Radiation-population monitoring of Pinus sylvestris L. in the zone of the Chernobyl power plant

Cytogenetic and genetic effects in populations of Pinus sylvestris L. suffered wiak, average, strong and sublethal radiation damage after the Chernobyl accident in 1986 were studied. The absorbed dozes for trees in these plantings were from 0.1 up to 20 Gy. It was shown that the amount of cells with chromosome aberrations in sprouts of seeds of a crop of 1993, are comparable with effects marked at once after accident in 1986. In 1997 and in 1998 the amount of cells with chromosome aberrations in sprouts of seeds in majority inspected plantings decreased to control values. The effect of adaptation was detected, when seeds of Pinus sylvestris L., gathered in 1997 from inspected trees, were exposed to additional 4 Gy gamma-radiation.     

Response of ectomycorrhizalPinus banksiana andPicea glauca to heavy metals in soil

Pinus banksiana andPicea glauca inoculated or not with the ectomycorrhizal fungusSuillus luteus were grown in a sandy loam soil containing a range of Cd, Cu, Ni, Pb and Zn concentrations. Ectomycorrhizal colonization rates were significantly reduced on Pinus and Picea seedlings by the heavy metals, particularly Cd and Ni. Needle tissue metal concentrations were lower in ectomycorrhizal seedlings at low soil metal concentrations. However, at higher soil concentrations, heavy metal concentrations of needle tissue were similar in ectomycorrhizal and nonmycorrhizal plants. The growth of nonmycorrhizal seedlings exposed to heavy metals was reduced compared to those inoculated withSuillus luteus. Apparently ectomycorrhizal colonization can protect Pinus and Picea seedlings from heavy metal toxicity at low or intermediate soil concentrations of Cd, Cu, Ni, Pb and Zn.     

Photosynthesis-Rubisco relationships in foliage of Pinus sylvestris in response to nitrogen supply and the proposed role of Rubisco and amino acids as nitrogen stores

Trees - Relationships between photosynthetic capacity, and needle contents of N, Rubisco and amino acids were investigated in potted Pinus sylvestris L. trees. Three-year-old seedlings of P....     

Rapid evolution towards heavy metal resistance by mountain birch around two subarctic copper-nickel smelters

Adaptations to pollution among long‐lived trees have rarely been documented, possibly because of their long reproductive cycles and the evolutionarily short timescales of anthropogenic pollution. Here, I present the results of a greenhouse experiment that suggest rapid evolutionary adaptation of mountain birch [Betula pubescens subsp. czerepanovii (Orlova) Hämet‐Ahti] to heavy metal (HM) stress around two copper–nickel smelters in NW Russia. The adaptation incurs a cost with reduced performance of adapted seedlings in pristine conditions. The industrial barrens around the studied smelters are extremely high‐stress sites with low seed germination and survival. It is likely that strong natural selection has eliminated all sensitive genotypes within one or two generations, with only the most tolerant individuals persisting and producing adapted seeds in the individual barrens. The results were similar from around both smelters, suggesting parallel evolution towards HM resistance.     

红花尔基自然保护区天然樟子松林种内种间竞争分析

运用Hegyi单木竞争指数分析了内蒙古红花尔基自然保护区天然樟子松（Pinus sylvestris var．mongolica Litv．）林内所有胸径大于2cm的樟子松、山杨（Populus davidiana Dode．）、白桦（Betula platyphylla Suk．）和山荆子（Malus baccata L．）的种内和种间竞争强度。结果表明,样地中主要的竞争木和对象木均为樟子松和白桦;樟子松的种内竞争强度（0．534）远大于种间竞争强度。随径级的增大,樟子松的种内竞争强度逐渐减小,且与胸径存在幂函数关系CI=A·D^-B。胸径达到30cm后,樟子松种内竞争强度变化不明显。作为竞争木,樟子松对其他树种产生了较大的竞争压力。     

The assessment of no adverse effect doses for plant populations chronically exposed to radionuclides of uranium and thorium decay series

Dose rates cause no adverse effects on natural populations of Pinus sylvestris L. and Vicia cracca L. inhabiting territories contaminated by uranium mill tailings and radium production wastes (Vodny settlement, Komi Republic) were determined. A significant increase in embryonic lethal mutation frequency in V. cracca legumes and decrease in seedlings survival rate as compared with control values were registered at dose rate equal to 1.67 mGy/day, that is 280 times higher than the one calculated for the reference site. The adverse effects in P. sylvestris expressed in increased frequency of chromosome aberrations in meristematic root tips and decreased reproductive capacity of seeds were determined at absorbed dose rate equal to 0.083 mGy/day. Data obtained show that the decrease in plant reproductive capacity in case of chronic exposure of radionuclides of uranium and thorium decay series can observe at lower weighted absorbed dose rates than in case of environmental contamination by artificial radionuclides.     

