Establishment of cryopreserved gene banks of European chestnut and cork oak

Cryopreservation of selected genotypes of European chestnut and cork oak was carried out in two laboratories in a project involving conservation of field collections. Plant material was selected on the basis of disease resistance (chestnut), growth habit, phytosanitary performance and cork quality (cork oak). The cryopreservation technique comprised of vitrification of shoot apices isolated from in vitro stock shoot cultures (chestnut) and somatic embryos (cork oak). Forty-three out of 46 chestnut genotypes assayed survived the freezing process, but only 63% recovered their capacity to produce new shoots. After completion of multiplication and rooting steps, the surviving shoots produced plants that were morphologically identical to those derived from non-supercooled material. All 51 cork oak genotypes withstood freezing and were able to produce new somatic embryos through a process of secondary embryogenesis. Multiplication and germination of the recovered embryos enabled production of plants that were morphologically identical to those derived from non-supercooled material. In light of the results obtained, long-term cryopreservation of these species is feasible, thereby ensuring conservation of valuable genotypes during field evaluation.     

Nutritional evaluation of “Braquiarão” grass in association with “Aroeira” trees in a silvopastoral system

This study aimed at evaluating the influence of a silvopastoral system (SPS) composed of “Aroeira” trees (Myracrodruon urundeuva) on the nutritional value of Brachiaria brizantha cv. Marandu forage grass over the rainy season. The experiment was conducted in the SPS located in the “Cerrado” biome in Lagoa Santa County, Minas Gerais state, Brazil, in the agricultural year of 2004–2005. It was evaluated the influence of “Aroeira”, whose density in the area was 140 trees/ha, on the following parameters of the B. brizantha cv. Marandu forage grass: forage mass harvest above stubble height, morphogenic attributes, bromatological composition and nutritional value. The effects of “Aroeira” trees on the micro-climatic conditions were also evaluated. The experiment adopted a randomized block design with five blocks (cutting seasons), two treatments (a shaded pasture—B. brizantha cv. Marandu shaded by M. urundeuva—and a pasture in full sun—B. brizantha cv. Marandu in full sun), and three replicates. The tree component, composed of the tree species M. urundeuva in the experimental density adopted, increased dry matter and crude protein of the forage mass harvest above stubble height, without affecting the concentration of neutral detergent fibre or acid detergent fibre, thus causing dry matter and organic matter in vitro degradability as well as effective degradability of the forage grass to remain the same as those obtained in full sun. Therefore, the results of this study show the possibility of using trees and grass to increase both animal production and environmental services.     

Site preference and occurrence patterns of <Emphasis Type="Italic">Picea jezoensis</Emphasis> and <Emphasis Type="Italic">Abies sachalinensis</Emphasis> on decayed logs in natural coniferous forests in Hokkaido,northern Japan

The objectives of this study were to investigate differences in the site preferences of seedlings of Picea jezoensis and Abies sachalinensis on decayed logs, and to examine the occurrence patterns of seedlings and saplings of the two species and whether they occur together or separately on logs. We characterized the habitats of 1–2-year-old seedlings of the two species on logs and examined the relationship of the two species on logs by growth stages in two plots. One plot had been disturbed about 50 years ago whereas the other had not for a long time. Although the thickness of moss and the litter layer in the habitats of 1–2-year-old seedlings were significantly different between the two species, the two species could occur together. In one study plot, seedlings and saplings of the two species occurred together. The initial occurrence pattern of the seedlings affected the occurrence patterns of the saplings on logs. The occurrence patterns of the seedlings and saplings of the two species on logs seemed to be affected by the abundance of seed trees. In the other study plot saplings of the two species occurred separately, but one species was not always competitively superior to the other species. Disturbance history affected the occurrence patterns of the saplings of the two species on decayed logs at the two study plots. Consequently, it is concluded that seed dispersal and the abundance of available logs, which are usually affected by disturbance, are significant factors in the natural regeneration of conifers in Hokkaido.     

Effects of water stress induced by sodium chloride and mannitol on proline accumulation,photosynthetic abilities and growth characters of eucalyptus (<Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis> Dehnh.)

The aim of this study was to investigate the biochemical, physiological and morphological responses of eucalypts to iso-osmotic salt and water deficit stress. Chlorophyll a, chlorophyll b, total chlorophyll (TC), total carotenoids, maximum quantum yield of PSII (F _v /F _m ), photon yield of PSII and non-photochemical quenching, in severely osmotic-stressed plantlets (−1.20 MPa) decreased significantly when compared to control plantlets, leading to low net photosynthetic rate (P _n ) and growth reduction. In addition, the photosynthesis and growth parameters of salt stressed plantlets declined to a greater degree than those cultivated in water-deficit conditions. On the other hand, the proline content of the osmotic-stressed leaves increased significantly, especially under iso-osmotic salt stress conditions. The TC content of iso-osmotic stressed leaves was significantly degraded, with low F _v /F _m , leading to P _n reduction and growth inhibition. Those parameters in salt stressed plantlets were significantly lower and to a greater degree than in plantlets subjected to water-deficit. The basic knowledge of biochemical, physiological and morphological changes in responses to iso-osmotic salt and water-deficit stresses will be further applied as effective indices for salt and water-deficit tolerant screening in large population of eucalyptus breeding program.     

