Functions of lumican and fibromodulin: lessons from knockout mice

Lumican and fibromodulin are collagen-binding leucine-rich proteoglycans widely distributed in interstitial connective tissues. The phenotypes of lumican-null (Lum ^–/–), Fibromodulin-null (Fmod ^–/–) and compound double-null (Lum ^–/– Fmod ^–/–) mice identify a broad range of tissues where these two proteoglycans have overlapping and unique roles in modulating the extracellular matrix and cellular behavior. The lumican-deficient mice have reduced corneal transparency and skin fragility. The Lum ^–/– Fmod ^–/– mice are smaller than their wildtype littermates, display gait abnormality, joint laxity and age-dependent osteoarthritis. Misaligned knee patella, severe knee dysmorphogenesis and extreme tendon weakness are the likely cause for joint-laxity. Fibromodulin deficiency alone leads to significant reduction in tendon stiffness in the Lum ^+/+ Fmod ^–/– mice, with further loss in stiffness in a lumican gene dose-dependent way. At the level of ultrastructure, the Lum ^–/– cornea, skin and tendon show irregular collagen fibril contours and increased fibril diameter. The Fmod ^–/– tendon contains irregular contoured collagen fibrils, with increased frequency of small diameter fibrils. The tendons of Lum ^–/– Fmod ^–/– have an abnormally high frequency of small and large diameter fibrils indicating a de-regulation of collagen fibril formation and maturation. In tissues like the tendon, where both proteoglycans are present, fibromodulin may be required early in collagen fibrillogenesis to stabilize small-diameter fibril-intermediates and lumican may be needed at a later stage, primarily to limit lateral growth of fibrils Published in 2003.     

Physical and Genetic Maps of thedeafwaddlerRegion on Distal Mouse Chr 6

Thedeafwaddler(dfw) mutation, displaying motor ataxia and profound deafness, arose spontaneously in a C3H/HeJ colony and was mapped previously to distal mouse Chr 6. In this study, a high-resolution genetic map was generated by positioning 10 microsatellite markers and 5 known genes on a 968-meioses intersubspecific backcross segregating fordfw[(CAST/Ei–+/+ × C3HeB/FeJ–dfw/dfw) × C3HeB/FeJ–dfw/dfw], giving the following marker order and sex-averaged distances:D6Mit64–(0.10 + 0.10 cM)–Pang–(1.24 + 0.36 cM)–Itpr1–(0.62 + 0.25 cM)–D6Mit108–(0.52 + 0.23 cM)–D6Mit54–(0.21 + 0.15 cM)–D6Mit23, D6Mit107, D6Mit328–(0.72 + 0.27 cM)–D6Mit11–(0.21 + 0.15 cM)–dfw–(0.93 + 0.31 cM)–Gat4, D6Mit55–(0.10 + 0.10 cM)–D6Mit63–(0.31 + 0.18 cM)–Syn2–(0.62 + 0.25 cM)–D6Mit44(Rho). Female and male genetic maps are similar immediately surrounding thedfwlocus, but show marked differences in other areas. A yeast artificial chromosome-based physical map suggests that the closest markers flanking thedfwlocus,D6Mit11(proximal) andGat4, D6Mit55(distal), are contained within 650–950 kb. The human homologues of the flanking lociItpr1(proximal) andSyn2(distal) map to chromosome 3p25–p26, suggesting that the human homologue of thedfwgene is located within this same region.     

Biooxidation of ferrous iron by immobilized Acidithiobacillus ferrooxidans in poly(vinyl alcohol) cryogel carriers

PVA-cryogels entrapping about 10⁹ cells of Acidithiobacillus ferrooxidans per ml of gel were prepared by freezing-thawing procedure, and the biooxidation of Fe²⁺ by immobilized cells was investigated in a 0.365 l packed-bed bioreactor. Fe²⁺ oxidation fits a plug-flow reaction model well. A maximum oxidation rate of 3.1 g Fe²⁺ l^–1 h^–1 was achieved at the dilution rate of 0.4 h^–1 or higher, while no obvious precipitate was determined at this time. In addition, cell-immobilized PVA-cryogels packed in bioreactor maintained their oxidative ability for more than two months under non-sterile conditions. Nomenclature: C _A0 – Concentration of Fe²⁺ in feed stream (g l^–1) C _A – Concentration of Fe^{2 +} in outlet stream (g l^{– 1}) D – Dilution rate of the packed-bed bioreactor (h^–1) F – Volumetric flow rate of iron solution (l h^–1) F _A0 – Mass flow rate of Fe²⁺ in the feed stream (g h^–1) K – Kinetic constant (l l^–1 h^–1) r _A – Oxidation rate of Fe²⁺ (g l^–1 h^–1) V – Volume of packed-bed bioreactor (l) X _A – Conversion ratio of Fe²⁺ (%)     

