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Rapid biochemical test for seed germinability.

Sugar and aminoacids were investigated in sterile distilled water after Pinus pinea seeds had been soaked in it during 24 hours at 37 degrees C. Germination viability decreased with the ageing of the seeds and was accompanied by sugar and aminoacids increase in the exudate. The sugar content from seeds with loss of germinability was about 50 to 80 times greater than that from seeds with good germinability. Paper chromatography showed that there was an increase in mono, di, oligosaccharides and amino-acids in seeds without germinability, but these substances were only found as trace quantities in viable seeds. The methods already described by TKAYANAGI and MURAKAMI to determine germinability could be applied with some modifications to Pinus pinea seeds. The presence of sugar in the exudate could be detected by urine sugar test after 24 hours at 37 degrees C. It was necessary to concentrate the exudate till a final volume of 2 ml.

Amino Acids↗

[Restoration of pine plantations after effect of ionizing radiation in the region of the accident at the Chernobyl Atomic Energy Station].

The main results of the 12-year radiation-genetic monitoring of radiobiological, cytogenetic, and genetic parameters in the Pinus sylvestris L. forest plantations from the zone of the accident at the Chernobyl nuclear power plant presented. Acute ionizing irradiation at doses > 1 Gy was shown to induce the formation of morphoses and depressed growth; at doses > 2 Gy, the reproductive ability of the trees declined. The radiobiological parameters showed a linear (or close to linear) dose-effect relationship. Acute irradiation at a dose of 0.5 Gy induced cytogenetic and genetic effects that were significantly higher than the corresponding control values. The relationship between the cytogenetic effects and the absorbed dose was exponential. The dependence of the mutation frequency at specific loci on the absorbed dose was described by a nonlinear curve. The results of cytogenetic analysis of sprouts obtained from seeds annually (1986-1998) collected in zones of slight, moderate, and strong damage of Pinus sylvestris L. are presented.

Chromosome Aberrations↗

[Niche characteristics of dominant populations in natural forest in north Guangdong].

Two communities of the natural evergreen forest in Luoba Nature Reserve, North Guangdong, each represented by a 2500 m2 plot, were surveyed with Tree-layer-frequency method. Based on these data, the niche characteristics of 12 dominant populations, i.e., Castanopsis fargesii, Pinus massoniana, Schima superba, Liquidambar formosana, Diospyros morrisiana, Myrica rubra, Castanopsis lamontii, Adinandra milletii, Castanopsis eyrei, Elaeocarpus silvestris, Daphniphyllum calycinum, Neolitsea chuii, were described and analyzed, with stand layer as the resource state and tree abundance as the resource state descriptor of niche. The results indicate that most of the shade-tolerant tree species have higher niche breadth values, while the intolerant tree species have the lower ones, with zero value for Pinus massoniana and Liquidambar fomosana, which is in accordance with the declining status of the two species in the community. All the dominant populations, both shade-tolerant and intolerant, showed some adaptation to the community environment. 54.5% of the population-links had a niche similarity over 0.5, and 51.5% of them had a niche overlap over 0.2 in community 1. Niche characterization combined with conventional Tree-layer-frequency method could better determine the status and regenerating potential of tree species in the community. On the other hand, measuring tree abundance in different stand layers as the resource niche could demonstrate tree's requirement for the habitat. Variations in tree composition result in varied niche breadth, which lead to the variations of niche similarity and niche overlap values.

Algorithms↗

Airborne pollen calendar of Izmir - Turkey.

A qualitative and quantitative evaluation of pollen concentration in the atmosphere of Izmir metropolitan area is presented. Investigations were undertaken from November 1998 to October 1999 using gravimetric method. The aeroplynological studies revealed that at lower level (1.60 m) pollen from 29 woody and 30 herbaceous taxa were determined, whereas at higher level (20 m) pollen from 26 woody and 24 herbaceous taxa were recorded. At lower level, dominating woody species were Pinus, Quercus, Oleaceae, Cupressaceae/Taxaceae; herbaceous species were Gramineae, Chenopodiaceae/ Amaranthaceae, Cruciferae and Plantago. At higher level, Pinus, Cupressaceae/ Taxaceae, Gramineae, Chenopodiaceae/Amaranthaceae, Cruciferae and Plantago pollens were dominant. Morus, Oleaceae, Quercus, Chenopodiaceae/Amaranthaceae, Gramineae and Plantago showed highest concentration and their allergic degree was very high. Annual pollen calendar of the area is shown.

