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Joint effects of elevated levels of ultraviolet-B radiation, carbon dioxide and ozone on plants.

There is growing interest regarding the joint effects of elevated levels of surface ultraviolet B (UV-B) radiation, carbon dioxide (CO2) and ozone (O3) on plants. Our current knowledge of this subject is too limited to draw any specific conclusions, although one might state that such effects are likely to be highly species dependent and may be more than additive, additive or less than additive. There are a number of uncertainties associated with the experimental protocols used and the conclusions reached in many studies. Nevertheless, in North America, there appear to be genotypes of three monocot crop species (Avena sativa L., Oryza sativa L. and Sorghum vulgare L.); six dicot crops (Cucumis sativus L., Lactuca sativa L., Lycopersicon esculentum Mill., Phaseolus vulgaris L., Pisum sativum L. and Solanum tuberosum L.) and two conifer species (Pinus ponderosa Dougl. and Pinus taeda L.) that may be considered sensitive to the joint effects of elevated levels of UV-B, CO2 and O3. However, to provide a more reliable assessment or validation of the predictions, future research must consider the concept of plant response surfaces and describe them more fully in numerical terms. Achieving that objective will require close cooperation among a number of scientists representing geographic locations with known spatial and temporal differences in UV-B, CO2 and O3 to conduct experiments under their site-specific conditions, using common plant materials and experimental protocols.

Acclimatization↗

Deladenus siricidicola, bedding (Neotylenchidae) parasitism evaluation in adult Sirex noctilio, Fabricius, 1793 (Hymenoptera: Siricidae).

This work aimed to evaluate the Deladenus siricidicola, Bedding (Neotylenchidae) parasitism in adults Sirex noctilio, Fabricius, 1793 (Hymenoptera: Siricidae). Timber was sampled by cutting out pieces of 0.80 m in length in Sirex noctilio attacked and Deladenus siricidicola inoculated Pinus taeda. Longs were 15-20 cm in diameter, according to the tree age. Samples were packed in gauze-cages, for daily observations, till S. noctilio adults emergence. The emerged insects were transported, in plastic containers to the laboratory, where they were sectioned and dissected under stereoscopic microscopy to observ the nematode occurrence. From the initially proposed ten units, nine of them were evaluated in a total of 1,810 emerged adult insects, being 1,441 males and 369 females. Nematode parasitism was shown in 267 males and 74 females, in a total of 341 infected insects (18.84%).

Analysis of Variance↗

Efficacy tests and determination of optimal spray timing values to control nantucket pine tip moth (Lepidoptera: Tortricidae) infestations.

The Nantucket pine tip moth, Rhyaciona frustrana (Comstock), a common regeneration pest of loblolly pine, Pinus taeda L., has been shown to reduce tree volume yields through larval feeding. Chemical applications can be effective in protecting trees from the growth losses associated with this feeding, and optimum spray timing values are commonly used to reduce the number of necessary applications and to increase insecticide efficacy. Optimal spray timing values for the Georgia Piedmont were obtained for the following four insecticides available for use in loblolly pine plantations: permethrin (Pounce), lambda-cyhalothrin (Warrior T), spinosad (SpinTor 2 SC), and Bacillus thuringiensis variety kurstaki Berliner (Foray 48B). Optimal timing values were similar between the first and second generations for each of these compounds. All of the insecticides used in this study significantly reduced tip moth damage below the control treatment levels. Lambda-cyhalothrin was the most efficacious and had the longest spray timing window. B. thuringiensis was the least effective and had the shortest timing window. Spinosad and permethrin were similar in efficacy and spray timing values. This information is applicable to regions where there are three tip moth generations per year, as found in the southern Piedmont region and the coastal plain of Virginia and most of North Carolina.

Animals↗

Bait matrix for delivery of chitin synthesis inhibitors to the formosan subterranean termite (Isoptera: Rhinotermitidae).

