Inheritance of RFLP loci in a loblolly pine three-generation pedigree

Summary A high-density restriction fragment length polymorphism (RFLP) linkage map is being constructed for loblolly pine (Pinus taeda L.). Loblolly pine cDNA and genomic DNA clones were used as probes in hybridizations to genomic DNAs prepared from grandparents, parents, and progeny of a three-generation outbred pedigree. Approximately 200 probes were evaluated for their ability to detect polymorphic loci between DNAs prepared from the two parent trees, 20–1010 and 11–1060, and cut with four different restriction enzymes: BamHI, DraI, EcoRI, and HindIII. More than half of the probes detecting single- or low-copy number sequences (56%) revealed polymorphisms between the two parents with at least one restriction enzyme. If necessary, an additional hybridization to DNAs prepared from the four grandparent trees was conducted to determine the zygosity of parent trees. Ten of these probes were hybridized to progeny DNAs from this cross and, as expected, the markers were inherited as simple codominant Mendelian alleles. Four of the ten probes detected segregation of three alleles at one locus, and four probes detected more than one independently segregating locus. RFLPs can be used immediately to assess genetic diversity in conifer populations and to efingerprint genotypes in tree improvement programs.     

Aluminum-mycorrhizal interactions in the physiology of pitch pine seedlings

Aluminum (Al) in the rhizosphere adversely affects plant nutrition and growth. Although many conifer species, and pitch pine (Pinus rigida) in particular, have evolved on acidic soils where soluble Al is often high, controlled environment studies often indicate that Al interferes with seedling growth and nutrient relations. Under normal field conditions, conifer roots grow in a symbiotic relationship with ectomycorrhizal fungi, and this association may modulate the effects of Al on root physiology. To investigate the influence of mycorrhizal infection on Al toxicity, pitch pine seedlings were grown with or without the ectomycorrhizal symbiont Pisolithus tinctorius and were exposed to low levels of Al in sand culture. Aluminum at 50 μM reduced nonmycorrhizal seedling growth and increased foliar Al concentrations, but did not alter photosynthetic gas exchange or other aspects of seedling nutrition. Nonmycorrhizal seedlings exposed to 200 μM Al exhibited decreased growth, increased transpiration rates, decreased water use efficiency, increased foliar Al and Na levels, and reduced foliar P concentrations. Seedlings inoculated with P. tinctorius exhibited unaltered growth, physiological function, and ionic relations when exposed to Al. The fungal symbiont evidently modulated ionic relations in the rhizosphere, reducing Al-P precipitation reactions, Al uptake, and subsequent root and shoot tissue Al exposure.     

Reproductive and genetic characteristics of rare,disjunct pitch pine populations at the northern limits of its range in Canada

Pitch pine, Pinus rigida Mill., is a rare species in Canada, existing as a disjunct population in the St. Lawrence River Valley in eastern Ontario and two northern outlier stands in southern Quebec along Canada's southern border with the United States. Reproductive and genetic characteristics of these small, scattered stands were investigated to develop a foundation for management and restoration in the event of range expansion northwards under anticipated climate warming. Seed yields and seed quality appear to be comparable to other eastern conifers, and to pitch pine at the center of its geographic range. For seed and seedling growth traits, most of the variation was attributable to differences among trees within stands and, to a lesser extent, among stands within a population; whereas the population effect was non-significant. For reproductive traits, such as numbers of filled and empty seeds per cone, reproductive efficiency, and inbreeding estimates, high levels of variation (ranging from 26% to 33%) were found among stands, suggesting that stand structural features, such as stand size and tree density within stands, play an important role in pollination environment and overall reproductive success. Estimates of genetic diversity at 32 allozyme gene loci indicate that these small, isolated stands have maintained relatively high levels of genetic diversity compared with populations at the center of its geographic range, and also relative to other widely dispersed eastern conifers. The relatively high levels of viable seed production and genetic diversity in native pitch pine populations indicate that native Canadian populations may be suitable seed sources for species restoration and range expansion in Canada.     

