Comparison of Site I auxin binding and a 22-kilodalton auxin-binding protein in maize

Several properties of a 43-kilodalton (kDa) auxin-binding protein (ABP) having 22-kDa subunits are shared by a class of auxin binding designated Site I. The spatial distribution of the ABP in the maize (Zea mays L.) mesocotyl corresponds with the distribution of growth induced by naphthalene-1-acetic acid and with the distribution of Site I binding as previously shown by J.D. Walton and P.M. Ray (1981, Plant Physiol. 68, 1334–1338). The greatest abundance of both ABP and Site I activity is at the apical region of the mesocotyl. The ABP and Site I activity co-migrate in isopycnic centrifugation with the endoplasmic-reticulum marker, cytochrome-c reductase. Red light, at low and high fluence, far-red and white light were used to alter the elongation rate of apical 1-cm sections of etiolated maize mesocotyls, the amount of auxin binding, and the abundance of the ABP. Relative changes in auxin binding and the ABP were correlated, but the growth rate was not always correlated with the abundance of the ABP.Abbreviations ABP auxin-binding protein - ER endoplasmic reticulum - FR far-red light - kDa kilodalton - NAA naphthalene-1-acetic acid - PM plasma membrane - R red light - SDS-PAGE sodium dodecylsulfate-polyacrylamide gel electrophoresis     

Indole-3-lactic acid is a weak auxin analogue but not an anti-auxin

Indole-3-lactic acid (ILA) is a naturally occurring indole derivative, preferably detected in soil bacteria and fungi and only in low amounts in plants. T-DNA gene 5 of Agrobacterium tumefaciens was found to be involved in the synthesis of ILA in transformed plant tissues, but the physiologic relevance for ILA production in plants is unclear. The related molecular structure of ILA to the natural auxin indole-3-acetic acid (IAA) makes ILA a good candidate for an auxin analogue. We examined the possible auxin activity of ILA on elongation, proliferation, and differentiation in Pisum sativum L. Results presented in this paper indicate that there are no or only weak effects of ILA toward the activity of auxins when used in the physiologic concentration range. Furthermore, no antagonistic effects of ILA were found. Biochemical analysis using the equilibrium dialysis binding system resulted in no high affinity ILA binding to an enriched protein fraction containing auxin-binding protein (ABP₄₄), whereas 1-naphthaleneacetic acid exhibited high affinity auxin binding.Abbreviations IAA indoleacetic acid - ILA indole-3-lactic acid - T-DNA transferred DNA - ABP auxin-binding protein - NAA naphthaleneacetic acid - MS Murashige and Skoog - MES 2-(N-morpholino)ethanesulfonic acid - BAP 6-benzylaminopurine     

Modelling of auxin-binding protein 1 suggests that its C-terminus and auxin could compete for a binding site that incorporates a metal ion and tryptophan residue 44

Sequence comparison indicates that auxin-binding protein 1 (ABP1) belongs to a family of proteins with the core β-barrel structure of the vicilins. Previous modelling within this family correctly predicted metal-ion binding and oligomeric properties of oxalate oxidase. ABP1 also contains a putative metal-ion-binding cluster of amino acids, adjacent to a tryptophan side chain, leading to a proposed auxin-binding site that incorporates metal-ion interaction with the auxin carboxylate. Modelling implicates W44 (Zea mays ABP1) in auxin binding, rather than W136 or W151. Reduced sequence similarity for the C-terminal region prevents model building. It is proposed that one of these C-terminal tryptophans, along with a neighbouring negatively charged side chain, occupies the binding pocket in the absence of auxin, thereby linking auxin binding to conformational change and C-terminal involvement in signalling. Received: 10 December 1999 / Accepted: 4 August 2000     

Partial Purification of a Soluble Gibberellin-Binding Protein from Mung Bean Hypocotyls

A soluble binding protein specific for GA₄, GA₇ and GA₉ waspartially purified from mung bean hypocotyls, and its characteristicswere examined. Affinity chromatography using immobilized GA₃coupled to Sepharose 4B via the C-7 carboxyl group was veryeffective for purification of the protein. The molecular weightof the protein in its native state was estimated to be 150–200kDa by gel-permeation chromatography. This protein may be aheterooligomer consisting of two subunits (23 kDa and 35 kDa).The optimum pH for binding of GA₄ to the protein was around6.0 and the apparent dissociation constant (K_d) was 310^-7 M. (Received April 24, 1992; Accepted December 16, 1992)     

