Steven E. McKeand

Average effect of a mutation in lignin biosynthesis in loblolly pine.

Abstract Cinnamyl alcohol dehydrogenase (CAD, E.C. 1.1.1.195) is a monolignol biosynthetic enzyme that catalyzes the final step of lignin subunit biosynthesis in higher plants. Recently, a mutant allele of the cad gene, cad-n1, encoding for the CAD enzyme, was discovered in loblolly pine. By reducing the expression of the cad gene, this mutant has a decreased lignin content and major changes in the lignin composition in wood. In this study, we found that the substitution of a wild-type allele by cad-n1 was associated with a significant effect on 2nd-year shoot elongation in a half-sib family of loblolly pine (designated family 7–1037). The average effect of cad-n1 appeared to increase with tree growth and was greater for stem radial growth than height growth. An increase of 14.1% in de-barked volume in year 4 was associated with cad-n1. Co-segregation analysis indicated that the cad locus itself might represent a gene that governs stem growth in pine. The significance of the mutation cad-n1 for tree growth and wood processing is discussed.

Genomic Estimated Breeding Values Using Genomic Relationship Matrices in a Cloned Population of Loblolly Pine

Replacement of the average numerator relationship matrix derived from the pedigree with the realized genomic relationship matrix based on DNA markers might be an attractive strategy in forest tree breeding for predictions of genetic merit. We used genotypes from 3461 single-nucleotide polymorphism loci to estimate genomic relationships for a population of 165 loblolly pine (Pinus taeda L.) individuals. Phenotypes of the 165 individuals were obtained from clonally replicated field trials and were used to estimate breeding values for growth (stem volume). Two alternative methods, based on allele frequencies or regression, were used to generate the genomic relationship matrices. The accuracies of genomic estimated breeding values based on the genomic relationship matrices and breeding values estimated based on the average numerator relationship matrix were compared. On average, the accuracy of predictions based on genomic relationships ranged between 0.37 and 0.74 depending on the validation method. We did not detect differences in the accuracy of predictions based on genomic relationship matrices estimated by two different methods. Using genomic relationship matrices allowed modeling of Mendelian segregation within full-sib families, an important advantage over a traditional genetic evaluation system based on pedigree. We conclude that estimation of genomic relationships could be a powerful tool in forest tree breeding because it accurately accounts both for genetic relationships among individuals and for nuisance effects such as location and replicate effects, and makes more accurate selection possible within full-sib crosses.

Loblolly pine tissue culture: Laboratory, greenhouse, and field studies

Conifer tissue culture had its beginnings in the late 1930s (10) and shoot regeneration cultures were first noted in 1950 (4). Since that time many species (7, 17), especially those using embryonic materials for starting expiants, have been cultured. Among some of the most studied species, Pinus radiata (2), Pseudotsuga menziesii (6), Pinus pinaster (8), Picea abies (22), and Pinus taeda (18), much laboratory data are accumulating. To date, little field data have been reported on the performance of tissue-cultured conifers (14), but several conifers are now established in field plantings (21) and data should be forthcoming.

Leaf-level gas-exchange uniformity and photosynthetic capacity among loblolly pine (Pinus taeda L.) genotypes of contrasting inherent genetic variation