Isolation of functional RNA from plant tissues rich in phenolic compounds.

A method for the isolation of RNA from different tissues of trees (seedlings, saplings, and adult trees) is described. Using this procedure it is possible to remove large amounts of disturbing polyphenolic compounds from nucleic acids. The method involves an acetone treatment of the freeze-dried and powdered plant material, the use of high salt concentrations in the extraction buffer and an aqueous two-phase system. These steps were combined with the conventional phenol/chloroform extraction and CsCl centrifugation. The method has been successfully applied to the isolation and purification of RNA from pine (Pinus sylvestris L. and Pinus mugo Turr.), Norway spruce (Picea abies L.), and beech (Fagus sylvatica L.). The functional quality of RNA extracted by this procedure has been characterized by its uv spectrum, by agarose gel electrophoresis with ethidium bromide staining, Northern blot hybridization, and in vitro translation.     

Intraspecific variation in Heterobasidion annosum for mortality rate on Pinus sylvestris and Picea abies seedlings grown in pure culture

One-month-old Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) seedlings were inoculated in vitro with Heterobasidion annosum strains, four each of the P-, S- and F-intersterility groups. Variation among strains and between the IS groups in virulence, expressed in mortality rate, was detected during twelve months after inoculation. Most of the strains were more virulent on spruce than on pine, and mortality of spruce seedlings was significantly higher. The P strains displayed similar virulence on both hosts, while S strains caused higher mortality of spruce seedlings and significantly lower mortality of pine seedlings. Strains of the F group were less virulent, but killed significantly (P < 0.05) more spruce than pine seedlings. In the interspecific analyses with two hosts, the isolates and IS groups accounted for most of the explained variation in the host mortality.     

The effect of industrial pollution on Zinc,Cadmium and Copper concentration in the xylem rings of Scot's pine (Pinus sylvestris L.) and in the soil

Summary The concentrations of heavy metals, including zinc, cadmium, and copper were studied in the xylem rings of Pinus sylvestris trees and in the soil of three heavily polluted sites and one non-polluted site. There was a high correlation between the concentration of heavy metals in the soil and recently produced xylem rings. However, the initiation of industrial pollution did not abruptly increase heavy metal concentrations in the xylem rings. There was a high correlation between the zinc content of trees growing near a zinc smelter and the values obtained by other investigators for pine trees growing on the same site.     

Genetic effects of acute and chronic ionizing irradiation on Pinus sylvestris L., inhabiting the Chernobyl' meltdown area]

The main results of the 12-year radiation-genetic monitoring of radiobiological, cytogenetic, and genetic parameters in Pinus sylvestris forest plantation from the Chernobyl meltdown area are presented. The acute ionizing irradiation at doses > 1 Gy was shown to induce 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-dependence relationship. The 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 seedlings obtained from seeds annually collected in zones of slight, moderate, and strong damage of Pinus sylvestris L. are presented.     

Effect of air traffic pollution on seed quality characteristics of Pinus brutia

This study was undertaken to examine to what degree traffic air pollution affect the reproductive material (the seeds) of Pinus brutia. The study focused on the investigation of the seeds morphological characteristics and physiological attitudes as well as heavy metal concentrations of P. brutia trees grown close the ring-road of Thessaloniki (Greece's second biggest city), polluted by heavy traffic conditions, and how these characteristics are changed with the distance from the pollution source. A sampling procedure was established for seed collection from various distances from the pollution source. Seeds were properly prepared, measured for their morphological characteristics and heavy metal concentrations, as well as for their physiological behaviour (germination and viability). Analysis of the results showed that even though there were no significant differences in seed heavy metal concentrations, and only few in seed morphological characteristics, related to the distance from the traffic pollution source, there was a significant alteration of physiological behaviour for the seeds originated closed to the traffic. All the seed collected close to the ring-road (sample distances 0 and 30 m from the road) were unable to germinate, even those subjected to pre-treatments and repeated germination trials over longer duration, while those collected from distances over 100 m from the ring-road showed a common germination behaviour.     