The effects of annual fertilization and complete competition control on fascicle morphology of different aged loblolly pine stands

The goal of this study was to determine how increased nutrient availability affects foliage morphology of loblolly pine (Pinus taeda L.) without introducing the confounding influence of light availability. Morphology of fascicles from the terminal leader (radius, length, specific needle area, density, and needles per fascicle) and terminal leader traits (length, foliated length, total leaf area, and total fascicles) were measured for different aged (ranged from 5-year-old to 12-year-old) loblolly pine stands growing on the Piedmont and lower Coastal Plain of Georgia, USA. A factorial combination of annual fertilization and complete interspecific competition control was applied as stand level treatments. Competition control and stand age generally did not affect fascicle morphology. In contrast, annual fertilization significantly increased fascicle length (5%), needles per fascicle (4%), and total leaf area of the terminal (18%), while decreasing specific needle area (4%). Fertilization also increased terminal leader growth (5%) and total number of fascicles (6%). Therefore, loblolly pine foliage morphology does have plasticity to respond to increased nutrient availability. However, the magnitude of these morphological changes is small compared to changes in total canopy leaf area.     

Short-term effects of thinning and liming on forest soils of pitch pine and Japanese larch plantations in central Korea

The influences of thinning (50% of standing density) and liming (Ca+Mg, 2 Mg ha⁻¹) on soil chemical properties were investigated for 2 years (2001, 2002) in 40-year-old pitch pine (Pinus rigida Mill.) and 44-year-old Japanese larch (Larix leptolepis Gord.) plantations established on similar soils. In general, soil properties varied significantly among plantations and treatments. For both plantations, thinning significantly increased soil organic C (SOC) concentrations whereas there were no significant changes in soil pH and Ca and Mg concentrations. In addition, thinning increased total soil N and Na concentrations for the pitch pine plantation and available P concentration for the Japanese larch plantation in the second year after the treatment. Liming did not affect soil chemical characteristics for the pitch pine plantation except for Na concentration. However, for the Japanese larch plantation, liming significantly increased soil pH and K, Ca and Mg concentrations and decreased SOC and total soil N concentrations. For both plantations, soil Al concentration did not change after thinning and liming and decreased exponentially with increased pH values. The increases in SOC and total soil N concentrations after thinning were possibly due to increases in decomposition of organic matter and root death. Although differences were not statistically significant, soil available P concentration tended to increase at early stages of liming for both plantations. These results suggested that thinning and liming seemed to regulate soil chemical properties for pitch pine and Japanese larch plantations established on similar soils.     

Effect of carbon dioxide, fertilization and irrigation on loblolly pine branch morphology

 Foliage and wood parameters of branches of 12-year-old loblolly pine (Pinus taeda L.) trees were characterized after 21 months of exposure to fertilizer, irrigation and elevated CO₂ treatments. Branches of loblolly pine trees were enclosed in plastic chambers and exposed to ambient, ambient +175 and ambient +350 umol mol^–1 CO₂ concentrations. Measurements of foliage and wood at the fascicle, flush and branch levels were made at the end of the 21 month study period. The +350 CO₂ treatment did not significantly increase fascicle radius or length but did increase the number of fascicles on the first flush. Fertilization significantly increased fascicle radius and length, while irrigation significantly increased number of fascicles and flush length of first flush. The +350 CO₂ treatment also significantly increased flush length of the first flush. Significant interaction of fertilization and irrigation with CO₂ was observed for fascicle length. Significant interactions of fertilization and irrigation were also observed for flush length, number of fascicles and fascicle length. Observed increases in fascicle radius, fascicle length, number of fascicles and flush length may have been responsible for the significantly higher flush leaf area observed for the all three treatments. Also, a combination of fertilization and irrigation increased leaf area by 82% compared to that in the control when averaged across CO₂ treatments. At the branch level +350 CO₂ treatment significantly increased shoot length but not the number of flushes on the branch. In general with the exception of bark density and total number of needle scales, neither fertilization nor irrigation had any significant effect on other branch level parameters. Results from this study indicate that with ‘global change’ an increase in CO₂ alone may increase leaf area via an increase in flush length and number of fascicles. Combining increases in CO₂ with fertilization and irrigation could greatly enhance leaf area which when coupled to observed increases in net photosynthesis as a result of elevated CO₂ could greatly increase productivity of loblolly pine trees. Received: 22 August 1996 / Accepted: 5 March 1997     