Environmental Monitoring↗

[Ecological distribution of macrofungi in Qingliang Mountain Natural Reserve, Anhui Province].

The north side of Qingliang Mountain Natural Reserveis is very rich of macrofungi resources. There are 160 species, belonging to 72 genera, 38 families, 10 orders and 2 classes, and distributing in the evergreen broadleaf forest, Pinus massoniana forest, evergreen-decidous mixed broadleaf forest, and Pinus taiwanensis forest. According to their economic value, these macrofungi may be grouped into two types: edible (72 species) and medicinal (55 species); and according their ecological habitat, they may be grouped into lignicolous (68 species), entomophilous (84 species), and ectomycorrhizal (21 species). Our study showed a certain regularity of their distribution, i.e., it varied with the ecological conditions of forests, tree species, soil types and altitude. Many economical macrofungi in this region could serve as one of the important natural resources, and had great potential usage on the edibility, medicinal and forestation.

Conservation of Natural Resources↗

[Dynamics of biomass and net primary productivity in succession of south subtropical forests in southwest Guangdong].

Coniferous forest (Pinus massoniana community), pine-borad leaved mixed forest (Pinus massoniana + Castanopsis kawakamii + Schima superba + Liquidambar formosana) and evergreen broad-leaved forest (Ixonanthes chinensis + Artocarpus styacifolius + Ormosia glaberrima + Cryptocarya concinna) are the three main communities representing 3 major stages in a secondary succession series in Heishiding Nature Reserve, Guangdong Province. Their biomass and net primary productivity (NPP) were studied by using harvest method (for trees and lianas) and clear cut method (for shrub and herb). The biomass and NPP were 246.697 t.hm-2 and 14.715 t.hm-2.yr-1 for the coniferous forest, 287.367 t.hm-2 and 17.179 t.hm-2.yr-1 for the pine-broad leaved mixed forest, and 357.976 t.hm-2 and 18.730 t.hm-2 yr-1 for the evergreen broad-leaved forest, respectively. These results indicated that these three stages were very close in the succession process, and that coniferous forest and mixed forest were more mature, while broad-leaved forest was relatively young. Therefore, under the conditions of no or only minor disturbance, their biomass and NPP showed an increasing trend with the succession of the forest communities in Heishiding.

Biomass↗

[Niche characteristics of plant populations in Quercus aliena var. acuteserrata stands in Qinling Mountains].

Based on the data collected from 31 plots and using Levins, Hurlbert and Pianka formulas, this paper calculated and analyzed the niche breadths and overlaps of 24 tree and 29 shrub populations in Quercus aliena var. acuteserrata stands in Mt. Qinling, Shaanxi. The results showed that the Levins and Hurlbert's niche breadths of Quercus aliena var. acuteserrata , Pinus armandi, Toxicodendron vernicifluum, Populus davidiana and Pinus tabulaeformis in arbor layer were 3.695, 1.695, 1.325, 0.840, 0.702 and 0.036, 0.299, 0.568, 1.721, 2.701 respectively, and those of Corrylus heterophylla, Lespedeza formosa and Smilax vaginata in shrub layer were 1.833, 1.466, 0.984 and 0.111, 0.300, 0.594, respectively. In general, the niche overlaps between the species with bigger niche breadth and other species were higher, and those of the species having similar environmental requirements were also higher.

Biodiversity↗

[Effects of different vegetation restoration of degraded red soil on earthworm population dynamics].

This study was conducted at the long-term experimental plots in Ecological Experimental Station of Red Soil in Yujiang County (28 degrees 15'30''N, 116 degrees 55'30''E ), Jiangxi Province, subtropical China. Earthworm population was investigated seasonally from May 1999 to February 2000, under different vegetations including four artificial woodlands [deciduous broadleaf woodland (Quercus chenii, Qc), evergreen broadleaf woodland (Schima superba, Ss), coniferous woodland (Pinus massonina, Pm) and mixed woodland (Schima superba-Pinus massonina, Sm)], two grasslands [gently-disturbed grassland (G1), undisturbed grassland (G2)] and control wasteland (CK). The results indicated that the population structure was very simple. Only Drawinda gisti characterized by pioneer was found. The seasonal averages of density and biomass were in the order of G2 > G1 > Qc > Ss > Pm > Sm > CK, and those of G2, G1 and Qc were significantly higher than those of the latters (P < 0.05). Seasonal fluctuations were obvious with dry-hot summer depressing the earthworm population sharply, leading to the aestivation of earthworm. Based on the variation coefficients of density and biomass, Qc had the highest ecosystem stability, followed by Sm and Ss, and G1, G2, and Pm had the lowest stability. The overall differentiation of earthworm population could be drawn through canonical discriminant analysis. There were significant correlations between earthworms and some soil properties (P < 0.01). Overall, the differentiation of earthworm population was driven by the quantity and quality of soil organic matter returned by the vegetations. Additionally, based on earthworm population, the importance of selecting appropriate vegetation types during the restoration of degraded red soil was emphasized.

