Effects of nitrogen limitation on water relations of jack pine (Pinus banksiana Lamb.) seedlings

The water relations of shoots of young jack pine (Pinus banksiana Lamb.) seedlings were examined 6 and 15 weeks after the initiation of four different dynamic nitrogen (N) treatments using a pressure-volume analysis. The N treatments produced a wide range of needle N concentrations from 12 to 32 mg g^?1 dry mass and a 10-fold difference in total dry mass at 15 weeks. Osmotic potential at full turgor did not change over the range of needle N concentrations observed. Osmotic potential at turgor-loss point, however, declined as N concentrations decreased, indicating an increased ability of N-deficient jack pine plants to maintain turgor. The increase could be attributed largely to an increase in cell wall elasticity, suggesting that elasticity changes may be a common, significant adaptation of plants to environmental stresses. Dry mass per unit saturated water almost doubled as needle N level dropped from 32 to 12 mg g^?1 and was inversely correlated to the bulk modulus of elasticity. This suggests that cell wall elasticity is determined more by the nature of its cross-linking matrix than by the total amount of cell wall material present. Developmental change was evident in the response of some water relation variables to N limitation.     

Differential expression during drought conditioning of a root-specific S-adenosylmethionine synthetase from jack pine (Pinus banksiana Lamb.) seedlings

Differential screening of a cDNA library constructed from root mRNA from jack pine (Pinus banksiana Lamb.) seedlings exposed to two cycles of drought conditioning identified a S-adenosylmethionine synthetase (sam-s) cDNA. A cDNA encoding the entire open reading frame of SAM-S was identified and characterized. Analysis of the full-length sam-s cDNA revealed that it was 1675 bp, encoded an open reading frame of 393 amino acids and had a predicted protein mass of 43 kDa. Jack pine sam-s was found to be highly similar to several other plant sam-s genes. RNA gel blot analysis showed that sam-s mRNA abundance increased following two cycles of drought conditioning and remained abundant after 3 d of rewatering. Expression of this gene appears to be root-specific. Quantitative slot blot analysis showed that two cycles of drought conditioning caused a 6-fold increase in sam-s mRNA abundance whereas heat shock, cold stress and anoxia did not result in the accumulation of sam-s mRNA. SAM-S enzyme activity increased 2-fold following two cycles of drought conditioning. The increase in the rate of SAM-s enzyme activity was also correlated with changes in rates of ethylene and betaine synthesis, biosynthetic pathways that utilize SAM as a substrate. Ethylene evolution and betaine abundance increased following two cycles of drought conditioning.     

Environmental stress and the relationships among allozyme heterozygosity,biomass and biomass components in jack pine (Pinus banksiana Lamb.)

The relationships among allozyme heterozygosity, biomass, and seven components of biomass were studied in fifteen open-pollinated families of jack pine grown on four soil substrates. Most of these relationships were curvilinear, and were found under sub-optimal environments. None of the eight characters showed a consistent relationship with allozyme heterozygosity on the four media. Allozyme heterozygosity (phenotype) and heterozygosity (heterosis) used in conventional breeding techniques appeared to respond similarly to environmental variation. The intensity of expression of the relationship between allozyme heterozygosity and quantitative traits may be partially under environmental control. This conclusion is consistent with previous findings on the expression of heterosis in different environments.     

Differences in foliage quality of young jack pine (Pinus banksiana Lamb.) on burned and clearcut sites: effects on jack pine budworm (Choristoneura pinus pinus Freeman)