The efficacy of three chitin synthesis inhibitors, diflubenzuron, hexaflumuron, and chlorfluazuron, incorporated into a novel bait matrix to kill the Formosan subterranean termite, Coptotermes formosanus Shiraki, was evaluated in the laboratory. The bait matrix was significantly preferred by C. formosanus over southern yellow pine wood in a two-choice feeding test. Bait formulations containing 250 ppm of the three chitin synthesis inhibitors were presented to termite nests with 2,500 individuals (80% workers and 20% soldiers) in the presence of alternative food sources consisting of cardboard and southern yellow pine, Pinus taeda L., wood. None of the bait formulations were significantly repellent or feeding deterrent to the termite workers evidenced by the lack of full consumption of alternative food sources. All nests presented with the bait formulations died within 9 wk, whereas the control nests (bait with no chitin synthesis inhibitors) remained alive 6 mo after the end of the study. No significant differences in consumption were observed among the chitin synthesis inhibitor treatments. Importance of this study for the improvement of current bait technology is discussed.

Animals↗

Nutritional ecology of the formosan subterranean termite (Isoptera: Rhinotermitidae): feeding response to commercial wood species.

The feeding preferences of the Formosan subterranean termite, Coptotermes formosanus Shiraki, were tested in three separate experiments on 28 different wood species. Experiment 1 was a multiple-choice test designed to test relative preferences among 24 wood species commercially available in New Orleans, LA. Experiment 2 was a similar study designed to test relative preferences among 21 wood species shown or reported to be unpalatable to the Formosan subterranean termite. Experiment 3 was a no-choice test to examine the feeding deterrence of the 10 least preferred wood species. Preference was determined by consumption rates. Birch (Betula alleghaniensis Britton), red gum (Liquidambar styraciflua L.), Parana pine [Araucaria angustifolia (Bert.) 1, sugar maple (Acer saccharum Marsh.), pecan (Carya illinoensis Wangenh.), and northern red oak (Quercus rubra L.) were the most preferred species by C. formosanus in order of consumption rate. All of these species were significantly more preferred than southern yellow pine (Pinus taeda L.), widely used for monitoring. Sinker cypress [ = old growth bald cypress, Taxodium distichum (L.)], western red cedar (Thuja plicata Donn), Alaskan yellow cedar (Chamaecyparis nootkatensis D. Don), eastern red cedar (Juniperus virginiana L.), sassafras [Sassafras albidum (Nutt.)], Spanish cedar (Cedrella odorata L.), Honduras mahogany (Swietenia macrophyla King), Indian rosewood (Dalbergia latifolia Roxb.), Honduras rosewood (D. stevensonii Standl.), and morado (Machaerium sp.) induced significant feeding deterrence and mortality to C. formosanus. The last eight species produced 100% mortality after 3 mo.

Animals↗

Use of haploid mixtures and heteroduplex analysis enhance polymorphisms revealed by denaturing gradient gel electrophoresis.

PCR-based codominant genetic markers were developed by using primer sequences designed from cDNA clones of loblolly pine (Pinus taeda L.). Such markers offer certain advantages relative to simple-sequence repeat (SSR), also known as short-tandem repeat (STR) markers, and include the ability to quantify and map DNA polymorphisms in expressed genes. However, detecting these DNA polymorphisms is more problematic because many DNA polymorphisms in genes involve base substitutions rather than insertions or deletions. Denaturing gradient gel electrophoresis (DGGE) is a sensitive and efficient method for detecting sequence differences among PCR fragments. This paper demonstrates the application of DGGE to genetically map expressed genes in loblolly pine. Also, heteroduplex DNA fragments, formed during the amplification of DNA from heterozygotes and from mixes of haploid DNAs from megagametophytes, enhanced and strengthened genetic interpretations and genotypic classifications.

Alleles↗

Differential staining of tannin in sections of epoxy-embedded plant cells.

A staining procedure is described for the light microscopic localization of ergastic tannins in epoxy sections of plant cells embedded for study by transmission electron microscopy. Callus and cell suspensions of Pseudotsuga menziesii and Pinus taeda fixed in glutaraldehyde:acrolein and then OsO4, followed by epoxy embedding, were sectioned 0.5 mum thick, stained on a glass slide with ethanolic Sudan black B at 60 C as described by Bronner, and then mounted in Karo syrup. Tannin deposits stained brownish-orange and were easily distinguished from lipid bodies of similar size, which stained dark blue to black, and from starch grains, which were unstained. The significance of this differential polychromasia was confirmed by transmission electron microscopy. This staining procedure should prove valuable in the cytoplasmic evaluation of the plant cell ergastics (especially tannins) via light microscopy whether or not electroc microscopic examination is intended.