Animals↗

Sensitization to common allergens, especially pollens, among children with respiratory allergy in the Trakya region of Turkey.

Asthma and allergic rhinitis are common problems in children and the causative pollen allergens vary according to the geographical area. The aim of this study was to investigate patterns of sensitization to common inhalant allergens, especially pollens, in Turkish children living in the Trakya region and to determine differences between rural and urban areas. Allergen skin testing was prospectively performed on 539 children aged between 4 and 17 years with respiratory allergy. The reaction was considered to be positive if the mean wheal diameter was at least 3 mm greater than that of the negative controls. We detected positive skin reactions in 420 (77.9%) children. Two hundred and eighty-one (52.1%) mite, 277 (51.4%) pollen, 174 (32.3%) mold, 65 (12.1%) animal dander, 12 (2.2%) cockroach and 6 (1.1%) latex skin sensitivities were detected. Among the pollen allergies 173 were cereal pollen (32.1%), 170 grass pollen (31.5%) and 144 tree pollen allergies (26.7%). The most common positive skin test among the pollens was to cultivated wheat (Titicum vulgare) (n = 116, 21,5%), followed by rye grass (Lolium perenne) and orchard grass (Dactylis glomerata). Positive skin reactions to Alternaria, to Candida albicans, and to all pollens except Ulmus competris, Pinus sylvetris, Platanus vulgaris and Tilia platyphyllos, were higher in children with allergic rhinitis than in those with asthma. In children from rural areas, allergic skin reactivity was found to be more common against Candida albicans, sheep dander and all pollens except Corylus avellana, Fraxinus excelsior, Populus alba, Pinus sylvetris, Platanus vulgaris and Chenopodium album, than in urban children. Although Trakya is close to Greece and other Mediterranean countries, this study suggests that the pollens, which sensitize children, are not similar.

Adolescent↗

[Methanol oxidation by the NAD-specific dehydrogenase of methane-assimilating yeasts].

The activity of NAD-specific dehydrogenase (NAD-DH) capable of methanol oxidation was found in cell-free extracts of methanol assimilating yeast cultures Pichia pinus and Candida boidinii. The optimum pH of the incubation mixture for the enzyme is 7.8--7.9; the value of Km of the enzyme for methanol is 1.4-10(-4) M in P. pinus and 1.04-10(-3) M in C. boidinii. NAD-DH is absent from the cells cultivated on glucose, and is induced with growth on a medium containing methanol.

Ascomycota↗

[Population dynamics of endangered plant species Abies chensiensis].

In order to know the endangered status and causes of Abies chensiensis in Qinlin Mountains, a field investigation on 18 plots was conducted on its age structure, life table and fecundity, and its population dynamics were predicted by time sequence model. The analysis on the age structure of Abies chensiensis populations showed that there were fewer young individuals, but middle-aged and old individuals were relatively rich. The population D in Abies chensiensis-Indigofera amblyantha-Carex lanceolata association showed a relatively stable development tendency, while other four populations (A, B, C and E) in Abies chensiensis-Pinus tabulaeformis-Sinarundinaria nitida-Carex lanceolata association, Abies chensiensis-Quercus aliena var. acutserrata-Litsea pungens-Carex lanceolata association, Abies chensiensis-Betula albo-sinensis-Sinarundinaria nitida-Duchesnea indica association, and Abies chensiensis-Pinus tabulaeformis-Smilax stans-Carex lanceolata association all showed an obviously declining tendency. The analysis on the life tables and survival curves showed that the survival curve of Abies chensiensis populations belonged to Deevey III, and the death peak of different populations was in the period of 60--100 years old. The number difference among populations reflected the population habitat. Time sequence prediction indicated the numbers of old individuals would be increased at the beginning, and decreased finally in 20, 40, and 80 years. It was difficult to maintain the population stability. Analysis on 10 ecological factors showed that tree coverage, soil organisms and air humidity influenced population positively, and human disturbance and sunlight influenced population negatively. In situ conservation should be taken as the most important management countermeasure, and natural regeneration should be promoted. At the same time, artificial population should be expanded.

