Ethylene Production by Fe-deficient Roots and its Involvement in the Regulation of Fe-deficiency Stress Responses by Strategy I Plants

Species that showed marked morphological and physiological responsesby their roots to Fe-deficiency (Strategy I plants) were comparedwith others that do not exhibit these responses (Strategy IIplants). Roots from Fe-deficient cucumber (Cucumis sativusL.‘Ashley’), tomato (Lycopersicon esculentumMill.T3238FER) and pea (Pisum sativumL. ‘Sparkle’) plantsproduced more ethylene than those of Fe-sufficient plants. Thehigher production of ethylene in Fe-deficient cucumber and peaplants occurred before Fe-deficient plants showed chlorosissymptoms and was parallel to the occurrence of Fe-deficiencystress responses. The addition of either the ethylene precursorACC, 1-aminocyclopropane-1-carboxylic acid, or the ethylenereleasing substance, Ethephon, to several Fe-sufficient StrategyI plants [cucumber, tomato, pea, sugar beet (Beta vulgarisL.),Arabidopsis(Arabidopsis thaliana(L.) Heynh ‘Columbia’), plantago(Plantago lanceolataL.)] promoted some of their Fe-deficiencystress responses: enhanced root ferric-reducing capacity andswollen root tips. By contrast, Fe-deficient roots from severalStrategy II plants [maize (Zea maysL. ‘Funo’), wheat(Triticum aestivumL. ‘Yécora’), barley (HordeumvulgareL. ‘Barbarrosa’)] did not produce more ethylenethan the Fe-sufficient ones. Furthermore, ACC had no effecton the reducing capacity of these Strategy II plants and, exceptin barley, did not promote swelling of root tips. In conclusion,results suggest that ethylene is involved in the regulationof Fe-deficiency stress responses by Strategy I plants.Copyright1999 Annals of Botany Company. Arabidopsis (Arabidopsis thaliana(L.) Heynch), barley (Hordeum vulgareL.), cucumber (Cucumis sativusL.), ethylene, iron deficiency, maize (Zea maysL.), pea (Pisum sativumL.), plantago (Plantago lanceolataL.), ferric-reducing capacity, sugar beet (Beta vulgarisL.), tomato (Lycopersicon esculentumMill.), wheat (Triticum aestivumL.).     

The humic acid-induced changes in the root concentration of nitric oxide,IAA and ethylene do not explain the changes in root architecture caused by humic acid in cucumber

The increase in root growth is one of the major effects of humic substances, but the mechanisms involved in humic acid-mediated changes in root growth, morphology and architecture are poorly known. Probably, humic substances may act on plant development through an action on the hormonal balance within the plant, either directly or indirectly by affecting the root uptake of some nutrients. In this study we investigate in cucumber plants the effects of a purified sedimentary humic acid (PHA), without detectable concentrations of the main phytoregulators in its structure, on root architecture, and its relationships with the functional action of three phytoregulators, indole-acetic acid, ethylene and nitric oxide, which are also affected by the root application of this humic acid. The results obtained using inhibitors of auxin transport or action, inhibitors of ethylene biosynthesis or action, and a scavenger of nitric oxide indicate that the increase in the root concentration of these phytoregulators caused by the root application of PHA does not play an essential role in the expression of the main changes on root architecture caused by PHA in developed cucumber plants. Other factors, which could act in coordination or independently of those phytoregulators affected by PHA root application, must be involved in the whole action on this humic acid on root architecture in cucumber.     

Radiomimetic Effect of Cisplatin on Cucumber Root Development: the Relationship between Cell Division and Cell Growth

