Anu Lavola

Nutrient availability and the effect of increasing UV-B radiation on secondary plant compounds in Scots pine

Growth and pigment composition of plants can be affected by both UV-radiation and availability of nutrients. Therefore, the UV-induction in a plant as a protective response against UV-B radiation may depend on the UV-B dose encountered and the nutritional state of a plant. To test this, young Scots pine (Pinus sylvestris) seedlings were grown under two nutrient levels (relative nutrient addition fates, RAR, of 4 and 6%) and exposed to six different daily doses of UV-radiation (UV-A and UV-B) for one growing season. Both the growth and the accumulation of photosynthetic pigments were positively affected by the availability of nutrients, while the accumulation of condensed tannins and its precursor, (+)-catechin decreased significantly under higher nutrient level, which may imply some trade-off between primary and secondary metabolites. However, nutrient level did not markedly affect the total accumulation of flavonols. Since there was no decrease in the growth of the seedlings and the content of photosynthetic pigments in needles under increased UV-B levels, the seedlings had apparently good protection against UV-B radiation. Protection was probably dependent on the UV-induction of flavonoid compounds in needles, which differed between the two nutrient levels used. Under moderate nutrient availability (RAR 4%), the accumulation of flavonols and also, chlorophylls was highest at the ambient or near to ambient levels of UV-radiation, the levels that pines are generally adapted in natural conditions. As the availability of nutrients and thereby, the resources in seedlings increased (RAR 6%), the UV-B levels higher than ambient increased specifically the accumulation of diacylated flavonols when dose responses were detected. Furthermore, the enhancement of UV-A radiation caused an accumulation of the dihydroflavonols, ampelopsin and taxifolin, under high nutrient availability. Since the flavonoid responses were very compound-specific, and in some cases were also dose-dependent and different between the two nutrient levels used, it can be suggested that the induction of secondary compounds in the flavonoid pathway is multi-step regulated by both environmental factors, UV-radiation and nutrients. Already the present variation in the ambient range of UV-B radiation at northern latitudes can influence the chemical composition of Scots pine seedlings and enhancement in nutrients may increase their responsiveness, particularly to high UV-intensities.

Sex-related differences in growth and carbon allocation to defence in Populus tremula as explained by current plant defence theories

Plant defence theories have recently evolved in such a way that not only the quantity but also the quality of mineral nutrients is expected to influence plant constitutive defence. Recently, an extended prediction derived from the protein competition model (PCM) suggested that nitrogen (N) limitation is more important for the production of phenolic compounds than phosphorus (P). We aimed at studying sexual differences in the patterns of carbon allocation to growth and constitutive defence in relation to N and P availability in Populus tremula L. seedlings. We compared the gender responses in photosynthesis, growth and whole-plant allocation to phenolic compounds at different combination levels of N and P, and studied how they are explained by the main plant defence theories. We found no sexual differences in phenolic concentrations, but interestingly, slow-growing females had higher leaf N concentration than did males, and genders differed in their allocation priority. There was a trade-off between growth and the production of flavonoid-derived phenylpropanoids on one hand, and between the production of salicylates and flavonoid-derived phenylpropanoids on the other. Under limited nutrient conditions, females prioritized mineral nutrient acquisition, flavonoid and condensed tannin (CT) production, while males invested more in above-ground biomass. Salicylate accumulation followed the growth differentiation balance hypothesis as low N mainly decreased the production of leaf and stem salicylate content while the combination of both low N and low P increased the amount of flavonoids and CTs allocated to leaves and to a lesser extent stems, which agrees with the PCM. We suggest that such a discrepancy in the responses of salicylates and flavonoid-derived CTs is linked to their clearly distinct biosynthetic origins and/or their metabolic costs.

Combination treatment of elevated UVB radiation, CO2 and temperature has little effect on silver birch (Betula pendula) growth and phytochemistry.

Elevations of carbon dioxide, temperature and ultraviolet-B (UBV) radiation in the growth environment may have a high impact on the accumulation of carbon in plants, and the different factors may work in opposite directions or induce additive effects. To detect the changes in the growth and phytochemistry of silver birch (Betula pendula) seedlings, six genotypes were exposed to combinations of ambient or elevated levels of CO2 , temperature and UVB radiation in top-closed chambers for 7 weeks. The genotypes were relatively similar in their responses, and no significant interactive effects of three-level climate factors on the measured parameters were observed. Elevated UVB had no effect on growth, nor did it alter plant responses to CO2 and/or temperature in combined treatments. Growth in all plant parts increased under elevated CO2 , and height and stem biomass increased under elevated temperature. Increased carbon distribution to biomass did not reduce its allocation to phytochemicals: condensed tannins, most flavonols and phenolic acids accumulated under elevated CO2 and elevated UVB, but this effect disappeared under elevated temperature. Leaf nitrogen content decreased under elevated CO2 . We conclude that, as a result of high genetic variability in phytochemicals, B. pendula seedlings have potential to adapt to the tested environmental changes. The induction in protective flavonoids under UVB radiation together with the positive impact of elevated CO2 and temperature mitigates possible UVB stress effects, and thus atmospheric CO2 concentration and temperature are the climate change factors that will dictate the establishment and success of birch at higher altitudes in the future.

