Pascal Gantet

Genetic control of root development in rice, the model cereal

Cereals possess a fibrous root system that is mainly composed of crown roots that emerge postembryonically from the nodes of the stem. Because the root system is not directly accessible and consequently difficult to study, it remains a target for breeders to improve the ability of plants to exploit the mineral and water resources of the soil. Breeding for root architecture necessitates identifying the genetic determinants of root development. This research is now underway in cereals, particularly in rice, the monocot model species. In this review, we examine recent data identifying genes that govern root development in cereals, such as ARL1/CRL1 in rice and RTCS in maize which encodes a conserved lateral organ boundary domain transcription factor involved in crown root initiation and development in response to auxin. Finally, we discuss the detection and validation of root development quantitative trait loci.

Molecular genetics of rice root development

Plant roots have a large range of functions, including acquisition of water and nutrients, as well as structural support. Dissecting the genetic and molecular mechanisms controlling rice root development is critical for the development of new rice ideotypes that are better adapted to adverse conditions and for the production of sustainably achieved rice yield potential. Most knowledge regarding the gene networks involved in root development has been accumulated in the model dicotyledon plant species Arabidopsis thaliana. Rice, the model monocotyledon species, presents several singularities compared to A. thaliana, including a root architecture characterized by a fibrous root system comprising five types of embryonic and postembryonic roots. The anatomy and morphology of the rice root system, which is typical for a cereal, differs from that of A. thaliana, for instance, by the presence of a lysigenous cortex and additional cell layers compared to the dicotyledon model. Moreover, the structure and functions of the root apical meristem (RAM) of rice are distinct from those of A. thaliana. Recently, several rice root mutants have been identified via forward or reverse genetics, and these will aid in forming hypothesis to characterize either the divergence or conservation of genetic pathways relative to A. thaliana. Furthermore, these mutants will help to identify key genes in rice roots that may be missing in A. thaliana. This review summarizes both classical and recent data concerning the molecular genetics of rice root development, including root anatomy and morphology, RAM structure, RAM patterning, and root mutants.

Transcription factors involved in terpenoid indole alkaloid biosynthesis in Catharanthus roseus

Plants produce alkaloids, among others, to protect themselves against microbial infection, herbivore attack or ultraviolet irradiation. For man, alkaloid metabolism is the source of many natural products with useful applications, including pharmaceuticals. A major mechanism regulating alkaloid production in plant cells is the control of the transcription of the biosynthetic genes. Several transcription factors involved in the regulation of alkaloid biosynthesis genes have been isolated and studied. There are indications that the abundance and activities of transcription factors themselves are regulated by external signals. The aim of this review is to give an update on the transcriptional regulation of terpenoid indole metabolism in Catharanthus roseus.

Post-embryonic root organogenesis in cereals: branching out from model plants.

The root architecture of higher plants is amazingly diverse. In this review, we compare the lateral root developmental programme in cereals and Arabidopsis thaliana. In cereals, cells in the endodermis are recruited to form the new root cap and overlying cortical cells divide to facilitate the emergence of the lateral root primordium. The TIR1/ABF2 auxin receptors and the AUX/IAA, ARF, and LBD transcriptional regulatory proteins are conserved in cereals and Arabidopsis. Several elements of this regulatory network are common to lateral and crown roots in cereals. Also, the ground meristem from which crown roots differentiate shows similarities with the root pericycle. Studies in cereals promise to give complementary insights into the mechanisms regulating the development of post-embryonic roots in plants.

Transcription Factor Networks. Pathways to the Knowledge of Root Development

Plants are bipolar organisms in which the apical part generates vegetative and reproductive organs and the basal part generates the root system. Roots perform a variety of biological functions. They keep the plant upright but are also the site of nutrient and water uptake and constitute important

Regulation of the terpene moiety biosynthesis of Catharanthus roseus terpene indole alkaloids

Precursor feeding experiments have shown that the terpene moiety biosynthesis is the most rate-limiting step of terpenoid indole alkaloids (TIA) produced by Catharanthus roseus suspension cells. The biosynthesis of TIA terpene moiety is strictly regulated by hormones: auxin inhibits, jasmonate stimulates and cytokinin and ethylene enhance their biosynthesis. Biochemical analyses coupled to molecular approaches have outlined a regulatory cross-talk between two metabolite pathways leading to the biosynthesis of terpene precursors. Terpenes derivatives produced through the mevalonic acid (MVA) pathway regulate the activity of the 2-C-methyl-d-erythritol 4-phosphate (MEP) non-mevalonate pathway producing the precursor of the TIA terpene moiety. This cross-talk involves regulatory prenylated proteins and acts on the regulation of the expression of early steps of monoterpenoid biosynthesis (ESMB) genes. The expression of the genes involved in the TIA terpene moiety biosynthesis is regulated in part by a general TIA transcription factor, ORCA3, and other unidentified transcription factors. This review sums up the essential information obtained at the metabolic, physiological, biochemical and molecular levels of regulation of TIA terpene moiety biosynthesis in C. roseus. We propose a synthetic scheme of the general regulatory network emerging from these experimental data, and discuss the related hypothesis and prospect in elucidating other key events involved in the regulation of TIA terpene moiety biosynthesis.