Cisplatin [DDP, cis-dichlorodiammine platinum (II)], a strongcytostatic and antineoplastic agent, was tested on seedlingsof cucumber Cucumis sativus L. for its general effect on rootdevelopment and its particular effects on root cell divisionand cell growth. DDP was characterized as a radiomimetic compoundsince both DDP (1·3 x 10^-5 M) and -irradiation (2·5-10kGy) drastically and irreversibly stopped development of embryoniclateral root primordia (LRPs) in the radicle by inhibiting bothmitotic activity and cell growth. In 20% of the LRPs of DDP-treatedroots, cells did not divide at all. Dividing cells completedno more than two cell cycles. These effects were specific becausewhen DDP was available to the roots only at the onset of celldivision, cell proliferation and cell growth were similar tothat produced by constant incubation. Neither DDP nor -irradiationaffected non-meristematic cell elongation. It was concludedthat cell growth of meristematic cells is closely related tocell division. However, non-meristematic cell growth is independentof DNA damage. This suggests DDP as a tool to reveal these autonomousprocesses in plants development and to detect tissue compartmentsin mature plant embryos which contain potentially non-meristematiccells.Copyright 1993, 1999 Academic Press Cucumis sativus, cucumber, cisplatin, cell growth, cell elongation, cell division, lateral root, root development     

Levels of IAA, Cytokinins, ABA and Ethylene in Rice Plants as Affected by a Gibberellin Biosynthesis Inhibitor, Uniconazole-P.

Effects of uniconazole-P, a triazole-type growth retardant,on endogenous levels of IAA, cytokinins, ABA and ethylene inrice seedlings were investigated. Endogenous levels of IAA andABA were similar between control and uniconazole-P-treated riceshoots. Evolution of ethylene was promoted slightly, being 1.8times greater under 0.3 ppm uniconazole-P treatment than thatof control. The most obvious effect was the increase of trans-Zand trans-RZ in shoots. Shoots treated with uniconazole-P (10mg/m² nursery box) contained 3.4 times and 3 times more trans-Zand trans-RZ than control, respectively. No significant differencesof cytokinin levels were recognized in roots except for cis-RZ.The increase of ethylene and active forms of cytokinins, andthe decrease of gibberellin in the shoots may be the basis forphysiological phenomena caused by uniconazole-P, namely thepromotion of flowering in woody plants and the enhancement offemaleness in cucumber. (Received September 9, 1987; Accepted October 20, 1987)     

Proteoid Root Development of Phosphorus Deficient Lupin is Mimicked by Auxin and Phosphonate

White lupin (Lupinus albus L.) develops proteoid (cluster) rootsin response to phosphorus deficiency. Proteoid roots are composedof tight clusters of rootlets that initiate from the pericycleopposite protoxylem poles and emerge from every protoxylem polewithin the proteoid root axis. Auxins are required for lateralroot development, but little is known of their role in proteoidroot formation. Proteoid root numbers were dramatically increasedin P-sufficient (+P) plants by application of the syntheticauxin, naphthalene acetic acid (NAA), to leaves, and were reducedin P-deficient (-P) plants by the presence of auxin transportinhibitors [2,3,5-triiodobenzoic acid (TIBA) and naphthylphthalamicacid (NPA)]. While ethylene concentrations in the root zonewere 1.5-fold higher in -P plants, there was no effect on proteoidroot numbers of the ethylene inhibitors aminoethoxyvinvylglycine(AVG) and silver thiosulphate. Phosphonate, which interfereswith plant perception of internal P concentration, dramaticallyincreased the number of proteoid root segments in +P plants.Activities of phosphoenolpyruvate carboxylase (PEPC), malatedehydrogenase (MDH) and exuded acid phosphatase in proteoidroot segments were not different from +P controls when NAA wasapplied to +P lupin plants, but increased to levels comparableto -P plants in the phosphonate treatment. Addition of TIBAor NPA to -P plants reduced PEPC and MDH activity of -P proteoidroots to levels found in +P or -P normal root tissues, but didnot affect acid phosphatase in root exudates. These resultssuggest that auxin transport from the shoot plays a role inthe formation of proteoid roots during P deficiency. Auxin-stimulatedproteoid root formation is necessary, but not sufficient, tosignal the up-regulation of PEPC and MDH in proteoid root segments.In contrast, phosphonate applied to P-sufficient white lupinelicits the full suite of coordinated responses to P deficiencyCopyright2000 Annals of Botany Company Lupinus albus L., white lupin, proteoid roots, auxin, ethylene, phosphonate, phosphorus deficiency     

Induction of Drought Tolerance in Cucumber Plants by a Consortium of Three Plant Growth-Promoting Rhizobacterium Strains

