Vitality of tree fine roots: reevaluation of the tetrazolium test

Reduction of triphenyltetrazolium chloride (TTC) by tissue to the red-colored insoluble triphenylformazan (TF) is directly linked to the activity of the mitochondrial respiratory chain. Thus, only living tissues should reduce TTC to TF. However, TF production can be detected when the TTC test is applied to control tissues boiled in water. This artifact is mainly the result of the hot ethanol extraction step, which is used to disintegrate the cells and solubilize the TF. We observed that cell wall materials such as cellulose and pectin interact with TTC at temperatures above 60 degrees C, reducing the TTC to TF. By replacing the hot ethanol extraction step with an extraction procedure that involved grinding the boiled tissues and extracting the TF with ethanol at room temperature, the formation of TF was almost entirely eliminated. Application of the modified TTC assay to fine roots of Norway spruce from forest topsoil indicated that the extent of TTC reduction was related to root morphological class as: white fine roots > brown fine roots > black fine roots > boiled fine roots, corresponding to formation of 10.8, 6.1, 0.2 and 0.1 mM TFg(-1)DW, respectively. No significant differences in TTC reduction were recorded between fresh and frozen tissues (frozen in liquid N2) for any fine root class. Application of the modified TTC assay to seedling roots stressed either by drying or by exposure to the toxic metals Cd or Al resulted in significant decreases in TF formation that were related to both the duration of stress and the concentrations of toxic substances, indicating that the modified TTC assay can be used to assess the physiological condition of roots.     

Tree roots in a changing world

Globally, forests cover 4 billion hectares or 30% of the Earth's land surface, and 20%–40% of the forest biomass is made up of roots. Roots play a key role for trees: they take up water and nutrients from the soil, store carbon (C) compounds, and provide physical stabilization. Estimations from temperate forests of Central Europe reveal that C storage in trees accounts for about 110 t C ha₋₁, of which 26 t C ha₋₁ is in coarse roots and 1.2 t C ha₋₁ is in fine roots. Compared with soil C, which is about 65 t C ha₋₁ (without roots), the contribution of the root C to the total belowground C pool is about 42%. Flux of C into soils by plant litter (stemwood excluded) compared with the total soil C pool, however, is relatively small (4.4 t C ha₋₁ year₋₁) with the coarse and fine roots each contributing about 20%. Elevated CO₂ concentrations and N depositions lead to increased plant biomass, including that of roots. Recent analysis in experiments with elevated CO₂ concentrations have shown increases of the forest net primary productivity by about 23%, and, in the case of poplars, an increase of the standing root biomass by about 62%. The turnover of fine roots is also positively influenced by elevated CO₂ concentrations and can be increased in poplars by 25%–45%. A recently established international platform for scientists working on woody root processes, COST action E38, allows the exchange of information, ideas, and personnel, and it has the aim to identify knowledge gaps and initiate future collaborations and research activities.     

Transgenic sterility in Populus: expression properties of the poplar PTLF, Agrobacterium NOS and two minimal 35S promoters in vegetative tissues

Transgenic sterility is a desirable trait for containment of many kinds of transgenes and exotic species. Genetically engineered floral sterility can be imparted by expression of a cytotoxin under the control of a predominantly floral-tissue-specific promoter. However, many otherwise desirable floral promoters impart substantial non-floral expression, which can impair plant health or make it impossible to regenerate transgenic plants. We are therefore developing a floral sterility system that is capable of attenuating undesired background vegetative expression. As a first step towards this goal, we compared the vegetative expression properties of the promoter of the poplar (Populus trichocarpa Torr. & Gray) homolog of the floral homeotic gene LEAFY (PTLF), which could be used to impart male and female flower sterility, to that of three candidate attenuator-gene promoters: the cauliflower mosaic virus (CaMV) 35S basal promoter, the CaMV 35S basal promoter fused to the TMV omega element and the nopaline synthase (NOS) promoter. The promoters were evaluated via promoter::GUS gene fusions in a transgenic poplar hybrid (Populus tremula L. x P. alba L.) by both histochemical and fluorometric GUS assays. In leaves, the NOS promoter conveyed the highest activity and had a mean expression level 5-fold higher than PTLF, whereas the CaMV 35S basal promoter fused to the omega element and the CaMV 35S basal promoter alone directed mean expression levels that were 0.5x and 0.35x that of PTLF, respectively. Differential expression in shoots, leaves, stems and roots was observed only for the NOS and PTLF promoters. Strongest expression was observed in roots for the NOS promoter, whereas the PTLF promoter directed highest expression in shoots. The NOS promoter appears best suited to counteract vegetative expression of a cytotoxin driven by the PTLF promoter where 1:1 toxin:attenuator expression is required.     

Ertragsbildung von getreidereichen Fruchtfolgen und Getreide-monokulturen in einem extensiven und intensiven Anbausystem

Yield formation in cereal-rich crop rotations and monocultures in an extensive and intensive crop-management system
In a long duration trial, conducted from 1979/80 to 1992 at TU-Munich's research station in Roggenstein, the performance of monocultures of winter wheat, winter barley and winter rye, as well as numerous cereal-crop rotations were compared in an extensive and intensive crop-management system. The results obtained can be summarized as follows.
Over the course of 13 years, the influence of the immediately preceding crop on the yield of the main crops was of much greater significance than the rotation as a whole. With winter wheat, no yield differences could be observed between monoculture and cereal crop rotation (if the rotation did not include oats). Oats, rape, field bean, pea, potato and maize as preceding crops, however, in crop management systems, led to, on average, an increase in yield of 13 dt/ha from the following wheat. Winter barley yields were not significantly different in monoculture, cereal crop rotations and crop rotations containing 66% cereals. Furthermore, winter rye yields were the same in monocultures and cereal crop rotations. With all cereals, intensification of fertilizing and chemical plant protection led to a considerable increase in yield, but did not diminish the effects of the preceding crop. Hence, even with the use of modern agronomical techniques it is impossible to compensate for yield losses due to crop rotation.     

Terminal deoxynucleotidyl transferase activity in thymocytes of kittens

Activity of terminal deoxynucleotidyl transferase (TdT) was measured in the thymus of 8- to 14-week-old kittens. Microscopic evidence of involution or other morphologic changes were not observed in formalin-fixed hematoxylin and eosin-stained thymic tissue sections. Thymic tissue from young kittens (8 weeks old) had low TdT activity, as determined by enzymatic catalysis or immunoassay procedures. Markedly higher TdT activity was measured in thymic tissue of older kittens (13 to 14 weeks old). Feline TdT catalyzed the polymerization of deoxynucleotides at a higher rate in Mn2+ than in Mg2+. Results of protein immunoblotting experiments indicated that the major form of feline thymic TdT is a single, high molecular weight polypeptide in 13- to 14-week-old kittens. Detection of TdT activity in the feline thymus indicated that TdT may have the same diagnostic and therapeutic value in feline oncology as in human oncology.