Differentiation of Plant Cells During Symbiotic Nitrogen Fixation

Nitrogen-fixing symbioses between legumes and bacteria of the family Rhizobiaceae involve differentiation of both plant and bacterial cells. Differentiation of plant root cells is required to build an organ, the nodule, which can feed and accommodate a large population of bacteria under conditions conducive to nitrogen fixation. An efficient vascular system is built to connect the nodule to the root, which delivers sugars and other nutrients to the nodule and removes the products of nitrogen fixation for use in the rest of the plant. Cells in the outer cortex differentiate to form a barrier to oxygen diffusion into nodules, which helps to produce the micro-aerobic environment necessary for bacterial nitrogenase activity. Cells of the central, infected zone of nodules undergo multiple rounds of endoreduplication, which may be necessary for colonisation by rhizobia and may enable enlargement and greater metabolic activity of these cells. Infected cells of the nodule contain rhizobia within a unique plant membrane called the peribacteroid or symbiosome membrane, which separates the bacteria from the host cell cytoplasm and mediates nutrient and signal exchanges between the partners. Rhizobia also undergo differentiation during nodule development. Not surprisingly, perhaps, differentiation of each partner is dependent upon interactions with the other. High-throughput methods to assay gene transcripts, proteins, and metabolites are now being used to explore further the different aspects of plant and bacterial differentiation. In this review, we highlight recent advances in our understanding of plant cell differentiation during nodulation that have been made, at least in part, using high-throughput methods

Plant growth-promoting rhizobacteria and root system functioning

The rhizosphere supports the development and activity of a huge and diversified microbial community, including microorganisms capable to promote plant growth. Among the latter, plant growth-promoting rhizobacteria (PGPR) colonize roots of monocots and dicots, and enhance plant growth by direct and indirect mechanisms. Modification of root system architecture by PGPR implicates the production of phytohormones and other signals that lead, mostly, to enhanced lateral root branching and development of root hairs. PGPR also modify root functioning, improve plant nutrition and influence the physiology of the whole plant. Recent results provided first clues as to how PGPR signals could trigger these plant responses. Whether local and/or systemic, the plant molecular pathways involved remain often unknown. From an ecological point of view, it emerged that PGPR form coherent functional groups, whose rhizosphere ecology is influenced by a myriad of abiotic and biotic factors in natural and agricultural soils, and these factors can in turn modulate PGPR effects on roots. In this paper, we address novel knowledge and gaps on PGPR modes of action and signals, and highlight recent progress on the links between plant morphological and physiological effects induced by PGPR. We also show the importance of taking into account the size, diversity, and gene expression patterns of PGPR assemblages in the rhizosphere to better understand their impact on plant growth and functioning. Integrating mechanistic and ecological knowledge on PGPR populations in soil will be a prerequisite to develop novel management strategies for sustainable agriculture

Ethanol, at physiological concentrations, affects ethylene sensing in tomato germinating seeds and seedlings

Ethanol is known to accumulate in various plant organs under various environmental conditions. However, there are very scarce data about ethanol sensing by plants. We observed that ethanol accumulates up to 3.5 mM during tomato seed imbibition, particularly when seeds were stacked. Stacked seeds germinated less than spread out seeds suggesting ethanol inhibits germination. In support of this, exogenous ethanol at physiological concentrations, ranging from 1 to 10 mM, inhibited germination of wild type tomato seeds. However, the germination pattern over the whole ethanol concentration range tested was modified in an ethylene insensitive mutant, never-ripe (nr). The effects of exogenous ethanol were not linked to differences in ethylene production by imbibed seeds. But, we observed that exogenous ethanol at a concentration as low as 0.01 mM down regulated the expression of some ethylene receptors. Moreover, the triple response induced by ethylene in tomato seedlings was partially alleviated by 1 mM ethanol. Similar observations were made on Arabidopsis seeds. These results show there are interactions between ethylene sensing and ethanol in plants

Identification by phenotypic, transcriptomics and bioinformatics strategies of Arabidopsis thaliana genes targeted by the beneficial bacteria, Phyllobacterium brassicacearum STM196