Summary Suitability of young jack pine as a host for jack pine budworm was examined on similarly-aged trees growing on two areas previously burned in wildfires and on two previously clearcut areas in northwest Wisconsin. Nitrogen, monoterpenes, and moisture levels of foliage, and xylem water potential were measured and related to larval survival and pupal weight of caged jack pine budworm larvae. Nitrogen, monoterpenes, needle weight, and needle moisture were higher in trees growing on clearcut sites than on burned area trees. Surival of budworms to early and late instar, pupation, and adult eclosion was greater for larvae caged on clearcut-area trees than on burned-area trees. Female pupal weight differed between older (ca 10 years old) and younger (ca 8 years old) trees, but not between clearcut and burned areas. Mean female pupal weight was greatest on lownitrogen trees, where larval survival was lowest. Foliar nitrogen was consistently included as a significant predictor in budworm survival regressions. Regressions indicated larval survival and pupal weight may be associated with different tree- and foliage-related traits. Results suggest long-lasting effects of previous forest disturbance may subsequently affect herbivorous insects such as jack pine budworm.     

NaCl and Na2SO4 alter responses of jack pine (Pinus banksiana) seedlings to boron

Using sand culture, we examined the responses of 6-month-old jack pine (Pinus banksiana Lamb.) seedlings to boron and salinity (sodium chloride and sodium sulfate) treatments. During 4 weeks of treatments, 60 mM NaCl and 60 mM Na₂SO₄ significantly decreased survival, new shoot length, number of new roots, shoot to root dry weight ratio and transpiration rates. When applied in absence of the salts, B had little effect on the measured variables. However, when applied together with salts, B decreased seedling survival, increased needle injury and altered tissue elemental concentrations in jack pine seedlings. In 2 mM B treatment, B concentration was higher in the shoots than in the roots. However, when 2 mM B was present in NaCl and Na₂SO₄ treatments, shoot boron concentration declined and greater proportion of B accumulated in the roots. This shift corresponded to a decline in transpiration rates. In plants treated with NaCl, Na accumulated primarily in the shoots, while in Na₂SO₄-treated plants Na accumulated mostly in the roots. Based on the electrolyte leakage and needle necrosis data, Cl^– appears to be the major factor contributing to seedling injury and B aggravates the injurious effects of NaCl. We suggest that Cl^– may contribute to Na and B toxicity in jack pine by altering cell membrane permeability leading to increased Na concentration in the shoots.     

Role of aquaporins in root water transport of ectomycorrhizal jack pine (Pinus banksiana) seedlings exposed to NaCl and fluoride

Effects of ectomycorrhizal (ECM) fungus Suillus tomentosus on water transport properties were studied in jack pine (Pinus banksiana) seedlings. The hydraulic conductivity of root cortical cells (L_pc) and of the whole root system (L_pr) in ECM plants was higher by twofold to fourfold compared with the non‐ECM seedlings. HgCl₂ had a greater inhibitory effect on L_pc in ECM compared with non‐ECM seedlings, suggesting that the mercury‐sensitive, aquaporin (AQP)‐mediated water transport was largely responsible for the differences in L_pc between the two groups of plants. L_pc was rapidly and drastically reduced by the 50 mm NaCl treatment. However, in ECM plants, the initial decline in L_pc was followed by a quick recovery to the pre‐treatment level, while the reduction of L_pc in non‐ECM seedlings progressed over time. Treatments with fluoride reduced L_pc by about twofold in non‐ECM seedlings and caused smaller reductions of L_pc in ECM plants. When either 2 mm KF or 2 mm NaF were added to the 50 mm NaCl treatment solution, the inhibitory effect of NaCl on L_pc was rapidly reversed in both groups of plants. The results suggest that AQP‐mediated water transport may be linked to the enhancement of salt stress resistance reported for ECM plants.     

Microtubule pattern and the occurrence of pre-prophase bands in embryogenic cultures of black spruce (Pieca mariana Mill.) and non-embryogenic cultures of jack pine (Pinus banksiana Lamb.)

Summary The organization of microtubules during interphase and prophase in embryogenic cultures of black spruce (Picea mariana) was investigated by indirect immunofluorescence. Somatic embryos of black spruce possessed an extensively branched and interconnecting network of fine interphase cortical microtubules. The development of pre-prophase bands (PPBs) in embryogenic black spruce cultures was compared with that in non-embryogenic cell cultures of jack pine (Pinus banksiana). PPBs in both species were initially arranged as a very broad array of microtubules, later (early to mid-prophase) becoming narrower and more intensely fluorescent. The occurrence of pre-prophase bands in relation to the number of phragmoplasts (i.e. PPB index) of black spruce somatic embryos was significantly higher (p<0.01) than that found for jack pine cells.     