呼伦贝尔沙地樟子松年轮生长对气候变化的响应

以内蒙古呼伦贝尔地区沙地樟子松为样本,建立了樟子松树木年轮宽度年表,应用相关分析和响应函数分析等年轮气候学方法,研究了樟子松径向生长对气候变化的响应。结果表明,樟子松年轮宽度与4月和6—9月平均温度呈显著负相关关系(P<0.05);与各月降水量多呈正相关关系,特别是与当年5—8月的月降水量呈显著正相关关系(P<0.05);树轮年表与前一年10月至当年10月的PDSI均呈显著正相关关系(P<0.05),其中与5月份PDSI的相关性最高。响应函数分析表明,年表与当年6—7月的平均气温、上一年10月和当年5—7月份的降雨存在显著的相关性,与5—7月份PDSI存在较显著的正相关性;综合来看,呼伦贝尔沙地樟子松生长同时受降水和温度的影响,其径向生长与气候因子间的关系属于降水敏感型,为区域降水重建提供了科学基础。     

Effect of industrial air pollution on wild plant seed germination and seedling growth

Germination of the following wild plant seeds was studied: bilberry (Vaccinium myrtillus L.), cowberry (V. vitis-idaea L.), bog bilberry (V. uliginosum L.), black crowberry (Empetrum hermaphroditum Hagerup), bearberry (Arctostaphylos uva-ursi (L.) Spreng.), bunchberry (Chamaepericlymenum suecicum (L.) Aschers. et Graebn.), cottongrass (Eriophorum polystachion L.), goldenrod (Solidago lapponica With.), fireweed (Chamaenerion angustifolium (L.) Scop.), marsh cinquefoil (Comarum palustre L.), cloudberry (Rubus chamaemorus L.), and Scots pine (Pinus sylvestris L.). The seeds were collected at different distances from the source of industrial air pollution (Severonickel smelter complex, Murmansk region). The task was the assessment of potential possibility of restoring corrupted north forest plant communities via seed propagation. By the response of reproductive structures to stressor, investigated species were divided into 3 groups: tolerant (members of the genus Vaccinium and Ch. angustifolium); moderately tolerant (C. palustre, E. polystachion, A. uva-ursi, and R. chamaemorus); and sensitive (P. sylvestris, E. hermaphroditum, and S. lapponica). Laboratory seed germinability of Vaccinium species was high (> 90%) regardless of the levels of Ni and Cu accumulation in the seeds and the index of technogenic load, whereas this index in E. hermaphroditum and P. sylvestris seeds was significantly reduced with the increase in the heavy metal contents in the seeds. The greenhouse experiments with the seeds of three Vaccinium species collected in the background area and in the zone of the highest pollution and germinated on the forest litter from the same sites and observation for seedling development allow us to conclude that a potential possibility of these species to seed propagation are not limited by their seed viability even under conditions of the highest technogenic load. In sites of environment pollution, the high metal content in the upper soil layer is the main factor limiting plant seed propagation.     

不同降水条件下科尔沁沙地南缘疏林草地樟子松针叶δ13C和叶性状特征

通过比较不同自然降水年份(极端干旱和极端湿润)19年生疏林草地樟子松的针叶δ13C、比叶面积和干物质含量,结合土壤含水量和地下水埋深,探讨了极端降水对樟子松水分利用的影响.结果表明:干旱年份(2009)樟子松林土壤含水量显著低于湿润年份(2010),但樟子松当年生针叶的δ13C在两年间没有显著差异,且两年相同月份间亦无显著差异;干旱年份当年生针叶的比叶面积显著低于湿润年份,而不同年份间干物质含量的差异不显著.在两种极端降水条件下,樟子松的水分利用效率没有明显变化,主要通过改变当年生针叶的比叶面积来适应降水量的变化.对于地下水埋深高于3.0m的疏林草地樟子松人工林生态系统,极端干旱不会严重影响樟子松的存活和生长.     

