetic association between drought hardiness in seedlings and seedling growth potential in favorable moisture regimes. A calorimetric investigation of a subset of these families showed that dark respiration is generally reduced under moisture stress, but that families differ in the degree to which respiration traits respond to drought. In the third drought hardiness study, x-ray densitometry analysis of increment cores from sapling-age trees of the same 39 families in the nursery trial, revealed that families differed widely in their response (i.e., in earlywood growth and ring density) to past summer droughts at one field test site. Good progress was made in developing hypervariable microsatellite (SSR) genetic markers for use in Douglas-fir seed orchard studies. Preliminary analysis revealed as many as 10-12 different variants (alleles) at some loci in DNA samples from less than 20 individuals. Finally, a plan was approved to evaluate three alternative miniaturized seed orchard designs in Douglas-fir. The three orchard types will be compared for quantity and quality of seed production and for ease and efficiency of management. Alternative flower initiation, height control, and control pollination techniques will also be investigated.

Impacts
The Pacific Northwest Tree Improvement Research Cooperative provides research support to operational tree improvement programs in the Pacific Northwest. Studies on the genetics of adaptive traits, such as drought hardiness, help in evaluating the impacts of breeding on adaptability, and in developing breeding methods to ensure that improved varieties will be adapted to a wide variety of planting environments. Studies in seed orchards are aimed at improving the genetic quality of seed produced and the cost effectiveness of seed production.

Publications

Balduman, L.M., Aitken, S.N., Harmon. M. and Adams, W.T. 1999. Genetic variation in cold hardiness of Douglas-fir in relation to parent tree environment. Canandian Journal of Forest Research 29:62-72.
Lomas, M.C. 1999. Physiology and genetics of drought hardiness in coastal Douglas-fir seedlings. M.S. Thesis. Oregon State Univ., Corvallis. 80 p.
O' Neill, G.A. 1999. Genetics of fall, winter, and spring cold hardiness in coastal Douglas-fir seedlings. Ph.D. Thesis. Oregon State Univ., Corvallis. 84 p.

Progress 01/01/98 to 12/31/98

Outputs
Much of the work this year focused on studies of genetics of drought hardiness in Douglas-fir. Data collection on a seedling drought study was completed, where 39 full-sib families from British Columbia were grown in raised nursery beds in Corvallis for three years. Application of severe drought in the second growing season had only minor influence on seedling growth (relative to a well-watered control), although 20% of the foliage and 18% of xlem conduits were damaged (xylem cavitation), on average, by the drought. Families varied widely in percent cavitation in the second growth ring (5-59%), suggesting this trait may be a useful indicator of drought hardiness in seedlings. An even more severe drought was applied to the seedlings in the third growing season in an attempt to ellicit a dramatic growth response to drought. Early indications are that we were quite successful. Andrew Bower used x-ray densitometry to retrospectively assess the influence of summer drought on annual growth ring components of trees in young Douglas-fir progeny tests. The results indicate that several growth ring components (particularly latewood traits) respond consistently to summer drought, and may be useful for evaluating sensitivity of families to drought in sapling-age trees. Work has begun on a study to use microsatellite (SSR) genetic markers to investigate factors influencing pollen contamination in Douglas-fir seed orchards. The cooperative is also developing a study plan to investigate the feasibility of micro-orchards for seed production in advanced-generation Douglas-fir breeding programs.

Impacts
(N/A)

Publications

Anekonda, T., Adams, W.T. and Aitken, S.N. 1998. Influence of second flushing on genetic assessment of cold hardiness in coastal Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco). Forest Ecology and Management 111:119-126.
Adams, T., Anekonda, T., Bower, A., Lomas, C. and Temel, F. 1998. Pacific Northwest Tree Improvement Research Cooperative Annual Report 1997-98. Forest Research Laboratory, Oregon State Univ., Corvallis. 24 p.
Bower, A.D. 1998. Response of annual growth ring components to soil mositure deficit in young, plantation grown Douglas-fir in coastal British Columbia. M.S. Thesis. Oregon State Univ., Corvallis. 88 p.

