Metabolism of gibberellin A19 is under photoperiodic control in Populus, Salix and Betula, but not in daylength-insensitive Populus overexpressing phytochrome A

Application experiments have suggested that short‐day‐induced cessation of elongation growth in trees is caused by photoperiodic regulation of the conversion of gibberellin GA₁₉ to GA₂₀. In the present study we examined further the photoperiodic control of GA metabolism in trees with focus on the conversion of GA₁₉ in Salix pentandra, hybrid aspen (Populus tremula × tremuloides) and silver birch (Betula pendula) using [17,17‐²H₂]‐GA₁₉ or unlabelled GAs in application studies. GA₂₀ and GA₁ were able to restore growth also in hybrid aspen and silver birch under short days (SD), whereas GA₁₉ had no or only a very small activity. Contrary to hybrid aspen and S. pentandra, the activity of GA₂₀ in silver birch was significantly lower than that of GA₁. Gas chromatography‐mass spectrometry (GC‐MS) analysis revealed a smaller turnover of [²H₂]‐GA₁₉ in SD than in long days (LD) in hybrid aspen. No such difference in turnover of [²H₂]‐GA₁₉ was observed in photoperiod‐insensitive hybrid aspen overexpressing PHYA. Application of unlabelled GAs to seedlings of S. pentandra, hybrid aspen and silver birch under SD followed by quantification of metabolites by GC‐MS analysis, showed that applied GA₁₉ was not readily converted to GA₂₀ and GA₁. Although the sensitivity to GAs is also known to decrease under SD, the present data are in favour of a photoperiodic regulation of the metabolism of GA₁₉in vivo in the woody species S. pentandra, hybrid aspen and silver birch. The data might also suggest that silver birch differs from S. pentandra and hybrid aspen by exhibiting a possible photoperiodic control also of the conversion of GA₂₀ to GA₁.     

Batatasin‐III and the allelopathic capacity of Empetrum nigrum

Batatasin‐III (3,3‐dihydroxy‐5‐methoxybibenzyl) is a phenolic compound associated with the allelopathic effect of the evergreen dwarf shrub Empetrum nigrum, and has been referred to as the causal factor for the species being successful in dominating extensive ecosystems. Yet, only a few plant species have been tested for their response to batatasin‐III, and little is known about whether environmental factors modify this allelopathic effect. In this study, we tested the inhibitory effect of purified batatasin‐III through bioassays on 24 vascular plant species and, for certain species, we tested if this effect depended on growth substrate (mineral vs organic substrate), pH, and fertilization. Moreover, we tested if batatasin‐III predicted the allelopathic effect of E. nigrum by analyzing the inhibitory effect of E. nigrum leaves and humus in relation to their batatasin‐III content. Our results confirmed batatasin‐III as a stable compound capable of inhibiting germination and/or mean root elongation in all of the tested species, but this effect was modified by growth substrate. Surprisingly, the measured batatasin‐III content of E. nigrum leaves (mean value 19.7 ± 10.8 (SE) mg g^?1) and humus (mean value of 1 ± 1.5 (SE) μg g^?1) did not predict the inhibitory effect on mean root elongation. Although batatasin‐III was found to be phytotoxic to all the tested species, we conclude that this substance alone should not be used as a proxy for the allelopathic effect of E. nigrum.     

Investigation of apoptosis in cultured cells infected with equine herpesvirus 1

Many viruses alter different stages of apoptosis of infected cells as a strategy for successful infection. Few studies have addressed mechanisms of equine herpesvirus 1 (EHV-1) strain-induced cell death. We investigated the effect of an abortigenic strain (AR8 strain) on heterologous Madin–Darby bovine kidney cells and homologous equine dermis (ED) cells cell lines. We compared morphologic and biochemical features of early and late apoptosis at different postinfection times. We investigated translocation of phosphatidylserine to the cell surface, nuclear fragmentation and changes in the cytoskeleton using flow cytometry and annexin V/propidium iodide staining, DNA laddering, terminal deoxynucleotidyl transferase UTP nick-end labeling assay and immunofluorescence staining of cytokeratin 18 cleavage. AR8 EVH-1 strain interfered with apoptosis in both cell lines, particularly during the middle stage of the replication cycle; this was more evident in ED cells. Although this antiapoptotic effect has been reported for other alpha herpesviruses, our findings may help elucidate how EHV-1 improves its infectivity during its cycle.     

Ecotype-dependent control of growth,dormancy and freezing tolerance under seasonal changes in <Emphasis Type="Italic">Betula pendula</Emphasis> Roth

Woody plants in the temperate and boreal zone undergo annual cycle of growth and dormancy under seasonal changes. Growth cessation and dormancy induction in autumn are prerequisites for the development of substantial cold hardiness in winter. During evolution, woody plants have developed different ecotypes that are closely adapted to the local climatic conditions. In this study, we employed distinct photoperiodic ecotypes of silver birch (Betula pendula Roth) to elucidate differences in these adaptive responses under seasonal changes. In all ecotypes, short day photoperiod (SD) initiated growth cessation and dormancy development, and induced cold acclimation. Subsequent low temperature (LT) exposure significantly enhanced freezing tolerance but removed bud dormancy. Our results suggested that dormancy and freezing tolerance might partially overlap under SD, but these two processes were regulated by different mechanisms and pathways under LT. Endogenous abscisic acid (ABA) levels were also altered under seasonal changes; the ABA level was low during the growing season, then increased in autumn, and decreased in winter. Compared with the southern ecotype, the northern ecotype was more responsive to seasonal changes, resulting in earlier growth cessation, cold acclimation and dormancy development in autumn, higher freezing tolerance and faster dormancy release in winter, and earlier bud flush and growth initiation in spring. In addition, although there was no significant ecotypic difference in ABA level during growing season, the rates and degrees of ABA alterations were different between the ecotypes in autumn and winter, and could be related to ecotypic differences in dormancy and freezing tolerance.     

