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Characterisation of Pt MYB1, an R2R3-MYB from pine xylem.

A cDNA encoding a member of the R2R3-MYB family of transcription factors was cloned from a library constructed from differentiating Pinus taeda (loblolly pine) xylem RNA. This MYB family member, Pinus taeda MYB1 (PtMYB1), was most abundantly expressed in differentiating xylem, as assessed by both ribonuclease protection assays, and by northern blot analysis with poly(A)-enriched RNA. Similar to other plant R2R3-MYB family members, recombinant Pt MYB1 protein was able to bind to AC elements in electrophoretic mobility shift assays (EMSAs). AC elements are DNA motifs rich in adenosine and cytosine that have been implicated in the xylem-localised regulation of genes encoding lignin biosynthetic enzymes. Pt MYB1 not only bound to AC elements, but was also able to induce AC-element-dependent shifts in the electrophoretic mobility of a plant promoter that contains three AC elements, the minimal PHENYLALANINE AMMONIA-LYASE 2 (PAL2) promoter from Phaseolus vulgaris. Transcriptional activation assays conducted using yeast showed that Pt MYB1 also activated transcription, and that it did so in an AC-element-dependent fashion. Pt MYB1 also activated transcription from the minimal PAL2 promoter in plant cells in an AC-element-dependent fashion, as revealed by transient transcriptional activation assays with microprojectile-bombarded tobacco NT-1 cells. Taken together, these finding are consistent with the hypothesis that Pt MYB1 may regulate transcription from cis -acting AC elements in pine xylem.

Adenosine Triphosphate↗

Volatiles associated with preferred and nonpreferred hosts of the Nantucket pine tip moth, Rhyacionia frustrana.

Ovipositing female Nantucket pine tip moth, Rhyacionia frustrana, prefer loblolly pine, Pinus taeda L., to slash pine, Pinus elliottii Engelm, except during the first spring following planting of seedlings. Host discrimination by R. frustrana increases as seedlings develop, suggesting that changes in the chemical composition of seedlings may mediate the moth's host preferences. Volatile compounds from slash and loblolly pine seedlings were collected using solid-phase microextraction (SPME) during the first year following planting. Four collection periods coincided with adult emergence and oviposition during each of four annual generations of R. frustrana in the Georgia Coastal Plain. Infestation of slash pine peaked during the second tip moth generation and was similar to the loblolly pine infestation level. By the fourth tip moth generation, slash pine infestation levels had declined and diverged considerably from those of loblolly pine. Significant differences in relative quantities of beta-pinene, alpha-phellandrene, limonene, beta-phellandrene, bornyl acetate, beta-caryophyllene, and an unidentified sesquiterpene occurred between slash and loblolly pine during the fourth generation. However, no strong correlation was observed between any individual compound and host damage that could readily explain the temporal changes in R. frustrana host preference. Gas chromatographic-electroantennographic detection (GC-EAD) analyses of standards identified 19 different seedling-associated compounds that elicited antennal responses from R. frustrana females, indicating that a blend of terpenoids may mediate host discrimination.

Animals↗

Growth and photosynthetic responses of four Virginia Piedmont tree species to shade.

To determine the effects of shade on biomass, carbon allocation patterns and photosynthetic response, seedlings of loblolly pine (Pinus taeda L.), white pine (Pinus strobus L.), red maple (Acer rubrum L.), and yellow-poplar (Liriodendron tulipifera L.) were grown without shade or in shade treatments providing a 79 or 89% reduction of full sunlight for two growing seasons. The shade treatments resulted in less total biomass for all species, with loblolly pine showing the greatest shade-induced growth reduction. Yellow-poplar was the only species to show increased stem height growth in the 89% shade treatment. The shade treatments increased specific leaf area of all species. Quantum efficiency, dark respiration and light compensation point were generally not affected by the shade treatments. Quantum efficiency, dark respiration, maximum photosynthesis and light compensation point did not change consistently between the first and second growing seasons. We conclude that differences in shade tolerance among these species are not the result of changes in the photosynthetic mechanism in response to shade.

Journal Article↗

Paternal chloroplast inheritance patterns in pine hybrids detected with trnL-trnF intergenic region polymorphism.