La présence dans la rhizosphère des plantes de "Plant Growth Promoting Rhizobacteria" (PGPR) augmente la croissance de la plante. Cependant, les éléments génétiques de la plante qui sont requis lors de cette interaction bénéfique sont encore largement méconnus. Une meilleure connaissance des voies de signalisations ciblées par une PGPR permettrait d'améliorer l'utilisation des PGPR en agriculture car ils seraient possible de sélectionner des variétés de plante plus sensibles à l'effet bénéfique des PGPR. Au sein du LSTM, l'équipe utilise le modèle biologique formé d'une part par la plante modèle Arabidopsis thaliana et, d'autre part, par la bactérie PGPR Phyllobacterium brassicacearum STM196 découverte au contact des racines du colza. A l'aide d'approches phénotypiques, transcriptomiques et bioinformatiques, nous avons découvert certaines des cibles génétiques de STM196 chez Arabidopsis. L'implication de la protéine à F-box EBF2 dans l'allongement des poils racinaires induit par la bactérie a été mise en évidence. De plus, nous avons identifié un rôle des facteurs de transcription ZAT10 et WRKY46 dans l'accumulation du H202 et dans la répression ed l'activité de la PAL induits par STM196. Enfin, en collaboration avec des bioinformaticiens, nous avons développé un outil en ligne appelé "Virtual Lab Book on Arabidopsis" (base de données et interface Web). A l'avenir, cet outil devrait faciliter l'identification de nouvelles cibles génétiques de STM196 grâce à la méta-analyse de nos expériencesThe presence in the plant rhizosphere of Plant Growth Promoting Rhizobacteria (PGPR) leads to an increase of plant growth. However, the plant genetic elements that are required during this beneficial interaction are still largely unknown. A better knowledge of the plant signaling components that are targeted by PGPR would help to improve the use of PGPR in agriculture as it would be possible to select plant varieties more sensitive to the beneficial effect of PGPR. In the LSTM, the team uses a biological model formed by the model plant Arabidopsis thaliana and the PGPR Phyllobacterium brassicacearum STM196 discovered in the rhizosphere of oil-seed rape. With the help of phenotypic, transcriptomics and Bioinformatics approaches, we had the opportunity to discover genetic targets of STM196 in Arabidopsis. The involvement of the F - box protein EBF2 in the root hair elongation triggered by the bacteria has been highlighted. Moreover, we have confirmed a role for ZAT10 and WRKY46 transcription factors in the H202 accumulation and PAL activity repression induced by STM196. Finally, in collaboration with bioinformaticians, we have developed an online tool called "Virtual Lab Book on Arabidopsis" (database and Web interface). In the future, this tool should help in the identification of new genetic targets of STM196 through the meta-analysis of our experiments.MONTPELLIER-BU Sciences (341722106) / SudocSudocFranceF

Lotus japonicus Metabolic Profiling. Development of Gas Chromatography-Mass Spectrometry Resources for the Study of Plant-Microbe Interactions

Symbiotic nitrogen fixation (SNF) in legume root nodules requires differentiation and integration of both plant and bacterial metabolism. Classical approaches of biochemistry, molecular biology, and genetics have revealed many aspects of primary metabolism in legume nodules that underpin SNF. Functional genomics approaches, especially transcriptomics and proteomics, are beginning to provide a more holistic picture of the metabolic potential of nodules in model legumes like Medicago truncatula and Lotus japonicus. To extend these approaches, we have established protocols for nonbiased measurement and analysis of hundreds of metabolites from L. japonicus, using gas chromatography coupled with mass spectrometry. Following creation of mass spectral tag libraries, which represent both known and unknown metabolites, we measured and compared relative metabolite levels in nodules, roots, leaves, and flowers of symbiotic plants. Principal component analysis of the data revealed distinct metabolic phenotypes for the different organs and led to the identification of marker metabolites for each. Metabolites that were enriched in nodules included: octadecanoic acid, asparagine, glutamate, homoserine, cysteine, putrescine, mannitol, threonic acid, gluconic acid, glyceric acid-3-P, and glycerol-3-P. Hierarchical cluster analysis enabled discrimination of 10 groups of metabolites, based on distribution patterns in diverse Lotus organs. The resources and tools described here, together with ongoing efforts in the areas of genome sequencing, and transcriptome and proteome analysis of L. japonicus and Mesorhizobium loti, should lead to a better understanding of nodule metabolism that underpins SNF

Development and properties of genetically encoded pH sensors in plants.