Regulation of Shoot and Root Development through Mutual Signaling

Plants adjust their development in relation to the availability of nutrient sources. This necessitates signaling between root and shoot. Aside from the well-known systemic signaling processes mediated by auxin, cytokinin, and sugars, new pathways involving carotenoid-derived hormones have recently been identified. The auxin-responsive MAX pathway controls shoot branching through the biosynthesis of strigolactone in the roots. The BYPASS1 gene affects the production of an as-yet unknown carotenoid-derived substance in roots that promotes shoot development. Novel local and systemic mechanisms that control adaptive root development in response to nitrogen and phosphorus starvation were recently discovered. Notably, the ability of the NITRATE TRANSPORTER 1.1 to transport auxin drew for the first time a functional link between auxin, root development, and nitrate availability in soil. The study of plant response to phosphorus starvation allowed the identification of a systemic mobile miRNA. Deciphering and integrating these signaling pathways at the whole-plant level provide a new perspective for understanding how plants regulate their development in response to environmental cues.

Histidine-containing phosphotransfer domain extinction by RNA interference turns off a cytokinin signalling circuitry in Catharanthus roseus suspension cells

We previously reported that cytokinins (CK) induce the fast and specific transcription of CrRR1, a gene encoding a type A response regulator in Catharanthus roseus cell cultures. Here, we characterized the CrHPt1 gene that encodes a histidine‐containing phosphotransfer domain. CrHPt1 was silenced through RNA interference (RNAi) to test its possible implication in the CK signalling pathway. In transgenic lines stably transformed with an intron‐spliced construct, the degradation of CrHPt1 transcripts abolishes the CK inductive effect on CrRR1 transcription. These result give a new in vivo functional argument for the crucial role of HPt proteins in the CK signalling pathway leading to the expression of the genes encoding type A response regulators. They also show that RNAi is a powerful strategy to turn off the CK signalling circuitry.

An in planta, Agrobacterium-mediated transient gene expression method for inducing gene silencing in rice (Oryza sativa L.) leaves.

BackgroundLocalized introduction and transient expression of T-DNA constructs mediated by agro-infiltration of leaf tissues has been largely used in dicot plants for analyzing the transitivity and the cell-to cell movement of the RNAi signal. In cereals, however, the morphology of the leaf and particularly the structure of the leaf epidermis, prevent infiltration of a bacterial suspension in cells by simple pressure, a method otherwise successful in dicots leaves. This study aimed at establishing a rapid method for the functional analysis of rice genes based on the triggering of RNA interference (RNAi) following Agrobacterium-mediated transient transformation of leaves.ResultsUsing an agro-infection protocol combining a wound treatment and a surfactant, we were able to obtain in a reliable manner transient expression of a T-DNA-borne uidA gene in leaf cells of japonica and indica rice cultivars. Using this protocol to transiently inhibit gene expression in leaf cells, we introduced hairpin RNA (hpRNA) T-DNA constructs containing gene specific tags of the phytoene desaturase (OsPDS) and of the SLENDER 1 (OsSLR1) genes previously proven to trigger RNAi of target genes in stable transformants. SiRNA accumulation was observed in the agro-infected leaf area for both constructs indicating successful triggering of the silencing signal. Accumulation of secondary siRNA was observed in both stably and transiently transformed leaf tissues expressing the HpRNA OsSLR1 construct. Gene silencing signalling was investigated in monitoring the parallel time course of OsPDS-derived mRNA and siRNA accumulation in the agro-infiltrated leaf area and adjacent systemic sectors. The sensitive RT-Q-PCR method evidenced a consistent, parallel decrease of OsPDS transcripts in both the agroinfiltred and adjacent tissues, with a time lag for the latter.ConclusionsThese results indicate that the method is efficient at inducing gene silencing in the agro-infected leaf area. The transfer of low amounts of siRNA, probably occurring passively through the symplastic pathway from the agro-infected area, seemed sufficient to trigger degradation of target transcripts in the adjacent tissues. This method is therefore well suited to study the cell-to-cell movement of the silencing signal in a monocot plant and further test the functionality of natural and artificial miRNA expression constructs.

Analysis of the expression of the AGL17-like clade of MADS-box transcription factors in rice.

In plants, MADS-box transcription factors are key regulators of floral and fruit development, organ dehiscence and stress responses. Nevertheless, the functions of most of them are still unknown. In Arabidopsis thaliana, the AGL17-like clade of MADS-box transcription factors comprises four members. AGL17 is involved in floral induction, whereas AGL44/ANR1 is involved in the adaptive development of roots in response to nitrate. AGL21 is primarily expressed in the roots and AGL16 in the leaves, suggesting that these transcription factors may be involved in the control of vegetative development. In Oryza sativa, the AGL17-like clade comprises five members, OsMADS23, OsMADS25, OsMADS27, OsMADS57 and OsMADS61. In a first attempt to characterize their functions, we used promoter::Gus reporter gene fusions and RT-qPCR to study the expression patterns of these genes and their regulation by different external stimuli. The OsMADS23, OsMADS25, OsMADS27 and OsMADS57 promoters were active in the roots central cylinder. In addition, the OsMADS57 promoter was active in leaves, whereas the OsMADS61 promoter was only active in the leaf tips and the stem base. OsMADS25 and OsMADS27 transcripts accumulated in response to osmotic stress, whereas the expression levels of OsMADS25, OsMADS27 and OsMADS57 were slightly induced by nitrate. Each of these five genes was responsive to various hormonal treatments. These distinct expression patterns indicate that these five genes have specific and non-redundant functions that likely differs from those of their A. thaliana homologs.