Identification of the site of inhibition of oncogenic ras-p21-induced signal transduction by a peptide from a ras effector domain

We have previously found that a peptide corresponding to residues 35–47 of the ras-p21 protein, from its switch 1 effector domain region, strongly inhibits oocyte maturation induced by oncogenic p21, but not by insulin-activated cellular wild-type p21. Another ras–p21 peptide corresponding to residues 96–110 that blocks ras–jun and jun kinase (JNK) interactions exhibits a similar pattern of inhibition. We have also found that c-raf strongly induces oocyte maturation and that dominant negative c-raf strongly blocks oncogenic p21-induced oocyte maturation. We now find that the p21 35–47, but not the 96–110, peptide completely blocks c-raf-induced maturation. This finding suggests that the 35–47 peptide blocks oncogenic ras at the level of raf; that activated normal and oncogenic ras–p21 have differing requirements for raf-dependent signaling; and that the two oncogenic-ras-selective inhibitory peptides, 35–47 and 96–110, act at two different critical downstream sites, the former at raf, the latter at JNK/jun, both of which are required for oncogenic ras-p21 signaling.     

Vascular expression of the grp1.8 promoter is controlled by three specific regulatory elements and one unspecific activating sequence

The bean grp1.8 full-length promoter is specifically active in vascular tissue during normal development of tobacco. Deletion of a negative regulatory element resulted in ectopic activity of the promoter in cortical cells of hypocotyls, roots and stems. A 169 bp fragment (–205 to –36) of the grp1.8 promoter conferred vascular-specific expression to CaMV 35S minimal promoters whereas a 141 bp fragment (–205 to –64) strongly activated these minimal promoters both in vascular and cortical cells. These experiments defined a new regulatory element (VSE) that is essential for vascular-specific expression and is located between –64 and –36. The 141 bp grp1.8 promoter sequence had enhancer-like properties as it was active in both orientations. A 24 bp sequence (bp –119 to –96, corresponding to the SE1 regulatory element) enhanced expression from several minimal promoters strongly but unspecifically, whereas a 26 bp sequence (–98 to –73, corresponding to the RSE regulatory element) induced vascular-specific expression. Thus, the grp1.8 promoter is regulated by a combinatorial mechanism that can integrate the action of different, non-additively acting regulatory elements into vascular-specific expression.     

Biofiltration of waste gases with the fungi Exophiala oligosperma and Paecilomyces variotii

Two biofilters fed toluene-polluted air were inoculated with new fungal isolates of either Exophiala oligosperma or Paecilomyces variotii, while a third bioreactor was inoculated with a defined consortium composed of both fungi and a co-culture of a Pseudomonas strain and a Bacillus strain. Elimination capacities of 77 g m^–3 h^–1 and 55 g m^–3 h^–1 were reached in the fungal biofilters (with removal efficiencies exceeding 99%) in the case of, respectively, E. oligosperma and Paecilomyces variotii when feeding air with a relative humidity (RH) of 85%. The inoculated fungal strains remained the single dominant populations throughout the experiment. Conversely, in the biofilter inoculated with the bacterial–fungal consortium, the bacteria were gradually overgrown by the fungi, reaching a maximum elimination capacity around 77 g m^–3 h^–1. Determination of carbon dioxide concentrations both in batch assays and in biofiltration studies suggested the near complete mineralization of toluene. The non-linear toluene removal along the height of the biofilters resulted in local elimination capacities of up to 170 g m^–3 h^–1 and 94 g m^–3 h^–1 in the reactors inoculated, respectively, with E. oligosperma and P. variotii. Further studies with the most efficient strain, E. oligosperma, showed that the performance was highly dependent on the RH of the air and the pH of the nutrient solution. At a constant 85% RH, the maximum elimination capacity either dropped to 48.7 g m^–3 h^–1 or increased to 95.6 g m^–3 h^–1, respectively, when modifying the pH of the nutrient solution from 5.9 to either 4.5 or 7.5. The optimal conditions were 100% RH and pH 7.5, which allowed a maximum elimination capacity of 164.4 g m^–3 h^–1 under steady-state conditions, with near-complete toluene degradation.     