Our previous work showed that a consortium of three plant growth-promoting rhizobacterium (PGPR) strains (Bacillus cereus AR156, Bacillus subtilis SM21, and Serratia sp. XY21), termed as BBS for short, was a promising biocontrol agent. The present study investigated its effect on drought tolerance in cucumber plants. After withholding watering for 13 days, BBS-treated cucumber plants had much darker green leaves and substantially lighter wilt symptoms than control plants. Compared to the control, the BBS treatment decreased the leaf monodehydroascorbate (MDA) content and relative electrical conductivity by 40% and 15%, respectively; increased the leaf proline content and the root recovery intension by 3.45-fold and 50%, respectively; and also maintained the leaf chlorophyll content in cucumber plants under drought stress. Besides, in relation to the control, the BBS treatment significantly enhanced the superoxide dismutase (SOD) activity and mitigated the drought-triggered down-regulation of the expression of the genes cAPX, rbcL, and rbcS encoding cytosolic ascorbate peroxidase, and ribulose-1,5-bisphosphate carboxy/oxygenase (Rubisco) large and small subunits, respectively, in cucumber leaves. However, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity was undetected in none of the culture solutions of three BBS constituent strains. These results indicated that BBS conferred induced systemic tolerance to drought stress in cucumber plants, by protecting plant cells, maintaining photosynthetic efficiency and root vigor and increasing some of antioxidase activities, without involving the action of ACC deaminase to lower plant ethylene levels.     

Effects of Tiller Separation and Root Pruning on the Growth of Lolium perenne L.

Experiments have been carried out to assess the role of inter-tillerinteractions within plants of Lolium perenne L. cv. S24 as factorswhich could play a part in determining the growth-rate of thewhole plant. In order to do this, the effect of tiller separationon plant growth was studied as well as the influence of theremoval of different parts of the root and shoot system. Wheneverplants were supplied with nutrients by means of culture solutions,separation of tillers brought about a marked increase in theamount of dry matter produced. Root pruning and in particularthe removal of root apices brought about similar increases ingrowth. Separation of tillers in soil-grown plants did not bringabout increased growth. These results are interpreted as indicatingthat separation of tillers or the initial removal of root apicesin plants supplied with nutrients by means of culture solution,promotes root branching which brings about an increased netassimilation rate arising from an increase supply of a growth-promotingsubstance, probably a cytokinin, from the root apices. The resultsare not taken as indicative of an intertiller interaction whichrestricts the growth of intact plants.     

The influence of the parasitic angiosperm Striga gesnerioides on the growth and photosynthesis of its host, Vigna unguiculata

Cowpea plants (Vigna unguiculata) infected with the root hemiparasiticangiosperm Striga gesnerioides accumulated less biomass thanuninfected plants over a growth period of 60 d. The allometricrelationship between shoot and root dry weight was similar inparasitized plants relative to control plants, as was the proportionof dry matter partitioned into leaf, stem and root tissue. However,infected plants failed to make any significant investment ofdry matter in pods. The rate of photosynthesis of the youngestfully expanded leaf of parasitized plants was significantlylower than for control plants. The lower rates of photosynthesiswere not attributable to stomatal limitation, a loss of chlorophyllor to an accumulation of carbohydrate. The depression of photosynthesisin the young leaves was transient. As control leaves aged, photosynthesisdeclined. This also occurred in Striga infected plants, butto a lesser extent resulting in higher rates of photosynthesisin mature leaves when compared to those of uninfected plants.The foliar nitrogen content of parasitized plants was higherthan control plants consistent with the slower rate of photosyntheticdecline of older leaves. The data are discussed with respectto the influence of parasitic weeds on host growth and photosynthesis. Key words: Cowpea, hemiparasite, allometry, nitrogen     

Temperature effects on the reactive oxygen species formation and antioxidant defence in roots of two cucurbit species with contrasting root zone temperature optima