The inheritance patterns of the chloroplast genomes of shortleaf pine ( Pinus echinata Mill.), loblolly pine ( Pinus taeda L.) and slash pine ( Pinus elliottii Engelm.) were investigated through the trnL-trnF intergenic spacer polymorphism analysis. The DNA sequences of this spacer differ among these three closely related Pinus species. A modified 'cold' PCR-SSCP (single-strand conformation polymorphism) analysis of this spacer shows that the artificial hybrids (F1) from the shortleaf pine (seed parent) x loblolly pine (pollen parent) cross, exhibit the loblolly pine profile. Additionally, nine putative hybrids between shortleaf pine and loblolly pine, previously identified by the IDH (Isocitrate dehydrogenase) allozyme marker, presented the shortleaf pine profile indicating that shortleaf pine, not loblolly pine, sired all of the putative hybrids. Nondenatured polyacrylamide-gel electrophoresis of the trnL-trnF intergenic spacer demonstrated that the artificial hybrids (F1) from the cross, slash pine (seed parent) x shortleaf pine (pollen parent), present the shortleaf pine profile. Those results confirmed that the chloroplast genome is paternally inherited in these three species of the genus Pinus. The significance of the trnL-trnF intergenic region polymorphism and our modified 'cold' SSCP protocol for population genetic studies is discussed.

Journal Article↗

Aluminum sensitivity of loblolly pine and slash pine seedlings grown in solution culture.

To probe variation in Al sensitivity of two co-occurring pine species, seedlings from six full-sib families of loblolly pine (Pinus taeda L.) and slash pine (Pinus elliottii Engelm.) were grown in solution culture containing 4.4 mM (high-Al) or 0.01 mM (low-Al) AlCl(3) at pH 4 for 58 days. On average, both pine species had 41% less total dry weight in the high-Al treatment than in the low-Al treatment. Stem volume growth of slash pine was more sensitive to the high-Al treatment than that of loblolly pine. In both species, the high-Al treatment inhibited root dry weight more than shoot dry weight. Within-species variation in Al sensitivity among families was greater in loblolly pine (24 to 52% inhibition of seedling dry weight) than in slash pine (35 to 47% inhibition of seedling dry weight). Foliar Al concentration was positively correlated with Al sensitivity in slash pine but not in loblolly pine; however, in both species, the concentration of Al in roots was 20-fold greater than in foliage.

Journal Article↗

Effects of CO(2) enrichment on growth and root (15)NH(4) (+) uptake rate of loblolly pine and ponderosa pine seedlings.

We examined changes in root growth and (15)NH(4) (+) uptake capacity of loblolly pine (Pinus taeda L.) and ponderosa pine (Pinus ponderosa Douglas. Ex Laws.) seedlings that were grown in pots in a phytotron at CO(2) partial pressures of 35 or 70 Pa with NH(4) (+) as the sole N source. Kinetics of (15)N-labeled NH(4) (+) uptake were determined in excised roots, whereas total NH(4) (+) uptake and uptake rates were determined in intact root systems following a 48-h labeling of intact seedlings with (15)N. In both species, the elevated CO(2) treatment caused a significant downregulation of (15)NH(4) (+) uptake capacity in excised roots as a result of a severe inhibition of the maximum rate of root (15)NH(4) (+) uptake (V(max)). Rates of (15)NH(4) (+) uptake in intact roots were, however, unaffected by CO(2) treatment and were on average 4- to 10-fold less than the V(max) in excised roots, suggesting that (15)NH(4) (+) absorption from the soil was not limited by the kinetics of root (15)NH(4) (+) uptake. Despite the lack of a CO(2) effect on intact root absorption rates, (15)NH(4) (+) uptake on a per plant basis was enhanced at high CO(2) concentrations in both species, with the relative increase being markedly higher in ponderosa pine than in loblolly pine. High CO(2) concentration increased total (15)NH(4) (+) uptake and the fraction of total biomass allocated to fine roots (< 2 mm in diameter) to a similar relative extent. We suggest that the increased uptake on a per plant basis in response to CO(2) enrichment is largely the result of a compensatory increase in root absorbing surfaces.

Journal Article↗

Integrating within-crown variation in net photosynthesis in loblolly and slash pine families.

