Physiological attributes of three- and four-needle fascicles of loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.)

Key message

Fascicle types differed morphologically but had similar photosynthetic capacity on a surface area basis.

Abstract

In Pinus species, fascicles can develop with a different number of needles than what is typical. For example, Pinus taeda fascicles typically have three needles, but sometimes have two or four. Although differing fascicle morphology could be a response to changes in the environment designed to optimize carbon gain or minimize water loss, we are unaware of any work comparing physiological differences between fascicles with different numbers of needles. We compared the physiological and morphological characteristics of three- and four-needle fascicles of a loblolly pine clone with an abnormally high abundance of four-needle fascicles to better understand whether differences in needle morphology affected photosynthetic capacity or transpiration. Three- and four-needle fascicles had equal length, diameter, and volume, but four-needle fascicles had significantly greater surface area, mass, and tissue density. Equal fascicle total volume resulted in smaller per-needle volume in four-needle fascicles compared to three-needle fascicles. On a unit surface area basis, light-saturated net assimilation, stomatal conductance and transpiration were similar between the three- and four-needle fascicles although the maximum rate of carboxylation was significantly greater in four-needle fascicles. On a per-fascicle basis, four-needle fascicles had greater transpiration, stomatal conductance, and maximum rate of light-saturated net assimilation. Our results suggest that several factors, including increased tissue density and stomatal density, offset the reduction in needle volume in four-needle fascicles, resulting in similar levels of gas exchange per unit surface area in three- and four-needle fascicles.

     

Microsatellite versus AFLP analyses of pre-management introgression levels in loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.) and shortleaf pine (<Emphasis Type="Italic">P. echinata</Emphasis> Mill.)

Loblolly pine and shortleaf pine are known to naturally hybridize. In this study, we used 42 microsatellite markers and isocitrate dehydrogenase isozyme to create genetic profiles of 202 loblolly and shortleaf pine trees grown from seed collected in the 1950s for the Southwide Southern Pine Seed Source Study. Estimated Φ_PT was low in both loblolly (0.061) and shortleaf (0.080) pines, indicating that most of the diversity is accounted for within seed sources. However, both loblolly and shortleaf pines showed significant correlations between seed sources’ genetic and geographic distances, with R ² of 0.43 and 0.17, respectively. The hybridization rate was 4.0%, with more hybrids west of the Mississippi River (8.1%) than east of the river (2.1%). Additionally, about the same proportion of both species (4.5% of loblolly and 3.3% of shortleaf pine) were identified as hybrids. These results are consistent with prior studies on these two species but do contrast with the results from an amplified fragment length polymorphism (AFLP) analysis of the same samples. For example, the AFLP study concluded that 6.3% of the trees were hybrids, or 1.4 times higher than determined by this study. Of the 12 hybrids identified in the AFLP study, six were not identified as hybrids here, and of the eight hybrids identified here, only four were identified in the AFLP study. Although similar in overall results, we suggest that the microsatellite analysis is more convincing than the AFLP analysis because microsatellites provide more information per genetic locus than do AFLPs.     

Genetic control of growth and shoot phenology in juvenile loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.) clonal trials

Southern pine genetic improvement programs have selected for faster early growth which has often increased yields over unimproved material, and some of this improvement is likely attributable to variation in growth phenology among genotypes. However, the genetics of shoot growth phenology traits are not well characterized. Loblolly pine cuttings and seedlings from parents originating in the Atlantic coastal plain (ACP) and Florida and grown on sites established in Palatka, FL and Cuthbert, GA were assessed for shoot phenology and growth traits during the second year and for growth in year 6. Individual-tree clonal repeatability in different growth and shoot phenology traits varied from 0.09 to 0.79 in cuttings, and was lower in Palatka than Cuthbert. Non-additive components of heritability were lower, with a few exceptions, than additive effects. Additive and genotypic correlations across sites were high (>0.6) for all traits measured in cuttings and for most seedling traits, suggesting low genotype × environment interactions between these two sites. Compared with progeny from crosses between ACP parents, progeny of Florida parents started growth earlier in the season and ended later. Strong genetic correlations were observed between second-year phenology traits and sixth-year height and diameter. This suggests some two-year traits could be useful for early selection of high-performing genotypes.     