Suboptimal root zone temperature (14°C) was imposed on chilling-sensitive cucumber (Cucumis sativus L.) and chilling-tolerant figleaf gourd (Cucurbita ficifolia Bouché) plants. Exposure of roots to low temperature for up to 10 days caused a strong growth inhibition in cucumber compared with figleaf gourd. Physiological analysis showed that generation of reactive oxygen species (ROS) such as hydrogen peroxide and superoxide anion was significantly induced in cucumber plants as fast as 1 day after low root zone temperature treatment. In addition to the significant induction of antioxidant superoxide dismutase activity, low root zone temperature also increased the mitochondrial electron transport allocated to alternative pathway while decreased cytochrome pathway salicylhydroxamic acid-resistant respiration. However, these defense responses could not compensate for the ROS production, resulting in membrane lipid peroxidation and loss of root cell viability in the low root zone temperature treated cucumber roots. In contrast, 14°C root zone temperature had no significant effects on figleaf gourd plant growth, antioxidant enzymes, ROS levels and alternative respiratory pathway. Hence, difference in ROS metabolism would be associated with the remarkable difference in adaptability of cucumber and figleaf gourd plants in response to suboptimal root zone temperature condition.     

NO and IAA Key Regulators in the Shoot Growth Promoting Action of Humic Acid in Cucumis sativus L.

Previous studies have reported that a purified sedimentary humic acid (PHA) was able to increase the concentration of nitric oxide (NO), indole-acetic acid (IAA) and ethylene in cucumber roots. Here, we investigated if these effects are functionally related to the ability of PHA to improve shoot growth. The effect of specific inhibitors of NO, IAA and ethylene functionality and signaling on PHA-induced shoot growth was studied. Likewise, the effect of these inhibitors on the synthesis and activity of the phytoregulators concerned by PHA action in cucumber roots was also explored. The results show that shoot growth promoted by PHA is due to an increase of IAA concentration in the root through both a NO-dependent and a NO-independent pathway. In addition, the increased ethylene production in the root is regulated by an IAA-dependent pathway. Finally, results also showed that the increase of ABA concentration in the root is regulated through both IAA- and ethylene-dependent pathways. In summary, the shoot growth promoting action of PHA involves a complex hormonal network. On one hand, the PHA action is functionally linked to increments in NO and IAA concentration in roots. And on the other hand, PHA action also increases ethylene and ABA root concentration mediated by NO-IAA dependent pathways.     

Genetic Analysis of the Effects of Polar Auxin Transport Inhibitors on Root Growth in Arabidopsis thaliana

Polar auxin transport inhibitors, including N-1-naphthylphthalamicacid (NPA) and 2,3,5-triiodobenzoic acid (TIBA), have variouseffects on physiological and developmental events, such as theelongation and tropism of roots and stems, in higher plants.We isolated NPA-resistant mutants of Arabidopsis thaliana, withmutations designated pir1 and pir2, that were also resistantto TIBA. The mutations specifically affected the root-elongationprocess, and they were shown ultimately to be allelic to aux1and ein2, respectively, which are known as mutations that affectresponses to phytohormones. The mechanism of action of auxintransport inhibitors was investigated with these mutants, inrelation to the effects of ethylene, auxin, and the polar transportof auxin. With respect to the inhibition of root elongationin A. thaliana, we demonstrated that (1) the background levelof ethylene intensifies the effects of auxin transport inhibitors,(2) auxin transport inhibitors might act also via an inhibitorypathway that does not involve ethylene, auxin, or the polartransport of auxin, (3) the hypothesis that the inhibitory effectof NPA on root elongation is due to high-level accumulationof auxin as a result of blockage of auxin transport is not applicableto A. thaliana, and (4) in contrast to NPA, TIBA itself hasa weak auxin-like inhibitory effect. (Received April 12, 1996; Accepted September 2, 1996)     

Effects of root anaerobiosis on ethylene production, epinasty, and growth of tomato plants

Experiments were performed to determine the source(s) of ethylene-causing epinasty in flooded tomato plants (Lycopersicon esculentum Mill.). Simultaneous measurements were made of ethylene synthesized by the roots and shoots of tomato plants exposed to either aerobic or anaerobic atmospheres in the root zone. When the root zone was made anaerobic by a flowing stream of N₂ gas, petiole epinasty and accelerated ethylene synthesis by the shoots were observed. In soil-grown plants, ethylene synthesis by the root-soil complex increased under anaerobic conditions; but when grown in inert media under the same conditions, ethylene synthesis by roots remained constant or declined during the period of rapid epinastic growth by the petioles. Other characteristic symptoms of flooding, e.g. reduced growth and chlorosis, were also observed in plants with anaerobic roots. Pretreatment of plants with AgNO₃, an inhibitor of ethylene action, completely prevented epinasty, demonstrating that ethylene is the agent responsible for waterlogging symptoms. These results indicate that deprivation of O₂ to the roots is the primary effect of soil flooding, and that this is sufficient to cause increased ethylene synthesis in the shoot. The basis of the observed root-shoot communication is unknown, but root-synthesized hormones or specific ethylene-promoting factors may be involved.     

