Root isoprene formation alters lateral root development.

Isoprene is a C5 volatile organic compound, which can protect aboveground plant tissue from abiotic stress such as short-term high temperatures and accumulation of reactive oxygen species (ROS). Here, we uncover new roles for isoprene in the plant belowground tissues. By analysing Populus x canescens isoprene synthase (PcISPS) promoter reporter plants, we discovered PcISPS promoter activity in certain regions of the roots including the vascular tissue, the differentiation zone and the root cap. Treatment of roots with auxin or salt increased PcISPS promoter activity at these sites, especially in the developing lateral roots (LR). Transgenic, isoprene non-emitting poplar roots revealed an accumulation of O2- in the same root regions where PcISPS promoter activity was localized. Absence of isoprene emission, moreover, increased the formation of LRs. Inhibition of NAD(P)H oxidase activity suppressed LR development, suggesting the involvement of ROS in this process. The analysis of the fine root proteome revealed a constitutive shift in the amount of several redox balance, signalling and development related proteins, such as superoxide dismutase, various peroxidases and linoleate 9S-lipoxygenase, in isoprene non-emitting poplar roots. Together our results indicate for isoprene a ROS-related function, eventually co-regulating the plant-internal signalling network and development processes in root tissue.

Effect of mechanical damage and wound healing on the viscoelastic properties of stems of flax cultivars (Linum usitatissimum L. cv. Eden and cv. Drakkar).

As plant fibres are increasingly used in technical textiles and their composites, underlying principles of wound healing in living plant fibres are relevant to product quality, and provide inspiration for biomimetic healing in synthetic materials. In this work, two Linum usitatissimum cultivars differing in their stem mechanical properties, cv. Eden (stems resistant to lodging) and cv. Drakkar (with more flexible stems), were grown without wound or with stems previously wounded with a cut parallel or transversal to the stem. To investigate wound healing efficiency, growth traits, stem biomechanics with Dynamic Mechanical Analysis and anatomy were analysed after 25-day recovery. Longitudinal incisions formed open wounds while transversal incisions generated stem growth restoring the whole cross-section but not the original stem organisation. In the case of transversal wound healing, all the bast fibre bundles in the perturbed area became lignified and pulled apart by parenchyma cells growth. Both Linum cultivars showed a healing efficiency from 79% to 95% with higher scores for transversal healing. Morphological and anatomical modifications of Linum were related to mechanical properties and healing ability. Alongside with an increased understanding of wound healing in plants, our results highlight their possible impact on textile quality and fibre yield.

Effect of Trichoderma-enriched organic charcoal in the integrated wood protection strategy.

The gradual elimination of chromium from wood preservative formulations results in higher Cu leaching and increased susceptibility to wood decay fungi. Finding a sustainable strategy in wood protection has become of great interest among researchers. The objective of these in vitro studies was to demonstrate the effect of T-720-enriched organic charcoal (biochar) against five wood decay basidiomycetes isolated from strongly damaged poles. For this purpose, the antagonistic potential of Trichoderma harzianum (strain T-720) was confirmed among other four Trichoderma spp. against five brown-rot basidiomycetes in dual culture tests. T-720 was genetically transformed and tagged with the green fluorescent protein (GFP) in order to study its antagonistic mechanism against wood decay basidiomycetes. It was also demonstrated that T-720 inhibits the oxalic acid production by basidiomycetes, a well-known mechanism used by brown-rot fungi to detoxify Cu from impregnated wood. Additionally, this study evaluated the effect of biochar, alone or in combination with T-720, on Cu leaching by different preservatives, pH stabilization and prevention of wood decay caused by five basidiomycetes. Addition of biochar resulted in a significant Cu binding released from impregnated wood specimens. T-720-enriched biochar showed a significant reduction of wood decay caused by four basidiomycetes. The addition of T-720-enriched biochar to the soil into which utility poles are placed may improve the efficiency of Cr-free wood preservatives.

Integrated control of wood destroying basidiomycetes combining Cu-based wood preservatives and Trichoderma spp.