Segregation of allozymes in megagametophytes of viable seed from a natural population of jack pine,Pinus banksiana Lamb.

Summary Segregation ratios of allozymes in haploid female gametophytes obtained from viable seed were studied in a natural population of jack pine, Pinus banksiana. Stability of these ratios was assessed for three levels of the sexually reproductive crown as well as for four years of natural fertilization. Analyses of observed segregation ratios of four of five polymorphic isozyme loci showed good correspondence to the overall 11 ratios expected for simple Mendelian inheritance. Allozymes of glucose-6-phosphate dehydrogenase did not segregate in the expected 11 ratio. In addition, there were significant deviations from the expected segregation ratio for all the loci at some sampling positions on individual trees. Heterogeneity of segregation among trees, strata and years could be the result of pollen pool heterogeneity, segregation distortion and/or recessive lethal and semi-lethal gene combinations resulting in early embryo abortion. These types of segregation deviations in viable seed can affect the estimation of allele frequencies from bulked samples of a small number of individuals, the inference of heterozygosity/homozygosity of parental trees, and estimates of selfing rates.     

Growth of mycorrhizal jack pine (Pinus banksiana) and white spruce (Picea glauca) seedlings planted in oil sands reclaimed areas

The effectiveness of ectomycorrhizal inoculation at the tree nursery seedling production stage on growth and survival was examined in jack pine (Pinus banksiana) and white spruce (Picea glauca) planted in oil sands reclamation sites. The seedlings were inoculated with Hebeloma crustuliniforme strain # UAMH 5247, Suillus tomentosus strain # UAMH 6252, and Laccaria bicolor strain # UAMH 8232, as individual pure cultures and in combinations. These treatments were demonstrated to improve salinity resistance and water uptake in conifer seedlings. The field responses of seedlings to ectomycorrhizal inoculation varied between plant species, inoculation treatments, and measured parameters. Seedling inoculation resulted in higher ectomycorrhizal colonization rates compared with non-inoculated control, which had also a relatively small proportion of roots colonized by the nursery contaminant fungi identified as Amphinema byssoides and Thelephora americana. Seedling inoculation had overall a greater effect on relative height growth rates, dry biomass, and stem volumes in jack pine compared with white spruce. However, when examined after two growing seasons, inoculated white spruce seedlings showed up to 75 % higher survival rates than non-inoculated controls. The persistence of inoculated fungi in roots of planted seedlings was examined at the end of the second growing season. Although the inoculation with H. crustuliniforme triggered growth responses, the fungus was not found in the roots of seedlings at the end of the second growing season suggesting a possibility that the observed growth-promoting effect of H. crustuliniforme may be transient. The results suggest that the inoculation of conifer seedlings with ectomycorrhizal fungi could potentially be carried out on a large scale in tree nurseries to benefit postplanting performance in oil sands reclamation sites. However, these practices should take into consideration the differences in responses between the different plant species and fungal strains.     

Phenylalanine ammonia-lyase-induced freezing tolerance in jack pine (Pinus banksiana) seedlings treated with low, ambient levels of ultraviolet‐B radiation