Mineral nutrition and auxin-induced growth of Triticum coleoptile sections

A revised method has been described for assaying auxin by thegrowth of Triticum coleoptile sections. With additions of Ca(NO₃)₂10^–4 and MgSO₄ 10^–5 mol liter^–1 the sensitivityand accuracy have been increased. This is mainly due to Ca.The coleoptiles obviously suffer from Ca-deficiency. The importanceof a strict time schedule for manipulations is emphasiced. Theduration of the tests is limited to 6 hr.Indole-3-acetic acidcan be determined in concentrations down to 10^–9mol liter^–1;from 10^–10 to 10^–9mol liter^–1 the log-logrelation between concentration and growth is linear. Above 10^–7mol liter^–1 elongation takes place under an abnormal increasein elastic extension, indicating that growth is limited by someunknown wallstabilizing factor. The interrelation between auxin,an anti-auxin and Ca are discussed. (Received May 8, 1973; )     

The Effect of Triacontanol on Peroxidase, IAA, and Plant Growth in Pisum sativum var.'Alaska' and 'Little Marvel'

The present study investigates the effects of triacontanol (CH₃(CH₂)₂₈CH₂OH),on plant growth (root and stem), peroxidase activity (apicalmeristem tissue), and auxin destruction (apical meristem tissue)in ‘Little Marvel’ dwarf (LM) and ‘Alaska’peas (AP). Triacontanol inhibited root growth in LM comparedto untreated controls. However, root growth in AP tissue wasenhanced by 1.0 mg I^–1 triacontanol and inhibited by allother treatments, in comparison to untreated controls. Wateruptake in triacontanol-treated AP plants was greater than inuntreated controls, with the converse being the case for LM.Triacontanol treatment caused an increase in peroxidase activityin both LM and AP plants compared to untreated controls. Interms of (1–¹⁴C)IAA destruction, GA₃ + 0.01 mg 1^–1triacontanol caused appreciable auxin breakdown (40%) in LMtissue, with GA₃ + 0.1 mg 1^–1 triacontanol giving a 43%decrease compared to untreated controls. In AP tissue, 10 µMGA3 increased auxin destruction by 188% whereas 0.1 mg I^–1triacontanol caused a 20% decrease compared to untreated controls.The effects of triacontanol on root and stem growth, peroxidaseactivity, and auxin destruction appear to be cultivar-specific,with respect to LM and AP varieties of peas.     

Photolytic Decarboxylation of Carboxyl-14C-labelled Indol-3yl-acetic Acid in Leaves of the Apple Tree

The nature and rate of degradation of carboxyl-¹⁴C-labelledindol-3y1-acetic acid (IAA-[l-¹⁴C]) were studied in apple leaves.The labelled auxin was applied to the cut surface of the growingshoot after the apical part had been removed. The respiratoryCO₂ absorbed by chromatographic paper as Na₂CO₃ then freed byphosphoric acid was quantitatively measured by an internal gascounter. It was found that the concentration of ¹⁴CO₂ evolvedby leaves was 77 times higher in daylight than in darkness.The ratio of ¹⁴CO₂/CO₂ obtained from respiration from the uppersurface of leaf blades was two and seven times higher than thatfrom the lower surface after 15 and 30 h of daylight, respectively.No such differences were noticed in darkness. Similarly, thetotal radioactivity of leaf tissues tripled in daylight, presumablybecause of photosynthetic incorporation of radioactive CO₂ evolvedduring decomposition of LAA. These facts demonstrate the photolyticcharacter of auxin decarboxylation in apple leaves. Prolongeddarkness seemed to provoke a large metabolite withdrawal fromleaves and, to some extent, to protect auxin against decarboxylation.     

Phytotoxicity of Phthalate Plasticisers: 2. SITE AND MODE OF ACTION

The effects of phthalate esters on chlorophyll a₂ fluorescencein radish plants (Raphanus sativus L. cv. Cherry Belle) wereexamined Fluorescence yield was increased in those plants exposedto an aerial concentration of 120 ng dm^–3 dibutyl phthaiatc(DBP) at a rate of 3.0 dm³ min^–1 for 13 d. Comparisonof fluorescence enhancement ratios and F_red/F_ox, suggests thatDBP inhibits photosynthesis in radish plants at a site afterQ_A. Both DBP and diisobutyl phthalate (DIBP) strongly inhibiteduncoupled (PS2+PS1) electron transport rates in thylakoids isolatedfrom spinach. At a chlorophyll concentration of 10 µgcm^–3 the concentrations of DBP and DIBP exhibiting 50%inhibition were 44 mmol m^–3 and 42 mmol m^–3 respectively.Basal electron transport rates were also inhibited, with 87mmol m^–3 of DBP or DIBP producing 50% inhibition. Measurementof photosystcm 1 activity suggested that the main site of actionof these phthalates was localized at a site near the reducingside of photosystem 2. Key words: Phthalate, plasticiser, chlorophyll, fluorescence, photosynthesis, inhibition     

