Sari Kontunen-Soppela

Emissions of volatile organic compounds and leaf structural characteristics of European aspen (Populus tremula) grown under elevated ozone and temperature

Northern forest trees are challenged to adapt to changing climate, including global warming and increasing tropospheric ozone (O(3)) concentrations. Both elevated O(3) and temperature can cause significant changes in volatile organic compound (VOC) emissions as well as in leaf anatomy that can be related to adaptation or increased stress tolerance, or are signs of damage. Impacts of moderately elevated O(3) (1.3x ambient) and temperature (ambient + 1 degrees C), alone and in combination, on VOC emissions and leaf structure of two genotypes (2.2 and 5.2) of European aspen (Populus tremula L.) were studied in an open-field experiment in summer 2007. The impact of O(3) on measured variables was minor, but elevated temperature significantly increased emissions of total monoterpenes and green leaf volatiles. Genotypic differences in the responses to warming treatment were also observed. alpha-Pinene emission, which has been suggested to protect plants from elevated temperature, increased from genotype 5.2 only. Isoprene emission from genotype 2.2 decreased, whereas genotype 5.2 was able to retain high isoprene emission level also under elevated temperature. Elevated temperature also caused formation of thinner leaves, which was related to thinning of epidermis, palisade and spongy layers as well as reduced area of palisade cells. We consider aspen genotype 5.2 to have better potential for adaptation to increasing temperature because of thicker photosynthetic active palisade layer and higher isoprene and alpha-pinene emission levels compared to genotype 2.2. Our results show that even a moderate elevation in temperature is efficient enough to cause notable changes in VOC emissions and leaf structure of these aspen genotypes, possibly indicating the effort of the saplings to adapt to changing climate.

Impacts of increasing ozone on Indian plants

Increasing anthropogenic and biogenic emissions of precursor compounds have led to high tropospheric ozone concentrations in India particularly in Indo-Gangetic Plains, which is the most fertile and cultivated area of this rapidly developing country. Current ozone risk models, based on European and North American data, provide inaccurate estimations for crop losses in India. During the past decade, several ozone experiments have been conducted with the most important Indian crop species (e.g. wheat, rice, mustard, mung bean). Experimental work started in natural field conditions around Varanasi area in early 2000s, and the use of open top chambers and EDU (ethylene diurea) applications has now facilitated more advanced studies e.g. for intra-species sensitivity screening and mechanisms of tolerance. In this review, we identify and discuss the most important gaps of knowledge and future needs of action, e.g. more systematic nationwide monitoring for precursor and ozone formation over Indian region.

Elevation of night-time temperature increases terpenoid emissions from Betula pendula and Populus tremula

Volatile organic compounds (VOCs) are expected to have an important role in plant adaptation to high temperatures. The impacts of increasing night-time temperature on daytime terpenoid emissions and related gene expression in silver birch (Betula pendula) and European aspen (Populus tremula) clones were studied. The plants were grown under five different night-time temperatures (6, 10, 14, 18, and 22 °C) while daytime temperature was kept at a constant 22 °C. VOC emissions were collected during the daytime and analysed by gas chromatography–mass spectrometry (GC-MS). In birch, emissions per leaf area of the C11 homoterpene 4,8-dimethy1-nona-1,3,7-triene (DMNT) and several sesquiterpenes were consistently increased with increasing night-time temperature. Total sesquiterpene (SQT) emissions showed an increase at higher temperatures. In aspen, emissions of DMNT and β-ocimene increased from 6 °C to 14 °C, while several other monoterpenes and the SQTs (Z,E)-α-farnesene and (E,E)-α-farnesene increased up to 18 °C. Total monoterpene and sesquiterpene emission peaked at 18 °C, whereas isoprene emissions decreased at 22 °C. Leaf area increased across the temperature range of 6–22 °C by 32% in birch and by 59% in aspen. Specific leaf area (SLA) was also increased in both species. The genetic regulation of VOC emissions seems to be very complex, as indicated by several inverse relationships between emission profiles and expression of several regulatory genes (DXR, DXS, and IPP). The study indicates that increasing night temperature may strongly affect the quantity and quality of daytime VOC emissions of northern deciduous trees.