International audienceFluorescent proteins (FPs) have given access to a large choice of live imaging techniques and have thereby profoundly modified our view of plant cells. Together with technological improvement of imaging, they have opened the possibility to monitor physico-chemical changes within cells. For this purpose, a new generation of FPs has been engineered. For instance, pHluorin, a point mutated version of green fluorescent protein, allows to get local pH estimates. In this paper, we will describe how genetically encoded sensors can be used to measure pH in the microenvironment of living tissues and subsequently discuss the role of pH in (i) exocytosis, (ii) ion uptake by plant roots, (iii) cell growth, and (iv) protein trafficking

PGPR-Arabidopsis interactions is a useful system to study signaling pathways involved in plant developmental control

Using their 1-amino cyclopropane-1-carboxylic acid (ACC) deaminase activity, many rhizobacteria can divert ACC from the ethylene biosynthesis pathway in plant roots. To investigate the role of this microbial activity in plant responses to plant growth-promoting rhizobacteria (PGPR), we analyzed the effects of acdS knock-out and wild-type PGPR strains on two phenotypic responses to inoculation—root hair elongation and root system architecture—in Arabidopsis thaliana. Our work shows that rhizobacterial AcdS activity has a negative effect on root hair elongation, as expected from the reduction of ethylene production rate in root cells, while it has no impact on root system architecture. This suggests that PGPR triggered root hair elongation is independent of ethylene biosynthesis or signaling pathway. In addition, it does indicate that AcdS activity alters local regulatory processes, but not systemic regulations such as those that control root architecture. Our work also indicates that root hair elongation induced by PGPR inoculation is probably an auxin-independent mechanism. These findings were unexpected since genetic screens for abnormal root hair development mutants led to the isolation of ethylene and auxin mutants. Our work hence shows that studying the interaction between a PGPR and the model plant Arabidopsis is a useful system to uncover new pathways involved in plant plasticity

The ethylene pathway contributes to root hair elongation induced by the beneficial bacteria Phyllobacterium brassicacearum STM196.

International audienceIn Arabidopsis roots, some epidermal cells differentiate into root hair cells. Auxin regulates root hair positioning, while ethylene controls cell elongation. Phyllobacterium brassicacearum STM196, a beneficial strain of plant growth promoting rhizobacteria (PGPR) isolated from the roots of field-grown oilseed rape, stimulates root hair elongation in Arabidopsis thaliana seedlings. We investigated the role of ethylene in the response of root hair cells to STM196 inoculation. While we could not detect a significant increase in ethylene biosynthesis, we could detect a slight activation of the ethylene signalling pathway. Consistent with this, an exhaustive survey of the root hair elongation response of mutants and transgenic lines affected in the ethylene pathway showed contrasting root hair sensitivities to STM196. We propose that local ethylene emission contributes to STM196-induceed root hair elongation

Zinc availability modulates plant growth and 1 immune responses via AZI1

Zinc is an essential micronutrient for all living organisms and is involved in a plethora of processes including growth and development, and immunity. However, it is unknown if there is a common genetic and molecular basis underlying multiple facets of zinc function. Here we used natural variation in Arabidopsis thaliana to study the role of zinc in regulating growth. We identify allelic variation of the systemic immunity gene AZI1 as a key for determining root growth responses to low zinc conditions. We further demonstrate that this gene is important for modulating root growth depending on the zinc and defence status. Finally, we show that the interaction of the immunity signal azelaic acid and zinc level to regulate root growth is conserved in rice. This work demonstrates that there is a common genetic and molecular basis for multiple zinc dependent processes and that nutrient cues can determine the balance of plant growth and immune responses in plants

The NRT2.5 and NRT2.6 genes are involved in growth promotion of Arabidopsis by the plant growth-promoting rhizobacterium (PGPR) strain Phyllobacterium brassicacearum STM196

International audienceThe Phyllobacterium brassicacearum STM196 strain stimulates Arabidopsis thaliana growth and antagonizes high nitrate inhibition of lateral root development. A previous study identified two STM196-responsive genes, NRT2.5 and NRT2.6 (Mantelin etal., 2006, Planta 223: 591603). We investigated the role of NRT2.5 and NRT2.6 in the plant response to STM196 using single and double Arabidopsis mutants. The single mutants were also crossed with an nrt2.1 mutant, lacking the major nitrate root transporter, to distinguish the effects of NRT2.5 and NRT2.6 from potential indirect effects of nitrate pools. The nrt2.5 and nrt2.6 mutations abolished the plant growth and root system architecture responses to STM196. The determination of nitrate content revealed that NRT2.5 and NRT2.6 do not play an important role in nitrate distribution between plant organs. Conversely, NRT2.5 and NRT2.6 appeared to play a role in the plant response independent of nitrate uptake. Using a nitrate reductase mutant, it was confirmed that the NRT2.5/NRT2.6-dependent plant signalling pathway is independent of nitrate-dependent regulation of root development. Our findings demonstrate that NRT2.5 and NRT2.6, which are preferentially expressed in leaves, play an essential role in plant growth promotion by the rhizospheric bacterium STM196