Molecular analysis of auxin-specific signal transduction

The auxin-binding protein (ABP1) of maize has been purified, cloned and sequenced. Homologues have been found in a wide range of plants and at least seven ABP sequences from four different species are now known. We have developed a range of anti-ABP antibodies and these have been applied to analysis of the structure, localization and receptor function of ABP. ABP1 is a glycoprotein with two identical subunits of apparent M _r =22 kDa. The regions recognised by our five monoclonal antibodies (MAC 256–260) and by polyclonal antisera from our own and other laboratories have been specified by epitope mapping and fragmentation studies. All polyclonal anti-ABP sera recognise two or three dominant epitopes around the single glycosylation site. Two monoclonals (MAC 256, 259) are directed at the endoplasmic reticulum (ER) retention sequence KDEL at the C-terminus. Early biochemical data pointed to six amino acids likely to be involved in the auxin binding site. Inspection of the deduced sequence of ABP1 showed a hexapeptide (HRHSCE) containing five of these residues. Antibodies were raised against a polypeptide embracing this region and recognised ABP homologs in many species, suggesting that the region is highly conserved. This is confirmed by more recent information showing that the selected polypeptide contains the longest stretch of wholly conserved sequence in ABP1. Most strikingly, the antibodies show auxin agonist activity against protoplasts in three different electrophysiological systems-hyperpolarization of tobacco transmembrane potential; stimulation of outward ATP-dependent H⁺ current in maize; modulation of anion channels in tobacco. The biological activity of these antibodies indicates that the selected peptide does form a functionally important part of the auxin binding site and strongly supports a role for ABP1 as an auxin receptor. Although ABP contains a KDEL sequence and is located mainly in the ER lumen, the electrophysiological evidence shows clearly that some ABP must reach the outer face of the plasma membrane. One possible mechanism is suggested by our earlier demonstration that the ABP C-terminus recognised by MAC 256 undergoes an auxin-induced conformational change, masking the KDEL epitope and it is of interest that this C-terminal region appears to be important in auxin signalling [22]. So far we have been unable to detect the secretion of ABP into the medium of maize cell (bms) cultures reported by Jones and Herman [7]. However, recent silver enhanced immunogold studies on maize protoplasts have succeeded in visualizing ABP at the cell surface, as well as auxin-specific clustering of the signal induced within 30 minutes. The function of ABP in the ER, as well as the mechanisms of auxin signal transduction both at plasma membrane and gene levels remain to be elucidated.     

The Stimulatory Effect of Indole-3-Acetic Acid on the Uptake of Amino-Acids by Tissue 1Helianthus annuus

Indole-3-acetic acid was observed to bring about a prompt andmarked increase in the amount of ¹⁴C accumulated by segmentsof sunflower hypocotyl from solutions of labelled glutamic acid,glycine, and lysine. The curve relating magnitude of effectto indole-3-acetic acid concentration followed the comparablecurves for water uptake and extension growth. The accumulation of ¹⁴C was related to the external concentrationof glutamic acid by a curve which departed only slightly fromlinearity. The percentage increase in ¹⁴C accumulation broughtabout by auxin did not decline to any appreciable extent withincreasing external concentration of glutamic acid. Under nitrogen the amount of ¹⁴C taken up from solutions oflabelled glutamic acid in 1·75 hour was cut down by approximatelyone-third, and the auxin effect was abolished. The Q₁₀ for ¹⁴Caccumulation between 16° C. and 26° C. was 1·2in the absence of indole-3-acetic acid, and was 1·3 inits presence. When net water uptake was eliminated by the addition of mannitolto the external solution, ¹⁴C accumulation in auxin-free mediawas not depressed. The percentage increase in ¹⁴C accumulationbrought about by auxin, however, was markedly reduced. The fate of the ¹⁴C accumulated was investigated by means ofchromatography on resin columns and on filter paper. About 30–40percent, of the ¹⁴C was in the form of glutamic acid after approximatelya hours' treatment. No marked difference in the level of glutamicacid was observed between auxin-treated and control segments.The effect of auxin was more evident on the amounts of otherradioactive derivatives, as yet unidentified. It was observed that, not only was the amount of CO₂ evolvedin respiration higher in the presence of indole-3-acetic acid,but that this CO₂ was richer in ¹⁴C, i.e. in auxin-treated tissueglutamic acid formed a larger proportion of the substrate respired. The possible implications of these observations are discussed.It is pointed out that indole-3-acetic acid may have achievedits effect by stimulating a transfer process, by lessening adiffusion resistance, or by promoting a process or processeswhich, by removing free amino-acids within the cell, maintainan inward diffusion gradient.     