Plant Cells↗

Effects of nutrition on age determination using eye lens weights.

Populations of hispid cotton rats (Sigmodon hispidus) were studied in 6 young loblolly pine (Pinus taeda) plantations in South Carolina, USA. Each plantation had 2, 1-ha, live-trapping grids; 1 to sample a Sigmodon population which continually received supplemental food, the other to sample a control population. After 15 months of live-trapping, the Sigmodon were killed and their eye lenses removed, dried and weighed. Birth dates were calculated for 425 Sigmodon which were first captured when young. Patterns of eye lens growth for food-supplemented and control animals were compared using the Richards' growth functions and were significantly different. For animals less than 165 days of age, the functions predicted greater eye lens weights for control animals than food-supplemented animals. The reverse was true for animals older than 165 days of age. Because of the influence of nutrition on eye lens growth patterns, the assumption that laboratory animals or known-age animals from one population can be used to estimate the ages of animals from another population is not always valid.

Age Factors↗

NADPH: protochlorophyllide oxidoreductases in white pine (Pinus strobus) and loblolly pine (P. taeda). Evidence for light and developmental regulation of expression and conservation in gene organization and protein structure between angiosperms and gymnosperms.

NADPH: protochlorophyllide oxidoreductase (pchlide reductase, EC 1.6.99.1) catalyzes the light-dependent reduction of protochlorophyllide in higher plants. Cloned cDNAs encoding two distinct pchlide reductases were isolated from a lambda gt11 library constructed from poly(A)+ RNA prepared from the cotyledons of dark-grown white pine (Pinus strobus) seedlings and a nuclear gene (lpcr) analogous to one of these cDNAs has been characterized from loblolly pine (P. taeda). The pine gene encodes an approximately 43 kDa precursor polypeptide consisting of a 334-amino acid mature protein and a 66-amino acid transit peptide. The deduced primary structures for the pine proteins are highly homologous to those reported from monocots and dicots. The coding portion of the pine lpcr gene is interrupted by four introns. The placement of these introns within the pine lpcr gene is identical to that observed in pea (Pisum sativum), suggesting conservation in gene organization between dicot and gymnosperm species. Western blot analysis using polyclonal antiserum against oat pchlide reductase detected in extracts of dark-grown pine cotyledons a single immunoreactive protein, which declined in abundance during a 48 h period of illumination with white light. Cotyledons of dark-grown seedlings were also found to accumulate high levels of pchlide reductase mRNA; however, little or no change in the steady-state levels of mRNA encoding pchlide reductase was observed in these tissues following illumination. Stem tissue of dark-grown seedlings did not contain significant levels of pchlide reductase mRNA, whereas stems of light-grown plants of the same age accumulated substantial amounts of the message. These results suggest that light and the developmental age of the tissue affect regulation of lpcr expression in pine.

Amino Acid Sequence↗

Major gene detection for fusiform rust resistance using Bayesian complex segregation analysis in loblolly pine.

The presence of major genes affecting rust resistance of loblolly pine was investigated in a progeny population that was generated with a half-diallel mating of six parents. A Bayesian complex segregation analysis was used to make inference about a mixed inheritance model (MIM) that included polygenic effects and a single major gene effect. Marginalizations were achieved by using Gibbs sampler. A parent block sampling by which genotypes of a parent and its offspring were sampled jointly was implemented to improve mixing. The MIM was compared with a pure polygenic model (PM) using Bayes factor. Results showed that the MIM was a better model to explain the inheritance of rust resistance than the pure PM in the diallel population. A large major gene variance component estimate (> 50% of total variance), indicated the existence of major genes for rust resistance in the studied loblolly pine population. Based on estimations of parental genotypes, it appears that there may be two or more major genes affecting disease phenotypes in this diallel population.

Basidiomycota↗

Improved somatic embryo maturation in loblolly pine by monitoring ABA-responsive gene expression.