Effect of shade stress on growth,morphology, and carbon dynamics of loblolly pine branches

Summary Shade treatments were applied to the terminal portions of branches in loblolly pine trees to test whether portions of branches were autonomous with respect to carbohydrates. The shade treatments reduced light by 50% and 72% compared with full sun conditions resulting in significant reductions in net photosynthesis. Branch growth (branch diameter and length, branch and needle biomass) decreased significantly within the shaded portion of the branch. Branch and needle morphology were also affected by shading. Lateral branches subtending terminal shoots were labelled with ¹⁴C 2 times during the growing season. No movement of ¹⁴C into the terminal portions of the branch was evident during the first growth flush. However, during the second flush of growth small but statistically significant amounts of ¹⁴C were imported into the terminal portion of the shaded branches from subtending laterals. It was concluded that loblolly pine shoots were usually autonomous with respect to carbohydrate supply, but that carbohydrate movement into the terminal shoot from subtending foliage could occur under conditions of very high stress.     

Transformation and regeneration of loblolly pine: shoot apex inoculation with Agrobacterium

Loblolly pine (Pinus taeda L.) is the mostimportant tree species in US commerce and has much to gain through geneticengineering. This species can be transformed using particle bombardment andAgrobacterium; however, the regeneration of plants fromtransgenic tissues has been difficult and the recovery of transgenic plants hasbeen rare. A shoot-based and genotype-independent transformation methodemploying Agrobacterium tumefaciens was used to facilitaterecovery of plants and permit the transformation of elite germplasm. Shootsfrom4–6 week old seedlings and adventitious shoots from culture wereinoculated with A. tumefaciens EHA101 (pGUS3), or EHA105(pSSLa.3), subjected to selection and regenerated. Shoots that survivedexhibited expression of the uidA gene (GUS) in a patterncharacteristic of the either the CaMV35S promoter (pGUS3), or the larch RbcSpromoter (pSSLa.3) transferred. Recovered plants were screened using PCRamplification. Southern DNA analyses and amplification of the T-DNA borderjunction confirmed genomic integration of both transferreduidA and nptII genes. In this proofofconcept study, the overall recovery of P. taeda shoots wasfair (10–20%), while recovery of intact rooted plants was poor (>1%)due to difficulty in rooting. Recovery of intact rooted plants from inoculatedshoots of P. eldarica and P. radiatawas more efficient (10–30%). The addition of a shoot multiplication stepand effective rooting protocols will improve the efficiency of this genotypeindependent transformation method in P. taeda, and inotherPinus spp.     

Correlations among indices of forest soil nutrient availability in fertilized and unfertilized loblolly pine plantations

Summary Various laboratory indices of N and P availability in forest soils correlated poorly among themselves and with on-site ion exchange resin (IER) estimates in both unfertilized and N+P fertilized loblolly pine plantations. IER nutrient availability estimates had greatest within-site variability than laboratory indices. Net nitrification was minimal in laboratory incubation of the mineral soil despite high rates of ammonification. In contrast, IER NO₃–N values were usually of the same magnitude as IER NH₄–N values. In both fertilized and unfertilized stands, at least one N availability index was negatively correlated with at least one P index. Soil N and P availabilities were generally higher on fertilized plots than on unfertilized plots 3.5 years after fertilization, and IER estimates showed the greatest number of plots with increased levels. The greater ability of the IER method to distinguish between fertilized and unfertilized plots indicated the method was affected by on-site factors that the laboratory methods do not assess.     