Variation in leaf-level gas exchange among widely planted genetically improved loblolly pine (Pinus taeda L.) genotypes could impact stand-level water use, carbon assimilation, biomass production, C allocation, ecosystem sustainability and biogeochemical cycling under changing environmental conditions. We examined uniformity in leaf-level light-saturated photosynthesis (A(sat)), stomatal conductance (g(s)), and intrinsic water-use efficiency (A(sat)/g(s) or δ) among nine loblolly pine genotypes (selected individuals): three clones, three full-sib families and three half-sib families, during the early years of stand development (first 3 years), with each genetic group possessing varying amounts of inherent genetic variation. We also compared light- and CO(2)-response parameters between genotypes and examined the relationship between genotype productivity, gas exchange and photosynthetic capacity. Within full-sib, half-sib and clonal genotypes, the coefficient of variation (CV) for gas exchange showed no consistent pattern; the CV for g(s) and δ was similar within clonal (44.3-46.9 and 35.5-38.6%) and half-sib (41.0-49.3 and 36.8-40.9%) genotypes, while full-sibs showed somewhat higher CVs (46.9-56.0 and 40.1-45.4%). In contrast, the CVs for A(sat) were generally higher within clones. With the exception of δ, differences in gas exchange among genotypes were generally insignificant. Tree volume showed a significant positive correlation with A(sat) and δ, but the relationship varied by season. Individual-tree volume and genotype volume were positively correlated with needle dark respiration (R(d)). Our results suggest that uniformity in leaf-level physiological rates is not consistently related to the amount of genetic variation within a given genotype, and δ, A(sat) and R(d) were the leaf-level physiological parameters that were most consistently related to individual-tree and genotype productivity. An enhanced understanding of molecular and environmental factors that influence physiological variation within and between loblolly pine genotypes may improve assessments of genotype growth potential and sensitivity to global climate change.

Phenological variation in height and diameter growth in provenances and families of loblolly pine

The phenology of 5- and 6-year old loblolly pine (Pinus taeda L.) trees was studied over two different growing seasons (1993 and 94) in southwest Georgia. These trees were from 7–9 open-pollinated families from each of four different provenances planted at two locations. The provenances were: Atlantic Coastal Plain (eastern SC), Gulf Hammock (north FL), Lower Gulf (south AL, MS) and Upper Gulf (north AL, MS). Provenances did not vary as to when height growth started in spring, but showed very significant differences for the date of growth cessation in fall. The Gulf Hammock source grew the most and also had the longest height growth period, while the Upper Gulf source was first to stop height growth and had the least annual height increment. Provenances were also significantly different for the date of cessation of diameter growth (a difference of 22 days between Gulf Hammock and Upper Gulf), and the order of cessation was the same as for height. Families within provenances were significantly different for date of cessation of height growth and diameter growth. When family means were considered across provenances, there was a correlation of 0.69 (p-value = 0.0001) between annual height increment and date of height growth cessation. There was a weaker association between faster growth and a longer growing season within provenances.

Genotype by environment interaction for index traits that combine growth and wood density in loblolly pine

Abstract Genotype×environment interactions of individual traits have been assessed in numerous experiments with forest trees. However, since breeding programs rarely aim at the improvement of a single trait, the impact of G×E on index or composite traits must also be assessed. In a study with 12-year-old loblolly pine families in the southeastern U.S., G×E variance was of relatively little importance compared to genetic variance for wood density but was of greater significance for several growth traits. An index that combined stem volume and wood density to improve dry weight but maintain wood density constant (restricted selection index) resulted in substantially greater G×E variance compared to either of the component traits. The interaction variance of an index trait is shown to be a function of the index coefficients and the G×E variances and covariances for its constituent traits. As a result, for some conditions it surpasses the magnitude of G×E variance for each component trait.

Rootstock effects in grafted conifers: A review

The literature on rootstock effects (on scions) in conifers was reviewed, specifically: graft success, compatibility, size, reproduction, phenology, crown and needle characters, mineral contents, organic compounds, water relations, disease resistance and wood properties. Scions usually had higher graft success and less incompatibility on more closely related rootstocks although there were exceptions. Even intergeneric grafts have succeeded on occasion. Although there were marked rootstock effects on growth and reproduction, the effects did not follow a pattern with increasing relationship. It is also likely that some crown characters and the nutrient content of scions can be manipulated by the use of rootstocks. For many characters, a specific rootstock may give a desired result only for a limited number of scion types (species, cultivars or clones). With some exceptions, the review shows that the subject has not been comprehensively studied. Many of the studies were either short-term, inadequately replicated, or poorly designed to allow firm conclusions about rootstock effects. The physiological and biochemical mechanisms, which cause the changes seen in morphology, are not well understood. Further research and more comprehensive study of rootstock effects on scion biology are recommended.