Bioactive polyphenols in leaves, stems, and berries of Saskatoon (Amelanchier alnifolia Nutt.) cultivars.

The Saskatoon berry is currently cultivated in many parts of the world for its suitability for various food products and due to its high content of nutrients and polyphenols. To determine the phytochemical profile of a Saskatoon plant, polyphenols from leaves, stems, and berries were screened from four cultivars grown in Finland using HPLC-DAD and HPLC-ESI/MS. The phenolic composition and concentrations varied among plant parts and cultivars. The main berry components were cyanidin-based anthocyanins (63% of the phenols), quercetin-derived flavonol glycosides, and hydroxycinnamic acids. The total anthocyanin content varied between 258.7 and 517.9 mg/100 fresh weight among cultivars. Protocatechuic acid was found for the first time in Saskatoon berries. The leaves consisted of quercetin- and kaempferol-derived glycosides (41% of the phenols), hydroxycinnamic acids (36%), catechins, and some neolignans. Quercetin 3-galactoside and 3-glucoside, (-)-epicatechin, and chlorogenic acid were the main phenolics in the leaves of all cultivars. The stem components were flavanone and flavonol glycosides (55% of the phenols), catechins (38%), and hydroxybenzoic acids. Concentrations of the main compound, eriodictyol 7-glucoside, varied among cultivars from 3.3 to 6.5 mg/g of stem dry weight. Very high proanthocyanidin contents were found in stems and leaves (10-14% of dry biomass), whereas berries contained a low amount of proanthocyanidins (3% of dry biomass). The findings reveal that leaves and stems of Saskatoon cultivars possess high amounts of various phenolic compounds that may offer new functional raw materials for a wide range of food and health products.

Interactive effects of supplemental UV-B and temperature in European aspen seedlings: Implications for growth, leaf traits, phenolic defense and associated organisms

Past studies reveal opposite effects of elevated UV-B and temperature on plant growth and concentrations of UV-B absorbing compounds, yet few studies have dealt with the combined and interactive effects of these two climate change factors on woody dioecious plants. We investigated the interactive effects of UV-B and temperature treatments on growth, leaf traits and phenolic concentrations in Populus tremula L. (European aspen) seedlings. We also considered the consequences of these effects on their associated organisms: herbivorous insects, rust pathogens, the presence of endophytic fungi and whether or not the responses differ between genders and genotypes. Supplemental temperature and UV-B were modulated to +2 °C and +30.77% above ambient conditions, respectively. Warming increased growth, photosynthesis and foliar nitrogen concentration but reduced leaf thickness and phenolic concentrations. On the other hand, supplemental UV-B increased total phenolic glycosides, mainly flavonols and phenolic acids, and partially counteracted the positive effects of warming on growth. Fast growing genotypes were less susceptible to the growth-reducing effect of combined UVB + T, less infected with rust disease and less prone to insect damage probably due to their higher salicylate and lower nitrogen concentrations. Under ambient temperature, the males of European aspen were taller and had bigger leaves than the females, while under elevated temperature, females grew bigger and, under UV-B, had more tremulacin than males. The multiple interactive effects of UV-B and temperature on growth, leaf traits and phenolic compounds, highlight the importance of multifactor experiments as a realistic predictor of plant responses to climate change.

UV‐B and temperature enhancement affect spring and autumn phenology in Populus tremula

Perennial plants growing at high latitudes synchronize growth and dormancy to appropriate seasons by sensing environmental cues. Autumnal growth cessation, bud set and dormancy induction are commonly driven by the length of photoperiod and light quality, and the responses are modified by temperature. However, although ultraviolet (UV)-B radiation is well known to affect plant growth and development, information on the effects on bud phenology is scarce. We examined the separate and combined effects of enhanced temperature and UV-B on autumnal bud set and spring bud break in female and male clones of Populus tremula in an outdoor experiment in Joensuu, Eastern Finland. Enhancements of UV-B and temperature were modulated to +30% and +2 °C, respectively, from June to October 2012. Enhanced UV-B accelerated bud set, while increased temperature delayed it. For both UV-B and temperature, we found sex-related differences in responsiveness. Temperature increase had a stronger delaying effect on bud maturation in male compared with female clones. Also, male clones were more responsive to UV-B increase than female clones. Increasing autumnal temperature enhanced bud break in spring for both sexes, while UV-B enhanced bud break in male clones. In conclusion, we found that UV-B affected phenological shifts in P. tremula, and that temperature and UV-B affected genders differently.