Recovery from winter depression of photosynthesis in pine and spruce

Summary Recovery from winter depression of photosynthesis was studied in Pinus sylvestris, Pinus conforta and Picea abies by means of chlorophyll fluorescence and gas exchange measurements. During the winter 1986–1987 the fluorescence yield was low and no variable fluorescence was detectable before the end of March. In the field recovery of variable fluorescence/maximum fluorescence (F_v/F_m) during spring was slow for all three species studied. The temperature dependence of recovery was confirmed from measurements of the potential rate of recovery of F_v/F_m at different temperatures in the laboratory. At 20° C, F_v/F_m increased from 0.1 to 0.8 within 3 days. Recovery of F_v/F_m was paralleled by an increase in apparent photon yield. No significant differences could be demonstrated between the studied tree species in potential rate of recovery in the laboratory or in actual recovery in the field.     

Induced defenses of loblolly pine, Pinus taeda: potential impact on Dendroctonus frontalis within-tree mortality

Loblolly pine (Pinus taeda L.) produces an induced defensive hypersensitive response in inner bark colonized by the southern pine beetle, Dendroctonus frontalis Zimm. (Coleoptera: Scolytidae), and its associated fungi. Adult beetles forced to colonize the induced response tissue in a laboratory study constructed galleries that were the same length as adults boring in normal phloem. However, each female laid fewer eggs in the induced tissue. Larval and pupal mortality were also higher in this tissue when compared to surrounding phloem. Beetles colonizing trees in response to pheromone baits constructed less gallery and laid fewer eggs in induced tissue than in surrounding normal phloem. These results suggest that the lesions produced by the induced defense system in conifers may not only contain the growth of fungi inoculated into trees during the attack phase of beetle colonization, but may also affect survival of bark beetle progeny.

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Résumé Une réaction hypersensible de défense a été induite dans les couches profondes de l'écorce de P. taeda colonisée par D. frontalis et le champignon qui lui est associé. Au laboratoire, des adultes, contraints de coloniser les tissus où une réaction hypersensible e été induite, ont foré des galeries de même longueur que celles creusées dans du phloème sain. Cependant les femelles ont pondu moins d'oeufs dans les tissus induits; les mortalités larvaires et nymphales étaient aussi plus fortes dans ce tissu que dans le phloème voisin. Les scolytes, ayant colonisé les arbres après attraction par des pièges à phéromones, forent moins de galeries et pondent moins d'oeufs dans le tissu induit que dans le phloème sain voisin. Ces résultats suggèrent que les lésions, provoquées par le système de défense induit des conifères, peuvent non seulement limiter la croissance du champignon inoculé dans l'arbre au cours de la colonisation des scolytes, mais aussi affecter la survie des descendants dans l'écorce.

     

Interactive effects of soil nitrogen and atmospheric carbon dioxide on root/rhizosphere carbon dioxide efflux from loblolly and ponderosa pine seedlings

We measured CO₂ efflux from intact root/rhizosphere systems of 155 day old loblolly (Pinus taeda L.) and ponderosa (Pinus ponderosa Dougl. ex Laws.) pine seedlings in order to study the effects of elevated atmospheric CO₂ on the below-ground carbon balance of coniferous tree seedlings. Seedlings were grown in sterilized sand culture, watered daily with either 1, 3.5 or 7 mt M NH ₄ ⁺ , and maintained in an atmosphere of either 35 or 70 Pa CO₂. Carbon dioxide efflux (mol CO₂ plant^–1 s^–1) from the root/rhizosphere system of both species significantly increased when seedlings were grown in elevated CO₂, primarily due to large increases in root mass. Specific CO₂ efflux (mol CO₂ g root^–1 s^–1) responded to CO₂ only under conditions of adequate soil nitrogen availability (3.5 mt M). Under these conditions, CO₂ efflux rates from loblolly pine increased 70% from 0.0089 to 0.0151 mol g^–1 s^–1 with elevated CO₂ while ponderosa pine responded with a 59% decrease, from 0.0187 to 0.0077 mol g^–1 s^–1. Although below ground CO₂ efflux from seedlings grown in either sub-optimal (1 mt M) or supra-optimal (7 mt M) nitrogen availability did not respond to CO₂, there was a significant nitrogen treatment effect. Seedlings grown in supra-optimal soil nitrogen had significantly increased specific CO₂ efflux rates, and significantly lower total biomass compared to either of the other two nitrogen treatments. These results indicate that carbon losses from the root/rhizosphere systems are responsive to environmental resource availability, that the magnitude and direction of these responses are species dependent, and may lead to significantly different effects on whole plant carbon balance of these two forest tree species.     

