Effect of open-air fumigation with sulphur dioxide and ozone on phyllosphere and endophytic fungi of conifer needles

The phyllosphere microbial populations inhabiting the needles of three conifer species, Scots pine (Pinus sylvestris L.), Sitka spruce (Picea sitchensis L.) and Norway spruce (Picea abies (L.) Karst.), exposed to SO₂ and O₃, in an open-air fumigation experiment were analysed over a 3 year period using serial dilution after washing, direct plating and a fluorescein diacetate (FDA) enzyme assay. Total fungal populations ranged from 10² to 10⁵ colonyforming units (CPU) g^?1 fresh weight of needles. The dominant fungi isolated from needles varied with tree species and isolation technique; Aureobasidium pullulans (de Bary) Arnaud was most common on Scots pine and Norway spruce and white yeasts on Sitka spruce using the dilution plating method. However, direct plating of needle segments onto culture media indicated that Sclerophoma pythiophila (Corda) Hohnel was dominant on Scots pine and A. pullulans on Sitka and Norway spruce. Green needles of Sitka spruce were found to be endophytically colonized by Rhizosphaera kalkhoffii Bubak, but seldom by Lophodermium piceae (Fuckel) Hohn during extensive sampling in 1990. Statistical analyses revealed significant differences (P<0.05) between plots in the 3 year mean of the total fungal populations or the fungal biomass (FDA assay) on all three tree species. Differences between plots were also observed for a number of dominant component species. Data were also analysed for treatment effects. A significant effect of SO₂ treatment was observed on the total fungal populations on Sitka spruce (P<0.05) which were reduced markedly by the low-SO₂ treatment, while the O₃ treatment caused a significant increase in total fungal numbers on Scots pine (P<0.05). The FDA activity on needles of both Scots pine and Sitka spruce was noticeably higher in the 0₃-only treatment plot, but the overall O₃ effect was not significant. Treatment effects were also detected on the occurrence of component species. The serial dilution method revealed an SO₂ effect (P<0.05) of a reduction in the occurrence of pink yeasts on Sitka spruce and an O₃ effect (P<0.05) of an increase in the occurrence of S. pythiophila on Sitka spruce (P<0.01) but a decrease of Epicoccum nigrum Link and Cladosporium spp. on Scots pine. The direct-plating method revealed an SO₂ effect of an increase in S. pythiophila on Norway spruce (P<0.05). Ozone treatment caused a significant increase in the isolation of a black strain of A. pullulans on Norway spruce (P<0.05). Endophytic colonization of Sitka spruce needles by R. kalkhoffii was found to be increased on two occasions by O₃ exposure.     

Scarification,fertilization and herbicide treatment effects on planted conifers and soil fertility

The influences of soil surface modification (blade scarification and plastic mulching), fertilization and herbicide application on soil nutrient and organic carbon content and tree growth and foliar nutrient status were examined after seven years in a study located within the Great Lakes-St. Lawrence forest region of Canada. Plots had been planted with white pine (Pinus strobus L.) and white spruce (Picea glauca [Moench] Voss) seedlings. Light (PAR), soil moisture and temperature were monitored and recorded throughout the growing season. Forest floor and soil mineral (0–20 cm layer) samples were collected from all experimental plots, except those which had plastic mulching. Foliar samples were collected in autumn and analysed for N, P and K and storage compounds. Seedling mortality was 20% higher in unscarified plots. Combined silvicultural treatments increased productivity as much as 14 times, but scarification reduced soil carbon and nutrient capital 2–3 fold. Herbicide application reduced soil carbon by at least 20 %. Foliar nutrient, protein, starch and lipid contents in autumn were little affected by treatment. The future management of such stands in Canada probably will include more shelterwood harvesting and crop rotations, silvicultural systems that are more closely aligned with natural forest succession.     