Abies↗

Vitamin D and its metabolites in the pollen of pine. Part 5: Steroid hormones in the pollen of pine species.

Unconjugated vitamin D and its metabolites were investigated in the pollen of Pinus nigra Ar. and Pinus sylvestris L. by TLC, HPLC and competitive radiochemical determination of 25-hydroxycholecalciferol (25-OHD3). It was found that vitamin D (D2, D3) was present in the pollen in amounts about 2 micrograms/10 g and 25-OHD3, 24,25-dihydroxycholecalciferol [24,25-(OH)2D3] and 1,25-dihydroxycholecalciferol [1,25-(OH)2D3] between 0.1 and 3 micrograms/10 g of pollen, dependent on pollen species and methods.

Chromatography, High Pressure Liquid↗

The incidence of pine pollen reactivity in an allergic atopic population.

In the past there has been controversy concerning the allergenicity of pine pollen. Most reports have suggested that pine pollen has not been a significant allergen due to its low antigenicity and relatively large size. While little investigation has been done, there are some older studies supporting pine pollen as an allergen and its use in individual cases for immunotherapy. To our knowledge, no extended testing or screening had been done. A total of 200 patients presenting with symptoms during the pine pollen season were tested to Pinus radiata. Five (3.2%) of the 155 pollen-sensitive patients exhibited skin test sensitivity to Pinus radiata. Based on this data it is concluded that the incidence of skin test reactivity to pine pollen is low.

Humans↗

Single-copy, species-transferable microsatellite markers developed from loblolly pine ESTs.

Microsatellites, or simple sequence repeats (SSRs), are usually regarded as the "markers of choice" in population genetics research because they exhibit high variability. The development cost of these markers is usually high. In addition, microsatellite primers developed for one species often do not cross-amplify in related species, requiring separate development for each species. However, microsatellites found in expressed sequence tags (ESTs) might better cross-amplify as they reside in or near conserved coding DNA. In this study, we identified 14 Pinus taeda (loblolly pine) EST-SSRs from public EST databases and tested for their cross-species transferability to P. contorta ssp. latifolia, P. ponderosa, and P. sylvestris. As part of our development of a P. contorta microsatellite set, we also compared their transferability to that of 99 traditional microsatellite markers developed in P. taeda and tested on P. contorta ssp. latifolia. Compared to traditional microsatellites, EST-SSRs had higher transfer rates across pine species; however, the level of polymorphism of microsatellites derived from ESTs was lower. Sequence analyses revealed that the frequencies of insertions/deletions and base substitutions were lower in EST-SSRs than in other types of microsatellites, confirming that EST-SSRs are more conserved than traditional SSRs. Our results also provide a battery of 23 polymorphic, robust microsatellite primer pairs for lodgepole pine.

Alleles↗

Additional virulence genes and sonication enhance Agrobacterium tumefaciens-mediated loblolly pine transformation.

Additional virulence (vir) genes in Agrobacterium tumefaciens and sonication were investigated for their impact on transformation efficiency in loblolly pine (Pinus taeda L.). Mature zygotic embryos of loblolly pine were co-cultivated with disarmed A. tumefaciens strain EHA105 containing either plasmid vector pCAMBIA1301 or vector pCAMBIA1301 with an additional 15.8-kb fragment carrying extra copies of the Vir B, Vir C, and Vir G regions from the supervirulent plasmid pTOK47. pCAMBIA1301 contains hygromycin resistance and the beta-glucuronidase (GUS) reporter gene. Expression of GUS was observed after 3-6 days of co-cultivation, with peak expression at approximately 21 days. The highest numbers of GUS-expressing areas were visible up to 21 days after co-cultivation, declining rapidly thereafter. Both transient and stable transformation efficiencies increased when the explants were sonicated before co-cultivation and/or the additional virB, virC, and virG genes were included with the pCAMBIA1301 plasmid T-DNA. Use of the plasmid with additional vir genes and sonication dramatically enhanced the efficiency of Agrobacterium-mediated gene transfer not only in transient expression but also in the recovery of hygromycin-resistant lines. Stably transformed cultures and transgenic plants were produced from embryos transformed with A. tumefaciens EHA105 carrying pCAMBIA1301 or pCAMBIA1301+pTOK47 in the three families of loblolly pine. The presence of the introduced GUS and hygromycin phosphotransferase genes in the transgenic plants was confirmed by polymerase chain reaction and Southern hybridization analyses.

Agrobacterium tumefaciens↗

Improving loblolly pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology, germination, and gene expression.

