Stimulation of root hair elongation in Arabidopsis thaliana by low phosphorus availability

Low phosphorus availability stimulates root hair elongation in many plants, which may have adaptive significance in soil phosphorus acquisition. We investigated the effect of low phosphorus on the elongation of Arabidopsis thaliana root hairs. Arabidopsis thaliana plants were grown in plant culture containing high (1000 mmol m^?3) or low (1 mmol m^?3) phosphorus concentrations, and root hair elongation was analysed by image analysis. After 15d of growth, low-phosphorus plants developed root hairs averaging 0.9 mm in length while high-phosphorus plants of the same age developed root hairs averaging 0.3 mm in length. Increased root hair length in low-phosphorus plants was a result of both increased growth duration and increased growth rate. Root hair length decreased logarithmically in response to increasing phosphorus concentration. Local changes in phosphorus availability influenced root hair growth regardless of the phosphorus status of the plant. Low phosphorus stimulated root hair elongation in the hairless axr2 mutant, exogenously applied IAA stimulated root hair elongation in wild-type high-phosphorus plants and the auxin antagonist CM PA inhibited root hair elongation in low-phosphorus plants. These results indicate that auxin may be involved in the low-phosphorus response in root hairs.     

Roles of phosphatidylinositol 3-kinase in root hair growth

The root hair is a model system for understanding plant cell tip growth. As phosphatidylinositol 3-phosphate [PtdIns(3)P] has been shown in other plant cell types to regulate factors that affect root hair growth, including reactive oxygen species (ROS) levels, cytoskeleton, and endosomal movement, we hypothesized that PtdIns(3)P is also important for root hair elongation. The enzyme that generates PtdIns(3)P, phosphatidylinositol 3-kinase (PI3K), was expressed in root hair cells of transgenic plants containing the PI3K promoter:beta-glucuronidase reporter construct. To obtain genetic evidence for the role of PtdIns(3)P in root hair elongation, we attempted to isolate Arabidopsis (Arabidopsis thaliana) mutant plants that did not express the gene VPS34 encoding the PI3K enzyme. However, the homozygous mutant was lethal due to gametophytic defects, and heterozygous plants were not discernibly different from wild-type plants. Alternatively, we made transgenic plants expressing the PtdIns(3)P-binding FYVE domain in the root hair cell to block signal transduction downstream of PtdIns(3)P. These transgenic plants had shorter root hairs and a reduced hair growth rate compared with wild-type plants. In addition, LY294002, a PI3K-specific inhibitor, inhibited root hair elongation but not initiation. In LY294002-treated root hair cells, endocytosis at the stage of final fusion of the late endosomes to the tonoplast was inhibited and ROS level decreased in a dose-dependent manner. Surprisingly, the LY294002 effects on ROS and root hair elongation were similar in rhd2 mutant plants, suggesting that RHD2 was not the major ROS generator in the PtdIns(3)P-mediated root hair elongation process. Collectively, these results suggest that PtdIns(3)P is required for maintenance of the processes essential for root hair cell elongation.     

Regulation of root development in <Emphasis Type="BoldItalic">Arabidopsis thaliana</Emphasis> by phytohormone-secreting epiphytic methylobacteria

In numerous experimental studies, seedlings of the model dicot Arabidopsis thaliana have been raised on sterile mineral salt agar. However, under natural conditions, no plant has ever grown in an environment without bacteria. Here, we document that germ-free (gnotobiotic) seedlings, raised on mineral salt agar without sucrose, develop very short root hairs. In the presence of a soil extract that contains naturally occurring microbes, root hair elongation is promoted; this effect can be mimicked by the addition of methylobacteria to germ-free seedlings. Using five different bacterial species (Methylobacterium mesophilicum, Methylobacterium extorquens, Methylobacterium oryzae, Methylobacterium podarium, and Methylobacterium radiotolerans), we show that, over 9 days of seedling development in a light-dark cycle, root development (hair elongation, length of the primary root, branching patterns) is regulated by these epiphytic microbes that occur in the rhizosphere of field-grown plants. In a sterile liquid culture test system, auxin (IAA) inhibited root growth with little effect on hair elongation and significantly stimulated hypocotyl enlargement. Cytokinins (trans-zeatin, kinetin) and ethylene (application of the precursor ACC) likewise exerted an inhibitory effect on root growth but, in contrast to IAA, drastically stimulated root hair elongation. Methylobacteria are phytosymbionts that produce/secrete cytokinins. We conclude that, under real-world conditions (soil), the provision of these phytohormones by methylobacteria (and other epiphytic microbes) regulates root development during seedling establishment.     