Association genetics of growth and adaptive traits in loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.) using whole-exome-discovered polymorphisms

In the USA, forest genetics research began over 100 years ago and loblolly pine breeding programs were established in the 1950s. However, the genetics underlying complex traits of loblolly pine remains to be discovered. To address this, adaptive and growth traits were measured and analyzed in a clonally tested loblolly pine (Pinus taeda L.) population. Over 2.8 million single nucleotide polymorphism (SNP) markers detected from exome sequencing were used to test for single-locus associations, SNP-SNP interactions, and correlation of individual heterozygosity with phenotypic traits. A total of 36 SNP-trait associations were found for specific leaf area (5 SNPs), branch angle (2), crown width (3), stem diameter (4), total height (9), carbon isotope discrimination (4), nitrogen concentration (2), and pitch canker resistance traits (7). Eleven SNP-SNP interactions were found to be associated with branch angle (1 SNP-SNP interaction), crown width (2), total height (2), carbon isotope discrimination (2), nitrogen concentration (1), and pitch canker resistance (3). Non-additive effects imposed by dominance and epistasis account for a large fraction of the genetic variance for the quantitative traits. Genes that contain the identified SNPs have a wide spectrum of functions. Individual heterozygosity positively correlated with water use efficiency and nitrogen concentration. In conclusion, multiple effects identified in this study influence the performance of loblolly pines, provide resources for understanding the genetic control of complex traits, and have potential value for assisting breeding through marker-assisted selection and genomic selection.     

Somatic embryogenesis in loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.): improving culture initiation with abscisic acid and silver nitrate

Loblolly pine (Pinus taeda L.) culture initiation was improved by the addition of abscisic acid (ABA) (3.7 µM), silver nitrate (20 µM), and guanosine 3,5-cyclic monophosphate, 8-bromo-, sodium salt (10 µM) to the medium and by raising cytokinin levels in the presence of 50 mg/l activated carbon (AC). Basal medium contained modified 1/2-P6 salts, 50 mg/l AC, Cu and Zn added to compensate for adsorption by AC, 1.5% maltose, 2% myo-inositol, 500 mg/l casamino acids, 450 mg/l glutamine, 2 mg/l -naphthaleneacetic acid (NAA), 0.55 mg/l 6-benzylaminopurine (BA), 0.53 mg/l kinetin, and 2 g/l Gelrite. Across 32 open-pollinated families initiation ranged from 0 to 53.4%, with an average of 17.9%. Further optimization of cytokinins to 0.63 mg/l BA and 0.61 mg/l kinetin along with the removal of ABA maintained initiation at 18.2% across 19 families. Survival of 2001 new initiations was tracked for 4–6 months. Survival averaged 28.8%. A test of 68 new initiations tracked closely for 4 months demonstrated that at least 80% of the cultures lost did not grow after transfer to the multiplication media, suggesting that many new initiations abort during the initiation process.Abbreviations ABA Abscisic acid - AC Activated carbon - BA 6-Benzylaminopurine - 8-Br-cGMP Guanosine 3,5-cyclic monophosphate, 8-bromo-, sodium salt - NAA -Naphthaleneacetic acid Communicated by G.C. Phillips     

Characterization of the tissue-specific expression of phenylalanine ammonia-lyase gene promoter from loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis>) in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis>

We isolated the 5′ flanking region of a gene for phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) from Pinus taeda, PtaPAL. To investigate the tissue-specific expression of the PtaPAL promoter, histochemical assay of GUS activity was performed using the transgenic tobacco expressing the PtaPAL promoter-GUS. The region of −897 to −420 in PtaPAL promoter showed high activities in the secondary xylem and response to bending stress. To characterize the cis-regulatory functions of the promoters for enzymes in phenylpropanoid biosynthesis, we examined the activity of chimeric promoters of PtaPAL and a 4-coumarate CoA ligase, Pta4CLα. The chimeric promoter showed similar activity as the Pta4CLα promoter. Electrophoretic mobility shift assays implicated −897 to –674 of PtaPAL promoter containing cis-elements of the expression in xylem of Pinus taeda. The results suggested that AC elements of PtaPAL have multiple functions in the expression under the various developmental stages and stress conditions in the transgenic tobacco. The nucleotide sequence data reported will appear in the EMBL, GenBank, and DDBJ Nucleotide Sequence Databases under the accession number AB449103 (PtaPAL promoter sequence).     