Radiation and genetic consequences of ionizing irradiation on population of Pinus sylvestris L. within the zone of the Chernobyl NPP

Main results of the nineteen year radiation-genetic monitoring of radiobiological, of cytogenetic, and of genetic parameters of Pinus sylvestris L. of forest plantation from the Chernobyl meltdown area are presented. The acute ionizing irradiation at doses >1 Gy induces the formation of morphoses and of depressed growth; at doses >2 Gy, the reproductive ability of the trees declined. The radiobiological parameters shown linear or close to it dose-dependence relationship. The acute irradiation at dose of 0.5 Gy induces 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 mutation frequency at specific loci on the absorbed dose was described by a nonlinear curve. The results of the cytogenetic analysis of seedlings obtained from seeds annually collected in zones of slight, of moderate, and of strong damage of Pinus sylvestris L. are presented.     

Variability of the cytological parameters of Pinus sylvestris L. seeds from the unique Hrenovskoy pine forest

Hrenovskoy pine forest is a unique island stand at the boundary of the species range of Scots pine Pinus sylvestris L. This object is of exceptional economic value, because it serves as a forest-seed base for the Voronezh oblast and some other regions of Russia; therefore, the stand and seed qualities have to be monitored constantly. The results of the first cytogenetic study of the seed progeny of P. sylvestris from the Morozov Grove, a high-quality stand in a reserved site within the Hrenovskoy pine forest, are reported. The studies have been performed in order to obtain a more correct assessment of seed quality based not only on their germination and energy of germination (traditionally used by forest breeders), but also on their genomic stability. The latter may be estimated by the stability of chromosome number in the somatic cells of seedlings and the regularity of mitotic divisions, because they also characterize the state of the generative system of parental forms and may serve as an integrated estimate of the stand development homeostasis. Therefore, the chromosome number, mitotic and nucleolar activities, and the number and spectrum of pathological mitoses (PMs) have been determined. Seedlings have been obtained from 240 seeds (collected from 12 trees) that resulted from free pollination. The cytological analysis of the rootlets of these seedlings has not detected any deviations from the chromosome number typical of the species P. sylvestris L. (2n = 24). However, considerable variation has been found in each family with respect to the mitotic index (MI) (from 4.2 +/- 0.36 to 8.1 +/- 0.39%) and the number of PMs (from 0.5 to 2.1%); micronuclei have also been found in each family (from 0.01 to 0.05%). In general, the phenotypic characteristics and the variation pattern of cytological parameters of the progeny of the trees studied in the Hrenovskoy pine forest, together with the high germination rate of seeds (90-98%), indicate that the current state of pines in the Hrenovskoy pine forest can be considered to be satisfactory. However, the presence of micronuclei in all variants and the decrease in MI in some individual plants (which was, on average, 6.1 +/- 0.03% in the sample studied, i.e., significantly lower than that of P. sylvestris from the Usmanskii pine forest (7.3 +/- 1.2%), where the conditions are optimal for this species) may serve as the bioindicators of the deterioration of both the state of trees themselves and the ecological conditions in the Hrenovskoy pine forest. Therefore, the population structure of this valuable autochtonic stand should be reproduced urgently.     

Use of stable isotopes to quantify nitrogen, potassium and magnesium dynamics in young Scots pine (Pinus sylvestris)

Two-yr-old Scots pine ( Pinus sylvestris ) seedlings were grown in sand culture for 1 yr with a generous supply of a balanced nutrient solution. Trees were repotted into clean sand in February 1998 and given either a reduced or adequate nutrient supply containing enriched ¹⁵N, ⁴¹K and ²⁶Mg to label nutrient uptake during spring 1998. Trees doubled their biomass during the experiment. Whole-tree net photosynthesis was reduced by 43% after 95 d in trees that received the lower nutrient supply ( P < 0.001), although differences in biomass between the two treatments were less pronounced. Remobilization contributed 83, 82 and 52% of the N, K and Mg, respectively, used to support growth of new tissues in trees that received reduced nutrient supply. Those receiving the higher nutrient supply still obtained 44–59% of nutrients used for spring growth of new tissues from remobilization. Current nutrient supply had no significant effect on the amount of N or Mg remobilized to new tissues but K remobilization was less in trees that received the lower nutrient supply ( P = 0.025). The importance of remobilization in young trees and problems associated with quantifying internal cycling of nutrients are discussed.     