Progress 01/01/97 to 12/31/97

Outputs
A primary goal of the PNWTIRC is to conduct quantitative genetics and breeding research on Douglas-fir growth and adaptability traits. A paper on spring cold hardiness in Douglas-fir was published in 1997 and a manuscript on "Adaptive Significance of Second Flushing in Coastal Douglas-fir" was prepared. Greg O'Neill began writing his Ph.D. dissertation on seedling cold hardiness, addressing the potential for early testing cold hardiness traits in Douglas-fir. Fatih Temel completed his M.S. thesis addressing the persistence stem defects from age 12 to 24 in Douglas-fir progeny tests. Genetic correlations between these traits at the two ages were strong. Andy Bower successfully defended his M.S. thesis entitled, "Response of Annual Growth Ring Components to Soil Moisture Deficit in Young, Plantation Grown Douglas-fir in Coastal British Columbia." Drought treatments were applied during the spring and summer in the "Seedling Drought Hardiness Study," and responses to drought in terms of growth, wood anatomy, and physiological traits assessed. This study has the main objective to understand genetic variation in drought hardiness and develop criteria for screening improved families. Wood density profiles from additional sapling trees in British Columbia progeny tests are currently being analyzed to evaluate genetic variation in annual growth ring components sensitive to summer drought. Preparation of new study plans on micro-orchard and pollen contamination research was initiated during 1997.

Impacts
(N/A)

Publications

AITKEN, S.N. and ADAMS, W.T. 1997. Spring cold hardiness under strong genetic control in Oregon populations of coastal Douglas-fir. Canadian Journal of Forest Research 27(11):1773-1780.
TEMEL, F. 1997. Persistence of age-age correlations of stem defects in Douglas-fir (Psuedotsuga menziesii var. menziesii (Mirb.) Franco). M.S. Thesis. Oregon State Univ., Corvallis. 52 pp.
ADAMS, W.T., ANEKONDA, T.S., O'NEILL, G., BOWER, A. and TEMEL, F. 1997. Pacific Northwest Tree Improvement Research Cooperative Annual Report 1996-97. Forest Research Laboratory, Oregon State Univ.,

Progress 01/01/96 to 12/30/96

Outputs
A primary objective of the Pacific Northwest Tree Improvement Research Cooperative is to understand the quantitative genetics of adaptability traits in Douglas-fir. Four papers resulting from the "Field Cold Hardiness Study" were published in 1996. A second study on cold hardiness is investigating the potential for early testing cold hardiness traits at the seedling stage. Data analysis on a third project, "Adaptive Significance of Second Flushing in Coastal Douglas-fir", was completed. A new major study on "Genetic Variation in Seedling Drought Physiology of Coastal Douglas-fir" was initiated. The major goal of this project is to assess physiological mechanisms with the aim of developing criteria for screening improved families for drought hardiness. In addition, wood density profiles for trees sampled in British Columbia progeny tests are currently being analyzed to determine whether annual growth ring variables reflect summer moisture stress and can be used to assess drought hardiness in older, field grown trees. In addition to the adaptability studies, progeny-test trees scored for stem defects 12 years ago were recently reassessed to evaluate the degree to which these effects persist with age and the validity of early selection for improved stem form. Work was also begun on searching the literature for flowering and pollen studies in conifers, a potential new area of the cooperator's research.

Impacts
(N/A)

Publications

ADAMS, W.T., AITKEN, S. N., BALDUMAN, L. and O'NEILL, G. 1996. Pacific Northwest Tree Improvement Research Cooperative Annual Report 1995-96. Forest Research Laboratory, Oregon State Univer., Corvallis. 28 pp.
AITKEN, S.N. and ADAMS, W.T. 1995. Screening for cold hardiness in coastal Douglas-fir. p. 321-324 In: Eucalypt Plantations: Improving Fibre Yield..., B.M.Potts, et al., eds. Proc., CRC-IUFRO Conf., 2/19-24, Hobart, Australia.
AITKEN, S.N. and ADAMS, W.T. 1995. Impacts of genetic selection for height growth on annual developmental... P.AD-1:1-8. In: Evolution of Breeding... Proc., IUFRO Conf., 7/31-8/4, Limoges, France.
AITKEN, S.N. and ADAMS, W.T. 1996. Family variation for cold hardiness in two populations of coastal Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.)Franco. Forest Ecology and Management 80(1-3):187-195.
AITKEN, S.N. and ADAMS, W.T. 1996. Genetics of fall and winter cold hardiness ofcoastal Douglas-fir in Oregon. Can. J. For. Res. 26:1828-1837.
O'NEILL, G. A., ADAMS, W.T. and AITKEN, S.N. 1996. Susceptibility to autumn frost... In Dieters, M.
J., et al. (eds.). Tree Improvement for Sustain. Trop. For. Proc. QFRI-IUFRO Conf., 10/27-11/1, Queensland, Australia.

