Eugene Diatloff

The Arabidopsis NRT1.1 transporter participates in the signaling pathway triggering root colonization of nitrate-rich patches

Localized proliferation of lateral roots in NO3−-rich patches is a striking example of the nutrient-induced plasticity of root development. In Arabidopsis, NO3− stimulation of lateral root elongation is apparently under the control of a NO3−-signaling pathway involving the ANR1 transcription factor. ANR1 is thought to transduce the NO3− signal internally, but the upstream NO3− sensing system is unknown. Here, we show that mutants of the NRT1.1 nitrate transporter display a strongly decreased root colonization of NO3−-rich patches, resulting from reduced lateral root elongation. This phenotype is not due to lower specific NO3− uptake activity in the mutants and is not suppressed when the NO3−-rich patch is supplemented with an alternative N source but is associated with dramatically decreased ANR1 expression. These results show that NRT1.1 promotes localized root proliferation independently of any nutritional effect and indicate a role in the ANR1-dependent NO3− signaling pathway, either as a NO3− sensor or as a facilitator of NO3− influx into NO3−-sensing cells. Consistent with this model, the NRT1.1 and ANR1 promoters both directed reporter gene expression in root primordia and root tips. The inability of NRT1.1-deficient mutants to promote increased lateral root proliferation in the NO3−-rich zone impairs the efficient acquisition of NO3− and leads to slower plant growth. We conclude that NRT1.1, which is localized at the forefront of soil exploration by the roots, is a key component of the NO3−-sensing system that enables the plant to detect and exploit NO3−-rich soil patches.

Root-targeted biotechnology to mediate hormonal signalling and improve crop stress tolerance

Since plant root systems capture both water and nutrients essential for the formation of crop yield, there has been renewed biotechnological focus on root system improvement. Although water and nutrient uptake can be facilitated by membrane proteins known as aquaporins and nutrient transporters, respectively, there is a little evidence that root-localised overexpression of these proteins improves plant growth or stress tolerance. Recent work suggests that the major classes of phytohormones are involved not only in regulating aquaporin and nutrient transporter expression and activity, but also in sculpting root system architecture. Root-specific expression of plant and bacterial phytohormone-related genes, using either root-specific or root-inducible promoters or grafting non-transformed plants onto constitutive hormone producing rootstocks, has examined the role of root hormone production in mediating crop stress tolerance. Root-specific traits such as root system architecture, sensing of edaphic stress and root-to-shoot communication can be exploited to improve resource (water and nutrients) capture and plant development under resource-limited conditions. Thus, root system engineering provides new opportunities to maintain sustainable crop production under changing environmental conditions.

CESA5 Is Required for the Synthesis of Cellulose with a Role in Structuring the Adherent Mucilage of Arabidopsis Seeds

Imbibed Arabidopsis (Arabidopsis thaliana) seeds are encapsulated by mucilage that is formed of hydrated polysaccharides released from seed coat epidermal cells. The mucilage is structured with water-soluble and adherent layers, with cellulose present uniquely in an inner domain of the latter. Using a reverse-genetic approach to identify the cellulose synthases (CESAs) that produce mucilage cellulose, cesa5 mutants were shown to be required for the correct formation of these layers. Expression of CESA5 in the seed coat was specific to epidermal cells and coincided with the accumulation of mucilage polysaccharides in their apoplast. Analysis of sugar composition showed that although total sugar composition or amounts were unchanged, their partition between layers was different in the mutant, with redistribution from adherent to water-soluble mucilage. The macromolecular characteristics of the water-soluble mucilage were also modified. In accordance with a role for CESA5 in mucilage cellulose synthesis, crystalline cellulose contents were reduced in mutant seeds and birefringent microfibrils were absent from adherent mucilage. Although the mucilage-modified5 mutant showed similar defects to cesa5 in the distribution of sugar components between water-soluble and adherent mucilage, labeling of residual adherent mucilage indicated that cesa5 contained less cellulose and less pectin methyl esterification. Together, the results demonstrate that CESA5 plays a major and essential role in cellulose production in seed mucilage, which is critical for the establishment of mucilage structured in layers and domains.

Characterization of anion channels in the plasma membrane of Arabidopsis epidermal root cells and the identification of a citrate-permeable channel induced by phosphate starvation.

Organic-acid secretion from higher plant roots into the rhizosphere plays an important role in nutrient acquisition and metal detoxification. In this study we report the electrophysiological characterization of anion channels in Arabidopsis (Arabidopsis thaliana) root epidermal cells and show that anion channels represent a pathway for citrate efflux to the soil solution. Plants were grown in nutrient-replete conditions and the patch clamp technique was applied to protoplasts isolated from the root epidermal cells of the elongation zone and young root hairs. Using SO42− as the dominant anion in the pipette, voltage-dependent whole-cell inward currents were activated at membrane potentials positive of −180 mV exhibiting a maximum peak inward current (Ipeak) at approximately −130 mV. These currents reversed at potentials close to the equilibrium potential for SO42−, indicating that the inward currents represented SO42− efflux. Replacing intracellular SO42− with Cl− or NO3− resulted in inward currents exhibiting similar properties to the SO42− efflux currents, suggesting that these channels were also permeable to a range of inorganic anions; however when intracellular SO42− was replaced with citrate or malate, no inward currents were ever observed. Outside-out patches were used to characterize a 12.4-picoSiemens channel responsible for these whole-cell currents. Citrate efflux from Arabidopsis roots is induced by phosphate starvation. Thus, we investigated anion channel activity from root epidermal protoplasts isolated from Arabidopsis plants deprived of phosphate for up to 7 d after being grown for 10 d on phosphate-replete media (1.25 mm). In contrast to phosphate-replete plants, protoplasts from phosphate-starved roots exhibited depolarization-activated voltage-dependent citrate and malate efflux currents. Furthermore, phosphate starvation did not regulate inorganic anion efflux, suggesting that citrate efflux is probably mediated by novel anion channel activity, which could have a role in phosphate acquisition.

Enhanced root growth of the brb (bald root barley) mutant in drying soil allows similar shoot physiological responses to soil water deficit as wild-type plants

The genetics, molecular biology and nutrient uptake of plant root hair mutants have been studied in detail, but their physiological responses to soil drying have not. Thus, the root hairless brb (bald root barley) barley (Hordeum vulgare L.) mutant and its wild type (WT) were grown in drying soil. Well-watered, pre-tillering plants showed no genotypic differences in daily transpiration and leaf elongation rate, and the ratio of day to night leaf elongation (D/N, a sensitive indicator of water stress). After withholding water for 25 days, root hydraulic conductivity and xylem ABA concentration were similar between genotypes, but WT plants had more tillers and D/N was more than halved in brb. To avoid possible developmental and nutritional differences confounding responses to water deficit, pre-tillering plants were allowed to dry soils of high and low phosphorus (P) status. Although leaf area, leaf water potential and shoot fresh weight (FW) were similar in the two genotypes, root FW of brb was greater by 44 and 18% in a high and low P soil respectively. This adaptive response allowed brb to maintain similar shoot growth and transpiration as WT plants, despite decreased effective root surface area in the absence of root hairs.