Plant-derived smoke solutions stimulate the growth of Lycopersicon esculentum roots in vitro

Aqueous extracts of smoke, derived from Themeda triandra, a fire-climax grass, and Passerina vulgaris, a fynbos plant, stimulated the growth of primary root sections of tomato roots in suspension culture. The optimal dilution for both extracts was 1:2000. Several of the fractions obtained from TLC separation of the Themeda and the Passerina extracts significantly promoted primary root growth. The auxins naphthaleneacetic acid (NAA), indolebutyric acid (IBA) and indoleacetic acid (IAA) were found to stimulate the growth of the primary root axis, with IAA and NAA significantly promoting lateral root number. Similarly, the naturally occurring cytokinins, zeatin and its derivatives (zeatin-O-glucoside; dihydrozeatin and zeatin riboside) stimulated primary root length. Zeatin and dihydrozeatin promoted secondary root growth, but only at very low concentrations.     

The role of microbial signals in plant growth and development

Plant growth and development involves a tight coordination of the spatial and temporal organization of cell division, cell expansion and cell differentiation. Orchestration of these events requires the exchange of signaling molecules between the root and shoot, which can be affected by both biotic and abiotic factors. The interactions that occur between plants and their associated microorganisms have long been of interest, as knowledge of these processes could lead to the development of novel agricultural applications. Plants produce a wide range of organic compounds including sugars, organic acids and vitamins, which can be used as nutrients or signals by microbial populations. On the other hand, microorganisms release phytohormones, small molecules or volatile compounds, which may act directly or indirectly to activate plant immunity or regulate plant growth and morphogenesis. In this review, we focus on recent developments in the identification of signals from free-living bacteria and fungi that interact with plants in a beneficial way. Evidence has accumulated indicating that classic plant signals such as auxins and cytokinins can be produced by microorganisms to efficiently colonize the root and modulate root system architecture. Other classes of signals, including N-acyl-L-homoserine lactones, which are used by bacteria for cell-to-cell communication, can be perceived by plants to modulate gene expression, metabolism and growth. Finally, we discuss the role played by volatile organic compounds released by certain plant growth-promoting rhizobacteria in plant immunity and developmental processes. The picture that emerges is one in which plants and microbes communicate themselves through transkingdom signaling systems involving classic and novel signals.Key words: Arabidopsis, alkamides, auxins, quorum-sensing, cytokinins     

Auxin and ethylene are involved in the responses of root system architecture to low boron supply in Arabidopsis seedlings

Changes in root architecture are one of the adaptive strategies used by plants to compensate for nutrient deficiencies in soils. In this work, the temporal responses of Arabidopsis (Arabidopsis thaliana) root system architecture to low boron (B) supply were investigated. Arabidopsis Col-0 seedlings were grown in 10 μM B for 5 days and then transferred to a low B medium (0.4 μM) or control medium (10 μM) for a 4-day period. Low B supply caused an inhibition of primary root (PR) growth without altering either the growth or number of lateral roots (LRs). In addition, low B supply induced root hair formation and elongation in positions close to the PR meristem not observed under control conditions. The possible role of auxin and ethylene in the alteration of root system architecture elicited by low B supply was also studied by using two Arabidopsis reporter lines (DR5:GUS and EBS:GUS) and two Arabidopsis mutants with impaired auxin and ethylene signaling (aux1-22 and ein2-1). Low B supply increased auxin reporter DR5:GUS activity in PR tip, suggesting that low B alters the pattern of auxin distribution in PR tip. Moreover, PR elongation in aux1-22 mutant was less sensitive to low B treatment than in wild-type plants, which suggests that auxin resistant 1 (AUX1) participates in the inhibition of PR elongation under low B supply. From all these results, a hypothetical model to explain the effect of low B treatment on PR growth is proposed. We also show that ethylene, via ethylene-insensitive 2 (EIN2) protein, is involved in the induction of root hair formation and elongation under low B treatment.     

