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E Jeannette

Publications and source records attributed to E Jeannette.

7 recordsLinked to original sources

Variations in sucrose and ABA concentrations are concomitant with heteroblastic leaf shape changes in a rhythmically growing species (Quercus robur).

In rhythmically growing woody species such as common oak (Quercus robur L.), stem growth is discontinuous and a bud forms at regular intervals at the shoot apex. These buds are composed of different types of leaves: laminate, aborted lamina and scale. The change in heteroblastic leaf shape from laminate to aborted lamina leaves is regarded as one of the events marking shoot growth arrest. To better understand the determinism of heteroblastic leaf shape change and thus, of rhythmicity, we studied morphogenetic events during the early days of the second flush of growth in oak, as well as changes in sucrose metabolism and abscisic acid (ABA) concentrations in control plants expressing the heteroblastic leaf shape change and in defoliated plants showing no heteroblastic leaf shape change and producing only laminate leaves. In control plants, the leaf shape change was underway on Day 5 of the second flush with the differentiation of the first two aborted lamina leaves. Sucrose concentration in the apices of control plants decreased between Days 3 and 5 during differentiation of the aborted lamina leaves. An inverse pattern was observed in defoliated plants, suggesting that sucrose acts as a signal triggering heteroblastic leaf shape changes. During the same period, acid cell wall invertase activity was high in young stem and laminate leaves of control plants, whereas the activity remained constant and low in the apices. If the laminate leaves were removed, the increase in apical sucrose concentration was proportionally higher than the decrease in apical acid vacuolar invertase activity, suggesting that, in the absence of young leaves, sucrose is imported to the apex. The sucrose concentration in the apex is therefore likely to be affected by trophic competition with the expanding laminate leaves. The decrease in apex sucrose concentration may be one of the mechanisms driving heteroblastic leaf shape change. Differentiation of aborted lamina leaves was followed by a decrease in the organogenic activity of the shoot apical meristem (SAM) between Days 7 and 9. High concentrations of ABA are associated with differentiation of aborted lamina and scale leaves and with low SAM organogenic activity. Shoot apical meristem organogenic activity remained high and ABA concentration in the apex remained low in defoliated plants producing only photosynthetic leaves. These results suggest that (1) ABA is involved in the gradual conversion of embryonic leaves to abnormal leaves, thereby regulating heteroblastic leaf shape changes and (2) changes in ABA concentration influence the intensity of SAM organogenic activity. Heteroblastic development and therefore rhythmic growth could be the result of competition between apices and laminate leaves, with competition first involving sucrose and thereafter ABA.

Abscisic Acid↗

Abscisic acid plasmalemma perception triggers a calcium influx essential for RAB18 gene expression in Arabidopsis thaliana suspension cells.

Pretreatment of Arabidopsis thaliana suspension cells with impermeant calcium chelator EGTA inhibited the ABA-induced RAB18 gene expression. However, extracellular calcium alone, up to 10 mM, did not trigger RAB18 expression. Spectrofluorimetric extracellular Ca(2+) measurement with Fluo-3 showed a fast, within 1 min, Ca(2+) influx associated with outer plasmalemma ABA perception. In the presence of the Ca(2+) blockers Cd(2+) and Ni(2+), RAB18 expression was suppressed. Pimozide and fluspirilene inhibited Ca(2+) influx and ABA-induced RAB18 expression. Thus we demonstrated the involvement of specific Ca(2+) influx in the ABA signaling sequence leading to RAB18 expression.

Abscisic Acid↗

Abscissic acid specific expression of RAB18 involves activation of anion channels in Arabidopsis thaliana suspension cells.

The abscissic acid (ABA) transduction cascade following the plasmalemma perception was analyzed in intact Arabidopsis thaliana suspension cells. In response to impermeant ABA, anion currents were activated and K(+) inward rectifying currents were inhibited. Anion current activation was required for the ABA specific expression of RAB18. By contrast, specific inhibition of K(+) channels by tetraethylammonium or Ba(2+) did not affect RAB18 expression. Thus, outer plasmalemma ABA perception triggered two separated signaling pathways.

Abscisic Acid↗

Induction of RAB18 gene expression and activation of K+ outward rectifying channels depend on an extracellular perception of ABA in Arabidopsis thaliana suspension cells.