Progress 01/01/95 to 12/30/95

Outputs
The research focus of this project continued to be the genetics of adaptability in coastal Douglas-fir. Data analysis and report writing continued for the Field Cold Hardiness Study, with two papers published and two additional papers submitted to journals on the genetics of cold hardiness and relationships between cold hardiness and growth rates. Two new studies were initiated, "Genetics of cold hardiness in coastal Douglas-fir seedlings" being conducted by PhD student Greg O'Neill, and "Age-age correlations of stem defects in coastal Douglas-fir," the MS research of Fatih Temel. Study plans were developed for two projects to be initiated early in 1996. The first project, "Genetic variation in seedling drought physiology of coastal Douglas-fir," will assess genetic relationships between stem hydraulic conductivity, biomass allocation, gas exchange, growth rates and phenology, and ability of seedlings to survive severe drought or grow under moderate drought. MS student Andrew Bower will investigate genetic variation in annual growth ring traits (earlywood and latewood proportion and density) relative to annual climatic variation in the second project, "Family variation in drought hardiness of sapling age Douglas-fir.".

Impacts
(N/A)

Publications

Progress 01/01/94 to 12/30/94

Outputs
Research focused on the amount of genetic variation and degree of genetic control for adaptive traits in coastal Douglas-fir. Data collection for the Field Cold Hardiness Study was completed in 1994. Cold hardiness, phenology and growth were measured repeatedly over a two-year period in saplings of 80 families. There is considerable genetic variation for cold hardiness in Douglas-fir. Heritabilities for cold hardiness vary seasonally, and range from low in the fall to very high in the spring. Fall and spring hardiness are independent traits. Bud phenology is highly correlated with spring cold hardiness, but only weakly genetically linked with fall hardiness. Lisa Balduman, an MS student, neared completion of a related project entitled "Variation in adaptive traits: Genetic and Environmental Components." Data collection was completed for a study of the phenotypic and genetic effects of second flushing on fall cold hardiness in coastal Douglas-fir.

Impacts
(N/A)

Publications

Progress 01/01/93 to 12/30/93

Outputs
The Field Cold Hardiness Study, investigating the adaptation of Douglas-fir families to temperature regimes, was the primary focus of research in 1993. Data on fall, winter and spring cold hardiness, timing of budburst and budset, and duration of shoot elongation were collected for forty families from each of two breeding populations growing in operational progeny tests. A companion study, titled "Genetic response of Douglas-fir to annual climatic cycles," assessing the impact of thermoperiod on cold hardiness, dormancy and phenology in controlled environments is planned pending funding. A preliminary study was completed on genetic variation in stable carbon isotope discrimination in Douglas-fir. Discrimination against 13C has been shown to be related to water-use efficiency (WUE), but surprisingly, trees from drier source environments discriminated more against 13C (and thus had lower WUE) than trees from wetter source environments. A small study on genetic variation in drought hardiness and associated physiological mechanisms (including WUE, hydraulic architecture and biomass allocation) was initiated. Data analysis and report writing continued for the Provenance Architecture Study and the Early Testing Study.

Impacts
(N/A)

Publications

Progress 01/01/92 to 12/30/92

Outputs
A large study on the adaptation of Douglas-fir families to temperature regimes was inititated in 1992. The Field Cold Hardiness Study is designed to determine the degree of genetic control and amount of genetic variation for cold hardiness traits, to determine the relationships between different cold hardiness traits (e.g., cold hardiness assessed in different tissues or on different sampling dates), and to quantify the relationship among cold hardiness, shoot phenology and growth. This is a two-year study of forty families in each of two breeding populations in Oregon. To date, data have been collected on date of budset, shoot elongation, and fall cold hardiness. A companion study of the impact of thermoperiod on annual developmental cycle traits of seedlings is planned pending funding. Work has also been initiated to investigate genetic variation in drought hardiness. A preliminary study was initiated in November 1992, to determine if genetic variation exists for carbon isotope discrimination in Douglas-fir. Discrimination against 13C has been shown to be tightly linked to water-use efficiency. Data analysis and report writing continues for two studies: The Provenance Architecture Study, designed to examine differences in the genetic control of and interrelationships among traits in ecologically different breeding zones; and the Early Testing Study, investigating the ability to predict the field performance of families from seedlings.