Comparing EST-based genetic maps between <Emphasis Type="BoldItalic">Pinus sylvestris</Emphasis> and <Emphasis Type="BoldItalic">Pinus taeda</Emphasis>

A genetic map of Pinus sylvestris was constructed using ESTP (expressed sequence tag polymorphism) markers and other gene-based markers, AFLP markers and microsatellites. Part of the ESTP markers (40) were developed and mapped earlier in Pinus taeda, and additional markers were generated based on P. sylvestris sequences or sequences from other pine species. The mapping in P. sylvestris was based on 94 F₁ progeny from a cross between plus-tree parents E635C and E1101. AFLP framework maps for the parent trees were first constructed. The ESTP and other gene sequence-based markers were added to the framework maps, as well as five published microsatellite loci. The separate maps were then integrated with the aid of AFLPs segregating in both trees (dominant segregation ratios 3:1) as well as gene markers and microsatellites segregating in both parent trees (segregation ratios 1:1:1:1 or 1:2:1). The integrated map consisted of 12 groups corresponding to the P. taeda linkage groups, and additionally three and six smaller groups for E1101 and E635C, respectively. The number of framework AFLP markers in the integrated map is altogether 194 and the number of gene markers 61. The total length of the integrated map was 1,314 cM. The set of markers developed for P. sylvestris was also added to existing maps of two P. taeda pedigrees. Starting with a mapped marker from one pedigree in the source species resulted in a mapped marker in a pedigree of the other species in more than 40% of the cases, with about equal success in both directions. The maps of the two species are largely colinear, even if the species have diverged more than 70 MYA. Most cases of different locations were probably due to problems in identifying the orthologous members of gene families. These data provide a first ESTP-containing map of P. sylvestris, which can also be used for comparing this species to additional species mapped with the same markers.Communicated by C. Möllers     

Biochemical changes during seed development in <Emphasis Type="Italic">Pinus taeda</Emphasis> L.

Amino acids, polyamines, 3-indoleacetic acid (IAA), abscisic acid (ABA), buffer-soluble protein and starch contents and dry matter accumulation were analyzed in megagametophytes containing developing embryos during seed development in Pinus taeda. The highest total amino acids and polyamine contents occurred at the cotyledonary stage, followed by a significant decrease in the mature seed. Free polyamines exhibited higher levels than conjugated ones, with putrescine being the predominant type until the cotyledonary stage, and spermidine at the mature seed stage. IAA content increased continually from the globular stage reaching the maximum at the cotyledonary stage, followed by a decrease in the mature seed. The highest ABA level occurred at the globular stage, followed by a continuous reduction until stabilization at the pre-cotyledonary stage. Buffer-soluble protein and starch contents, and dry matter increased progressively during development, reaching their maximum values at the mature stage.     

An auxin-inducible gene from loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.) is differentially expressed in mature and juvenile-phase shoots and encodes a putative transmembrane protein

We have isolated a gene from loblolly pine, 5NG4, that is highly and specifically induced by auxin in juvenile loblolly pine shoots prior to adventitious root formation, but substantially down-regulated in physiologically mature shoots that are adventitious rooting incompetent. 5NG4 was highly auxin-induced in roots, stems and hypocotyls, organs that can form either lateral or adventitious roots following an auxin treatment, but was not induced to the same level in needles and cotyledons, organs that do not form roots. The deduced amino acid sequence shows homology to the MtN21 nodulin gene from Medicago truncatula. The expression pattern of 5NG4 and its homology to a protein from Medicago involved in a root-related process suggest a possible role for this gene in adventitious root formation. Homology searches also identified similar proteins in Arabidopsis thaliana and Oryza sativa. High conservation across these evolutionarily distant species suggests essential functions in plant growth and development. A 38-member family of genes homologous to 5NG4 was identified in the A. thaliana genome. The physiological significance of this redundancy is most likely associated with functional divergence and/or expression specificity of the different family members. The exact biochemical function of the gene is still unknown, but sequence and structure predictions and 5NG4::GFP fusion protein localizations indicate it is a transmembrane protein with a possible transport function.Electronic Supplementary Material Supplementary material is available in the online version of this article at Abbreviations ABA Abscisic acid - BA Benzylaminopurine - EST Expressed sequence tag - NAA 1-Naphthaleneacetic acid - GFP Green fluorescent protein - ORF Open reading frame     