Growth and age structure of the swan mussel Anodonta cygnea (L.) at different depths in lake Mattsee (Salzburg, Austria)

Unionid clams were collected at 1–2 m, 3–4 m and 6–7 m depth in lake Mattsee, a moderately mesotrophic lake, to investigate the effect of depth on clam growth and age structure. No significant differences in age structure of Anodonta cygnea were found (p=0.65). Three and ten years old clams were present at all depths, but in different percentages. Whereas at 1–2 m 13.3% of the collected clams were <4 years old, this percentage was 4.4% at 6–7 m and 7.1% at 3–4 m. A greater percentage (6.7%) of older mussels (9, 10 years) were collected at 6–7 m than at 1–2 m (2.2%). Growth declined with depth. Total length at a given age of clams at 1–2 m and 3–4 m did not differ (p=0.54), whereas differences were significant between clams at 1–2 m and 6–7 m (p<0.05) as well as between 3–4 m and 6–7 m (p<0.05). The Growth constant k was highest at 1–2 m depth.     

Photosynthetic acclimation and photoinhibition on exposure to high light in shade-developed leaves of Fagus crenata seedlings

The physiological response of leaves developed in low light (L) on Fagus crenata seedlings exposed to different levels of high light (H: high light, M: medium light) was studied. Measurements were conducted on potted seedlings in the F. crenata forest understory. The seedlings with leaves developed in L were transferred to H (L–H) and M (L–M) in summer. On exposure to high light, the photochemical efficiency of dark-adapted PSII (F_v/F_m) immediately decreased and was followed by a subsequent recovery in both L–H and L–M leaves. The mean value of F_v/F_m in L–H leaves was lower than that in L–M leaves through experiments, indicating that the degree of photoinhibition in L–H leaves was greater than that in L–M leaves. About 1 month after transfer, 37% and 5% of leaves had fallen in L–H and L–M seedlings, respectively. This result also indicated the greater photoinhibition in L–H leaves. Moreover, the photosynthetic capacity (P_Nmax) of L–H leaves decreased. In contrast, the P_Nmax of L–M leaves increased, although the P_Nmax was lower than that of M control leaves. An increase in the xanthophyll cycle pool (VAZ), indicating an increase of the photoprotective function, was found in both L–H and L–M leaves. Especially, the VAZ pool in L–M leaves was higher than that in M leaves by the end of experiments. L–M leaves may avoid photoinhibition effectively by the decrease in excess light with the increase of the P_Nmax or VAZ pool, compared to L–H leaves. Thus, the physiological acclimation on exposure to high light depended on the degree of high light. To achieve successful photosynthetic acclimation with slight photoinhibition, the variation of light intensity before and after exposure to high light would be an important factor because of the difference in excess light.     

Sequential kinetic parameter estimation of anaerobic fluidized bed bioreactor using an optimization technique

Mathematical model parameters for the methanogenic degradation of propylene glycol were estimated in a sequential manner by means of an optimization technique. Model parameters determined from an initial experimental data set using one bioreactor were then verified with the results from a second bioreactor. The proposed methodology is a useful tool to obtain model parameters for continuous flow reactors with completely mixed regime. Abbrevations: S – substrate concentration (mg COD l^–1); S _in – influent substrate concentration (mg COD l^–1); D _L – dilution rate (day^–1); – stoichiometric coefficients (ND); nx – number of microbial species (ND); X _S – fixed biomass concentration (mg biomass l^–1); X _L – suspended biomass concentration of (mg biomass l^–1); k _d – decay rate of biomass (day^–1); b _S – specific detachment rate of biofilm (day^–1); – specific growth rate of biomass (day^–1); _m – maximum specific growth rate of biomass (day^–1); K _S – half saturation constant (mg COD l^–1); K _I – inhibition constant (mg COD l^–1).     