Brassinosteroid-induced exaggerated growth in hydroponically grown Arabidopsis plants

The effects of root application of brassinolide (BL) on the growth and development of Arabidopsis plants ( Arabidopsis thaliana ecotype Columbia [L.] Heynh) were evaluated. Initially, all leaves were evaluated on plants 18, 22, 26 and 29 days old. The younger leaves were found to exhibit maximal petiole elongation and upward leaf bending in response to BL treatment. Therefore, based on these results leaves 6, 7 and 8 on 22–24-day-old plants were selected for all subsequent studies. Elongation along the length of the petiole in response to BL treatment was uniform with the exception of an approximately 4 mm region next to the leaf where upward curvature was observed. Both BL and 24-epibrassinolide (24-epiBL) were evaluated, with BL being more effective at lower concentrations than 24-epiBL. The exaggerated growth induced by 0.1 μ M BL was not observed in plants treated with 1 000-fold higher concentrations of GA₃, IAA, NAA or 2,4-D (100 μ M ). In addition, no exaggerated growth effects were observed when plants were treated with 200 ppm ethylene or 1 m M ACC. All treatments with BL, NAA, 2,4-D, IAA or ACC promoted ethylene and ACC production in wild type Arabidopsis plants, but only BL triggered exaggerated plant growth. BL also promoted exaggerated growth and elevated levels of ACC and ethylene in the ethylene insensitive mutant etr1-3 , showing that the effect of BR on growth is independent of ethylene. This work provides evidence that BR-induced exaggerated growth of Arabidopsis plants is independent of gibberellins, auxins and ethylene.     

AKT1 and TRH1 are required during root hair elongation in Arabidopsis

TRH1 is a member of the AtKT/AtKUP/AtHAK family of potassium carriers that is required for root hair elongation and AKT1 is an inward rectifying potassium channel expressed in the root epidermis, endodermis and cortex of Arabidopsis thaliana. Plants homozygous for the trh1-1 mutation form short root hairs. The Trh1(-) phenotype cannot be suppressed by growing plants homozygous for the trh1-1 mutation in the presence of high external KCl concentration. This indicates an absolute requirement for TRH1 in root hair tip growth. Plants homozygous for the akt1-1 mutation develop longer root hairs than the wild type when grown in 0 mM external potassium, but develop shorter hairs than the wild type when grown in higher concentrations [>10 mM] of potassium. These data indicate that both TRH1 and AKT1 are active in the root hair over a wide range of external potassium concentrations, but suggest they have different functions in the growing hair cell.     

Sunflower root growth regulation: the role of jasmonic acid and its relation with auxins

Jasmonates are lipid-derived hormones that act as signal molecules in abiotic and biotic stresses and influence several aspects of plant growth and development. In this work we have investigated the effect of jasmonic acid (JA) on the root architecture of Helianthus annuus seedlings and if JA and auxins interact to modulate the growth of the primary root (PR) and lateral roots (LR). The addition of μM concentrations of JA to the growing medium of sunflower seedlings decreased the growth of the PR and LR, and also reduced the number of LR. Moreover, treatment with ibuprofen, an inhibitor of JA synthesis, increased PR and LR root length causing a deep effect on root architecture. Hence, not only exogenous but also the endogenous JA regulates sunflower root growth. Microscopic analysis showed that the application of JA reduces the cortex cell length and the estimated cell production rate in root meristem while ibuprofen only affects the cell elongation. A possible interaction between JA and auxins to regulate root growth was further analyzed. We show that JA produced its phenotype even in the presence of reduced levels of auxin generated by treatment with an auxin transport inhibitor. Besides, the auxin produced its phenotype even when ibuprofen was applied. In conclusion, JA may induce primary and lateral root growth inhibition in sunflower by an auxin-independent pathway.     

低磷供应对拟南芥根系构型的影响

在人工气候箱中,采用Johnson培养基对拟南芥在低磷供应条件下根系构型的变化进行了研究,结果表明：拟南芥在磷饥饿诱导下,主根缩短,侧根密度、根毛的数量和长度显著增加,并且,根尖到第一侧根和第一根毛的距离也大大缩短。这些改变增加了根系比表面积,并且使得根系分布更加靠近土壤表层,有利于提高植物吸收土壤中有机磷的效率。低磷胁迫还导致拟南芥根系分生组织区细胞形状变异,柱细胞数量减少;主根生长和细胞伸长的动力学分析显示,磷饥饿促使拟南芥主根生长变缓,细胞长度随磷饥饿程度的加深迅速缩小。CycB1;1:GUS染色分析结果表明,低磷破坏拟南芥根系分生组织细胞分裂能力,这些结果说明磷胁迫同时抑制了细胞的伸长和分裂,从而引起拟南芥主根的缩短。     

N-acyl-L-homoserine lactones: a class of bacterial quorum-sensing signals alter post-embryonic root development in Arabidopsis thaliana