We examined photosynthetic characteristics of two fast- and two slow-growing half-sib families of both loblolly pine (Pinus taeda L.) and slash pine (Pinus elliottii var. elliottii Engelm.) on two sites in northern Florida to: (1) quantify variation in light-saturated net photosynthesis (Amax) associated with vertical crown position and foliage age; (2) quantify the amount and distribution of leaf area by foliage age class; and (3) determine whether photosynthetic indices, ranging from leaf-level through whole-crown Amax, were related to growth differences among species and families. In both species, leaf-level Amax was higher in more recently formed foliage both within the same year (where Amax in the third flush averaged 10 to 30% higher than Amax in the first flush) and between years (where Amax in current-year foliage averaged 20 to 40% higher than Amax in 1-year-old foliage). When expressed on a leaf area basis, Amax of current-year foliage was higher in slash pine than in loblolly pine, but Amax expressed on a mass basis did not differ between species. Loblolly pine had higher whole-tree leaf area than slash pine, whereas whole-tree Amax did not differ between species. When the mean values for fast-growing families were compared with the mean values for slow-growing families, there were no differences in leaf-level characteristics, whereas at the whole-tree level, fast-growing families had higher leaf area and whole-tree Amax than slow-growing families in both species. When comparisons were made among the individual fast- and slow-growing families, however, results were more variable. In both species, stem volume growth was strongly correlated with whole-tree Amax, with most of the strength of the correlation deriving from the relationship between volume growth and tree leaf area.

Genetic Variation↗

Loblolly pine and slash pine responses to acute aluminum and acid exposures.

In response to concerns about aluminum and HCl exposure associated with rocket motor testing and launches, survival and growth of full-sib families of loblolly pine (Pinus taeda L.) and slash pine (Pinus elliottii Engelm.) were evaluated in a nursery bed experiment. Each species was exposed to a single soil application of aluminum chloride (0.33 M AlCl(3), pH 2.5), hydrochloric acid (0.39 M HCl, pH 0.6) or water, with or without mycorrhizal inoculation with Pisolithus tinctorius (Coker and Couch). After 20 weeks without inoculation, survival in AlCl(3) and HCl treatments averaged 52% for loblolly pine and 72% for slash pine. Inoculation improved survival of loblolly pine, receiving HCl from 49 to 73%, and of those receiving AlCl3, from 55 to 90%. Inoculation also resulted in improved survival and growth of individual families in AlCl(3), but not in HCl treatments. Results illustrate the relative resistance of both pine species to the acute treatments supplied, the improvement in resistance associated with mycorrhizal inoculation and the importance of field testing, following hydroponic screening, to verify the resistance to soil-supplied stresses.

Aluminum Chloride↗

Nutritional ecology of the Formosan subterranean termite (Isoptera: Rhinotermitidae): growth and survival of incipient colonies feeding on preferred wood species.

The wood of 11 plant species was evaluated as a food source significantly impacting the growth and survival of incipient colonies of the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Colonies of C. formosanus feeding on pecan, Carya illinoensis (Wangenh.), and red gum, Liquidambar styraciflua L., produced significantly more progeny than colonies feeding on other wood species tested. Progeny of colonies feeding on pecan and American ash, Fraxinus americana L., had significantly greater survival than progeny of colonies feeding on other wood species. Colonies feeding on a nutritionally supplemented cellulose based matrix showed similar fitness characteristics as colonies feeding on the best wood treatments. These results indicate that differences observed in colony fitness can be partially explained by nutritional value of the food treatment, raising the possibility that wood from different tree species have different nutritional values to the Formosan subterranean termites. Colonies feeding on loblolly pine, Pinus taeda L., and ponderosa pine, Pinus ponderosa Laws., had significantly lower survival and produced significantly fewer workers and soldiers than colonies feeding on other wood species. Colony survival from 90 to 180 d of age and from 90 to 360 d of age was significantly correlated with the number of workers present at 90 d of colony age, indicating that colony survival depends on the presence of workers. Wood consumption in a multiple-choice study was significantly correlated with colony fitness value. This suggests that feeding preference of C. formosanus is at least partially influenced by the nutritional value of the food source.

Animal Nutritional Physiological Phenomena↗

An ancient bottleneck in the Lost Pines of central Texas.