During loblolly pine zygotic embryo development, increases in mRNAs for three ABA-responsive LEA-like genes coincided with the two developmental stage-specific peaks of endogenous ABA accumulation (Kapik et al. 1995). These ABA concentration profiles from zygotic embryo development were used to develop several tissue culture approaches that altered the exposure of somatic embryos to exogenous ABA. Elevating exogenous ABA at a time corresponding to mid-maturation improved the germination and resulted in more zygotic-like expression of selected genes in somatic embryos. Extending the time on maturation medium for a fourth month increased embryo yield, dry weight, and germination in high-and low-yield genotypes. Optimizing the amounts of embryogenic suspension, plated and exogenous ABA concentration increased from 22 to 66% in the early-stage bipolar embryos that developed to the cotyledonary stage.

Abscisic Acid↗

Genome histories clarify evolution of the expansin superfamily: new insights from the poplar genome and pine ESTs.

Expansins comprise a superfamily of plant cell wall-loosening proteins that has been divided into four distinct families, EXPA, EXPB, EXLA and EXLB. In a recent analysis of Arabidopsis thaliana and Oryza sativa expansins, we proposed a further subdivision of the families into 17 clades, representing independent lineages in the last common ancestor of monocots and eudicots. This division was based on both traditional sequence-based phylogenetic trees and on position-based trees, in which genomic locations and dated segmental duplications were used to reconstruct gene phylogeny. In this article we review recent work concerning the patterns of expansin evolution in angiosperms and include additional insights gained from the genome of a second eudicot species, Populus trichocarpa, which includes at least 36 expansin genes. All of the previously proposed monocot-eudicot orthologous groups, but no additional ones, are represented in this species. The results also confirm that all of these clades are truly independent lineages. Furthermore, we have used position-based phylogeny to clarify the history of clades EXPA-II and EXPA-IV. Most of the growth of the expansin superfamily in the poplar lineage is likely due to a recent polyploidy event. Finally, some monocot-eudicot clades are shown to have diverged before the separation of the angiosperm and gymnosperm lineages.

Arabidopsis↗

Wood formation from the base to the crown in Pinus radiata: gradients of tracheid wall thickness, wood density, radial growth rate and gene expression.

Wood formation was investigated at five heights along the bole for two unrelated trees of Pinus radiata. Both trees showed clear gradients in wood properties from the base to the crown. Cambial cells at the base of the tree were dividing 3.3-fold slower than those at the crown, while the average thickness of cell walls in wood was highest at the base. Cell wall thickness showed an overall correlation coefficient of >0.7 with wood density in both genotypes. Microscopic examination of developing tracheids showed that 33% of cells had formed secondary cell walls at the base of the tree, reducing to 3% at the crown. In total, 455 genes differentially expressed in developing xylem tissue from either the base or the crown were identified using modified differential display. RT-PCR analysis of 156 genes confirmed differential expression for 77%. Of the genes tested, 73% showed gradients in transcript abundance either up or down the bole of the tree, although the steepness of the gradients differed between genes. Genes involved in cell division and expansion tended to be more highly expressed in the crown of the tree, and two putative cell-cycle repressor genes were expressed 2-fold higher at the base. Conversely, transcripts of genes involved in secondary wall thickening were more abundant at the base of the tree. These results suggest that differences in the rate of cambial cell division, differences in the rate and duration of tracheid wall thickening, and differences in gene expression underpin the gradients of wood properties found in pines.

Cell Wall↗

Metabolic profiling: a new tool in the study of wood formation.

In the realm of plant genomics, metabolic profiling has become a valuable tool with which to assess the effect of genetic and/or environmental factors on plant development. This paper reports the first application of metabolic profiling on differentiating xylem tissue of loblolly pine. A protocol is presented for the analysis of loblolly pine xylem tissue. The effects of sample preparation, extraction, and derivatization on the corresponding metabolite profiles and yields have been investigated and are reported. Gas chromatography-mass spectroscopy has been used to quantify >60 polar and lipophilic metabolites from wood-forming tissue. It was possible to assign chemical structures to approximately half of these compounds. Comparison of six loblolly pine genotypes, three high cellulose (50-52%) and three medium (45-48%) cellulose, showed distinct metabolic profiles. Principal component analysis enabled the assignment of metabolic phenotypes using these large data sets. Metabolic phenotype clustering occurred in which the three high-cellulose genotypes were segregated from the medium-cellulose genotypes. These results demonstrate the use of metabolic profiling for the study of wood-forming tissue and as a tool in functional genomics.