Water status and growth of loblolly pine (Pinus taeda L.) callus

Water status of Pinus taeda L. callus supported on Murashige and Skoog (MS) liquid medium was characterized over an 8 week period using thermocouple psychrometry. Medium with 30 gl⁻¹ sucrose was used to produce a high water potential (Ψ_w) of −0.4 MPa (H), and the same medium was used to create a moderate Ψ_w of −0.7 MPa (M) by the addition of 10% polyethylene glycol (PEG, w/v, MW=8000). Calli were produced from cotyledon explants on H medium for 2 weeks and then transferred to either M or H medium. Callus absorption of PEG accounted for 40% of the callus dry weight and less than 7% of the callus fresh weight. Callus dry weight (without the PEG fraction) on M medium was 40% of that observed on H medium. Fresh weight on M medium was only 15% of that observed on H medium. The Ψ_w of both H and M media remained constant throughout the culture period. On H medium, callus Ψ_w and osmotic potential (Ψ_s) both increased 0.05 MPa/week with the callus Ψ_w approaching that of the external medium. On M medium, callus Ψ_w and Ψ_s both decreased more than 0.1 MPa/week with the callus Ψ_w decreasing greatly below that of the external medium. The latter was attributed to a rapidly produced osmotic shock induced upon callus transfer and/or PEG which caused less callus hydration and resulted in reduced growth. Callus turgor potential (Ψ_p) was estimated to be +0.02 to +0.09 MPa and turgor was maintained as callus Ψ_w increased or decreased. After 8 weeks, cell volumes from callus on M medium were 50 to 60% less than on H medium, suggesting that reduced cell volumes were related to turgor maintenance.     

In vitro propagation of loblolly pine via direct somatic organogenesis from mature cotyledons and hypocotyls

A high-efficiency two-step culture procedure for direct somaticorganogenesis in loblolly pine (Pinus taeda L.) resulting inthe formation of multiple shoot structures induced on cotyledons andhypocotyls of mature zygotic embryos is described. Mature zygoticembryos of eight genotypes of loblolly pine were used as explants toinduce direct somatic organogenesis with this two-step culture method,involving the induction and the differentiation of direct adventitiousshoots. After mature zygotic embryos of eight genotypes of loblolly pinewere cultured on induction medium containing 2,4-dichlorophenoxyaceticacid (2,4-D) or -naphthaleneacetic acid (NAA), 6-benzyladenine(BA), and kinetin for 2–3 weeks, embryos were transferred todifferentiation medium. Adventitious shoot regeneration via directsomatic organogenesis with the frequency of 8.7–27.8% wasobtained from mature zygotic embryo cultures of the genotypes tested.The highest mean number of 32.6 adventitious shoots per mature zygoticembryo was produced from genotype La. The tissue culture protocol of invitro shoot regeneration via direct somatic embryogenesis was optimizedafter examining the periods of the induction culture, chillingtreatment, glutamine concentration, and basic medium levels. Rooting wasachieved on TE medium supplemented with 0.5 mg/l indole-3-butyric acid(IBA), 0.5 mg/l gibberellic acid (GA₃), and 1 mg/l6-benzyladenine (BA), and regenerated plantlets were established insoil. These results suggested that adventitious shoot regeneration viadirect somatic organogenesis could be useful for clonal micropropagationof some genotypes of loblolly pine and for establishing a transformationsystem of this coniferous species.     