Gene expression responses of paper birch (Betula papyrifera) to elevated CO2 and O3 during leaf maturation and senescence

Gene expression responses of paper birch (Betula papyrifera) leaves to elevated concentrations of CO(2) and O(3) were studied with microarray analyses from three time points during the summer of 2004 at Aspen FACE. Microarray data were analyzed with clustering techniques, self-organizing maps, K-means clustering and Sammons mappings, to detect similar gene expression patterns within sampling times and treatments. Most of the alterations in gene expression were caused by O(3), alone or in combination with CO(2). O(3) induced defensive reactions to oxidative stress and earlier leaf senescence, seen as decreased expression of photosynthesis- and carbon fixation-related genes, and increased expression of senescence-associated genes. The effects of elevated CO(2) reflected surplus of carbon that was directed to synthesis of secondary compounds. The combined CO(2)+O(3) treatment resulted in differential gene expression than with individual gas treatments or in changes similar to O(3) treatment, indicating that CO(2) cannot totally alleviate the harmful effects of O(3).

Vertical profiles reveal impact of ozone and temperature on carbon assimilation of Betula pendula and Populus tremula

Rising temperature and tropospheric ozone (O(3)) concentrations are likely to affect carbon assimilation processes and thus the carbon sink strength of trees. In this study, we investigated the joint action of elevated ozone and temperature on silver birch (Betula pendula) and European aspen (Populus tremula) saplings in field conditions by combining free-air ozone exposure (1.2 × ambient) and infrared heaters (ambient +1.2 °C). At leaf level measurements, elevated ozone decreased leaf net photosynthesis (P(n)), while the response to elevated temperature was dependent on leaf position within the foliage. This indicates that leaf position has to be taken into account when leaf level data are collected and applied. The ozone effect on P(n) was partly compensated for at elevated temperature, showing an interactive effect of the treatments. In addition, the ratio of photosynthesis to stomatal conductance (P(n)/g(s) ratio) was decreased by ozone, which suggests decreasing water use efficiency. At the plant level, the increasing leaf area at elevated temperature resulted in a considerable increase in photosynthesis and growth in both species.

Differential gene expression in senescing leaves of two silver birch genotypes in response to elevated CO2 and tropospheric ozone

Long-term effects of elevated CO(2) and O(3) concentrations on gene expression in silver birch (Betula pendula Roth) leaves were studied during the end of the growing season. Two birch genotypes, clones 4 and 80, with different ozone growth responses, were exposed to 2x ambient CO(2) and/or O(3) in open-top chambers (OTCs). Microarray analyses were performed after 2 years of exposure, and the transcriptional profiles were compared to key physiological characteristics during leaf senescence. There were genotypic differences in the responses to CO(2) and O(3). Clone 80 exhibited greater transcriptional response and capacity to alter metabolism, resulting in better stress tolerance. The gene expression patterns of birch leaves indicated contrasting responses of senescence-related genes to elevated CO(2) and O(3). Elevated CO(2) delayed leaf senescence and reduced associated transcriptional changes, whereas elevated O(3) advanced leaf senescence because of increased oxidative stress. The combined treatment demonstrated that elevated CO(2) only temporarily alleviated the negative effects of O(3). Gene expression data alone were insufficient to explain the O(3) response in birch, and additional physiological and biochemical data were required to understand the true O(3) sensitivity of these clones.

Preservation of transgenic silver birch (Betula pendula Roth) lines by means of cryopreservation

The aim of the study was to develop a preservation method for transgenic silver birch (Betula pendula Roth) lines based on cryopreservation. Specific attention was paid to transgene stability and functioning. Vegetative buds collected from one- or two-year-old silver birches representing four transgenic lines and two wild-type lines were used as explants. Generally, the average regeneration of either transgenic or wild-type, cryoperserved and non-cryopreserved control buds was excellent, and varied from 72 to 100 percent. The regeneration percentage of cryopreserved buds was, however, significantly lower than that of non-cryopreserved control buds when estimated two weeks after thawing, but the differences were no longer significant four weeks after thawing. Growth of the plants in the greenhouse was more dependent on the clone than on the cryopreservation treatment. The studied transgenic lines have three (line E/5) to nine (line R/3.2) copies of transferred neomycin phosphotransferase genes that were also found to be stable after cryopreservation. In general, the neomycin phosphotransferase transcript levels did not change due to cryopreservation. The results indicate that it is possible to apply the cryopreservation technique to preserve valuable transgenic lines of a forest tree, silver birch. The method presented here leads to high regeneration percentages combined with transgene stability and functioning.