Rhizospheric streptomycetes as potential biocontrol agents of <Emphasis Type="Italic">Fusarium</Emphasis> and <Emphasis Type="Italic">Armillaria</Emphasis> pine rot and as PGPR for <Emphasis Type="Italic">Pinus taeda</Emphasis>

Pinus taeda is one of the main timber trees in Brazil, occupying 1.8 million ha with an annual productivity of 25–30 m³ ha⁻¹. Another important species is Araucaria angustifolia, belonging to the fragile Rainforest biome, which for decades has been a major source of timber in Brazil. Some diseases that affect the roots and/or the stem of these trees and cause “damping-off” of the seedlings, with economic and environmental losses for the forest sector, are caused by the plant pathogenic fungi Fusarium sp. or Armillaria sp. This research project intended to isolate actinobacteria from the Araucaria rhizosphere, which present an antagonistic effect against these fungi. After the selection of the best pathogen inhibitors, morphologic characteristics, enzyme production, and their effect on the growth of Pinus taeda were studied. The actinobacteria were tested for their antagonistic capacity against Fusarium sp. in Petri plates with PDA as substrate. The inhibition zone was measured after 3, 5, 7, and 10 days. Of all the isolates tested, only two of them maintained inhibition zones up to 4 mm for 10 days. The inhibition of Armillaria sp. was tested in liquid medium and also in Petri dishes through the evaluation of the number of the fungal rhizomorphs in dual culture with the actinobacteria. It was found that all five isolates were able to inhibit the rhizomorph production, with the best performance of the isolate A43, which was capable of inhibiting both fungi, Fusarium and Armillaria. In a greenhouse experiment, the effect of five isolates on the growth of Pinus taeda seedlings was tested. Plant height, stem diameter, root and shoot dry matter were determined. The Streptomyces isolate A43 doubled plant growth. These results may lead to the development of new technologies in the identification of still unknown bacterial metabolites and new management techniques to control forest plant diseases.     

Effects of antibiotics on the elimination of <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> from loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis>) zygotic embryo explants and on transgenic plant regeneration

Three antibiotics were evaluated for their effects on the elimination of Agrobacterium tumefaciens during the genetic transformation of loblolly pine ( Pinus taeda L.) using mature zygotic embryos as targets. Agrobacterium tumefaciens strains, EHA105, GV3101, and LBA 4404, all harbouring the plasmid pCAMBIA1301, which carries the selectable marker gene, hygromycin phosphotransferase ( hpt) controlled by the cauliflower mosaic virus 35S promoter and terminator, and the uidA reporter gene (GUS) driven by the cauliflower mosaic virus 35S promoter and the terminator of nopaline synthase gene, were used in this study. Exposure to 350 mg l^-1 carbenicillin, claforan, and timentin respectively for up to 6 weeks did not eliminate the Agrobacterium, while antibiotics at 500 mg l^-1 eradicated them from the co-cultivated zygotic embryos. All three antibiotics increased callus growth and shoot regeneration at 350 and 500 mg l^-1 each, but reduced callus growth and shoot regeneration at 650 mg l^-1 when compared with controls. Putative transgenic calli were selected for continued proliferation and differentiation on 4.5 mg l^-1 hygromycin-containing medium. Transformed calli and transgenic plants produced on a selection medium containing 4.5 mg l^-1 hygromycin were confirmed by GUS histochemical assays, by polymerase chain reaction (PCR), and by Southern blot analysis. These results are useful for future studies on optimizing genetic transformation procedures in loblolly pine.     

Soil CO<Subscript>2</Subscript> efflux in loblolly pine (<Emphasis Type="Italic">Pinus taeda</Emphasis> L.) plantations on the Virginia Piedmont and South Carolina Coastal Plain over a rotation-length chronosequence

