Identification of novel drought-related mRNAs in common bean roots by differential display RT-PCR.

Drought is a major constraint for the production of common bean (Phaseolus vulgaris L.). To identify molecular responses to water deficit, we performed a differential display RT-PCR (DDRT) analysis using roots of bean plants grown aeroponically and submitted to dehydration. This allowed us to visualise 1200 DDRT bands, 8.7% of which showed a clear regulation by dehydration, and to clone 42 cDNAs, called PvD1 to PvD42. Among them, 20 early-dehydration-responsive cDNAs were selected by reverse northern that were induced or repressed before detectable water status changes and induction of ABA-regulated genes. Northern analysis for 16 PvD clones confirmed these early regulations and allowed us to identify four late dehydration-responsive genes. Their putative involvement in signalling, protein turn-over and translocation, chaperones as well as root growth modulations in response to water stress is discussed.

[MicroRNAs: a new class of gene expression regulators]. / Les microARN: une nouvelle classe de régulateurs de l'expression génique.

MicroRNAs (miRs) are small non coding RNA, about 21-25 nucleotides in length, that direct post transcriptional regulation of gene expression through interaction with homologous mRNAs. Hundreds miR genes have been identified in animals and 40 in plants. Many of them are conserved between related species, and in some cases across phyla. Two mechanisms for regulation of gene expression by miRs have been reported. As described for lin-4 and let-7 miR of C.elegans, miRs can inhibit translation, which seems to represent the major mode of regulation in animals, or can direct cleavage of target mRNAs, which seems to represent the major mode of regulation in plants.

Disruption of AtOCT1, an organic cation transporter gene, affects root development and carnitine-related responses in Arabidopsis.

In animals, organic cation/carnitine transporters (OCTs) are involved in homeostasis and distribution of various small endogenous amines (e.g. carnitine, choline) and detoxification of xenobiotics such as nicotine. Here, we describe the characterization of AtOCT1, an Arabidopsis protein that shares most of the conserved features of mammalian plasma membrane OCTs. Transient expression of an AtOCT1::GFP fusion protein in onion epidermal cells and Arabidopsis protoplasts supported localization in the plasmalemma. AtOCT1 functionally complemented the Deltacit2/Deltaagp2p yeast strain that is defective in plasma membrane carnitine transport. Disruption of AtOCT1 in an Arabidopsis oct1-1 knockout mutant affected both the expression of carnitine-related genes and the developmental defects induced by exogenous carnitine. RT-PCR and promoter-uidA fusion analysis showed that AtOCT1 was expressed in vascular tissues of various organs and at sites of lateral root formation. Correlating with this expression pattern, oct1-1 seedlings grown in vitro exhibited a higher degree of root branching than the wild-type, showing that the disruption of AtOCT1 affected root development under certain conditions.

Characterization of the expression of Phaseolus vulgaris OCT1, a dehydration-regulated gene that encodes a new type of phloem transporter.

A cDNA coding for a putative organic cation transporter (OCT) was isolated from Phaseolus vulgaris roots by differential display RT-PCR and the corresponding full-length cDNA (named PvOCT1) was subsequently obtained by RACE-PCR. Hydropathy profiles of the deduced amino acid sequence (547 residues) predicted the existence of twelve membrane-spanning domains, which are highly conserved in the major facilitator superfamily (MFS). Three specific domains, which characterize organic ion transporters in animals, can also be observed in the predicted protein. In the non-stressed plants, northern analysis showed that PvOCT1 is strongly expressed in roots and stems, while in situ hybridization revealed the presence of PvOCT1 transcripts in phloem cells. In roots PvOCT1 transcript levels transitorily increased after one hour of dehydration and then dramatically decreased. This decrease was associated with enhanced abundance of PvNeED1 mRNA encoding the enzyme thought to catalyze the limiting step of abscisic acid biosynthesis.