Changes in leaf trichomes and epicuticular flavonoids during leaf development in three birch taxa

BACKGROUND AND AIMS: Changes in number of trichomes and in composition and concentrations of their exudates throughout leaf development may have important consequences for plant adaptation to abiotic and biotic factors. In the present study, seasonal changes in leaf trichomes and epicuticular flavonoid aglycones in three Finnish birch taxa (Betula pendula, B. pubescens ssp. pubescens, and B. pubescens ssp. czerepanovii) were followed. METHODS: Trichome number and ultrastructure were studied by means of light, scanning and transmission electron microscopy, while flavonoid aglycones in ethanolic leaf surface extracts were analysed by high-pressure liquid chromatography. KEY RESULTS: Density of both glandular and non-glandular trichomes decreased drastically with leaf expansion while the total number of trichomes per leaf remained constant, indicating that the final number of trichomes is established early in leaf development. Cells of glandular trichomes differentiate before those of the epidermis and produce secreted material only during the relatively short period (around 1-2 weeks) of leaf unfolding and expansion. In fully expanded leaves, glandular trichomes appeared to be at the post-secretory phase and function mainly as storage organs; they contained lipid droplets and osmiophilic material (probably phenolics). Concentrations (mg g(-1) d. wt) of surface flavonoids decreased with leaf age in all taxa. However, the changes in total amount ( microg per leaf) of flavonoids during leaf development were taxon-specific: no changes in B. pubescens ssp. czerepanovii, increase in B. pendula and in B. pubescens ssp. pubescens followed by the decline in the latter taxon. Concentrations of most of the individual leaf surface flavonoids correlated positively with the density of glandular trichomes within species, suggesting the participation of glandular trichomes in production of surface flavonoids. CONCLUSIONS: Rapid decline in the density of leaf trichomes and in the concentrations of flavonoid aglycones with leaf age suggests that the functional role of trichomes is likely to be most important at the early stages of birch leaf development.     

Comparative analysis of leaf trichome structure and composition of epicuticular flavonoids in Finnish birch species

The morphology, ultrastructure, density and distribution of trichomes on leaves of Betula pendula, B. pubescens ssp. pubescens, B. pubescens ssp. czerepanovii and B. nana were examined by means of light, scanning and transmission electron microscopy. The composition of flavonoids in ethanolic leaf surface extracts was analysed by high pressure liquid chromatography. All taxa examined contained both glandular and non-glandular trichomes (short and/or long hairs) but differed from each other in trichome ultrastructure, density and location on the leaf. Leaves of B. pubescens were more hairy than those of B. pendula, but the latter species had a higher density of glandular trichomes. Of the two subspecies of B. pubescens, leaves of ssp. pubescens had more short hairs on the leaf surface and four times the density of glandular trichomes of leaves of ssp. czerepanovii, whereas, in the latter subspecies, short hairs occurred largely on leaf veins, as in B. nana. The glandular trichomes were peltate glands, consisting of medullar and cortical cells, which differed structurally. Cortical cells possessed numerous small, poorly developed plastids and small vacuoles, whereas medullar cells had several large plastids with well-developed thylakoid systems and fewer vacuoles. In B. pubescens subspecies, vacuoles of the glandular cells contained osmiophilic deposits, which were probably phenolic, whereas in B. pendula, vacuoles of glandular trichomes were characterized by the presence of numerous myelin-like membranes. The composition of epicuticular flavonoids also differed among species. The two subspecies of B. pubescens and B. nana shared the same 12 compounds, but five of these occurred only in trace amounts in B. nana. Leaf surface extracts of B. pendula contained just six flavonoids, three of which occurred only in this species. In summary, the structure, density and distribution of leaf trichomes and the composition of epicuticular flavonoids represent good taxonomic markers for Finnish birch species.     

The nitrogen economy of mountain birch seedlings: implications for winter survival

1 Seedlings of mountain birch Betula pubescens ssp. czerepanovii were grown outdoors, under different environmental conditions, during their first growing season at Abisko, northern Sweden. Winter survival of seedlings was studied in relation to their growth and nitrogen (N) acquisition rate during the previous growing season.
2 Effects of fertilization and soil temperature on seedling growth and N acquisition were analysed in a factorial experiment including seven fertilizer levels and two temperature treatments. Effects of shading and neighbours ( B. p. ssp. czerepanovii and Empetrum hermaphroditum ) on seedling growth and N economy were evaluated in another experiment including five different neighbour interaction treatments.
3 An increase in either soil temperature or fertilization rate caused the birch seedlings to take up more N and grow faster. The relative growth rate and rate of N accumulation during the 12-week growing season were closely related to winter survival: at a relative growth rate of 2.0% and 3.8% day⁻¹, the winter survival of mountain birch seedlings was estimated to be 5% and 95%, respectively. This range corresponded to a relative N accumulation rate between 2.4% and 4.3% day⁻¹.
4 The relative N accumulation rate was clearly reduced by shading and by the plant–plant interactions studied. The effects of shading and birch neighbours but not of Empetrum on the rate of N accumulation could be explained by lower soil temperature.
5 Nutrient supply, soil temperature, vegetation shade and, presumably, allelopathy affect the N acquisition of first-year mountain birch seedlings, and thus also influence their winter survival. Soil temperature might be the major influence on the survival rate, due to its strong influence on the root N uptake rate.     