Preliminary compositional nutrient diagnosis norms for cowpea (Vigna unguiculata (L.) Walp.) grown on desert calcareous soil

This study calculated the compositional nutrient diagnosis (CND) norms of cowpea (Vigna unguiculata (L.) Walp.), as well as identified significant nutrient interactions of this crop growing in an irrigated calcareous desert soil. Three genotypes were distributed in rows in a 2-ha field. The soil showed high heterogeneity in its chemical properties. For statistical analysis, 86 foliar composite samples from healthy plants were used. Preliminary CND norms were developed using a cumulative variance ratio function and the ² distribution function. Means and standard deviations of row-centered log ratios V_X of five nutrients (N, P, K, Ca, and Mg) and a filling value R, which included all nutrients not chemically analyzed. Preliminary CND norms are: V_N^*=0.174±0.095, V_P^*=–2.172±0.234, V_K^*=–0.007±0.267, V_Ca^*=–0.022±0.146, V_Mg^*=–1.710±0.132, and V_R5^*=3.728±0.084. These CND norms are associated with dry bean yields higher than 1.88 t ha^–1, and are associated with the following foliar concentrations: 26.2 g N kg^–1, 2.5 g P kg^–1, 22.9 g K kg^–1, 21.6 g Ca kg^–1, and 4 g Mg kg^–1. Cowpea plants growing in desert calcareous soils took up lower amounts of N, P, and K than those considered as optimum in a previous report. Six interactions were strongly indicated for cowpea through principal component analyses: positive for Ca–Mg, and negative for N–Ca, N–Mg, Ca–P, Mg–P, and K–P. Furthermore, two interactions were identified using simple correlations, negative N–P and positive K–Ca.     

Deletion analysis of the Brassica napus cruciferin gene cru 1 promoter in transformed tobacco: promoter activity during early and late stages of embryogenesis is influenced by cis-acting elements in partially separate regions

To define sequences in the cruciferin gene cru1 promoter of importance for expression, tobacco (Nicotina tabacum L.) plants were transformed with constructs in which the cru1 promoter, in front of the intact cru1 structural gene, was truncated at –1216, –974, –736, –515, –306, –46 and –17 bp relative to the cap-site. Cru1 expression in tobacco seeds was studied by Northern analysis, Western analysis and in-situ hybridizations. Comparisons of the Northern analysis of RNA from tobacco seeds harvested at 18 d after pollination with the Western analysis of protein from mature seeds showed that the regions between –974 to –736 and –306 to –46 were important for the expression of cru1 at an early developmental stage, whereas the regions –736 to –515 and –515 to –306 were important for expression throughout embryogenesis. By investigating the mRNA levels in transgenic seeds at different stages of development, indications were obtained that the two latter regions exerted their effects during the later stages. The in-situ hybridization showed that cru1 mRNA was distributed in parenchyma cells throughout the embryo in seeds expressing constructs –974 and –736. Constructs –515 and –306 showed an expression restricted to the axis or axis and parts of the cotyledons. Sequence comparisons of the cru1 promoter with other storage-protein gene promoters, identified several motifs implicated in gene regulation. Gel retardation assays with synthetic oligonucleotides showed that a region present in both cru1 and BnC1 promoters, a CANNTG motif, an SEF3 motif, an abscisic-acid-responsive element and an RY-like motif interacted specifically in vitro with DNA-binding proteins present in nuclear extracts from seeds of Brassica napus L. harvested 40 d after pollination.Abbreviations ABA abscisic acid - DAP days after pollination This work was supported by grants from the Swedish Research Council for Forestry and Agriculture and from the Swedish Natural Science Research Council. Ms Ulla Pihlgren, Elisabeth Westergren and Elfi Öhren are acknowledged for expert technical assistance.     