Phenylalanine ammonia-lyase (PAL) induction in UVB-exposed plants leads to an increased synthesis of UV-absorbing phenols. As phenols, including anthocycanins, are linked to many protective mechanisms in plants, we tested the hypothesis that UVB-induced phenol accumulation, mediated by PAL, may confer freezing tolerance in jack pine ( Pinus banksiana Lamb) seedlings. The hypothesis was tested by applying UVB in the presence and absence of the PAL-inhibitor, 2-aminoindan-2-phosphonic acid (AIP). Jack pine seedlings were grown for 3 weeks with and without 10 µ M aqueous AIP. Each treatment was then divided into two groups. One group received near-ambient UVB (5.5 kJ m⁻² day⁻¹of biologically effective radiation) for up to 30 h. A second, control group of seedlings received no UVB. Anthocyanin concentration declined by > 99% in PAL-inhibited seedlings and other methanol-extractable UV-absorbing phenols declined by > 48%, relative to the controls. A 20-h exposure to UVB increased seedling freezing (−15°C) tolerance in the absence of the PAL-inhibitor, as shown by a 30% reduction in membrane injury, determined by electrolyte leakage measurements. In PAL-inhibited seedlings, by contrast, the same UVB pre-treatment increased freezing injury by 48%. A longer (30 h) UVB exposure was damaging to both AIP-treated and untreated seedlings. Root feeding with 10 µ M AIP during a 3-week exposure of older (6-month-old) seedlings similarly reduced phenol accumulation in UVB-exposed seedlings. The decline in phenol production in PAL-inhibited seedlings correlated with increased freezing injury. These results suggest a role for ambient UVB in seedling frost hardiness, mediated by a PAL-induced production of phenolic compounds.     

Population structure of a lodgepole pine (Pinus contorta) and jack pine (P. banksiana) complex as revealed by random amplified polymorphic DNA.

We studied the population structure of a lodgepole (Pinus contorta Dougl.) and jack pine (Pinus banksiana Lamb.) complex in west central Alberta and neighboring areas by assessing random amplified polymorphic DNA (RAPD) variability in 23 lodgepole pine, 9 jack pine, and 8 putative hybrid populations. Of 200 random primers screened, 10 that amplified 39 sharp and reproducible RAPDs were chosen for the study. None of the 39 RAPDs were unique to the parental species. RAPD diversity ranged from 0.085 to 0.190 among populations and averaged 0.143 for lodgepole pine, 0.156 for jack pine, 0.152 for hybrids, and 0.148 for all 40 populations. The estimated population differentiation based on G(ST) was 0.168 for hybrids, 0.162 for lodgepole pine, 0.155 for jack pine, and 0.247 across all 40 populations. Cluster analysis of genetic distances generally separated jack pine from lodgepole pine and hybrids, but no division could be identified that further separated lodgepole pine from hybrids. The observed weak to mild trend of "introgression by distance" in the complex and neighbouring areas was consistent with the view that introgressive hybridization between lodgepole and jack pines within and outside the hybrid zone may have been through secondary contact and primary intergradation, respectively.     

Relationship between net photosynthesis and nitrogen in Scots pine: Seasonal variation in seedlings and shoots

The relationship between light saturated net photosynthesis (A_max) and nitrogen concentration (N) was studied in needles of both Scots pine seedlings, grown at three relative growth rates (2,6 and 8%) controlled by nutrient addition rate, and Scots pine shoots collected from four sites with different fertility. In the seedlings, A_max was measured on 14 different dates starting at the beginning of the second growing season and ending when growth of the new shoot and the secondary needles had finished. In shoots from the natural stands A_max of the previous-year shoots was measured on 6 dates throughout the growing season.Both in seedlings and shoots, the correlation between A_max and N was poor, when data from all sampling dates were taken together. However, A_max was correlated with N in most instances when the age of the needles was considered and the data were examined either at weekly intervals (seedlings) or separately for each sampling date (shoots). The slope of the A_max vs N relationship varied greatly between sampling dates. In the seedlings the correlation between A_max and N was strongest by the time when the new needles were developing. In the shoots the correlation was significant from mid June until mid August, while no correlation was found in the beginning and at the end of the growing season.Our data indicate that in pine needles the photosynthesis-nitrogen relationship is more complex than in broadleaved species. Contrary to the broadleaved species, where the correlation is independent of sampling time, in this conifer the time of the year affects the correlation and there are phases during the growing season when the correlation is poor or nonexistent.Abbreviations A_max light saturated net photosynthesis - kPa kilopascal - N nitrogen concentration in the needles - PFD photon flux density (400–700 nm) - RGR relative growth rate     

