Resynchronization of the Circadian Corticosterone Rhythm After a Light/Dark Shift in Juvenile and Adult Mice

Most of the extensive literature concerning the resynchronization of circadian rhythms after a Zeitgeber shift is devoted to the dependence of resynchronization on the mode of the shift and the strength of the Zeitgeber, as well as on the circadian function investigated. Ontogenetic influences have rarely been investigated. Therefore, we studied the resynchronization of several circadian rhythms in juvenile and adult female laboratory mice. We present here the results concerning the corticosterone rhythm. The daily rhythms were determined as transverse profiles (2-h intervals) before as well as 3, 7, and 14 days after an 8-h phase delay of the light/dark cycle produced by a single prolongation of dark time. The corticosterone concentration in serum was determined radioimmunologically. In the control animals the daily patterns were bimodal, with main maxima at the end of the light time and secondary ones just after lights on. Ontogenetic differences were small. In adult mice the amplitude was slightly increased due to an increase in the maximum values, and the time of highest hormone concentrations was slightly phase advanced. In juvenile mice, a distinct daily pattern with a phase position in relation to the light/dark cycle corresponding to that of control animals was present on the 3rd day after the Zeitgeber shift. The daily mean as well as the minimum and maximum values increased initially and reached the values of control animals during the second week. In adult animals, a pronounced daily rhythm with the normal phase position was present only at the 7th postshift day. The amplitude, daily mean, and maximum values were decreased, and the minimum values were increased. The initial values were not reached even after 2 weeks. The results show that resynchronization was faster in juvenile mice compared with adult mice. As a possible cause for the observed age-related differences, a not yet stabilized phase-coupling between various circadian rhythms is supposed.     

Biotechnology of ornamental plants: when beauty joins science—preface from the editors

Plant Cell, Tissue and Organ Culture (PCTOC) -     

Genetic diversity within and between seedstock populations of several German autochthonous provenances and conventionally propagated nursery material of blackthorn (<Emphasis Type="Italic">Prunus spinosa</Emphasis> L.)

Genetic diversity within and between seedstocks of autochthonous provenances of common blackthorn (Prunus spinosa L.) from several locations in Germany was determined and compared with the diversity in conventionally propagated (German and Hungarian) seedstocks using a highly reproducible high-annealing-temperature random amplified polymorphic DNA (HAT-RAPD) protocol. Based on the distribution of 359 markers obtained with 11 primers we found relatively low genetic diversity in the studied autochthonous blackthorn populations (H ₀ 0.1182–0.1333), with the majority distributed within the populations (92.22%) and only 7.78% among them. Similar levels of diversity were also found in the conventional seedstocks. Accordingly, genetic differentiation among these populations is rather low (pairwise F _st 0.0284–0.1266). In one case, we were not able to differentiate between an autochthonous population and conventional (F _st 0.0284) one. We discuss the results with respect to German conservation laws and their practical implementation.     

Monogenic Recessive Mutations Causing Both Late Floral Initiation and Excess Starch Accumulation in Arabidopsis

A recessive Arabidopsis mutation, carbohydrate accumulation mutant1 (cam1), which maps to position 22.8 on chromosome 3, was identified by screening leaves of ethyl methanesulfonate-mutagenized M2 plants stained with iodine for altered starch content. Increased starch content in leaves of the cam1 mutant was observed at the onset of flowering. This mutant also had a delayed floral initiation phenotype with more rosette leaves than the parental line. In addition, activities of several enzymes associated with starch metabolism were altered in the cam1 mutant. The late-flowering mutant gigantea (gi) also manifested an elevated starch level in leaves. However, not all late-flowering mutants had increased leaf starch content. Double mutants cam1 adg1 (for ADP-glucose pyrophosphorylase), cam1 pgm (for phosphoglucomutase), and gi pgm had no observable starch in leaves but showed the late-flowering phenotype, demonstrating that the elevated starch content is not the cause of late floral initiation. The pleiotropic effects of cam1 and gi suggest that they may play regulatory roles in starch metabolism and floral initiation. These data suggest that starch accumulation and floral initiation may share a common regulatory pathway.     

Transformation of cauliflower (Brassica oleracea L. var. botrytis) — an experimental survey

The paper compares different approaches for the genetic transformation of cauliflower (Agrobacterium-mediated, PEG-mediated and/or electroporation). Transient expression of the neomycin phosphotransferase II (NPTII) gene could be detected after direct gene transfer. Stable transformation was achieved using both Agrobacterium-mediated and direct gene transfer. Expression as well as incorporation of the NPTII sequence could be demonstrated.     