Analysis of gene regulation in growing pollen tubes of angiosperm and gymnosperm species using microprojectile bombardment

A microprojectile based transient expression assay was used to investigate the functional conservation of gene regulatory mechanisms in the male gametophytes of an angiosperm ( Nicotiana tabacum ) and two gymnospermous ( Picea abies and Pinus pinaster ) species. The activities of two angiosperm gene promoters, which have previously shown to be either preferentially expressed in the male gametophyte ( lat52 ) or highly expressed in both the sporophyte and male gametophyte ( Act I), were analysed. The results showed that in P. abies and P. pinaster , activity of the Act 1 promoter was significantly higher than the activity of the lat52 promoter, while the converse was observed in N. tabacum . Detailed analysis of lat52 5'promoter deletions demonstrated that although the minimal -67 bp lat52 core promoter was active at low levels in all three species, upstream regulatory elements conserved among several pollen-expressed genes, including the PBI element, were not functional in P. abies and P. pinaster . These results suggest that both taxa-specific and conserved regulatory mechanisms operate to control gene expression during pollen germination and tube growth.     

Lack of influence of photoperiod on the metabolism of gibberellin A20 in Salix pentandra

The influence of photoperiod on the metabolism of GA₂₀ in Salix pentandra was studied by feeding [³H]-GA₂₀ to seedlings which had been grown previously under long day (LD) or short day (SD) conditions. After 48 h in LD or SD, metabolites were separated on sequential, silica gel partition columns and reversed-phase C₁₈ HPLC. The principal metabolite co-chromatographed with [³H]-GA₁ and this conversion was confirmed by feeding [²H]-GA₂₀, which was converted to [²H]-GA₁ as identified by gas chromatography-selected ion monitoring. Chromatographic evidence also indicated the minor conversion of [³H]-GA₂₀ to [³H]-GA₈ (via [³H]-GA₁) and trace conversion to [³H]-GA₂₉ (GAs A_1.8,20.29 are native in Salix). Ethyl acetate-insoluble [³H] metabolites were formed and could be cleaved by cellulase to release putative [³H]-GA₂₀ and [³H]-GA₁ suggesting the conversion to glucosyl conjugates of these GAs. Metabolism of [³H]-GA₂₀ was slightly more rapid in plants previously grown under LD than SD, an effect which reflected the generally increased shoot growth under LD. However, altering the photoperiod after [³H]-GA₂₀ addition had only a slight effect on the metabolism of [³H]-GA₂₀ in Salix seedlings. This indicates that the conversion of GA₂₀ to GA₁ is not a controlling step in the photoperiodic regulation of growth cessation in Salix.     

Stent implantation through a self-expanding stent

Jailing of a side-branch is a known complication of stent implantation, and makes access to the side-branch difficult, especially if the stent is of the self-expanding type. Although plain balloon angioplasty is feasible for the jailed side-branches, the use of newer devices (a stent, Rotablation or atherectomy) has not been described. We describe a novel way of treating a side-branch jailed by a self-expanding stent by using stent implantation through the strut of a self-expanding stent.     

Effects of different gibberellins on elongation growth under short day conditions in seedlings of Salix pentandra

Fifteen different gibberellins (GA's) were tested for their ability to induce elongation growth under short day conditions in seedlings of Salix pentandra L. GA's were applied either to the apex or they were injected into a mature leaf. GA₃ was highly active and also GA₄₊₇ and GA₄ showed high activity. GA₁, GA₂, GA₅, GA₉, GA₁₃, GA₂₀, GA₃₆ and GA₄₇ showed moderate activity. GA₁₆, GA₁₇, GA₂₇ and GA₄₁ exhibited low or no activity in doses up to 10 μg per plant. In general, a better growth response was obtained with an application to the apex than with an injection into the leaf.     

Experimental control of flowering and vivipary in timothy (Phleum pratense)

Environmental and hormonal control of flowering and vivipary in four Norwegian timothy ( Phleum pratense L.) cultivars has been studied in phytotron and by aseptic culture of inflorescence explants. The critical photoperiod for flowering increased with increasing temperature (12–18°C) and it was 13 to 15 h for the southern and 14 to 16.5 h for the northern cultivars. Diurnal temperature fluctuation significantly stimulated flower formation compared to the corresponding constant temperature treatment. Plants grown in 16-h photoperiod contained normal sexual flowers, but a high percentage of spikes developed in 12- or 14-h photoperiod contained viviparous plantlets. One- to four-weeks in continuous light before treatment with 12-h photoperiod increased the number of spikes per plant, but did not enhance the frequency of vivipary. Experiments with aseptic cultures showed that generative versus vegetative development of timothy inflorescence was affected by plant hormones. Kinetin stimulated the vegetative development and induced proliferation both in inflorescence initials and in spikelets isolated at heading time.