Effects of water stress on gas exchange of field grown <Emphasis Type="Italic">Zea mays</Emphasis> L. in Southern Italy: an analysis at canopy and leaf level

Zea mays is cultivated in the Mediterranean regions where summer drought may lead to photoinhibition when irrigation is not available. In this work the response of maize to water stress was evaluated by gas exchange measurements at the canopy and leaf level. Leaf gas exchange was assessed before, during and after water stress, while canopy turbulent fluxes of mass and energy were performed on a continuous basis. In the early growth period, a linear increment of net ecosystem photosynthetic rate (P _NE) to incoming of photosynthetic photon flux density (PPFD) was found and net leaf photosynthetic rate (P _NL) showed the tendency to saturate under high irradiance. During water stress, the relationship between P _NE and PPFD became curvilinear and both P _NE and P _NL saturated in a range between 1,000 and 1,500 μmol (photons) m⁻² s⁻¹. Leaf water potential (ψ_l) dropped from −1.50 to −1.88 MPa during water stress, indicating that leaf and canopy gas exchanges were limited by stomatal conductance. With the restoration of irrigation, P _NE, P _NL and ψ_l showed a recovery, and P _NE and P _NL reached the highest values of whole study period. Leaf area index (LAI) reached a value of 3.0 m² m⁻². The relationship between P _NE and PPFD remained curvilinear and P _NE values were lower than those of a typical well-irrigated maize crop. The recovery in P _NE and P _NL after stress, and ψ_l values during stress indicate that the photosynthetic apparatus was not damaged while soil moisture stress after-effects resulted in a sub-optimal LAI values, which in turn depressed P _NE.     

Relationship of aluminium and calcium to net CO2 exchange among diverse Scots pine provenances under pollution stress in Poland

Light-saturated net photosynthesis (A_sat), dark respiration (RD), and foliar nutrient content of eight European Scots pine (Pinus sylvestris L.) provenances were measured at experimental sites in western Poland. Two-year-old seedlings were planted in 1984 at two sites with similar soils in areas of contrasting air pollution. One site was near a point source of SO₂ and other pollutants, and another 12 km to the southeast in an area free of acute air pollution was treated as a control. The eight provenances were from a large north-tosouth latitudinal range (60 to 43° N). At the heavily polluted site Scots pine trees exhibited lower growth rates and crown dieback and deformation. Soil pH, Ca and Mg were at least 10 times lower, and Al 10 times higher at the polluted than the control site. In 1991, concentrations of Al, P, Ca, S, Mn, Fe, and Zn in oneyear old Scots pine foliage were higher and Mg lower at the polluted than control site. At both sites foliar Mg levels were within the range considered deficient (0.6 mg g^-1), and at the polluted site, Al concentrations were very high (670 to 880 g g^-1). In all provenances, RD of one-year-old needles was higher (by 22% on average) and A_sat was lower (by 37% on average) at the polluted than the control site. The ratio of A_sat: RD was half as great in all provenances at the polluted (4 to 6) than control site (8 to 11). Provenances of southern origin had greater increases in RD and water-use efficiency at the polluted site than other provenances. Within the polluted site alone, or across both sites, A_sat in Scots pine was negatively correlated to the Al: Ca ratio (p<0.001, r=–0.93). Across sites RD increased with needle N and Al (multiple regression, p<0.001). The data suggest that at the polluted site there is excessive soil Al and deficient Mg availability, low needle Mg and high Al concentrations and high Al: Ca ratios, and that these have resulted in reduced photosynthetic capacity and increased respiration.     