Nuclear DNA content variation in seeds from 22 half-sib families of jack pine (Pinus banksiana,Pinaceae)

In order to understand the amount of DNA content variation and its potential roles, both absolute DNA amount and cell cycle phases in 22 half-sib families of jack pine were examined using flow cytometry. When the variability due to differences in speed of germination was taken into account, embryos from superior families (classified on the basis of height growth during field trials) had significantly higher levels of all nuclei classes greater than 4C. Mean DNA contents per nucleus were significantly lower in embryos from superior families compared to inferior ones. Analysis of megagametophyte tissue showed that the mother trees of these embryos expressed a similar pattern. Absolute DNA values were also established on the emerging radicle and the hypocotyl + cotyledons region (HC) separately in five of the families. Nuclei isolated from the emerging radicles had significantly lower levels of DNA than those isolated from the HC region. For three of these families, absolute DNA values from nuclei of the hypocotyl + cotyledons region were established on individual embryos with varying cotyledon numbers. In all three families total DNA amount per nucleus decreased with increasing cotyledon number. A better understanding of differences observed in DNA content during germination, as well as in total DNA content per nuclei among different half-sib families of jack pine, may help in the identification of factors that influence growth and adaptation of this species.     

Growth and photosynthesis of Lycopersicon esculentum (L.) plants as affected by nitrogen deficiency

Fully expanded leaves of tomato (Lycopersicon esculentum) growing with either complete or nitrogen-deficient nutrient solution were analysed for leaf water status, gas exchange and chlorophyll fluorescence during the vegetative and reproductive phases. N-deficiency did not affect leaf water relations but did decrease light saturated photosynthetic rate as well as stomatal conductance in the vegetative stage. A lower variable to maximum fluorescence ratio (Fv/Fm) was found in N-limited plants which also showed an increase in leaf starch content and in starch to sucrose ratio. The inhibition of photosynthesis and the alteration of photosynthates partitioning were responsible for the growth reduction in N-stressed plants. During the reproductive phase the limitation of photosynthesis may be due to a large accumulation of starch which determines both a decrease in the carbon demand from the sinks and a decrease in CO2 conductance in the mesophyll. This revised version was published online in July 2006 with corrections to the Cover Date.     

Factors affecting transient gene expression in electroporated black spruce (Picea mariana) and jack pine (Pinus banksiana) protoplasts

Summary Methods were developed for transient gene expression in protoplasts of black spruce (Picea mariana) and jack pine (Pinus banksiana). Protoplasts were isolated from embryogenic suspension cultures of black spruce and from non-embryogenic suspensions of jack pine. Using electroporation, transient expression of the chloramphenicol acetyltransferase (CAT) gene was assayed and shown to be affected by the cell line used, by voltage, temperature, and by the plasmid concentration and conformation. Increasing the plasmid DNA concentration (0–150g ml^–1) resulted in higher levels of transient CAT expression. In jack pine, linearized plasmid gave 2.5 times higher levels of CAT enzyme activity than circular. Optimal voltage varied for each cell line of the two species within the range 200–350 V cm^–1 (960 F). A heat shock treatment of protoplasts for 5 min at 45 °C resulted in enhanced CAT gene expression for both species.NRCC No. 30491     

A mitochondrial DNA minisatellite reveals the postglacial history of jack pine (Pinus banksiana), a broad-range North American conifer

