The effect of two stilbene compounds, pinosylvin and resveratrol, on the growth of several fungi was evaluated in plate tests. Wood decay tests were carried out with birch and aspen samples impregnated with the two stilbenes. In plate experiments, resveratrol had an enhancing effect on growth at concentrations where pinosylvin was already enough to prevent the growth of most fungi studied. Pinosylvin impregnated at 0.2% (w/w) concentration significantly reduced the decay caused by all fungi except Phellinus tremulae. In contrast, a resveratrol content of 0.8%, did not protect the wood from decay. A pinosylvin-synthase-encoding gene from Pinus sylvestris was transferred into aspen ( Populus tremula) and two hybrid aspen clones ( Populus tremulax tremuloides) by Agrobacterium tumefaciens-mediated transformation. Transgenic plants accumulated pinosylvin synthase-specific mRNA and showed stilbene synthase enzyme activity in vitro. Transgenic aspen line H4 showed increased resistance to Phellinus tremulae, while two hybrid aspen transformants decayed faster than the control trees. However, we were unable to detect the accumulation of stilbenes in the transgenic plantlets.

A field trial of 15 transgenic birch lines expressing a sugar beet chitinase IV gene and the corresponding controls was established in southern Finland to study the effects of the level of sugar beet chitinase IV expression on birch resistance to fungal diseases. The symptoms caused by natural infections of two fungal pathogens, Pyrenopeziza betulicola (leaf spot disease) and Melampsoridium betulinum (birch rust), were analysed in the field during a period of 3 years. The lines that had shown a high level of sugar beet chitinase IV mRNA accumulation in the greenhouse also showed high sugar beet chitinase IV expression after 3 years in the field. The level of sugar beet chitinase IV expression did not significantly improve the resistance of transgenic birches to leaf spot disease. Instead, some transgenic lines were significantly more susceptible to leaf spot than the controls. The level of sugar beet chitinase IV expression did have an improving effect on most parameters of birch rust; the groups of lines showing high or intermediate transgene expression were more resistant to birch rust than those showing low expression. This result indicates that the tested transformation may provide a tool for increasing the resistance of silver birch to birch rust.

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Well watered (WW) or drought-stressed (DS) saplings of ozone-sensitive and ozone-tolerant (less sensitive) birch (Betula pendula Roth) clones were exposed for 43 d to 0 nl l −" or 100 nl l −" ozone. Relative growth rates of leaves, stem, and roots, leaf discolouration, stomatal conductance and induction of genes encoding stress-related proteins PR-10, PAL and a LEA-group protein BP8 were determined. In general, both ozone and drought stress, singly and in combination, increased transcript levels of PR-10 in both clones. This was related to lower induction of PAL (except in older leaves of the tolerant clone), and increased proportions of visibly injured and yellowed leaves in ozone-exposed plants. The clones differed in their stomatal conductance and growth responses. In the less sensitive clone 2, ozone did not affect growth rates, but high stomatal conductance was observed in WW ozoneexposed plants. The more sensitive clone 5 showed, on the contrary, reduced growth rates and low stomatal conductance in WW ozone plants. Interestingly, clone 2 was sensitive to drought stress alone, whereas clone 5 was highly sensitive to ozone and drought stress experienced together. The results show that appearance of visible injuries (necrotic flecks) and enhanced yellowing of leaves coincided with the induction of genes for stress proteins PR-10 and PAL. The short-term growth responses, however, seemed to be separate processes. Additionally, stomatal conductance was related to leaf injuries and growth rates in a complicated manner, emphasizing the complex nature of ozone sensitivity\tolerance mechanisms in birch.Key words : Betula pendula Roth (birch), clones, ozone, drought, stress proteins. Forest trees grow in a dynamic environment and are continuously exposed to several abiotic and biotic stress factors (Lefohn, 1992). Atmospheric ozone and drought are potential stress factors for forest trees in the USA (e.g. McLaughlin & Downing, 1996) and Europe including Scandinavia (e.g. Pearson & Mansfield, 1993 ;Karlsson et al., 1995 ; * To whom correspondence should be addressed. E-mail : Elina.Paakkonen!messi.uku.fi Pa$ a$ kko$ nen, 1996 ; Pa$ a$ kko$ nen, Holopainen & Ka$ renlampi, 1997 a). Physiological, structural, and biochemical responses of birch to ozone have recently been studied in several open-field and chamber experiments. Typical ozone responses in ozonesensitive birch clones were impaired shoot growth, accelerated leaf senescence, visible leaf injuries, ultrastructural chloroplast injuries, increased stomatal density and lower stomatal conductance (Pa$ a$ kko$ nen et al., 1993(Pa$ a$ kko$ nen et al., a,b, 1996 a ; Pa$ a$ kko$ nen, 1996). Lower net photosynthesis and decreased Rubisco and chlorophyll contents have 296 E. PaW aW kkoW nen and others also been observed during open-field fumigations . In the chamber exposures, on the other hand, increased activities of superoxide dismutase, peroxidase and glutathione reductase, induction of PAL (phenylalanine ammonia-lyase), accumulation of free pu...

Mycorrhizal fungi contain chitin in their cell walls and may be influenced by transgenic chitinases. This study examined the ability of a transgenic tree, silver birch (Betula pendula Roth), constitutively expressing the sugar beet chitinase IV gene, to form ectomycorrhizae with Paxillus involutus (Batsch) Fr. Eight transgenic lines showing varying levels of sugar beet chitinase IV expression and the non-transgenic control plants were inoculated by P. involutus in vitro , and the morphology of the mycorrhizae, mycorrhization efficiency and shoot and root fresh weights were studied. All the transgenic birch lines were able to form normal ectomycorrhizae containing distinctive mantles and Hartig nets. The level of sugar beet chitinase IV expression had no significant influence on mycorrhizal colonization. The only significant differences between transgenic and control plants were detected in weight parameters. According to these results, the expression of sugar beet chitinase IV in silver birch is not deleterious to formation of ectomycorrhizae between birch and P. involutus .