Genetic Polymorphism of Erythrocytic Enzymes in Yakut Populations

Polymorphism of the erythrocytic enzymes PGM1, ACP1, ESD, and GLO1 was found in Yakut populations. The allelic frequencies of the polymorphic systems studied varied within the following ranges: PGM1*1+, 0.5833–0.7791; PGM1*1–, 0.0345–0.1176; PGM1*2+, 0.1250–0.2813; ACP1*A, 0.1429–0.3382; ACP1*B, 0.6548–0.8571; ESD*2, 0.1250–0.4643; and GLO1*1, 0.0116–0.2845.     

Intrasexual Competition and Mating Behavior in <Emphasis Type="Italic">Ptomascopus morio</Emphasis> (Coleoptera: Silphidae Nicrophorinae)

In nicrophorine beetles, genus Nicrophorus care their larva using small vertebrate carrion, whereas genus Ptomaucopusreproduce with small vertebrate carrion but show no parental care. Aggression and sexual behavior were examined in Ptomascopus morio and Nicrophorus quadripunctatus. Nicrophorus quadripunctatus had intense female–female as well as male–male contests. In Ptomascopus morio, by contrast, female–female aggression was rarely observed. Male–male aggression (pushing, biting, male–male mounting) in Ptomascopus morio was observed when a resource for breeding was present, whether or not a female was present. The lack of female–female aggression, and male–male aggression when resources but not females are present, suggest that the mating system of Ptomascopus morio is resource defense polygyny. Large males of Ptomascopus morio were also found to exhibit mate choice, preferring large females over small females.     

Interannual and between species comparison of the lipids, fatty acids and sterols of Antarctic krill from the US AMLR Elephant Island survey area

Antarctic euphausiids, Euphausia superba, E. tricantha, E. frigida and Thysanoessa macrura were collected near Elephant Island ¦ during 1997 and 1998. Total lipid was highest in E. superba small juveniles (16 mg g⁻¹ wet mass), ranging from 12 to 15 mg in other euphausiids. Polar lipid (56–81% of total lipid) and triacylglycerol (12–38%) were the major lipids with wax esters (6%) only present in E. tricantha. Cholesterol was the major sterol (80–100% of total sterols) with desmosterol second in abundance (1–18%). 1997 T. macrura and E. superba contained a more diverse sterol profile, including 24-nordehydrocholesterol (0.1–1.7%), trans-dehydrocholesterol (1.1–1.5%), brassicasterol (0.5–1.7%), 24-methylenecholesterol (0.1–0.4%) and two stanols (0.1–0.2%). Monounsaturated fatty acids included primarily 18:1(n−9)c (7–21%), 18:1(n−7)c (3–13%) and 16:1(n−7)c (2–7%). The main saturated fatty acids in krill were 16:0 (18–29%), 14:0 (2–15%) and 18:0 (1–13%). Highest eicosapentaenoic acid [EPA, 20:5(n−3)] and docosahexaenoic acid [DHA, 22:6(n−3)] occurred in E. superba (EPA, 15–21%; DHA, 9–14%), and were less abundant in other krill. E. superba is a good source of EPA and DHA for consideration of direct or indirect use as a food item for human consumption. Lower levels of 18:4(n−3) in E. tricantha, E. frigida and T. macrura (0.4–0.7% of total fatty acids) are more consistent with a carnivorous or omnivorous diet as compared with herbivorous E. superba (3.7–9.4%). The polyunsaturated fatty acid (PUFA) 18:5(n−3) and the very-long chain (VLC-PUFA), C₂₆ and C₂₈ PUFA, were not present in 1997 samples, but were detected at low levels in most 1998 euphausiids. Interannual differences in these biomarkers suggest greater importance of dinoflagellates or some other phytoplankton group in the Elephant Island area during 1998. The data have enabled between year comparisons of trophodynamic interactions of krill collected in the Elephant Island region, and will be of use to groups using signature lipid methodology.     