N-acyl-homoserine lactones (AHLs) belong to a class of bacterial quorum-sensing signals important for bacterial cell-to-cell communication. We evaluated Arabidopsis thaliana growth responses to a variety of AHLs ranging from 4 to 14 carbons in length, focusing on alterations in post-embryonic root development as a way to determine the biological activity of these signals. The compounds affected primary root growth, lateral root formation and root hair development, and in particular, N-decanoyl-HL (C10-HL) was found to be the most active AHL in altering root system architecture. Developmental changes elicited by C10-HL were related to altered expression of cell division and differentiation marker lines pPRZ1:uidA, CycB1:uidA and pAtEXP7:uidA in Arabidopsis roots. Although the effects of C10-HL were similar to those produced by auxins in modulating root system architecture, the primary and lateral root response to this compound was found to be independent of auxin signalling. Furthermore, we show that mutant and overexpressor lines for an Arabidopsis fatty acid amide hydrolase gene (AtFAAH) sustained altered growth response to C10-HL. All together, our results suggest that AHLs alter root development in Arabidopsis and that plants posses the enzymatic machinery to metabolize these compounds.     

The effect of abscisic acid on root and shoot growth of cauliflower plants

Addition of abscisic acid (ABA) to the nutrient solution increased the root to shoot ratio of hydroponically-grown cauliflower plants by reducing the dry weight of the shoot and increasing that of the root. At concentrations higher than 10^–7 M, ABA increased root branching and root hair formation. Root extension was inhibited in plants kept continuously in solutions containing high ABA concentrations but following removal from the ABA solution root elongation was increased in comparison with plants given no ABA treatment. This elongation was greatest in plants with increased root branching caused by higher ABA concentrations.     

Exogenous auxin effects on growth and phenotype of normal and hairy roots of Pueraria lobata (Willd.) Ohwi

Pueraria lobata hairy roots have faster elongationand more branches than normal roots. The responses of hairy roots and normalroots to treatment with three auxins, indole-3-acetic acid (IAA),indole-3-butyric acid (IBA), and naphthalene acetic acid (NAA) were different.In normal roots, all three auxins strongly stimulated lateral root formation atall tested concentrations. Responses to IAA and IBA in primary root growth andlateral root elongation were similar and depended on concentration; promotionat0.1 M, no effect at 1.0 M, and inhibition at2.5 M. In hairy roots, lateral root formation varied inresponseto the different auxins, i.e. depressed by NAA, unaffected by IAA, and promotedby IBA. Primary root growth was slightly inhibited by IBA and was unaffected byIAA. However, mean lateral root length was reduced in response to IAA and IBA.Only NAA exerted strong inhibition on primary and lateral root elongation inboth root types. The similar free IAA and conjugated IAA content but quitedifferent basal ethylene production and biosynthesis in hairy and normal rootssuggested different mechanisms of response to exogenous auxins in the two roottypes.     

Phospholipase Dε and phosphatidic acid enhance Arabidopsis nitrogen signaling and growth

Activation of phospholipase D (PLD) produces phosphatidic acid (PA), a lipid messenger implicated in cell growth and proliferation, but direct evidence for PLD and PA promotion of growth at the organism level is lacking. Here we characterize a new PLD gene, PLDε , and show that it plays a role in promoting Arabidopsis growth. PLDε is mainly associated with the plasma membrane, and is the most permissive of all PLDs tested with respect to its activity requirements. Knockout (KO) of PLDε decreases root growth and biomass accumulation, whereas over-expression (OE) of PLDε enhances root growth and biomass accumulation. The level of PA was higher in OE plants, but lower in KO plants than in wild-type plants, and suppression of PLD-mediated PA formation by alcohol alleviated the growth-promoting effect of PLDε. OE and KO of PLDε had opposite effects on lateral root elongation in response to nitrogen. Increased expression of PLDε also promoted root hair elongation and primary root growth under severe nitrogen deprivation. The results suggest that PLDε and PA promote organism growth and play a role in nitrogen signaling. The lipid-signaling process may play a role in connecting membrane sensing of nutrient status to increased plant growth and biomass production.     

Effect of Competition and Treatment with Inhibitor of Ethylene Perception on Growth and Hormone Content of Lettuce Plants

We elucidated the effect of increased planting density (single and grouped competing plants) on concentrations of auxin, abscisic acid, and cytokinins in normal lettuce plants and in those with ethylene perception inhibited by 1-methylcyclopropene (1-MCP). An attempt was made to relate the changes in hormone concentration induced by competition and inhibition of ethylene sensitivity to growth responses of lettuce planting. The results showed changes in concentrations of auxins, cytokinins, and ABA in the response of lettuce to crowding. Accumulation of ABA in shoots was likely to contribute to inhibition of transpiration of the plants grown in the presence of neighbors. This assumption was supported by the results of application of an inhibitor of ABA synthesis (fluridone and carotenoid biosynthesis herbicide) resulting in increased transpiration of grouped, but not single plants. Increased planting density led to the decline in root auxins paralleled by inhibition of root growth. This effect was likely to be due to decreased auxin transport to the roots from the shoots suggested by accumulation of auxins in the shoots and inhibition of root growth by application of the auxin transport inhibitor [N-(1-naphtyl)phtalamic acid (NPA)]. Importance of the changes in hormone concentrations was confirmed by data showing that disturbance of auxin and cytokinin distribution detected in MCP-treated plants was accompanied by corresponding modification of the growth response.