We measured soil surface CO₂ efflux (F_s) in loblolly pine stands (Pinus taeda L.) located on the Virginia Piedmont (VA) and South Carolina Coastal Plain (SC) in efforts to assess the impact climate, productivity, and cultural practices have on F_s in the managed loblolly pine ecosystem. The effect of stand age on F_s was examined using a replicated chronosequence approach in which stands ranging from 1 to 25 years since planting were investigated. Soil CO₂ efflux was measured on both VA and SC sites for over a year using a closed dynamic system. Multiple linear regression was used to evaluate F_s correlates and examine the relationship between candidate explanatory variables and F_s. Soil temperature (top 10 cm) was the major correlate with F_s on both locations. We observed a positive age effect on F_s in VA stands and no relationship between age and F_s in SC stands. Annual soil C efflux declined with stand age in SC due to both reductions in soil temperatures as crown closure occurs and a diminishing heterotrophic C substrate pool. Annual estimated efflux ranges from 16.7 to 13.2 Mg C ha⁻¹ for 1 and 20-year-old stands, respectively. In contrast, annual soil C efflux increased with age in VA stands as a result of the positive relationship between stand age and F_s, which appears to be related to an increase in the contribution of root respiration to total F_s over time. In VA stands, efflux estimates range from 7.6 to 12.3 Mg C ha⁻¹ for 1 and 20-year-old stands, respectively. These results demonstrate the need to further consider the impact forest management and within-region variability have on soil C efflux over time when estimating C budgets.     

Multi-trait association mapping in sugar beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L.)

Association mapping promises to overcome the limitations of linkage mapping methods. The main objective of this study was to examine the applicability of multivariate association mapping with an empirical data set of sugar beet. A total of 111 diploid sugar beet inbreds was selected from the seed parent heterotic pool to represent a broad diversity with respect to sugar content (SC). The inbreds were genotyped with 26 simple sequence repeat markers chosen according to their map positions in proximity to previously identified quantitative trait loci for SC. For SC and beet yield (BY), the genotypic variances were highly significant (P < 0.01). Based on the global test of the bivariate mixed-model approach, four markers were significantly associated with SC, BY, or both at a false discovery rate of 0.025. All four markers were significantly (P < 0.05) associated with BY but only two with SC. The identification of markers associated with SC, BY, or both indicated that association mapping can be successfully applied in a sugar beet breeding context for detection of marker-phenotype associations. Furthermore, based on our results multivariate association mapping can be recommended as a promising tool to discriminate with a high mapping resolution between pleiotropy and linkage as reasons for co-localization of marker-phenotype associations for different traits.     

Plant regeneration in Stone pine (<Emphasis Type="Italic">Pinus pinea</Emphasis> L.) by somatic embryogenesis

Regeneration of plants by somatic embryogenesis (SE) was achieved in Stone pine (Pinus pinea), one of the most characteristic tree species of the Mediterranean ecosystem. The initial explants were megagametophytes containing zygotic embryos from five selected half-sib families collected at different dates over 2 consecutive years. Rates of extrusion and initiation of SE differed in both years. However, qualitative patterns were very similar: for most families, the responsive developmental window was from late cleavage polyembryony to early cotyledonary stage. The highest overall mean frequencies of extrusion and SE initiation (7 and 0.9%, respectively, for the five families and the eight 2006 collections) were obtained on a modified Litvay’s medium with 9 μM 2,4-D and 4.5 μM BAP, supplemented with L-glutamine and casein hydrolysate. Families showed large differences in frequencies of SE initiation from year to year. Only seven embryogenic lines were induced in 2005, representing three of the five families tested, whereas 34 lines from all the families were obtained in 2006. Proliferation of embryonal masses (EM) was significantly improved when they were subcultured after dispersing in liquid medium and collected on filter paper disks, instead of being subcultured as small clumps. This effect showed a significant interaction with genotype. Several preconditioning treatments and culture media combinations were tested for embryo development and maturation. The high proliferation rate of EM hampered somatic embryo development. However, up to 42 mature embryos from different lines of three of the five families were obtained, 23 of them germinated and seven converted into somatic seedlings.     

Species richness and seasonal abundance of ectomycorrhizal fungi in plantations of<Emphasis Type="Italic"> Eucalyptus dunnii</Emphasis> and<Emphasis Type="Italic"> Pinus taeda</Emphasis> in southern Brazil

The abundance and diversity of ectomycorrhizal fungi (EMF) was assessed based on the collection of basidiocarps during 12 months comprising the spring of 1995, and the summer, autumn, and winter of 1996, in three stands of young, middle-aged, and rotation age plantations of Pinus taeda and Eucalyptus dunnii, in the state of Santa Catarina, southern Brazil. A total of 3,085 collections yielded 34 presumed EMF taxa in ten genera, including mushroom-like and sequestrate species. Fruiting patterns of EMF differed with host and season, and host specificity was apparent in some. The overall relative importance (RI) and the Shannon diversity index (H) suggested that stands of E. dunnii had a more diverse aboveground EMF community than those of P. taeda. Overall, species of Scleroderma and Laccaria were not only the most abundant but also had the highest biomass values. The results show that a small number of species of abundant biomass and a larger number of species of less-abundant biomass characterize each forest class.     