Exchange of HCO 3 − for monovalent anions across the human erythrocyte membrane

Summary A stopped-flow rapid reaction apparatus was used for measuring changes in extracellular pH (pH _o ) of red cell suspensions under conditions wheredpH _o /dt was determined by the rate of HCO ₃ ^– /X ^– exchange across the membrane (X ^–=Cl^–, Br^–, F^–, I^–, NO ₃ ^– or SCN^–). The rate of the exchange at 37°C decreased forX ^– in the order: Cl^–>Br^–>F^–>I^–>NO ₃ ^– >SCN^–, with rate constants in the ratios 10.860.770.550.520.31. When HCO ₃ ^– is exchanged for Cl^–, Br^–, F^–, NO ₃ ^– or SCN^–, a change in the rate-limiting step of the process takes place at a transition temperature (T _T ) between 16 and 26°C. In I^– medium, however, no transition temperature is detected between 3 and 42°C. AlthoughT _T varies withX ^–, the activation energies both above and belowT _T are similar for Cl^–, Br^–, NO ₃ ^– and F^–. The values of activation energy are considerably higher whenX ^–=I^– or SCN^–. The apparent turnover numbers calculated for HCO ₃ ^– /X ^– exchange (except forX ^–=I^–) at the correspondingT _T ranged from 140 to 460 ions/site ·sec for our experimental conditions. These findings suggest that: (i) HCO ₃ ^– /X ^– exchange for allX ^– studied takes place via the rapid anion exchange pathway; (ii) the rate of HCO ₃ ^– /X ^– exchange is influenced by the specific anions involved in the 11 obligatory exchange; and (iii) the different transition temperatures in the Arrhenius diagrams of the HCO ₃ ^– /X ^– exchange do not seem to be directly related to a critical turnover number, but may be dependent upon the influence ofX ^– on protein-lipid interactions in the red blood cell membrane.     

Influence of operational parameters in the flocculation and production ofLactobacillus plantarum

Summary The influence of different operational parameters, such as the dilution rate (D) and the bleeding rate (B), in the production of a flocculent strain ofLactobacillus plantarum was studied. The effect of the dilution rate was demonstrated to be related to the lactic acid concentration inside the reactor. The effect of the bleeding rate was shown to be critical in the stabilization of the operation (due to a better pH control). It also allowed a continuous recovery of cells outside the reactor. Viability testing of the lactic starter cultures showed that operation with cell purge increased the viability of the starter cultures obtained.Nomenclature B Bleeding rate, h^–1 - D Dilution rate, h^–1 - F Feed flow rate, L h^–1 - I Feed velocity, m h^–1 - Specific growth rate, h^–1 - v Lactic acid specific productivity, g g^–1 h^–1 - P Product concentration (lactic acid), g L^–1 - P _out Product concentration leaving the system, g L^–1 - Q _b Bleeding flow rate, L h^–1 - R Recirculation velocity, m h^–1 - S Substract concentration, g L^–1 - t Time, h - T _p Time of ascensional flow (length of the column/total ascensional velocity), h - T _r Residence time (1/D), h - V Volume of the reactor, L - X Cell concentration, g L^–1 - X _out Cell concentration leaving the system, g L^–1     

Inhibitory effect of garlic on bacterial pathogens from spices

An unconventional technique for primary screening of bacterial susceptibility to garlic (Allium sativum Linn.), using a slice from its clove, was described. Aqueous extracts of garlic were found to possess a potent bacteriostatic principle against Gram-positive as well as Gram-negative foodborne bacterial pathogens. In agar medium, the minimum inhibitory concentrations (MICs) of garlic were 6–10 mg ml^–1 for Bacillus cereus, 30–40 mg ml^–1 for Staphylococcus aureus (excepting the isolate from garlic, where the MIC was 100 mg ml^–1), 20–30 mg ml^–1 for Clostridium perfringens, 10 mg ml^–1 for Escherichia coli (30 mg ml^–1 for the garlic isolate), 40–100 mg ml^–1 for Salmonella, and 10–40 mg ml^–1 for Shigella. It inhibited the growth of all these strains, which were resistant to some commonly used antibiotics. Most of the tested strains were resistant to penicillins, although sensitive to garlic. While the growth of B. cereus and Cl. perfringens was completely inhibited at 10 and 70 mg garlic, respectively, ml^–1 test broth, their respective enterotoxin production ceased at 10 and 50 mg garlic ml^–1.     