Is ethylene involved in regulation of root ferric reductase activity of dicotyledonous species under iron deficiency?

Most dicotyledonous species respond to Fe deficiency by developing some mechanisms known as Fe-deficiency responses. The role of ethylene in regulation of root ferric reductase activity of wild-type tomato (Lycopersicon esculentum L.) and its mutant Never ripe (Nr), bean (Phaseolus vulgaris L., cv. Bifeng 80-30), and cucumber (Cucumis sativus L., cv. Xintaimici) plants grown in nutrient solution without Fe supply was studied under controlled condition. The results show that: (i) the tomato mutant Nr, which is insensitive to ethylene, presented rapid increase in root ferric reductase activity after omitting Fe from the nutrient solution; (ii) the initial time for increase in root ferric reductase activity was earlier than that in ethylene production after onset of Fe deficiency in the three species; (iii) like cobalt (3 μM Co²⁺), an inhibitor for ethylene production, high concentration of zinc (50 μM Zn²⁺) and copper (5 μM Cu²⁺) also suppressed the increase in root ferric reductase activity of Fe-starved plants; (iv) under Fe-sufficient conditions, indol-3-butylric acid (IBA) stimulated root ferric reductase activity of cucumber and bean plants, and this stimulating effect could not be suppressed by aminoethoxyvinylglycine (AVG, an inhibitor for ethylene synthesis). These results suggested that ethylene might not be directly involved in the regulation of root ferric reductase activity of Fe-deficient dicotyledonous species.     

Quantitative Morphology of the Developing Cucumber Leaf

Leaves were harvested from cucumber plants (Cucumis sativuscv. Butchers' Disease Resister) grown at 27 C in controlledenvironments. Gompertz and logistic curves fitted lamina lengthdata well, but Gompertz were found to be more useful. The averageintervascular interval increased from approx. 0.1–0.4mm during leaf development. The average maximum number of cellsthrough or around which material had to pass between mesophyllcells and the vascular system was approx. 7 and varied little,supporting the hypothesis that veins develop so as not to allowthis number of cells to exceed a critical value. Cucumis sativus, cucumber leaf, growth analysis, Gompertz, cell number, vein, vein length, transport     

Ethylene accumulation in waterlogged Rumex plants promotes formation of adventitious roots

Accumulation of the gaseous plant hormone ethylene is very importantfor the induction of several responses of plants to flooding.However, little is known about the role of this gas in the formationof flooding-induced adventitious roots. Formation of adventitiousroots in Rumex species is an adaptation of these plants to floodedsoil conditions. The large air-spaces in these roots enablesdiffusion of gases between shoot and roots. Application of ethylene to non-flooded Rumex plants resultedin the formation of adventitious roots. In R. palustris Sm.shoot elongation and epinasty were also observed. The numberof roots in R. thyrsiflorus Fingerh. was much lower than inR. palustris, which corresponds with the inherent differencein root forming capacity between these two species. Ethyleneconcentrations of 1.5–2µI I_{– 1} induced a maximumnumber of roots in both species. Quantification of ethylene escaping from root systems of Rumexplants that were de-submerged after a 24 h submergence periodshowed that average ethylene concentrations in submerged rootsreached 1.8 and 9.1 µl I_–1 in R. palustris and R.thyrsiflorus, respectively. Inhibition of ethylene productionin R. palustris by L--(2-aminoethoxyvinyl)-glycine (AVG) or-aminobutyric acid (AIB) decreased the number of adventitiousroots induced by flooding, indicating that high ethylene concentrationsmay be a prerequisite for the flooding-induced formation ofadventitious roots in Rumex species. Key words: Adventitious roots, epinasty, ethylene, flooding, Rumex, shoot elongation     