Clonal production of loblolly pine ( Pinus taeda L.) through somatic embryogenesis has the potential to meet the increasing industrial demands for high-quality uniform raw materials. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Twenty-five newly initiated loblolly pine genotypes were followed through the process of liquid culture establishment, embryo maturation, germination, and retrieval from cryogenic storage. A maturation medium, capable of promoting the development of loblolly pine somatic embryos that can germinate, is presented that combines 1/2 P6 modified salts, 2% maltose, 13% polyethylene glycol 8000 (PEG), 5 mg/l abscisic acid (ABA), and 2.5 g/l Gelrite. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also described. A set of somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed. The quality of the resulting embryos was examined and compared to that of zygotic embryos using such parameters as morphology, dry weight, germination performance, and gene expression. All of the observations that were made support the conclusion that even with the new maturation medium somatic embryos grow approximately only halfway through the normal sequence of development and then prematurely discontinue growth.

Abscisic Acid↗

A promoter from the loblolly pine PtNIP1;1 gene directs expression in an early-embryogenesis and suspensor-specific fashion.

The PtNIP1;1 gene encodes an aquaglyceroporin that is expressed early in embryogenesis and appears to be expressed preferentially in the suspensor [V.T. Ciavatta et al. (2001) Plant Physiol 127:211-224]. An 899-bp fragment 5' to the PtNIP1;1 open reading frame (NIP(-899)) was cloned from loblolly pine (Pinus taeda L.) genomic DNA and fused to the beta-glucuronidase (GUS) reporter gene. The resulting plasmid, pNIP-GUS, was transformed into Norway spruce (Picea abies L.) embryogenic cultures by co-bombarding with a plasmid containing a bar gene construct as a selectable marker. The identity of lines selected on medium containing the herbicide Basta and showing beta-glucuronidase activity was confirmed by polymerase chain reaction as harboring GUS. Histochemical GUS assays of these lines revealed GUS activity in all cells of proembryogenic masses. During early embryogeny, GUS staining was intense in the suspensor region but not detectable in embryonal masses. GUS staining was absent by mid-embryogeny. By contrast, a control transgenic line, transformed with EuCAD-GUS, expressed GUS throughout embryo development. These results suggest that NIP(-899) contains elements that drive early embryogenesis-specific expression and suspensor-specific expression. This is the first example of a suspensor-specific promoter in conifers.

Aminobutyrates↗

Interactive effects of CO2 and O3 on a ponderosa pine plant/litter/soil mesocosm.

To study individual and combined impacts of two important atmospheric trace gases, CO2 and O3, on C and N cycling in forest ecosystems; a multi-year experiment using a small-scale ponderosa pine (Pinus ponderosa Laws.) seedling/soil/litter system was initiated in April 1998. The experiment was conducted in outdoor, sun-lit chambers where aboveground and belowground ecological processes could be studied in detail. This paper describes the approach and methodology used, and presents preliminary data for the first two growing seasons. CO2 treatments were ambient and elevated (ambient + 280 ppm). O3 treatments were elevated (hourly averages to 159 ppb, cumulative exposure > 60 ppb O3, SUM 06 approximately 10.37 ppm h), and a low control level (nearly all hourly averages <40 ppb. SUM 06 approximately 0.07 ppm h). Significant (P < 0.05) individual and interactive effects occurred with elevated CO2 and elevated O3. Elevated CO2 increased needle-level net photosynthetic rates over both seasons. Following the first season, the highest photosynthetic rates were for trees which had previously received elevated O3 in addition to elevated CO2. Elevated CO2 increased seedling stem diameters, with the greatest increase at low O3. Elevated CO2 decreased current year needle % N in the summer. For 1-year-old needles measured in the fall there was a decrease in % N with elevated CO2 at low O3, but an increase in % N with elevated CO2 at elevated O3. Nitrogen fixation (measured by acetylene reduction) was low in ponderosa pine litter and there were no significant CO2 or O3 effects. Neither elevated CO2 nor elevated O3 affected standing root biomass or root length density. Elevated O3 decreased the % N in coarse-fine (1-2 mm diameter) but not in fine (< 1 mm diameter) roots. Both elevated CO2 and elevated O3 tended to increase the number of fungal colony forming units (CFUs) in the AC soil horizon, and elevated O3 tended to decrease bacterial CFUs in the C soil horizon. Thus, after two growing seasons we showed interactive effects of O3 and CO2 in combination, in addition to responses to CO2 or O3 alone for a ponderosa pine plant/litter/soil system.

Atmosphere Exposure Chambers↗