The dispersion of Trogoderma granarium in a temperature gradient and comparison with other stored product beetles

The response of stored-product beetles to a temperature gradient was measured with particular emphasis on the initial distribution. When initially introduced at the centre, the following zones were preferred: for Trogoderma granarium 28–33°C (The peak response of females shifts toward the warm side if they are mixed with males, in comparison to the response of female population). Tribolium castaneum 25–34°, Oryzaephilus surinamensis 22–26°, Tenebrio molitor 23–28°, Sitophilus oryzae 20–24°, Callosobruchus maculatus 22–24°, Rhyzopertha dominica 22–28°. T. castaneum and T. molitor aggregated at the corners under isothermal conditions. Some of the species, especially C. maculatus, show hardly any dispersal either in a temperature gradient or under isothermal conditions.

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Zusammenfassung Die Reaktion von vorratsschädlichen Käfern gegenüber einem Temperaturgradienten wurde mit besonderer Berücksichtigung der Ausgangsverteilung gemessen. Bei ursprünglicher Einbringung in das Zentrum wurden folgende Vorzugsbereiche festgestellt; für Trogoderma granarium 28–33°. Der Reaktionsgipfel der Weibchen verschiebt nach der warmen Seite wenn sie mit Männchen gemischt sind, in Vergleichung zu der Responz der weiblichen Population, für Tribolium castaneum 25–34°, für Oryzaephilus surinamensis 22–26°, für Tenebrio molitor 23–28°, für Sitophilus oryzae 20–24°, für Callosobruchus maculatus 22–24° und für Rhyzopertha dominica 22–28°. T. castaneum und T. molitor sammelten sich unter isothermalen Bedingungen in den Ecken. Einige der Arten zeigten weder in einem Temperaturgefälle noch unter isothermalen Bedingungen irgendeine geordnete Verteilung.

     

Promoter analysis of the auxin-regulated tobacco glutathione S-transferase genes Nt103-1 and Nt103-35

We have analysed the promoter regions of two closely related auxin-regulated glutathione S-transferase genes. All active deletion constructs tested showed expression of the reporter gene -glucuronidase (gusA) in root tips of young seedlings and newly developing lateral roots. Auxin treatment greatly enhanced the level of expression. The Nt103-1 promoter region –370/–276 was found to be necessary, at least as a quantitative element to confer auxin-responsiveness to a reporter gene, and sequences responsible for the auxin-responsiveness must be located downstream of –370. The region –651/–370 contains sequence information necessary for uninduced expression. The Nt103-35 promoter manifested its auxin-responsiveness within the –504/–310 region. Electrophoretic mobility shift analysis, using nuclear extracts from tobacco leaves and suspension cells, identified a factor binding to a sequence (ap103, TGAGTCT) at position –560 of the Nt103-1 promoter, which shows homology to the mammalian AP-1 site. A second factor was found to bind a sequence (as103, ATAGCTAAGTGCTTACG) with homology to the CaMV 35S promoter as-1 element. The as103 element is present in both promoters and positioned around –360, so within the region determined to be indispensable for the response to auxin. A third factor was found binding to the –276/–190 region of both promoters. Combined, these data point to the relevance of a 90 bp region for auxin-induced activity of both tobacco genes. The ASF-1 like factor binding to the as103 element within this region might be involved in mediating the auxin response.     

Linkage of Genes Controlling Morphological Traits with Isozyme Markers of Rye Chromosomes

Data on linkage of 12 rye genes controlling morphological traits (el, Vs, ln, w, np, ct2, Hs, Ddw, cb, mn, vi1, mp) with one or several isozyme markers of individual rye chromosomes (2R–7R) are presented. Linkage of the following gene pairs was established: chromosome 2R: Est3/5–el, el–-Glu, Sod2–el, Sod2–Vs; chromosome 3R: ln–Got4; chromosome 4R: w–Got1, np–Got1; chromosome 5R: Est4–ct2, Est6/9–ct2, ct2–Est2, ct2–Aco2, Est2–Hs, Aco2–Hs, Est2–Ddw, Aco2–Ddw; chromosome 6R:Lap2–cb, cb–Aco1, Est10–mn; chromosome 7R: Acph2/3–vi1, Got2–vi1, mp–Acph2/3. The reasons for mapping a very small number of genes in rye in spite of high intraspecific variability of this species are discussed. An approach is suggested to improve this situation by simultaneous identification and mapping of all diverse spontaneous mutations maintained in heterozygous state in various rye cultivars.