Genetic Parameter Estimates for Growth Traits from Diallel Tests of Loblolly Pine Throughout the Southeastern United States

Abstract Variation in heritability and in genetic correlation estimates were evaluated for juvenile tree height and volume for six testing areas of loblolly pine (Pinus taeda L.) in the southeastern United States. Variance components and their functions (heritability and type B genetic correlations) were estimated from 265 six-parent disconnected diallel series, tested in almost 1000 trials (4 tests per diallel series). Original data were collected at age 6 years from about one million trees (265 diallel series x 30 crosses x 36 trees per cross/site x 4 sites) planted in field tests. Genetic tests were from the second cycle of breeding in the North Carolina State University - Industry Cooperative Tree Improvement Program. The overall unbiased individual-tree narrow-sense heritability for height was 0.19 and for volume was 0.16. The broad-sense heritabilities for height (0.24) and for volume (0.22) were higher than narrow-sense heritabilities due to the presence of non-additive genetic variance. There were moderate regional differences in these estimates, with tests in the Lower Gulf Coastal Plain tending to have the highest heritabilities for growth traits. There was very little association between site index and heritability, but heritabilities were higher on sites with the highest survival and highest test precision. Genotype x environment interactions were generally low both for half-sib and full-sib families, indicating that families can be operationally deployed to different sites with little concern about unpredictable performance.

PULPING AND BLEACHING OF PARTIALLY CAD-DEFICIENT WOOD

ABSTRACT Mutant loblolly pine trees that are partially deficient in cinnamyl alcohol dehydrogenase (CAD) have been studied as a possible new source of pulpwood. Young (4- and 6-year-old) partially CAD-deficient pine trees are ˜20% more easily delignified (pulping and bleaching) and provide similar pulp yields to that of similarly aged normal pines grown on the same plots. Bleached pulp from a 6-year-old partially CAD-deficient pine tree displayed better strength properties than the same age normal pine tree; this probably reflects the milder pulping conditions needed in the case of the partially CAD-deficient tree. Studies also were conducted on a limited number of 14-year-old trees from a different genetic background. In contrast to the results with young trees, no real differences in ease of delignification, pulp yields, bleached pulp strength properties, and wood specific gravities were observed with the 14-year-old trees. There would likely be no penalty if partially CAD-deficient trees were used for lumber products. The rapid growth of partially CAD-deficient trees could make them a valuable pulpwood.

Genetic effects on total phenolics, condensed tannins and non- structural carbohydrates in loblolly pine (Pinus taeda L.) needles

Carbon allocation to soluble phenolics (total phenolics, proanthocyanidins (PA)) and total non-structural carbohydrates (TNC; starch and soluble sugars) in needles of widely planted, highly productive loblolly pine (Pinus taeda L.) genotypes could impact stand resistance to herbivory, and biogeochemical cycling in the southeastern USA. However, genetic and growth-related effects on loblolly pine needle chemistry are not well characterized. Therefore, we investigated genetic and growth-related effects on foliar concentrations of total phenolics, PA and TNC in two different field studies. The first study contained nine different genotypes representing a range of genetic homogeneity, growing in a 2-year-old plantation on the coastal plain of North Carolina (NC), USA. The second study contained eight clones with different growth potentials planted in a 9-year-old clonal trial replicated at two sites (Georgia (GA) and South Carolina (SC), USA). In the first study (NC), we found no genetic effects on total phenolics, PA and TNC, and there was no relationship between genotype size and foliar biochemistry. In the second study, there were no differences in height growth between sites, but the SC site showed greater diameter (diameter at breast height (DBH)) and volume, most likely due to greater tree mortality (lower stocking) which reduced competition for resources and increased growth of remaining trees. We found a significant site × clone effect for total phenolics with lower productivity clones showing 27-30% higher total phenolic concentrations at the GA site where DBH and volume were lower. In contrast to the predictions of growth-defense theory, clone volume was positively associated with total phenolic concentrations at the higher volume SC site, and PA concentrations at the lower volume GA site. Overall, we found no evidence of a trade-off between genotype size and defense, and genetic potential for improved growth may include increased allocation to some secondary metabolites. These results imply that deployment of more productive loblolly pine genotypes will not reduce stand resistance to herbivory, but increased production of total phenolics and PA associated with higher genotype growth potential could reduce litter decomposition rates and therefore, nutrient availability.