Boron uptake by ectomycorrhizas of silver birch

Boron (B) is an essential micronutrient for plants but it is thought not to be essential for fungi. We studied whether the extraradical mycelia of Paxillus involutus in symbiosis with silver birch (Betula pendula) take up B and transport it to the host plant. We grew mycorrhizal plants in flat microcosms with a partitioning wall, below which there was only extraradical mycelium. A boric acid solution enriched in 10B was applied to these mycelia. Increased 10B/11B isotope ratios were subsequently measured in birch leaves, stems, and roots plus mycorrhizas in the upper compartment. Boron was therefore taken up by the mycorrhizal mycelia and transported to the host plant in this species combination.

Effects of long-term UV-exposure and plant sex on the leaf phenoloxidase activities and phenolic concentrations of Salix myrsinifolia (Salisb.)

The accumulation of flavonoids on the leaf surface is a well-characterized protective mechanism against UV-B radiation. Other protective mechanisms, such as the induction of antioxidative enzymes and peroxidase-mediated lignification may also be important. The effects of UV-B radiation have mainly been considered in short-term studies, whereas ecologically more relevant long-term field studies are still rare. Here we examined the effects of long-term exposure to enhanced UV-B radiation on the activities of two antioxidative enzymes, polyphenol oxidase (PPO; EC 1.10.2.2 and EC 1.14.18.1) and guaiacol peroxidase (POD; EC 1.11.1.7), as well as the phenolic concentrations in two sexes of the dioecious species, Salix myrsinifolia. After three consecutive growth seasons with enhanced UV-B radiation, we found that PPO activity was decreased by UV radiation in male plants, which might explain their lower UV-B tolerance when compared to female plants. In addition, male plants had higher specific activity than did female plants under ambient conditions, supporting the idea that males of S. myrsinifolia are generally more growth-oriented than females. By contrast, neither UV treatment nor sex had significant effects on the POD activities of willows. Gender differences in the concentrations of phenolic compounds are in line with the general concept that males are less well defended than females. We suggest that the inability to increase PPO and POD activity, along with lower accumulation of UV-B absorbing compounds under UV-B exposure, might be one of the reasons why males had thinner leaves and were less tolerant of UV-B than were females.

Interannual variation in UV-B and temperature effects on bud phenology and growth in Populus tremula

Warming affects phenological processes such as spring bud break and autumnal bud set, and also growth rates of trees. Recently, it has been shown that these physiological processes also may be influenced by the ultraviolet-B (UV-B) part of the solar spectrum, and there are reasons to expect that the two environmental factors induce interactive effects when acting in concert. In this study, our aim was to elucidate how experimental enhancements in temperature and UV-B, alone and in combination, affect growth and seasonal phenology of Eurasian aspen (Populus tremula) over several growing seasons (three years). Moreover, we tested how environmentally induced changes in phenology affect the growth achieved over each season, that is, the importance of a prolonged growing season for growth yield. The plants grew in an outdoor experiment with modulated enhancements of temperature and UV-B during the growing season. Both UV-B and temperature enhancement affected bud set dates, while bud break dates were only affected by temperature enhancement. Temperature delayed bud set in all years, but gradually less over years, while UV-B yielded earlier bud set the first year but showed a delayed response the following years. Bud break was always earlier under temperature enhancement. The experimentally induced extension of the growing season in both ends had a positive effect of growth throughout the three-year period. However, the reduced responsiveness of bud set to both enhancement treatments suggest that the plants gradually acclimated to the modified climate, a finding that should also be investigated for other tree species.

Variability in the composition of phenolic compounds in winter-dormant Salix pyrolifolia in relation to plant part and age

The phenolic phytochemicals of winter-dormant Salix pyrolifolia were determined from the vegetative buds, and the bark and wood of different-aged twigs by HPLC-DAD and UHPLC-QTOF-MS analyses. All the plant parts were composed of salicylate glucosides and the other Salix-specific, simple phenolic glucosides as well as of phenolic acids, flavonoids and the high molecular-weight condensed tannins. The flavonoid composition was most diverse in buds and they also contained a large amount of chlorogenic acid (5-caffeoylquinic acid IUPAC), while salicylate glucosides and simple phenolic glucosides predominated in bark. The wooden interior part of the twigs contained fewer components and the lowest concentrations of compounds. Salicortin was the main compound in winter-dormant S. pyrolifolia (over 10% of bark biomass), but the concentrations of picein, salireposide, isosalipurposide, catechin and condensed tannins were also high. The flavonoid composition was highly naringenin- and quercetin-biassed. The composition of phytochemicals was organ-specific and remained relatively similar between different-aged trees. However, there were compound-specific fluctuations in the concentrations of phytochemicals with the age of the trees and within plant parts. Generally, the one-year-old plants differed from the older trees in their high concentration of condensed tannins in all the plant parts studied and in the highest concentration of isosalipurposide in bark, while the total amounts of salicylate glucosides in plant parts, and of naringenin glucosides in buds, tended to be highest in 20 year-old-trees.