Production and rooting behaviour of <Emphasis Type="Italic">rol</Emphasis>B-transgenic plants of grape rootstock ‘Richter 110’ (<Emphasis Type="Italic">Vitis berlandieri × V. rupestris</Emphasis>)

Genetic improvement of grape rootstocks is aimed at protection against grape phylloxera and other soil-borne pests and diseases, good rooting and graft compatibility as well as adaptability to a wide range of soil and climatic conditions. Apart from the long evaluation period required, breeding is complicated by the high heterozygosity in grapes. As an alternative to traditional crossing, gene transfer permits addition of single traits, largely without affecting the genetic background of existing valuable cultivars. Here we report on the production and rooting behaviour of transgenic grape rootstock ‘Richter 110’ carrying the Agrobacterium rhizogenes rolB gene, which is known to promote rooting. Transformation was achieved by co-cultivation of somatic embryogenic callus with Agrobacterium tumefaciens LBA4404 harbouring plasmid pHrB. The T-DNA of pHrB contains the hpt gene, conferring hygromycin resistance, and the rolB gene under control of its own promoter. PCR using transgene-specific primers verified the presence of hpt in all 36 hygromycin resistant clones selected, while only 24 clones also possessed the rolB gene. Rooting behaviour was examined in vitro, using tip, node and internode explants, and in aeroponic culture in the greenhouse, using single-node cuttings. Compared to internodes of non-transgenic ‘Richter 110’, those of rolB-transgenic clones in general showed significantly higher rooting ability and, in contrast to the former, were able to root profusely even in the absence of auxin. Cuttings of three rolB-transgenic clones in aeroponic culture produced almost twice as many primary roots as those of the non-transgenic control.     

Age‐depending effects of narcosis and transmitter implantation on circadian body temperature and activity rhythms in mice

Summary

The effects of narcosis and of telemetry transmitter implantation on core temperature and locomotor activity were investigated in female laboratory mice of various age (3, 15 and 52 weeks old). Following surgery a transient hypothermia was observed. The body temperatures measured 30 min after beginning of narcosis were lower in juvenile and in presenile mice (29.6° ±0.8°C resp. 30.0° ±0.2°C) than in adult animals (31.9° ±0.3°C). The following temperature increase was fastest in juvenile mice. Normal body temperature was reached after 6h 20’ already. Adult and presenile mice needed 8h 30’ resp. 7h 30’. The temperature increase seemed to be independent from activity behaviour of the animals. No substantial differences could be obtained whether the transmitters had room or body temperature before implantation and whether the animals were warmed after surgery by an infrared bulb or not. Probably, the temperature increase depended mainly on the elimination rate of the drug.

Normal circadian core temperature and activity rhythms reappeared on average within 5–6 days in juvenile mice and a little faster in adult (4–5 days) as well as in presenile ones (3–4 days). However, interindividual differences in recovery time were more pronounced than age‐dependent variations.

Circadian core temperature and activity patterns were quite similar in all three age classes investigated. Ontogenetic differences concern, besides changes in daily mean values, mainly a temperature amplitude increasing with age, as well as a high percentage of ultradian components in the activity pattern of juvenile mice compared to older ones.

Telemetry systems are widely used for long‐term measurements of core temperature in laboratory animals (Clement et al., 1989; Refinetti and Menaker, 1992). In our investigations of ontogenetic changes of the circadian temperature and activity rhythms in mice we used an integrated telemetry and data acquisition system (Dataquest, Data Sciences Inc., USA). It comprises implantable wireless transmitters, telemetry receivers, a consolidation matrix and a data acquisition system. The aim of a preliminary study was to analyse the effects of narcosis and transmitter implantation. The time required to recover normal values of body temperature and of locomotor activity as well as normal circadian rhythms was determined, considering also ontogenetic variations.     

Revisiting the provenance delineation of a widespread shrub, <Emphasis Type="Italic">Frangula alnus</Emphasis>—the role of spatial,temporal and environmental patterns

Including population genetic aspects into the selection of planting material within the framework of conservation and restoration measures is of vital importance for the long-term persistence of populations. This is especially true facing climate change since genetic diversity and the spread of potentially beneficial alleles are important for the adaptability of populations. Therefore, knowledge about genetic variability within and between populations is a critical aspect when determining provenance regions. In our study, we investigated the population genetic structure of a widespread, insect-pollinated and mainly bird-dispersed shrub species, Frangula alnus, on the basis of seven microsatellites and two chloroplast DNA markers throughout Germany. The aim was to determine the spatial, temporal and ecological processes genetically structuring populations to critically revise existing provenance regions. Therefore, we conducted analyses on different spatial scales (country-wide, regional and local) using the two different marker sets in addition to environmental variables. We detected distinct patterns on all spatial scales which indicated influences of historic recolonization processes, regional differences of seed dispersal across the landscape, as well as small-scale spatial genetic structures attributable to local dispersal processes. No relation of underlying environmental gradients such as temperature or precipitation and genetic patterns was found. We conclude that different aspects of historic and more recent gene flow shape population genetic structures and that a thorough analysis on a variety of spatial, temporal and environmental scales is necessary to appropriately select planting material for conservation and restoration measures. Correspondingly, management advice regarding provenance delineations will be provided.     

Phytohormone-regulated β-amylase gene expression in rice

The expression of -amylase genes in rice (Oryza sativa) and its regulation by phytohormones gibberellic acid (GA) and abscisic acid (ABA) were examined. Upon germination -amylase is synthesizedde novo in aleurone cells and (GA) is not required. Exogenous addition of GA does not enhance the -amylase activity, while ABA inhibits the -amylase activity, mRNA accumulation, and the germination of rice seeds. GA can reverse ABA inhibition of -amylase expression, but not ABA inhibition of seed germination. Such regulation represents a new interaction of ABA and GA.