Copper-adapted Suillus luteus, a symbiotic solution for pines colonizing Cu mine spoils

Natural populations thriving in heavy-metal-contaminated ecosystems are often subjected to selective pressures for increased resistance to toxic metals. In the present study we describe a population of the ectomycorrhizal fungus Suillus luteus that colonized a toxic Cu mine spoil in Norway. We hypothesized that this population had developed adaptive Cu tolerance and was able to protect pine trees against Cu toxicity. We also tested for the existence of cotolerance to Cu and Zn in S. luteus. Isolates from Cu-polluted, Zn-polluted, and nonpolluted sites were grown in vitro on Cu- or Zn-supplemented medium. The Cu mine isolates exhibited high Cu tolerance, whereas the Zn-tolerant isolates were shown to be Cu sensitive, and vice versa. This indicates the evolution of metal-specific tolerance mechanisms is strongly triggered by the pollution in the local environment. Cotolerance does not occur in the S. luteus isolates studied. In a dose-response experiment, the Cu sensitivity of nonmycorrhizal Pinus sylvestris seedlings was compared to the sensitivity of mycorrhizal seedlings colonized either by a Cu-sensitive or Cu-tolerant S. luteus isolate. In nonmycorrhizal plants and plants colonized by the Cu-sensitive isolate, root growth and nutrient uptake were strongly inhibited under Cu stress conditions. In contrast, plants colonized by the Cu-tolerant isolate were hardly affected. The Cu-adapted S. luteus isolate provided excellent insurance against Cu toxicity in pine seedlings exposed to elevated Cu levels. Such a metal-adapted Suillus-Pinus combination might be suitable for large-scale land reclamation at phytotoxic metalliferous and industrial sites.     

不同降水条件下科尔沁沙地南缘疏林草地樟子松针叶δ13C和叶性状特征

通过比较不同自然降水年份(极端干旱和极端湿润)19年生疏林草地樟子松的针叶δ¹³C、比叶面积和干物质含量,结合土壤含水量和地下水埋深,探讨了极端降水对樟子松水分利用的影响.结果表明: 干旱年份(2009)樟子松林土壤含水量显著低于湿润年份(2010),但樟子松当年生针叶的δ¹³C在两年间没有显著差异,且两年相同月份间亦无显著差异;干旱年份当年生针叶的比叶面积显著低于湿润年份,而不同年份间干物质含量的差异不显著.在两种极端降水条件下,樟子松的水分利用效率没有明显变化,主要通过改变当年生针叶的比叶面积来适应降水量的变化.对于地下水埋深高于3.0 m的疏林草地樟子松人工林生态系统,极端干旱不会严重影响樟子松的存活和生长.     

Differential responses of three fungal species to environmental factors and their role in the mycorrhization of<Emphasis Type="Italic"> Pinus radiata</Emphasis> D. Don

Three ectomycorrhizal (ECM) isolates of Rhizopogon luteolus, R. roseolus and Scleroderma citrinum were found to differ markedly in their in vitro tolerance to adverse conditions limiting fungal growth, i.e. water availability, pH and heavy metal pollution. S. citrinum was the most sensitive, R. luteolus intermediate and R. roseolus the most tolerant species. Pinus radiata D. Don seedlings were inoculated in the laboratory and in a containerised seedling nursery with spore suspensions of the three ECM species. Colonisation percentage was considerably lower under nursery conditions, probably due to competition by native fungi. The effects of nursery ECM inoculation on seedling growth depended on the fungal species. Only R. roseolus-colonised plants showed a significantly higher shoot growth than non-mycorrhizal plants. All three fungi induced significantly higher root dry weights relative to control plants. Despite the low mycorrhizal colonisation, mycorrhization with all three species improved the physiological status of nursery-grown seedlings, e.g. enhanced root enzyme activity, shoot nutrient and pigment content, net photosynthesis rate and water use efficiency. Of the three fungal species, R. roseolus was the most effective; this species was also the most adaptable and showed the greatest range of tolerance to adverse environmental conditions in pure culture. It is, therefore, proposed as a promising fungal species for ECM inoculation of P. radiata in the nursery.