Response of understorey vegetation and Scots pine root systems to fertilization at multiple deficiency stress

The stump and root systems of Scots pine (Pinus sylvestris L.) and field-layer vegetation were sampled before and three growing seasons after drainage and fertilization of a low-shrub pine bog in SE Norway. Although the understorey vegetation roots responded significantly to nutrient application with higher concentrations of Ca and P, root biomass weights did not change. The fine and small pine roots responded with higher N, Ca, P and S concentrations, while those of Mn and Zn were significantly reduced. The NPK-application resulted in significantly higher pine root biomass. Relative to the total stores in the root zone the amounts of most elements in roots shifted to higher ratios at NPK-application. High figures for K, B and Mn indicate tight biochemical cycles of these elements. Compared to totals in above and below ground biomass, major parts of Fe and Pb were held by the roots. In contrast the field layer roots kept a very small per cent of total K, while the pine roots were low in Mn. The understorey vegetation was primarily restricted by P-deficiency, while the pine trees were also restricted by low supply of N. The field and the tree layer species seem to differ with respect to required nutrient concentrations in the root zone. These characteristics are important for direction and extent of successional changes after fertilization in low-shrub pine bog ecosystems.     

Uptake of 24Mg by excised pine roots: A preliminary study

Uptake of ²⁴Mg by excised roots of Pinus sylvestris L. during up to 4 h long incubations in 99.9 atom % ²⁴Mg (50 M) was measured by ICP-MS. A rapid initial uptake phase (30 min) was followed by a slower uptake. This was interpreted as a shift from a phase dominated by saturable ion exchange (free space uptake), to a non-saturable phase, during which the rate of uptake was 0.077±0.0.012 mol Mg g^–1 (d.wt.) h^–1. The metabolic uncoupler DNP (2,4-dinitrophenol) at 50 M decreased the Mg uptake rate by 35% only, but the effect of DNP was significant (p<0.01). Several problems related to a high variability in the experimental material were encountered, and further refinement of this approach in studies of plant Mg uptake is suggested.     

Two New Species of Aphelenchoididae (Nemata) from Declining Red Pines in Maryland

Aphelenchoides resinosi n. sp. and Ektaphelenchus joyceae n. sp. are described and illustrated from red pines of the Allegheny plateau of Maryland, USA. The new species were found in trees infested with Bursaphelenchus xylophilus. Primary diagnostic characters of A. resinosi females are constriction of the head, basal stylet knobs, tong postuterine sac, two incisures in the lateral field, and conical tail four to five anal body widths long with a simple terminal mucro. Diagnostic characters of the males are two pairs of subventral caudal papillae and spicule shape: Primary diagnostic characters of E. joyceae females are a slight constriction of the head, six similar lips, conical tail, and short postuterine sac. Diagnostic characters of the males are spicule size and shape, a single row of spermatocytes, and one pair of caudal papillae. Within-tree distributions of A. resinosi and E. joyceae are presented. A total of 70% of both red-needled and chlorotic-needled trees in the study were positive for A. resinosi and E. joyceae. Branch hierarchy was related to the percentage of samples positive for A. resinosi.     

THE IMPACT OF TREE SQUIRRELS (TAMIASCIURUS) ON LIMBER PINE SEED DISPERSAL ADAPTATIONS

Tree squirrels (Tamiasciurus) are important selective agents on conifer reproductive strategies (Smith 1970, 1975). Although this is well established for wind-dispersed pines, the impact of tree squirrels on bird-dispersed pines has been largely ignored. I assessed the impact of tree squirrels on the allocation of reproductive energy in the bird-dispersed limber pine (Pinus flexilis) by comparing its cone and seed traits from three sites in the Rocky Mountains where tree squirrels (Tamiasciurus) are present to those from three mountain ranges in the Great Basin where tree squirrels are absent. As predicted, differences between the two regions in individual cone and seed traits are consistent with the hypothesis that tree squirrels are important selective agents on these traits. In the absence of tree squirrels, limber pine allocates more than twice as much energy to kernel compared with that invested in putative seed defenses (cone, resin, and seed coat) as does limber pine where tree squirrels are present. Such a large difference is particularly striking, because tree squirrels may have become extinct in the Great Basin in only the last 12,000 yr. Although many factors influence the allocation of energy to cones and seeds, no single factor other than the presence of tree squirrels is compatible with the large and consistent differences between limber pine in the Rocky Mountains and Great Basin. These results show that tree squirrels are an important constraint on the evolution of cone and seed traits that promote the dispersal of seeds by birds.     