The production of new generation of wood preservatives (without addition of a co-biocide) in combination with an exchange of wood poles on identical sites with high fungal inoculum, has resulted in an increase of premature failures of wood utility poles in the last decades. Wood destroying basidiomycetes inhabiting sites where poles have been installed, have developed resistance against wood preservatives. The objective of the in vitro studies was to identify a Trichoderma spp. with a highly antagonistic potential against wood destroying basidiomycetes that is capable of colonizing Cu-rich environments. For this purpose, the activity of five Trichoderma spp. on Cu-rich medium was evaluated according to its growth and sporulation rates. The influence of the selected Trichoderma spp. on wood colonization and degradation by five wood destroying basidiomycetes was quantitatively analyzed by means of dry weight loss of wood specimens. Furthermore, the preventative effect of the selected Trichoderma spp. in combination with four Cu-based preservatives was also examined by mass loss and histological changes in the wood specimens. Trichoderma harzianum (T-720) was considered the biocontrol agent with higher antagonistic potential to colonize Cu-rich environments (up to 0.1% CuSO4 amended medium). T. harzianum demonstrated significant preventative effect on wood specimens against four wood destroying basidiomycetes. The combined effect of T. harzianum and Cu-based wood preservatives demonstrated that after 9 months incubation with two wood destroying basidiomycetes, wood specimens treated with 3.8 kg m-3 copper-chromium had weight losses between 55-65%, whereas containers previously treated with T. harzianum had significantly lower weight losses (0-25%). Histological studies on one of the wood destroying basidiomycetes revealed typical decomposition of wood cells by brown-rot fungi in Cu-impregnated samples, that were notably absent in wood specimens previously exposed to T. harzianum. It is concluded that carefully selected Trichoderma isolates can be used for integrated wood protection against a range of wood destroying basidiomycetes and may have potential for integrated wood protection in the field.

Superior wood for violins--wood decay fungi as a substitute for cold climate.

Violins produced by Antonio Stradivari during the late 17th and early 18th centuries are reputed to have superior tonal qualities. Dendrochronological studies show that Stradivari used Norway spruce that had grown mostly during the Maunder Minimum, a period of reduced solar activity when relatively low temperatures caused trees to lay down wood with narrow annual rings, resulting in a high modulus of elasticity and low density. The main objective was to determine whether wood can be processed using selected decay fungi so that it becomes acoustically similar to the wood of trees that have grown in a cold climate (i.e. reduced density and unchanged modulus of elasticity). This was investigated by incubating resonance wood specimens of Norway spruce (Picea abies) and sycamore (Acer pseudoplatanus) with fungal species that can reduce wood density, but lack the ability to degrade the compound middle lamellae, at least in the earlier stages of decay. Microscopic assessment of the incubated specimens and measurement of five physical properties (density, modulus of elasticity, speed of sound, radiation ratio, and the damping factor) using resonance frequency revealed that in the wood of both species there was a reduction in density, accompanied by relatively little change in the speed of sound. Thus, radiation ratio was increased from 'poor' to 'good', on a par with 'superior' resonance wood grown in a cold climate.

Transpiration, CO2 assimilation, WUE, and stomatal aperture in leaves of Viscum album (L.): Effect of abscisic acid (ABA) in the xylem sap of its host (Populus x euamericana).

Leaves of the mistletoe Viscum album (L.) show a high rate of transpiration, even when the host is under severe drought stress. The hypothesis that a strong control of ABA influx from the xylem sap of the host into the mistletoe prevents stomatal closure in mistletoe leaves was tested under the following conditions: sections of poplar twigs carrying a mistletoe were perfused with artificial xylem sap that contained different ABA concentrations and both transpiration and ABA levels were analysed in mistletoe leaves. Despite variation by a factor of 10(4), the ABA content of the host xylem did not affect ABA levels, leaf transpiration, CO(2) assimilation, WUE, or the degree of stomatal aperture in mistletoe leaves. These observations support the hypothesis of a strong control of ABA influx from the host of the xylem into the mistletoe, although degradation of ABA before it enters the mistletoe leaves cannot be excluded. This mechanism may ensure a water and nutritional status favourable for the mistletoe, even if the water status of the host is impaired. Despite the lack of short-term sensitivity of ABA levels in mistletoe leaves to even strong changes of ABA levels in the xylem sap of the host, ABA levels in mistletoe leaves were relatively high compared to ABA levels in the leaves of several tree species including poplar. Since significant transpiration of the mistletoe leaves was observed despite high ABA levels, a diminished sensitivity of the stomata of mistletoe leaves to ABA has to be concluded. The stomatal density of adaxial Viscum leaves of 89+/-23 stomata per mm is lower than those reported in a study performed at the end of the 19th century.

Unusual patterns in the distribution of calcium oxalate in spruce needles and their possible relationships to the impact of pollutants.

The patterns of distribution of calcium oxalate crystals in spruce needles have been investigated in healthy needles and those subjected to either acid precipitation (ph 2.5) or various ozone fumigations. In the acid treatment, deposits of Ca oxalate were lacking in the outer walls of the epidermal cells. Under the impact of ozone, unusual complexes of Ca oxalate crystals embedded in a matrix of callose and cellulose projected from the walls inwards into the cells, filling the lumina of epidermal and hypodermal cells completely. In mesophyll cells, the complexes projected more or less from the walls into the vacuoles. In other cases, crystals appeared within the vacuoles of mesophyll cells. In dead cells, precipitates of Ca oxalate or phosphate apparently indicate destruction of the cytoplasm after a major influx of Ca2+ . It is speculated, that the physiological significance of enhanced calcium leaching has been overestimated. With regard to ozone impact, it seems that the major problem of thereby enhanced membrane permeabilities may not be an increased efflux of mineral elements, but rather an increased influx of Ca into the symplast and subsequent disturbances in metabolism.