<Emphasis Type="Italic">Pseudomonas chlororaphis</Emphasis>: a salt-tolerant bacterial inoculant for plant growth stimulation under saline soil conditions

Saline soils constitute a serious production problem for vegetable crops as they are known to suppress plant growth. One of the possible measures to improve crop health in such conditions is to use salt-tolerant bacterial inoculants which can control diseases and promote plant growth. In the present work the ability of Pseudomonas chlororaphis isolate TSAU13 to promote cucumber and tomato plant growth and to improve fruit yield by protecting these plants against soil-borne pathogens in salinated soil were investigated. The bacterial strain stimulated shoot growth (up to 32%), dry matter (up to 43%), and the fruit yield of tomato and cucumber (up to 16%) compared to the uninoculated control plants under saline conditions. The strain was able to survive on the root of 2-month-old plants. 29% of the cucumber and 27% of the tomato plants which had grown in soil to which no Fusarium solani spores had been added were diseased, whereas in the presence of the pathogenic fungus 58% of the cucumber and 52% tomato plants had disease symptoms. P. chlororaphis TSAU13 showed statistically significant disease reduction in comparison to the Fusarium-uninfected and infected control plants. Those results showed that P. chlororaphis TSAU13 has a great biotechnological potential in improvement of vegetable production in commercial greenhouses under saline conditions.     

Chilli rhizosphere fungus Aspergillus spp. PPA1 promotes vegetative growth of cucumber (Cucumis sativus) plants upon root colonisation

A rhizosphere fungus was isolated from roots of chilli plants and identified as Aspergillus spp. PPA1. The fungus was tested for its ability to promote the growth of cucumber plants in a pot experiment. Cucumber seeds were sown in sterilised field soil amended with wheat grain inoculum (WGI) of PPA 1 at the rate of 0.5, 1 and 1.5% w/w, and plants were grown for 21 days in a net house. The treatment with PPA1 significantly increased shoot length, shoot fresh weight, shoot dry weight, root length, root fresh weight, root dry weight, plant length, leaf area and leaf chlorophyll content of cucumber plants compared to non-treated control. The growth promotion rate increased with the increasing concentration of inoculum of PPA1 applied to the soil. The fungus was re-isolated from the roots of cucumber plants at higher frequencies. These results suggest that Aspergillus spp. PPA1 is a root colonising plant-growth promoting fungus for cucumber.     

Anatomy, Growth and Survival of a Long-hypocotyl Mutant of Cucumis sativus Deficient in Phytochrome B

Plant dry matter (DM) partitioning, survival rates, stem anatomy,and stem water conductivity were investigated in wild-type (WT)and long-hypocotyl (lh) mutant seedlings of cucumber (Cucumissativus) grown as isolated individuals under natural radiation.The lh mutant is severely deficient in phytochrome B. Wild-typeseedlings accumulated more DM than lh seedlings over a 4-weekgrowth period in the glasshouse. Leaf and root DM were higherin the WT but stem DM was higher in the lh mutant. Stem DM perunit length was larger in WT than in lh mutant seedlings, evenwhen the two genotypes were compared at equal whole plant DM,which was achieved by growing the plants under different irradiance.In WT seedlings, the hypocotyl was shorter but thicker, withlarger average cell diameter than the lh mutant. In hypocotyltransverse sections the area occupied by load-bearing tissues(xylem and phloem fibres) and the number and diameter of xylemvessels were larger in WT than lh seedlings. Survival ratesof the lh mutant were normal in the glasshouse but very lowoutdoors due to hypocotyl fracture. The water conductivity ofhypocotyl sections was higher in WT than lh seedlings, but nosignificant differences in water conductivity were observedwhen the root remained attached to the hypocotyl. These resultssuggest (a) that compared to the WT, tall and slender lh plantsare more susceptible to mechanical stresses created by windimpact, and (b) that if the lh lesion affects the phyB geneonly, phytochrome B plays a role in the elicitation of anatomicaland morphological changes that specifically increase fitnessin open environments.Copyright 1994, 1999 Academic Press Cucumis sativus (cucumber), light phenotypes, phytochrome, photomorphogenesis, shade phenotypes, stem growth