Determinants of woody species richness in Scot pine and beech forests: climate, forest patch size and forest structure

We analysed patterns of woody species richness in Pinus sylvestris and Fagus sylvatica forests in Catalonia (NE Spain) from forestry inventory databank in relation to climate and landscape structure. Both types of forests are found within the same climatic range, although they have been managed following somewhat different goals. Overall, woody species richness significantly increased when conditions get closer to the Mediterranean ones, with milder temperatures. Differences between the two types of forests arose when comparing the relationship between richness and forest patch size. Woody species richness increased in pine forests with patch size, while the opposite trend was observed in beech forests. This pattern is explained by the different behaviour of structural canopy properties, since leaf area index and canopy cover showed a steeper increase with increasing forest patch size in Fagus forests than in Pinus ones. Accordingly, richness decreased with canopy cover in Fagus plots, but not in Pinus ones. We suggest that these differences would be related to management history, which may have enhanced the preservation of beech stands in larger forest landscape units.     

Spatial variation in sapwood area to leaf area ratio and specific leaf area within a crown of silver birch

Spatial variation in sapwood area to leaf area ratio (Huber value, HV) and specific leaf area (SLA) was examined in branches of closed-canopy trees of silver birch (Betula pendula Roth). HV increased basipetally within a crown and decreased with increasing branch order, but exhibited no significant radial trend along a primary branch. HV was primarily determined by branch position in a crown and branch diameter at the sampling point, being independent of the size of the tree and branch. Greater HV in the lower-crown branches is considered a means to mitigate differences in hydraulic transport capacity between the branches located in different canopy layers. Beside branch position and sampling location on a branch, SLA depended significantly on several other variables characterising tree and branch size. SLA increased basipetally within a crown and along a primary branch, but exhibited no significant trend with branch orders. Because height caused leaf area (A_L) to diminish more rapidly than leaf dry weight, A_L primarily determined the vertical variation in SLA.     

Patterns of cytokinins and leaf gas exchange among canopy layers of a mature sugar maple (<Emphasis Type="Italic">Acer saccharum</Emphasis>) stand

Leaf cytokinins (CKs) were profiled within four locations throughout the inner and outer layers of a mature sugar maple (Acer saccharum) canopy. Leaf CK was associated with leaf gas exchange activity and some corresponding microclimate variables. Both inner and outer layers in the upper canopy had higher concentrations of leaf CKs than the lower canopy layers and the difference was comprised primarily by riboside forms of CK. Transpiration (E) showed a similar pattern to leaf CK content, with significantly higher rates in the upper canopy. There was, however, no clear pattern discernable in stomatal conductance (gs), other than it tended to be higher in the outer canopy layers. The upper/outer canopy showed a significantly different environment than all other canopy positions with higher photosynthetically active radiation (PAR), ultra-violet light (UV-B) and leaf temperature. Simple linear regression analysis showed that the nucleotide CK group (including iPNT, cis- and trans-[9RMP]Z, [9RMP]DZ) was positively related to PAR. Exogenous applications of benzylaminopurine (BAP), showed that low concentrations of BAP reduced E and g _s, and indicated that CK may help regulate stomatal aperture. The similar patterns in E and CK content suggest that CKs and leaf gas exchange are functionally connected.     

Regulation of loblolly pine (Pinus taeda L.) arginase in developing seedling tissue during germination and post-germinative growth

After seed germination, hydrolysis of storage proteins provides a nitrogen source for the developing seedling. In conifers the majority of these reserves are located in the living haploid megagametophyte tissue. In the developing loblolly pine (Pinus taeda L.) seedling an influx of free amino acids from the megagametophyte accompanies germination and early seedling growth. The major component of this amino acid pool is arginine, which is transported rapidly and efficiently to the seedling without prior conversion. This arginine accounts for nearly half of the total nitrogen entering the cotyledons and is likely a defining factor in early seedling nitrogen metabolism. In the seedling, the enzyme arginase is responsible for liberating nitrogen, in the form of ornithine and urea, from free arginine supplied by the megagametophyte. In this report we investigate how the seedling uses arginase to cope with the large arginine influx. As part of this work we have cloned an arginase cDNA from a loblolly pine expression library. Analysis of enzyme activity data, accumulation of arginase protein and mRNA abundance indicates that increased arginase activity after seed germination is due to de novo synthesis of the enzyme. Our results suggest that arginase is primarily regulated at the RNA level during loblolly pine seed germination and post-germinative growth.