Fine genetic mapping and physical delimitation of the lesion mimic gene <Emphasis Type="Italic">spotted leaf 5</Emphasis> (<Emphasis Type="Italic">spl5</Emphasis>) in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Spotted leaf 5 (spl5), a lesion mimic mutant, was first identified in rice (Oryza sativa L.) japonica cv. Norin8 in 1978. This mutant exhibits spontaneous disease-like lesions in the absence of any pathogens and resistance to rice blast and bacterial blight; however, the target gene has not yet been isolated. In the present study, we employed a map-based cloning strategy to finely map the spl5 gene. In an initial mapping with 100 F₂ individuals (spl5/spl5) derived from a cross between the spl5 mutant and indica cv. 93-11, the spl5 gene was located in a 3.3-cM region on chromosome 7 using six simple sequence repeat (SSR) markers. In a high-resolution genetic mapping, two F₂ populations with 3,149 individuals (spl5/spl5) were derived from two crosses between spl5 mutant and two indica cvs. 93-11 and Zhefu802 and six sequence-tagged site (STS) markers were newly developed. Finally, the spl5 gene was mapped to a region of 0.048 cM between two markers SSR7 and RM7121. One BAC/PAC contig map covering these markers’ loci and the spl5 gene was constructed through Pairwise BLAST analysis. Our bioinformatics analysis shows that the spl5 gene is located in the 80-kb region between two markers SSR7 and RM7121 with a high average ratio of physical to genetic distance (1.67 Mb/cM) and eighteen candidate genes. The analysis of these candidate genes indicates that the spl5 gene represents a novel class of regulators controlling cell death and resistance response in plants.     

Population structure analysis and association mapping of bacterial blight resistance in <Emphasis Type="Italic">indica</Emphasis> rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) accessions

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo), is a serious disease in rice production worldwide. To understand the genetic diversity of bacterial blight resistance a population consisting of 175 indica accessions from nine countries was collected and detected their association between SSR (Simple Sequence Repeat) markers and resistance to six bacterial races. The resistance phenotypes of various rice accessions were evaluated through artificial inoculation under controlled conditions in 2013 and 2014. Association analysis showed that 17 SSR markers were significantly associated with resistance to four bacterial races and the phenotypic variations explained (PVE) ranged from 7.43 to 15.05%. Among the 17 associated SSR markers, two SSR markers located in previously reported genes regions, and 15 SSR markers were newly identified in this study. These results validated a new approach to map resistance genes of rice to bacterial blight. These markers could be used for marker-assisted selection (MAS) in rice bacterial blight resistance breeding programs.     

Seasonal changes in water source of four families of loblolly pine (<Emphasis Type="Italic">Pinus</Emphasis><Emphasis Type="Italic">taeda</Emphasis> L.)

Source water utilized by four families of loblolly pine (Pinus taeda L.) was assessed by comparing the H isotope composition ('D) of xylem sap and of soil water from four depths (0-20 cm, 20-40 cm, 1.2 m, and 2.1 m) across 1 year. Soil water 'D values varied with soil moisture content in the well-drained, sandy site and at each of the four soil depths. In September and November 1997 and May through November 1998, xylem sap 'D values closely matched the soil water 'D values of the upper soil horizons (0-20 and 0-40 cm) and, therefore, reflected significant water uptake from upper regions of the soil profile. In March 1998, xylem sap 'D values closely matched the soil water 'D values of the 1.2 m soil depth and, therefore, indicated that trees during this portion of the growing season were obtaining their water from deep in the soil profile. Analysis of source water use with a two-ended mixing model in the 3 months of collection that exhibited a range of soil water 'D values across the soil profile confirmed that trees utilized different sources of water depending upon season of the year. In September 1997 and November 1998, source water uptake was primarily from the upper soil profile while in March 1998, source water uptake was from deep in the soil profile. With few exceptions, we did not find striking differences in source water use between drought-hardy families and those that were locally adapted.