Jack pine (Pinus banksiana Lamb.) is a broadly distributed North American conifer and its current range was covered by the Laurentian ice sheet during the last glacial maximum. To infer about the history and postglacial colonization of this boreal species, range-wide genetic variation was assessed using a new and highly variable minisatellite-like marker of the mitochondrial genome. Among the 543 trees analysed, 14 distinct haplotypes were detected, which corresponded to different repeat numbers of the 32-nucleotide minisatellite-like motif. Several haplotypes were rare with limited distribution, suggesting recent mutation events during the Holocene. At the population level, an average of 2.6 haplotypes and a mean haplotype diversity (H) of 0.328 were estimated. Population subdivision of genetic diversity was quite high with G(ST) and R(ST) values of 0.569 and 0.472, respectively. Spatial analyses identified three relatively homogeneous groups of populations presumably representative of genetically distinct glacial populations, one west and one east of the Appalachian Mountains in the United States and a third one presumably on the unglaciated northeastern coastal area in Canada. These results indicate the significant role of the northern part of the US Appalachian Mountains as a factor of vicariance during the ice age. A fourth distinct group of populations was observed in central Québec where the continental glacier retreated last. It included populations harbouring haplotypes present into the three previous groups, and it had higher level of haplotype diversity per population (H = 0.548) and lower population differentiation (G(ST) = 0.265), which indicates a zone of suture or secondary contact between the migration fronts of the three glacial populations. Introgression from Pinus contorta Dougl. var. latifolia Engelm. was apparent in one western population from Alberta. Altogether, these results indicate that the mitochondrial DNA variation of jack pine is geographically highly structured and it correlates well with large-scale patterns emerging from recent phylogeographical studies of other tree boreal species in North America.     

Population dynamics of the white pine weevil, Pissodes strobi, infesting jack pine, Pinus banksiana, in Ontario, Canada

1. Eight generations of white pine weevil, Pissodes strobi (Peck) , infesting the terminal shoots of young jack pine trees, were sampled as larvae, pupae, and adults between 1988 and 1995.
2. The density of adult weevils increased rapidly for the first 3 years of the study, then declined in the next 4 years. Between-generation rates of change in density of weevils were related to mean number of weevils produced per terminal shoot of the host tree. Net increases in population density followed years in which a relatively high number of weevils emerged per terminal shoot and vice versa.
3. The mean number of weevils emerging per terminal shoot was determined by survival of weevils between the end of the larval stage and the successful emergence of adult weevils.
4. There was no clear relationship between survival of the post-feeding stages of the weevil and rates of parasitism or bird predation on these same stages. There was, however, a strong negative relationship between survival of weevils within the terminal shoot and the abundance of the facultative dipteran predator Lonchaea corticis Taylor.
5. The relationship among generational rates of change in populations, survival, and predation was evident irrespective of the age of the trees infested, suggesting that population dynamics of the weevil may be influenced as much by local predation pressure as by physical and biotic changes associated with growth of the young host trees.     

Differences in defence responses of Pinus contorta and Pinus banksiana to the mountain pine beetle fungal associate Grosmannia clavigera are affected by water deficit

We tested the hypotheses that responses to the mountain pine beetle fungal associate Grosmannia clavigera will differ between the evolutionarily co‐evolved host lodgepole pine (Pinus contorta var. latifolia) and the naïve host jack pine (Pinus banksiana) and that these responses will be influenced by water availability. G. clavigera inoculation resulted in more rapid stem lesion development in lodgepole than in jack pine; water deficit delayed lesion development in both species. Decreased hydraulic conductivity was observed in inoculated lodgepole pine seedlings, likely because of tracheid occlusion by fungal hyphae and/or metabolite accumulation. Drought but not inoculation significantly impacted bark abscisic acid levels. Jasmonic and salicylic acid were implicated in local and systemic responses of both species to G. clavigera, with salicylic acid appearing to play a greater role in jack pine response to G. clavigera than lodgepole pine. Water deficit increased constitutive levels and/or attenuated induced responses to G. clavigera for several monoterpenes in lodgepole but not jack pine. Instead, inoculation of well‐watered but not water deficit jack pine resulted in a greater number of xylem resin ducts. These findings reveal mechanisms underlying differences in G. clavigera‐induced responses between lodgepole and jack pine hosts, and how water availability modulates these responses.