Impacts
(N/A)

Publications

Progress 01/01/91 to 12/30/91

Outputs
Data collection was completed for two new studies in 1991. The Provenance Architecture Study was designed to examine differences in the genetic control of and interrelationships among traits in ecologically different breeding zones. Data were collected on growth, stem quality, wood quality, and phenology for 80 to 90 families in progeny tests on two sites in each of two breeding zones. Preliminary analyses indicate that there is significant genetic variation for all traits, and that the genetic relationships among traits do not differ greatly among breeding populations, with one exception: The relationship between volume growth and wood density was nonsignificant in one breeding zone and significant and unfavorable in the other. In the Cold Hardiness Study, shoot cuttings from all trees in the Provenance Architecture Study were laboratory tested for cold hardiness. There was significant genetic variation for cold hardiness on all sites, however, there was also strong family by site interaction in one breeding zone. The family mean correlation between growth and cold damage ranged from small or non-significant on some sites to strong and unfavorable (r=0.7) on one site. Further analysis of the ongoing Early Testing Study indicates that 30% to 40% of all families to be tested in a breeding program could be culled based on first-year performance in a greenhouse or nursery test, thereby reducing the size and cost of field progeny tests without reducing genetic gain.

Impacts
(N/A)

Publications

Progress 01/01/90 to 12/30/90

Outputs
The cooperative worked on three projects dealing with tree improvement in coastal Douglas-fir. Two on-going projects dealt with early testing and the genetics of wood quality. Planning was begun on a third project to address mechanisms of long-term adaptability. Results were obtained on three aspects of wood density (WD) in 15 year-old trees: the potential for early assessment, genetic relationships between WD and growth rhythm, and age trends in WD. Early testing appears very promising; WD assessed as early as age 1 was found to have a moderate genetic correlation with WD at age 15, with selection at age 1 estimated to be nearly 50% as effective as delaying selection until age 15. WD was genetically correlated with longer diameter growth periods (r = 0.67), but uncorrelated with date of budburst (r = 0.10). Despite the moderate correlation between WD and length of the diameter growth period, selection for WD is expected to have little adverse effect on adaptability because cambial phenology traits are weakly inherited. Age trends in relative earlywood WD were found to differ among families. The early plateau in age trend of some families may be an indication of earlier transition from juvenile to mature wood in these families.

Impacts
(N/A)

Publications

Progress 01/01/89 to 12/30/89

Outputs
The nursery phase of the early testing study was completed with the harvest of 12,000 Douglas-fir seedlings in 3 tests. Analysis of a portion of the data indicates that nursery tests can be effective for at least low-level culling of families for poor height growth, undesirable branch angle, and stem sinuosity in older trees. Results from the wood quality study show that although phenotypic variation in juvenile wood density (JWD) (15-year-old trees) is only about 1/3 to 1/2 that for growth traits in Douglas-fir narrow-sense heritability for JWD is high (.59); Thus, JWD can readily be improved by selection and breeding. A negative genetic correlation was found between JWD and bole volume (-.55), indicating it will be impossible to achieve maximal gains in bole volume growth without negatively affecting wood quality. Comparison of several selection indices which included measurement data on both JWD and bole volume, however, shows that good gains in bole volume can be achieved without loss of JWD, or while increasing JWD, when appropriate indices are employed. The Pilodyn wood tester was found to be an inexpensive, but effective, means of ranking families for JWD of outer growth rings.

Impacts
(N/A)

Publications

Progress 01/01/88 to 12/30/88

Outputs
The Cooperative now has three studies dealing with Douglas-fir tree improvement: 1) "Measurement strategies in young (10 to 15 year-old) progeny tests"; 2) Early testing for grwth, stem form, and adaptive traits"; and 3) "Genetic of wood quality". The newest finding in the first study is that the incidence of ramicorn (i.e., large, steep branches) is weakly inherited, but is still highly amenable to reduction through tree-breeding because it is highly variable among individual trees. However, ramicorn occurrence was found to be strongly and positively correlated genetically with tree height. This indicates it may be difficult to reduce ramicorns without also negatively influencing volume production. Preliminary analysis of the early testing data is encouraging because, although mean heights of 12 year-old families in field tests were found to be weakly correlated (r<.50) with seedling traits of the same families in nurseries, the correlations were consistent and were strong enough to be useful for early culling of families with poor genetic potential for growth. Two wood quality projects were initiated. The first investigates genetic control of wood density components (i.e., earlywood density, latewood density, and latewood proportion) in young Douglas-fir. The second examines the potential for early testing of wood density.