Searching for common responsive parameters for ozone tolerance in 18 rice cultivars in India: Results from ethylenediurea studies

Eighteen rice (Oryza sativa) cultivars were screened for ozone (O3) tolerance and for the most responsive parameters with ethylenediurea (EDU) treatments at two experimental sites experiencing high ambient O3 conditions in the Indo-Gangetic Plains (IGP) of India. EDU was applied at 15 day intervals until the final harvest phase as a foliar spray at 300 ppm in order to protect the plants from the adverse effects of O3. Antioxidant activity, malondialdehyde content (MDA), chlorophyll content, gas exchange, and chlorophyll fluorescence (Fv/Fm) at the vegetative and flowering phases and harvest-related parameters were studied, for a total of 24 parameters. Seven of the studied cultivars had higher than average grainweightplant(-1) in all site and treatment combinations and can be recommended for cultivation in areas suffering from high O3 concentrations. The most responsive parameters with EDU treatment in high O3 across all cultivars were superoxide dismutase (SOD) and catalase (CAT) activities, the contents of oxidised (GSSG) and reduced (GSH) glutathione and MDA, and shoot weight plant(-1). These results indicated that the O3 scavenging activity of EDU is mediated through an antioxidant defence system rather than a direct effect on physiological parameters, such as photosynthesis and stomatal conductance.

Ozone affects ascorbate and glutathione biosynthesis as well as amino acid contents in three Euramerican poplar genotypes

Ozone is an air pollutant that causes oxidative stress by generation of reactive oxygen species (ROS) within the leaf. The capacity to detoxify ROS and repair ROS-induced damage may contribute to ozone tolerance. Ascorbate and glutathione are known to be key players in detoxification. Ozone effects on their biosynthesis and on amino acid metabolism were investigated in three Euramerican poplar genotypes (Populus deltoides Bartr. × Populus nigra L.) differing in ozone sensitivity. Total ascorbate and glutathione contents were increased in response to ozone in all genotypes, with the most resistant genotype (Carpaccio) showing an increase of up to 70%. Reduced ascorbate (ASA) concentration at least doubled in the two most resistant genotypes (Carpaccio and Cima), whereas the most sensitive genotype (Robusta) seemed unable to regenerate ASA from oxidized ascorbate (DHA), leading to an increase of 80% of the oxidized form. Increased ascorbate (ASA + DHA) content correlated with the increase in gene expression in its biosynthetic pathway, especially the putative gene of GDP-l-galactose phosphorylase VTC2. Increased cysteine availability combined with increased expression of γ-glutamylcysteine synthetase (GSH1) and glutathione synthetase (GSH2) genes allows higher glutathione biosynthesis in response to ozone, particularly in Carpaccio. In addition, ozone caused a remobilization of amino acids with a decreased pool of total amino acids and an increase of Cys and putrescine, especially in Carpaccio. In addition, the expression of genes encoding threonine aldolase was strongly induced only in the most tolerant genotype, Carpaccio. Reduced ascorbate levels could partly explain the sensitivity to ozone for Robusta but not for Cima. Reduced ascorbate level alone is not sufficient to account for ozone tolerance in poplar, and it is necessary to consider several other factors including glutathione content.

Genome sequencing and population genomic analyses provide insights into the adaptive landscape of silver birch

Silver birch (Betula pendula) is a pioneer boreal tree that can be induced to flower within 1 year. Its rapid life cycle, small (440-Mb) genome, and advanced germplasm resources make birch an attractive model for forest biotechnology. We assembled and chromosomally anchored the nuclear genome of an inbred B. pendula individual. Gene duplicates from the paleohexaploid event were enriched for transcriptional regulation, whereas tandem duplicates were overrepresented by environmental responses. Population resequencing of 80 individuals showed effective population size crashes at major points of climatic upheaval. Selective sweeps were enriched among polyploid duplicates encoding key developmental and physiological triggering functions, suggesting that local adaptation has tuned the timing of and cross-talk between fundamental plant processes. Variation around the tightly-linked light response genes PHYC and FRS10 correlated with latitude and longitude and temperature, and with precipitation for PHYC. Similar associations characterized the growth-promoting cytokinin response regulator ARR1, and the wood development genes KAK and MED5A.