Development of needle retention in Scots pine (Pinus sylvestris) in 1957–1991 in northern and southern Finland

The needle trace method was used to study retrospectively the long-term latitudinal variation in needle retention in Scots pine (Pinus sylvestris L.) in Finland. The mean annual summer needle retention (ANR) along the main stem varied from 3.4 to 6.0 needle sets during the period 1957–1991. The lowest values were observed in southern and the highest in northern Finland. The length of the growing season, expressed as the thermal sum (threshold value +5 °C), was negatively correlated with the mean ANR (r=-0.96). The geographical needle retention pattern (NRP) for the period 1957–1991 showed a clearly increasing trend from 1957 to 1969 (southern Finland) and to 1975 (northern Finland); thereafter, the NRP tended to decrease close to its minimum value recorded in 1991. The general level of the NRP was approximately 5.0 needle sets in northern Finland and 3.5–4.0 needle sets in southern Finland. The NRP, with its 6–12 year cycle for southern Finland, was clearly periodical. Differences in the NRP among the ten stands in southern Finland were small, whereas the said periodicity was missing and the differences were high among the stands in northern Finland. The results indicate that variation in the number of needle sets, viz. defoliation of pines, is a normal phenomenon. The role of net carbon assimilation as a regulator of the number of needle sets is discussed.     

Aluminium causes nutrient imbalance and structural changes in the needles of Scots pine without inducing clear root injuries

The effects of aluminium chloride (AICI3) treatments (50 and 150 mg/l) on 3-year-old Scots pine (Pinus sylvestris L.) seedlings were studied in a sand culture during 2 growing periods in an open field experiment. Even by the end of the first growing period, a decline was observed in the concentrations of Ca, Mg and P within the needles, and of Ca and Mg in the roots. After the second growing period, increased N and K concentrations were observed in the needles of Al-treated seedlings. Both the needles and roots of Al-treated seedlings showed, after the second growing period, a decline in growth and increased concentrations of AI as the amount of AICI3 in the nutrient solution increased. Al-induced changes in needle structure were found to be symptomatic of a nutrient imbalance, particularly of Mg and P. Al-stress did not result in any observable changes in root anatomy or in the number of mycorrhizas. Scots pine proved to be rather resistant to Al-stress, indicating that direct Al-injuries are not likely in the field, though Al-stress may be a contributing factor in the formation of nutrient imbalances.     

Chemical composition and ultrastructural changes in Scots pine needles in a forest decline area in southwestern Finland

Tree decline and deaths have been observed among 15 to 20-year-old Scots pines (Pinus sylvestris L.) in a dry heath forest in southwestern Finland. The sudden decline in height growth, the dieback of leading shoots and the yellowing of needles in young shoots in the upper part of the tree are typical symptoms of the decline of these young pines. Needle ultrastructure and chemical composition of Scots pines with or without decline and fluctuations of them in different seasons were studied. Afflicted trees were found to suffer from a deficiency in calcium and magnesium with low concentration of foliar nitrogen and phosphorus observed in all the trees studied. Ultrastructural study revealed changes characteristic of different seasons and measured nutrient status of needles. A clear reduction of membrane system in chloroplasts, especially related to Mg deficiency, was observed in most samples. The symptoms related to N deficiency, the translucent appearance of the cytoplasm and chloroplast stroma, and the elongated chloroplasts, as well as swelling of mitochondria, indicating P deficiency, were also found in the needles sampled from this forest decline area. The present study showed that it is possible to detect specific nutrient deficiency symptoms in needle ultrastructure in field samples and for use as sensitive indicators of unbalanced nutrient status.     

The morphological background to imbibition in seeds ofPinus sylvestris L. of different provenances

An examination was made of the structure of the coats of Scots pine (Pinus sylvestris L.) seeds of different provenance and the contribution of this factor to differences in imbibition. The seed coat layers derived from the integument, the sarcotesta, sclerotesta and endotesta did little to restrict imbibition, even though the sclerotesta of the northern provenance seeds was composed of a double multicellular layer and the sarcotesta contained large numbers of pigmented, phenol-bearing cells. In addition to the micropyle, the sclerotesta was found to possess structural openings at the chalazal end and at the ridge joining the two halves of the seed, but being covered by the pigmented cells of the sarcotesta, these did not allow water to enter any more than did the micropyle itself. Imbibition was chiefly regulated by the lipophilic covers surrounding the endosperm, which are mainly of nucellar origin, especially by the megaspore membranes nearest to the endosperm, the outer and inner exine. The nucellar cap covering the micropylar end of the endosperm proved to be impermeable to water, and its edge extended between the exine layers, which further enhanced the importance of the endosperm covers as regulators of imbibition.