Important progress has been made regarding the characterization of the ABA signalling components using genetic and molecular approaches (Leung and Giraudat, 1998). However, we do not yet know the mechanism of ABA perception. Conflicting results concerning the site of ABA perception have been published. The prevailing view is that since ABA controls many responses, different sites of perception for ABA might exist. In order to establish the cellular localisation of the ABA receptors in Arabidopsis thaliana suspension cells, we developed two physiological tests based upon the capacity of impermeant ABA-BSA conjugate to mimic permeant free ABA effects. We show that purified ABA-BSA conjugate is able to trigger RAB18 gene expression and that this response is strictly due to the natural (+)-ABA enantiomer. The rate of RAB18 gene expression was independent of the level of ABA uptake by the cells. Using the voltage-clamp technique we show that ABA-BSA, similarly to ABA, evokes a membrane depolarization and activates time- and voltage-dependent outward rectifying currents (ORC). We demonstrate that these ORC are due to a K+ efflux as assessed by tail currents and specific inhibition by both tetraethylammonium (TEA) and Ba2+. These observations provide evidence in favour of an extracellular site for ABA perception.

Abscisic Acid↗

Chlamydomonas reinhardtii thioredoxins: structure of the genes coding for the chloroplastic m and cytosolic h isoforms; expression in Escherichia coli of the recombinant proteins, purification and biochemical properties.

Based on known amino acid sequences, probes have been generated by PCR and used for the subsequent isolation of cDNAs and genes coding for two thioredoxins (m and h) of Chlamydomonas reinhardtii. Thioredoxin m, a chloroplastic protein, is encoded as a preprotein of 140 amino acids (15,101 Da) containing a transit peptide of 34 amino acids with a very high content of Ala and Arg residues. The sequence for thioredoxin h codes for a 113 amino acid protein with a molecular mass of 11,817 Da and no signal sequence. The thioredoxin m gene contains a single intron and seems to be more archaic in structure than the thioredoxin h gene, which is split into 4 exons. The cDNA sequences encoding C. reinhardtii thioredoxins m and h have been integrated into the pET-3d expression vector, which permits efficient production of proteins in Escherichia coli cells. A high expression level of recombinant thioredoxins was obtained (up to 50 mg/l culture). This has allowed us to study the biochemical/biophysical properties of the two recombinant proteins. Interestingly, while the m-type thioredoxin was found to have characteristics very close to the ones of prokaryotic thioredoxins, the h-type thioredoxin was quite different with respect to its kinetic behaviour and, most strikingly, its heat denaturation properties.

Amino Acid Sequence↗

Expression of ADP-glucose pyrophosphorylase in maize (Zea mays L.) grain and source leaf during grain filling.

The time course of ADP-glucose pyrophosphorylase activity and of starch accumulation rate measured in grain, from pollination to maturity, in Zea mays L. plants grown outdoors, was coincident for 2 years. No such correlation was observed in the adjacent leaf, which, furthermore, presented large year-to-year differences in starch accumulation pattern. Analysis of the expression of ADP-glucose synthase at the protein levels, using antibodies directed against the Bt2 or Sh2 subunits, established that the variation of activity in the grain was explained by parallel changes in the content of both subunits. The cDNA for Bt2 and Sh2 subunits were used as probes to quantify the corresponding messenger. In grain, the time course of Bt2 and Sh2 mRNA accumulation anticipated, with a similar pattern, the specific peptide variations, which suggests a transcriptional control of expression. By contrast, the control of leaf activity by protein content was less obvious than in the grain, and changes in leaf enzyme specific activity were suggested during the first 20 d after pollination. A clone homologous to the grain Bt2 subunit cDNA was isolated from a maize leaf cDNA library, and a sequence comparison showed that the leaf clone (L2) was a partial cDNA representing one-third of the mature peptide. A 97% homology was observed between Bt2 and L2 in their coding region, but homology was poor in the 3' noncoding border. This result demonstrates that Bt2 and L2 arise from different genes presenting a tissue-specific expression pattern and provides an explanation for the earlier reported differences between leaf and grain in the size of peptide and mRNA for the Bt2-homologous subunit.

Amino Acid Sequence↗