Wound-induced oxidative responses in mountain birch leaves

AIMS: The aim of the study was to examine oxidative responses in subarctic mountain birch, Betula pubescens subsp. czerepanovii, induced by herbivory and manual wounding. METHODS: Herbivory-induced changes in polyphenoloxidase, peroxidase and catalase activities in birch leaves were determined. A cytochemical dye, 3,3-diaminobenzidine, was used for the in situ and in vivo detection of H2O2 accumulation as a response to herbivory and wounding. To localize peroxidase activity in leaves, 10 mm H2O2 was applied to the dye reagent. KEY RESULTS: Feeding by autumnal moth, Epirrita autumnata, larvae caused an induction in polyphenoloxidase and peroxidase activities within 24 h, and a concomitant decrease in the activity of antioxidative catalases in wounded leaves. Wounding also induced H2O2 accumulation, which may have both direct and indirect defensive properties against herbivores. Wound sites and guard cells showed a high level of peroxidase activity, which may efficiently restrict invasion by micro-organisms. CONCLUSION: Birch oxidases together with their substrates may form an important front line in defence against herbivores and pathogens.     

Birch family and environmental conditions affect endophytic fungi in leaves

We investigated whether variation in foliar endophyte frequency among mountain birch trees from different maternal families was due in part to genetic differences among trees. The effect of different environmental conditions on the susceptibility of these mountain birch families to foliar endophytes was tested. The 3-year study was conducted in two tree gardens (altitudinal difference 180 m) with ten families of mountain birch. The frequency of the commonest endophytic fungus in mountain birch leaves, Fusicladium sp., was significantly affected by mountain birch family, with the ranking of families relative to Fusicladium sp. frequency being consistent from environment to environment. Variation in the frequency of Melanconium sp. was difficult to interpret because of significant family × garden × year interaction. Fusicladium sp. and Melanconium sp. endophytes were most frequent in different tree individuals, families and gardens. We conclude that mountain birch trees show heritable variation in their foliar endophyte frequency, and thus one of the conditions necessary for foliar endophytes of mountain birch trees to be able to affect the evolution of their host plant is fulfilled. However, the effect of mountain birch family on the frequency of endophytic fungi varies depending on the endophytic genera in question and partially also on environmental conditions. Received: 28 March 1998 / Accepted: 14 October 1998     

Growth response of altitudinal ecotypes of mountain birch to temperature and fertilisation

High- and low-altitude ecotypes of mountain birch (Betula pubescens ssp. czerepanovii) showed clear differences in their responses to various experimental conditions, including two temperature regimes and four fertilisation rates. There was, however, no simple way to characterise the elevational ecotypes in terms of relative growth rate, nitrogen (N) productivity, or root N uptake rate. The leaf N concentration was generally higher in the high-altitude seedlings than in the low-altitude seedlings. At low temperature, high-altitude mountain birch maintained a relatively high growth rate by combining high root N uptake rate and high leaf N concentration with high N productivity. An increase in temperature and/or fertiliser rate resulted in a marginal increase in N productivity in the high-altitude seedlings but resulted in a strong increase in N productivity in the low-altitude seedlings. In parallel, increased temperature resulted in a pronounced decrease in leaf N concentration only in the low-altitude seedlings. Our results suggest that the weak growth response to increased temperature in high-altitude mountain birch is functionally related to high leaf N concentration. The high leaf N concentration of high-altitude mountain birch is genetically determined and has an adaptive value in a cold environment. This suggests that there is a trade-off between high N productivity at low temperature and a strong response of N productivity to temperature. Received: 21 March 1998 / Accepted: 1 December 1998     

Wood ants and a geometrid defoliator of birch: predation outweighs beneficial effects through the host plant

Soil amelioration by a wood-ant species and its consequences for the larval performance of autumnal moths feeding on mountain birch were studied at various distances from the nest mound. Soil nitrate and ammonium nitrogen did not show any clear relationship with distance. However, trees growing in the mound had over 20% more foliar nitrogen than more distant trees. When moth larvae were experimentally protected from predation, their survival rate and final weight tended to decrease with increasing distance. In a laboratory experiment with detached leaves, the relative growth rate of larvae was roughly 30% higher on leaves from trees located on the mound. Differences in larval performance refute the Plant Stress Hypothesis proposed by T.C.R. White and support P.W. Price's Plant Vigor Hypothesis. Predation by ants was examined along the same gradient in trees with and without a glue band that excluded ants from the canopy. Reduction in the daily survival rate of larvae attributable to ant predation was about 35% in trees growing in the mound and around 5% at a distance of 20 m. Other things being equal, about 25 times more larvae entering the penultimate instar would achieve the pupal stage outside the wood-ant territory than in the vicinity of the mound. While both the fertilizing and predatory influence of wood ants is clear, the domain of predation is much larger than the area where trees and their herbivores can exploit enhanced nutrient levels in and around ant mounds. The existence of undamaged green islands around ant mounds in otherwise totally defoliated mountain-birch forests cannot be explained by soil amelioration by wood ants but rather by their predatory activity. Received: 21 November 1996 / Accepted: 8 September 1997