     

Effects of tyrosine kinase and phosphatase inhibitors on microtubules in Arabidopsis root cells

To investigate the role of tyrosine phosphorylation/dephosphorylation processes in plant cells the morphology of Arabidopsis thaliana primary roots and the organization of cortical microtubules (MTs) were studied after inhibition of protein tyrosine kinases (PTKs) and tyrosine phosphatases (PTPs). It was found that all tested types of PTKs inhibitors (herbimycin A, genistein and tyrphostin AG 18) altered root hair growth and development, probably as a result of their significant influences on MTs organization in root hairs. The treatment also led to MTs reorientation and disruption in epidermis and cortex cells of both elongation and differentiation zones of primary roots. Enhanced tyrosine phosphorylation after treatment with a PTPs inhibitor (sodium orthovanadate) resulted in intense induction of root hair development and growth and caused a significant shortening of the elongation zone. It also led to changes of MTs orientation from transverse to longitudinal in epidermis and cortex cells of the elongation and differentiation zones of the root. From the data obtained we can suppose that tyrosine phosphorylation can be involved in the dynamics and organization of MTs in different types of plant cells.     

Factors increasing ethylene production enhance the sensitivity of root growth to auxins

Light inhibits root elongation, increases ethylene production and enhances the inhibitory action of auxins on root elongation of pea ( Pisum sativum L. cv. Weibulls Marma) seedlings. To investigate the role of ethylene in the interaction between light and auxin, the level of ethylene production in darkness was increased to the level produced in light by supplying 1-aminocyclopropane-1-carboxylic acid (ACC) or benzylaminopurine (BAP). Ethylene production was measured in excised root tips after treatment of intact seedlings for 24 h, while root growth was measured after 48 h. Auxin, at a concentration causing a partial inhibition of root elongation, did not increase ethylene production significantly. A 4-fold increase in ethylene production, caused either by light, 0.1 μ M ACC or 0.1 μ M BAP, inhibited root elongation by 40–50%. The auxins 2,4-dichlorophenoxyacetic acid and indolebutyric acid applied at 0.1 μ M inhibited root elongation by 15–25% in darkness but by 50–60% in light. Supply of ACC or BAP in darkness enhanced the inhibitory effects of auxins to about the same extent as in light. The inhibition caused by the auxins as well as by the BAP was associated with swelling of the root tips. ACC and BAP treatment synergistically increased the swelling caused by auxins. We conclude that auxin and ethylene, when applied or produced in partially inhibitory concentrations, act synergistically to inhibit root elongation and increase root diameter. The effect of light on the response of the roots to auxins is mediated by a light-induced increase in ethylene production.     

The effect of zinc on the biosynthesis of tryptophan,andol auxins and gibberellins in barley

The action of zinc on the growth of barley and the biosynthesis of indol compounds and gibberellin-like substances was investigated in a number of concentrations of zinc from doses stimulating growth to toxic doses. The seeds were soaked before sowing in solutions of zinc sulphate (5.10^?5 to 5.10^?1% Zn), and the plants cultivated for 7 days in water. Lower concentrations of zinc increased both plant growth and the biosynthesis of tryptophan and auxins. At the optimum concentration of 5.10^?3% Zn this increase in tryptophan amounted to 241% of the variant without zinc; in substances with an R_F corresponding to indolyacetic acid, the increase determined by the biological test, was 207% as against the variant without zinc. Higher concentrations of zinc inhibited growth, the tryptophan content was decreased to below that of the control without zinc and the auxin content also fell to below the control values. Zinc also influenced the content of gibberellin-like substances in the plants. At a concentration of 5.10^?3% Zn the increase in the growth activity in the gibberellic acid area of the chromatogram was 294% of the variant without zinc. At toxic concentrations of zinc, the content of gibberellin-like substances fell to below that of the controls. The finding that zinc acts simultaneously on the biosynthesis of auxins and gibberellins is also evidence for the common action of growth substances of various chemical types on plant growth.