A possible role of prolyl oligopeptidase during Linum usitatissimum (flax) seed development.

Involvement of prolyl oligopeptidases (POPs) in the control of several mammalian peptide hormone signalling pathways has been studied extensively in recent years. POPs are ubiquitous enzymes, but little attention has been paid to understanding their function in plants. Using a cDNA-AFLP approach, two flax (Linum usitatissimum) POP ESTs were identified as being specifically expressed in the early stages of flax seed development. This specific expression was confirmed using real time RT-PCR and in situ hybridisation approaches. Seed expression of Arabidopsis POP genes was measured and showed no specificity. Comparison between results obtained with flax and Arabidopsis is discussed in order to address a hypothetic function for POPs during seed formation. These results provide the first insights into POP gene expression and hypothetical function in plants.

In situ expression of two storage protein genes in relation to histo-differentiation at mid-embryogenesis in Medicago truncatula and Pisum sativum seeds.

The seed consists of several layers of specialized cell-types that divide and differentiate following a highly regulated programme in time and space. A cytological approach was undertaken in order to study the histo-differentiation at mid-embryogenesis in Medicago truncatula as a model legume, and in Pisum sativum using serial sections of embedded immature seed. Little published information is available about seed development in Medicago species. The observations from this study revealed a number of distinctive features of Medicago seed development and differentiation. Transfer cells, involved in nutrient transfer to the embryo, were clearly identified in the thin-walled parenchyma of the innermost integument. Histological Schiff-naphthol enabled carbohydrate accumulation to be followed in the different seed compartments, and revealed the storage protein bodies. Non-radioactive mRNA in situ hybridization, was carried out using mRNA probes from two highly expressed genes encoding the major vicilin and legumin A storage protein types. The timing of mRNA expression was related to that of the corresponding proteins already identified.

Dehydrin transcript fluctuations during a day/night cycle in drought-stressed sunflower.

To investigate environmental stimuli involved in the modulation of drought-induced gene expression, the influence of the day/night cycle on the expression of two dehydrin genes (HaDhn1 and HaDhn2) in leaves of sunflowers subjected to mild or severe drought stress has been studied. It was observed that the HaDhn1, but not HaDhn2, transcript oscillated in a diurnal fashion. In severely stressed plants, the peak of HaDhn1 mRNA accumulation occurred at midday.

Molecular and physiological responses to water deficit in drought-tolerant and drought-sensitive lines of sunflower. Accumulation of dehydrin transcripts correlates with tolerance.

To investigate correlations between phenotypic adaptation to water limitation and drought-induced gene expression, we have studied a model system consisting of a drought-tolerant line (R1) and a drought-sensitive line (S1) of sunflowers (Helianthus annuus L.) subjected to progressive drought. R1 tolerance is characterized by the maintenance of shoot cellular turgor. Drought-induced genes (HaElip1, HaDhn1, and HaDhn2) were previously identified in the tolerant line. The accumulation of the corresponding transcripts was compared as a function of soil and leaf water status in R1 and S1 plants during progressive drought. In leaves of R1 plants the accumulation of HaDhn1 and HaDhn2 transcripts, but not HaElip1 transcripts, was correlated with the drought-adaptive response. Drought-induced abscisic acid (ABA) concentration was not associated with the varietal difference in drought tolerance. Stomata of both lines displayed similar sensitivity to ABA. ABA-induced accumulation of HaDhn2 transcripts was higher in the tolerant than in the sensitive genotype. HaDhn1 transcripts were similarly accumulated in the tolerant and in the sensitive plants in response to ABA, suggesting that additional factors involved in drought regulation of HaDhn1 expression might exist in tolerant plants.

Tissue-specific expression of Arabidopsis AKT1 gene is consistent with a role in K+ nutrition.

AKT1, a putative inwardly directed K+ channel of Arabidopsis, restores long-term potassium uptake in a yeast mutant defective in K+ absorption. In this paper, the expression pattern of the gene encoding AKT1 is described. Northern blots indicate that AKT1 transcripts are preferentially accumulated in Arabidopsis roots. Owing to the difficulties in producing large quantities of Arabidopsis roots under hydroponic conditions, experiments were undertaken with Brassica napus, a related species. Potassium starvation experiments on B. napus plants show that changes in the K+ status of the organs do not modify AKT1 mRNA accumulation. Western blot analysis of B. napus proteins confirms the presence of AKT1 at the root plasma membrane. Tissue-specific expression directed by the Arabidopsis AKT1 gene promoter was investigated by analysis of beta-glucuronidase (GUS) activity in transgenic Arabidopsis containing an AKT1-GUS gene fusion. As determined by fluorimetric and histochemical tests, the AKT1 promoter directs preferential expression in the peripheral cell layers of root mature regions. The discrete activity found in leaves relates to leaf primordia and to small groups of cells, hydathodes, found on toothed margins of the Arabidopsis leaf lamina. These data are discussed with regard to a possible role of AKT1 in K+ nutrition of plants.

Organization and expression of the gene coding for the potassium transport system AKT1 of Arabidopsis thaliana.

We have isolated and sequenced the genomic clone coding for the potassium transport system AKT1 of Arabidopsis thaliana. Southern blot analysis indicated that the gene is present in one copy in the Arabidopsis genome. The coding sequence is interrupted by ten introns. Sequence comparisons of AKT1 polypeptide with the voltage-gated inward rectifying Arabidopsis K+ channel KAT1, and with voltage- or cyclic nucleotide-gated channels from insects and mammals, revealed a highly conserved domain found specifically in both plant polypeptides, and corresponding to about the last 50 amino acids of their C-terminal region. Northern blot analysis of AKT1 expression in Arabidopsis seedlings indicated that AKT1 is preferentially expressed in roots. No transcript was detected in extracts from heterotrophic suspension culture cells. Depleting K+ in the Arabidopsis seedling culture medium for 4 days led to a strong decrease in K+ tissue content (ca. 50%), but did not affect AKT1 transcript level.