Inheritance and detoxification enzyme levels in <Emphasis Type="Italic">Tetranychus urticae</Emphasis> Koch (Acari: Tetranychidae) strain selected with chlorpyrifos

Changes in insecticide susceptibilities and detoxifying enzyme activities were measured in a strain of Tetranychus urticae Koch following repeated exposure to the organophosphate insecticide, chlorpyrifos. Twelve consecutive selection at the LC₆₀ of the parental strain increased resistance from 8.58 to 91.45 fold. The interaction of some synergists [piperonyl butoxide, triphenyl phosphate and S-benzyl-O,O-diisopropyl phosphorothioate (IBP)] with chlorpyrifos was analyzed in the selected strain. Solely IBP showed a low synergistic effect with chlorpyrifos. The selected strain also demonstrated resistance against abamectin, propargite, clofentezine and fenpyroximate. The mode of resistance inheritance to chlorpyrifos was found to be incompletely dominant, and not sex-linked. Non-specific esterase enzyme activity was raised from 19.35 to 33.59 mOD/min/mg proteins during the selection period and it was observed that esterase band intensities visualized by polyacrylamide gel electrophoresis increased. This study has investigated the selection of resistance to chlorpyrifos and documented resistance to abamectin, propargite, clofentezine and fenpyroximate in Turkish T. urticae. Esterase enzymes may be playing a role in chlorpyrifos resistance while glutathione S-transferase (GST) and P450 enzymes do not appear to have any significant involvement.     

Forest monitoring with TerraSAR-X: first results

Several TerraSAR-X satellite images acquired in high resolution spotlight mode with different polarisations for two study sites in southern Germany were used to distinguish forest from other land cover classes (agriculture, built-up, water bodies) using logistic regression models. In general, we observed that the mean and particularly the standard deviation of the backscatter were viable measures to discriminate land cover classes. Both measures were lowest for water bodies and highest for built-up areas, with agricultural areas and forest in intermediate positions. Trees outside forest were not differentiable from forest with the applied method. The HH-polarised image was better suited for a classification of built-up areas, whereas the VV-polarised image was more appropriate when classifying agricultural areas. Consequently, the combination of the two differently polarised images leads to a significantly better model. Since forests in one study area were generally found on steeper slopes in comparison to other land cover classes, the inclusion of terrain slope further improved the classification, which resulted in an overall accuracy of 92–95%. Systematic differences in the parameter values of the explanatory variables for one class between the study areas may be caused by differing class probabilities. Thus, transferring the model of one study area to the image of another resulted in a 7–9% loss of accuracy.     

Five-year effects of cutting and herbicide treatments on control of <Emphasis Type="Italic">Rhododendron flavum</Emphasis> Don., and macronutrient pools in eastern beech (<Emphasis Type="Italic">Fagus orientalis</Emphasis> Lipsky) forests of Turkey

This experiment was carried out in a mature (>100-year-old) eastern beech forest in the Eastern Black Sea Region (BSR) of Turkey. The effects of experimental practices to control the woody plant Rhododendron flavum Don., including grubbing, manual cutting, and foliar and cut-stump spraying with imazapyr and triclopyr, were assessed on understory and forest floor biomass and nutrients, as well as soil bulk density and nutrient content. The cut plots had 5, 15, and 38 times greater competing vegetation biomass than did the plots treated with triclopyr, grubbing, and imazapyr, respectively. Because of the increased biomass, the amount of understory N, P, Ca, K, S, and Mg content on the cut plots was also substantially higher when compared to the grubbed, imazapyr-treated, and triclopyr-treated plots. Triclopyr and imazapyr applications brought about two- and threefold greater dead organic matter on the forest floor than did cutting and grubbing, respectively. As a result, forest floor macronutrient contents on the imazapyr- and triclopyr-treated plots were significantly greater than on the cut plots. Five years after treatments, soil bulk density at the 10–20 cm depth on the imazapyr-sprayed plots was about 14% higher than that on other treatments. Herbicides, especially imazapyr and grubbing were effective in controlling rhododendron. Macronutrients stored in the rhododendron were released to the forest floor after vegetation was controlled with herbicides.     

In vitro plant regeneration system for <Emphasis Type="Italic">Cassia siamea</Emphasis> Lam., a leguminous tree of economic importance

A method for rapid in vitro propagation of Cassia siamea Lam. using cotyledonary node explants, excised from 14-day old aseptic seedlings, has been established. Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) singly or in combination with auxins was used for regeneration studies. Among the single treatment of three cytokinins BA at 1.0 μM was found to be optimum for direct shoot regeneration as it induced an average of 8.20 ± 0.66 shoots per explant. The regeneration frequency further enhanced with the application of auxin along with optimal BA concentration. The highest frequency for shoot regeneration (90%), the maximum number of shoots per explant (12.20 ± 0.73) and the maximum shoot length (6.40 ± 0.07) cm were obtained on the medium consisted of MS + 1.0 μM BA + 0.5 μM NAA. Successful in vitro rooting was induced from cut end of the microshoots when placed on half-strength MS + IBA (2.5 μM). The regenerated shoots with well developed root system were successfully acclimatized and established in pots containing sterilized garden soil and garden manure (1:1) and grown under greenhouse conditions with 85% survival rate.