Cellulose↗

Rapid prediction of solid wood lignin content using transmittance near-infrared spectroscopy.

A rapid transmittance near-infrared (NIR) spectroscopic method has been developed to characterize the lignin content of solid wood. Using simple, multiple regression, and partial least-squares statistical analysis the lignin contents of wood wafers, taken from increment cores, and synthetic wood, prepared by blending milled wood lignin and holocellulose, were compared and quantified. Strong correlations were obtained between the predicted NIR results and those obtained from traditional chemical methods. In addition to the experimental protocol and method development, NIR results from wood samples with different particle sizes and various lignin contents are discussed.

Lignin↗

Rapid screening of wood chemical component variations using transmittance near-infrared spectroscopy.

A rapid transmittance near-infrared (NIR) spectroscopy method was developed to predict the variation in chemical composition of solid wood. The effect of sample preparation, sample quantity (single versus stacked multiple wood wafers), and NIR acquisition time on the quantification of alpha-cellulose and lignin content was investigated. Strong correlations were obtained between laboratory wet chemistry values and the NIR-predicted values. In addition to the experimental protocol and method development, improvements in calibration error associated with utilizing stacked multiple wood wafers as opposed to single wood wafers are also discussed.

Cellulose↗

Differences between lignin in unprocessed wood, milled wood, mutant wood, and extracted lignin detected by 13C solid-state NMR.

Solid-state 13C nuclear magnetic resonance (NMR) spectroscopy was applied to intact and isolated loblolly pine wood samples to identify potential structural changes induced by tree age, milling, lignin extraction, or naturally occurring mutations. Special attention was paid to ketone and aldehyde as well as nonpolar alkyl groups, which could be observed at low concentrations (<2 in 1000 C) using improved spinning-sideband suppression with gated decoupling. Carbonyl structures were present in intact wood, and there are more keto groups than aldehydes. Their concentrations increased from juvenile to mature wood and with milling time, whereas extraction did not alter the C=O fraction. Significant amounts of aldehyde and dihydroconiferyl alcohol residues were present in coniferyl aldehyde dehydrogenase-deficient wood, confirming solution-state NMR spectra of the corresponding lignin. These results demonstrate the utility of solid-state NMR as an assay for changes in the lignin structure of genetically modified plants.

Alcohols↗

Relationships between net photosynthesis and foliar nitrogen concentrations in a loblolly pine forest ecosystem grown in elevated atmospheric carbon dioxide.

We examined the effects of elevated carbon dioxide concentration ([CO2]) on the relationship between light-saturated net photosynthesis (A(sat)) and area-based foliar nitrogen (N) concentration (N(a)) in the canopy of the Duke Forest FACE experiment. Measurements of A(sat) and N(a) were made on two tree species growing in the forest overstory and four tree species growing in the forest understory, in ambient and elevated [CO2] FACE rings, during early and late summer of 1999, 2001 and 2002, corresponding to years three, five and six of CO2 treatment. When measured at the growth [CO2], net photosynthetic rates of each species examined in the forest overstory and understory were stimulated by elevated [CO2] at each measurement date. We found no effect of elevated [CO2] on N(a) in any of the species. The slope of the A(sat)-N relationship was 81% greater in elevated [CO2] than in ambient [CO2] when averaged across all sample dates, reflecting a differential CO2 effect on photosynthesis at the top and bottom of the canopy. We compared A(sat)-N relationships in trees grown in ambient and elevated [CO2] at two common CO2 concentrations, during late summer 2001 and both early and late 2002, to determine if the stimulatory effect of elevated [CO2] on photosynthesis diminishes over time. At all three sample times, neither the slopes nor the y-intercepts of the A(sat)-N relationships of trees grown in ambient or elevated [CO2] differed when measured at common CO2 concentrations, indicating that the responses of photosynthesis to long-term elevated [CO2] did not differ from the responses to a short-term increase in [CO2]. This finding, together with the observation that N(a) was unaffected by growth in elevated [CO2], indicates that these overstory and understory trees growing at the Duke Forest FACE experiment continue to show a strong stimulation of photosynthesis by elevated [CO2].

Atmosphere↗