The retreating edge hypothesis for species responding to climate change predicts severe bottlenecks and eventual extinction. The disjunct Lost Pines population at the westernmost edge of the widespread Pinus taeda range is well suited for testing this prediction. The occurrence of one or more genetic bottlenecks in the Lost Pines population was tested using 34 nuclear microsatellite markers and a control sample from the larger, more continuous east Texas P. taeda forests. The Lost Pines population has undergone drastic contractions in effective population size between 3000 and 30 000 years bp. These results were supported by: (i) detection of transient heterozygosity excess, (ii) a mode-shift indicator of allele frequencies, and (iii) a ratio of allele number to allele size range. No bottleneck was detected for the east Texas control using any of the three methods. The distribution of allele frequencies was skewed for the Lost Pines population compared to the control, indicating a loss of rare alleles. However, allelic diversity was similar between the Lost Pines population and its east Texas control; the mean allele number per locus was 5.29 and 5.38, respectively. It is proposed that the Lost Pines population was the western refugium for P. taeda during Pleistocene glaciation and that East Texas P. taeda forests descended from the bottlenecked Lost Pines population.

Climate↗

No clustering for linkage map based on low-copy and undermethylated microsatellites.

Clustering has been reported for conifer genetic maps based on hypomethylated or low-copy molecular markers, resulting in uneven marker distribution. To test this, a framework genetic map was constructed from three types of microsatellites: low-copy, undermethylated, and genomic. These Pinus taeda L. microsatellites were mapped using a three-generation pedigree with 118 progeny. The microsatellites were highly informative; of the 32 markers in intercross configuration, 29 were segregating for three or four alleles in the progeny. The sex-averaged map placed 51 of the 95 markers in 15 linkage groups at LOD > 4.0. No clustering or uneven distribution across the genome was observed. The three types of P. taeda microsatellites were randomly dispersed within each linkage group. The 51 microsatellites covered a map distance of 795 cM, an average distance of 21.8 cM between markers, roughly half of the estimated total map length. The minimum and maximum distances between any two bins was 4.4 and 45.3 cM, respectively. These microsatellites provided anchor points for framework mapping for polymorphism in P. taeda and other closely related hard pines.

Chromosome Mapping↗

A survival model for unthinned loblolly pine plantations that incorporates non-planted tree competition, site quality, and incidence of fusiform rust.

Future biomass yields are functionally related to the number of trees surviving at a given age. A stand level survival model was developed that incorporates competition of non-planted trees, site quality, and the incidence of fusiform rust (Cronartium quercuum [Berk.] Miyabe ex Shirai f. sp. fusiforme). The model consists of a system of two equations, one of which represents the number of surviving trees infected by fusiform rust while the other represents the number of trees not infected by fusiform rust. Data from unthinned loblolly pine (Pinus taeda L.) plantations in East Texas were used to fit and evaluate the survival model and illustrate its use. The model successfully predicted that the number of surviving loblolly pine trees decreased as the number of non-planted trees increased. The model also successfully predicted the transition of loblolly pine trees from an uninfected to an infected status by fusiform rust.

Basidiomycota↗

Nucleotide diversity and linkage disequilibrium in loblolly pine.

Outbreeding species with large, stable population sizes, such as widely distributed conifers, are expected to harbor relatively more DNA sequence polymorphism. Under the neutral theory of molecular evolution, the expected heterozygosity is a function of the product 4N(e)mu, where N(e) is the effective population size and mu is the per-generation mutation rate, and the genomic scale of linkage disequilibrium is determined by 4N(e)r, where r is the per-generation recombination rate between adjacent sites. These parameters were estimated in the long-lived, outcrossing gymnosperm loblolly pine (Pinus taeda L.) from a survey of single nucleotide polymorphisms across approximately 18 kb of DNA distributed among 19 loci from a common set of 32 haploid genomes. Estimates of 4N(e)mu at silent and nonsynonymous sites were 0.00658 and 0.00108, respectively, and both were statistically heterogeneous among loci. By Tajima's D statistic, the site frequency spectrum of no locus was observed to deviate from that predicted by neutral theory. Substantial recombination in the history of the sampled alleles was observed and linkage disequilibrium declined within several kilobases. The composite likelihood estimate of 4N(e)r based on all two-site sample configurations equaled 0.00175. When geological dating, an assumed generation time (25 years), and an estimated divergence from Pinus pinaster Ait. are used, the effective population size of loblolly pine should be 5.6 x 10(5). The emerging narrow range of estimated silent site heterozygosities (relative to the vast range of population sizes) for humans, Drosophila, maize, and pine parallels the paradox described earlier for allozyme polymorphism and challenges simple equilibrium models of molecular evolution.