Impacts
(N/A)

Publications

Progress 01/01/87 to 12/30/87

Outputs
One on-going cooperative study had been to develop efficient means of assessing growth and stem quality in young Douglas-fir progeny tests. Measurement methods and inheritance of four branching traits (branch diameter, length, angle and number per whorl) were assessed in 12-to-13-year-old trees from 90 open-pollinated families growing at three test sites in western Oregon. Inexpensive means of measuring these traits have been found; e.g., branch diameter in the central portion of the bole can be accurately assessed by measuring in the front face of the tree, the single largest branch at the whorl nearest breast height. Percent genetic gains for branching traits appear to be of similar magnitude to those expected for height and diameter in these tests. When the branching traits were adjusted for tree size, they were found to be only weakly intercorrelated, suggesting it would not be difficult to breed for and combination of traits desired. Furthermore, small and negative genetic correlation was observed between stem diameter and adjusted branch diameter, indication that knot size can be decreased in selection programs with little or no reduction in volume growth potential.

Impacts
(N/A)

Publications

Progress 01/01/86 to 12/30/86

Outputs
A large study on early testing in Douglas-fir was initiated. Early testing is a process for making genetic selections on the basis of seedling performance after one of two year's growth. Early testing allows the culling of poor performing families prior to the establishment of longer term field tests, so that field testing can be more efficient and less costly. Seed from 140 families was sown in three nurseries. Nursery performance of the families for a variety of traits will be compared to volume growth of the same families in 10-15 year-old field tests. Two graduate student projects on Douglas-fir and closely related to early testing were also initiated. One deals with the genetic control of shoot and cambial phenology, the feasibility of early selection for these traits, and their relationship to growth. The second examines genetic components of nursery competition and then implications for the management of families in nurseries. In another project, the efficiencies of measuring heights in 12-13 year-old progeny tests of Douglas-fir with a clinometer and a height-pole were compared. Height measurement with a clinometer took about 60% of the time and resulted in 94% of the genetic gain for height obtainable using a height-pole. Two Graduate Students Associated.

Impacts
(N/A)

Publications

Progress 01/01/85 to 12/30/85

Outputs
The Pacific Northwest Tree Improvement Research Cooperative has 16 members, including representatives from private industry and government agencies in the U.S. and Canada. Early results from a study to determine the best traits for assessing growth and stem quality in young (10-15 year old) Douglas-fir progeny tests are as follows: It is quicker and nearly as effective to measure DBH with calipers as with a D-tape; Good results in estimating average branch diameter at a whorl can be obtained by measuring only the two largest branches on the whorl face and computing their average; For purposes of parental or family selection, stem sinuosity can be measured efficiently by visually estimating, in one-half stem diameter units, stem displacement at the second interval below the leader. Large variation in the degree of sinuosity, along with a moderate individual-tree heritability (0.19) for this trait, indicates stem straightners could be readily improved in Douglas-fir breeding programs; Selecting for stem volume based on diameter alone was nearly as efficient (90%) as selection based on both diameter and height measurements, but would result in small increases in stem taper.

Impacts
(N/A)

Publications

Progress 01/01/84 to 12/30/84

Outputs
A cooperative study was initiated to provide information which will help in choosing traits and measurement methods for assessing growth and stem quality in young (10-20 years-old) Douglas-fir progeny tests. In Phase I of this study, 180 trees (20 families) in a 13-year-old progeny test plantation were intensively measured to develop measurement techniques and provide preliminary information on phenotypic and genotypic variation in seven traits. As an example, stem sinuosity was found to vary significantly (PC .05) among families (family heritability = .76) and most efficiently measured by scoring the frequency and displacement of crooks in the second interwhorl from the top of tree, and then combining them into a "sinuosity index". Of the three branch traits investigated, branch diameter and angle were both found to vary significantly among families and have fairly high family heritabilities (.57 and .65); but, no family differences were detected in number of branches per main whorl. In Phase II, measurement techniques are being evaluated and further refined by measuring 90 families growing on each of three test sites. Also provided is a good quantity of data for making reliable estimates of the genetic control of these traits.

Impacts
(N/A)

Publications