Animals↗

Single-copy, species-transferable microsatellite markers developed from loblolly pine ESTs.

Microsatellites, or simple sequence repeats (SSRs), are usually regarded as the "markers of choice" in population genetics research because they exhibit high variability. The development cost of these markers is usually high. In addition, microsatellite primers developed for one species often do not cross-amplify in related species, requiring separate development for each species. However, microsatellites found in expressed sequence tags (ESTs) might better cross-amplify as they reside in or near conserved coding DNA. In this study, we identified 14 Pinus taeda (loblolly pine) EST-SSRs from public EST databases and tested for their cross-species transferability to P. contorta ssp. latifolia, P. ponderosa, and P. sylvestris. As part of our development of a P. contorta microsatellite set, we also compared their transferability to that of 99 traditional microsatellite markers developed in P. taeda and tested on P. contorta ssp. latifolia. Compared to traditional microsatellites, EST-SSRs had higher transfer rates across pine species; however, the level of polymorphism of microsatellites derived from ESTs was lower. Sequence analyses revealed that the frequencies of insertions/deletions and base substitutions were lower in EST-SSRs than in other types of microsatellites, confirming that EST-SSRs are more conserved than traditional SSRs. Our results also provide a battery of 23 polymorphic, robust microsatellite primer pairs for lodgepole pine.

Alleles↗

Interactions of Hylastes species (Coleoptera: Scolytidae) with Leptographium species associated with loblolly pine decline.

Hylastes spp. (Coleoptera: Scolytidae) were evaluated as potential vectors of Leptographium spp. fungi. Bark beetles were trapped from stands ofloblolly pine, Pinus taeda L., exhibiting a range of decline symptoms in central Alabama. Under controlled conditions, field-collected adult Hylastes salebrosus Eichoff (Coleoptera: Scolytidae) and Hylastes tenuis Eichoff (Coleoptera: Scolytidae), which had been surface-sterilized and inoculated with Leptographium terebrantis Barras & Perry and Leptographium serpens (Goid.) Wingfield, transmitted the fungi into 100% of wounded and unwounded loblolly root sections with which they were confined. None of the sterilized and uninoculated beetles transmitted any Leptographium spp. to roots. Significantly more H. salebrosus and H. tenuis brood emerged from roots infected with Leptographium species than from sterile roots, indicating an enhancement of Hylastes reproduction.

Animals↗

Assessing plant response to ambient ozone: growth of ozone-sensitive loblolly pine seedlings treated with ethylenediurea or sodium erythorbate.

One-year-old seedlings from an ozone-sensitive half-sib family of loblolly pine (Pinus taeda L.) were transplanted into replicated plots in blocks in a large forest clearing near Nacogdoches, Texas. Seedlings were either non-treated (controls) or treated bi-weekly with foliar sprays of ethylenediurea (EDU), at 150, 300 or 450 ppm or sodium erythorbate (NaE), at 515, 1030, or 1545 ppm, for three growing seasons. Results from the final third year harvest indicated that both EDU and NaE increased all growth parameters, with significant differences only for EDU at 450 ppm. Both EDU and NaE would be useful for long-term studies on assessing the effects of ambient ozone on established native plants.

Air Pollutants↗

Abnormal lignin in a loblolly pine mutant.

Novel lignin is formed in a mutant loblolly pine (Pinus taeda L.) severely depleted in cinnamyl alcohol dehydrogenase (E.C. 1.1.1.195), which converts coniferaldehyde to coniferyl alcohol, the primary lignin precursor in pines. Dihydroconiferyl alcohol, a monomer not normally associated with the lignin biosynthetic pathway, is the major component of the mutant's lignin, accounting for approximately 30 percent (versus approximately 3 percent in normal pine) of the units. The level of aldehydes, including new 2-methoxybenzaldehydes, is also increased. The mutant pines grew normally indicating that, even within a species, extensive variations in lignin composition need not disrupt the essential functions of lignin.

Alcohol Oxidoreductases↗