Interactions of syndecan-1 and heparin with human collagens

Glycosaminoglycan (GAG)–collagen interactions play importantroles in cell adhesion and extracellular matrix assembly; however,the chemical bases for these interactions are not fully understood.We have used affinity co-electrophoresis (ACE) (Lee,M.K. andLander,A.D., Proc. Natl. Acad. Sci. USA, 88, 2768–2772,1991) to study the binding of the heparan sulphate proteoglycansyndecan-1 and heparin to human collagens. [³⁵S]Syndecan-1 [fromnormal murine mammary gland (NMuMG) epithelial cells] and low-M_r({small tilde}6 kDa) [¹²⁵I]heparin were subjected to electrophoresisthrough agarose gel lanes containing human collagens at variousconcentrations, and binding affinities were measured from shiftsin migration of the labelled materials. Results demonstratethat the affinities of each collagen for syndecan-1 and low-M_rheparin were similar, and followed the order: type V> >type IV     

Response of Glutamine Synthetase and Glutamate Synthase Isoforms to Nitrogen Sources in Rice Cell Cultures

As a model system with no photorespiration and no long distancetransport, rice cell cultures (Oryza saliva L. cv Sasanishiki)were used to investigate the effect of nitrogen sources on thelevels of isoforms of glutamine synthetase (GS) and glutamatesynthase (GOGAT). Isoforms of GS and GOGAT were analyzed byimmunoblotting methods and their activities in early growthphase of the cells. Cytosolic type GS (41 kDa subunit) and NADH-GOGATwere the major isoforms in the rice cells grown in normal R-2medium. However, contents of plastid type GS (44 kDa subunit)and Fd-GOGAT increased in response to NO_–³ supply. NADH-GOGATactivity also increased following the supply of NO_–³.In vitro translated products from poly(A)⁺RNA prepared fromthe cells showed that the precursor of plastid type GS (49 kDa)was detected at 48 h after the inoculation. Supply of NH₊⁴ resultedin an increase in NADH-GOGAT activity but had no effect on thelevels of Fd-GOGAT, of polypeptides of the plastid type GS orof the corresponding mRNAs. (Received May 30, 1990; Accepted August 23, 1990)     

Interaction between lysine 102 and aspartate 338 in the insect amino acid cotransporter KAAT1

KAAT1 is a lepidopteran neutral amino acid transporter belonging to the NSS super family (SLC6), which has an unusual cation selectivity, being activated by K⁺ and Li⁺ in addition to Na⁺. We have previously demonstrated that Asp338 is essential for KAAT1 activation by K⁺ and for the coupling of amino acid and driver ion fluxes. By comparing sequences of NSS family members, site-directed mutagenesis, and expression in Xenopus laevis oocytes, we identified Lys102 as a residue likely to interact with Asp338. Compared with wild type, the single mutants K102V and D338E each showed altered leucine uptake and transport-associated currents in the presence of both Na⁺ and K⁺. However, in K102V/D338E double mutant, the K102V mutation reversed both the inhibition of Na⁺-dependent transport and the block in K⁺-dependent transport that characterize the D338E mutant. K⁺-dependent leucine currents were not observed in any mutants with D338E. In the presence of the oxidant Cu(II) (1,10-phenanthroline)₃, we observed specific and reversible inhibition of K102C/D338C mutant, but not of the corresponding single cysteine mutants, suggesting that these residues are sufficiently close to form a disulfide bond. Thus both structural and functional evidence suggests that these two residues interact. Similar results have been obtained mutating the bacterial transporter homolog TnaT. Asp338 corresponds to Asn286, a residue located in the Na⁺ binding site in the recently solved crystal structure of the NSS transporter LeuT_Aa (41). Our results suggest that Lys102, interacting with Asp338, could contribute to the spatial organization of KAAT1 cation binding site and permeation pathway. residue interaction; oxidants; tertiary structure     

Overexpression of the Auxin Binding PROTEIN1 Modulates PIN-Dependent Auxin Transport in Tobacco Cells

Background

Auxin binding protein 1 (ABP1) is a putative auxin receptor and its function is indispensable for plant growth and development. ABP1 has been shown to be involved in auxin-dependent regulation of cell division and expansion, in plasma-membrane-related processes such as changes in transmembrane potential, and in the regulation of clathrin-dependent endocytosis. However, the ABP1-regulated downstream pathway remains elusive.

Methodology/Principal Findings

Using auxin transport assays and quantitative analysis of cellular morphology we show that ABP1 regulates auxin efflux from tobacco BY-2 cells. The overexpression of ABP1can counterbalance increased auxin efflux and auxin starvation phenotypes caused by the overexpression of PIN auxin efflux carrier. Relevant mechanism involves the ABP1-controlled vesicle trafficking processes, including positive regulation of endocytosis of PIN auxin efflux carriers, as indicated by fluorescence recovery after photobleaching (FRAP) and pharmacological manipulations.

Conclusions/Significance

The findings indicate the involvement of ABP1 in control of rate of auxin transport across plasma membrane emphasizing the role of ABP1 in regulation of PIN activity at the plasma membrane, and highlighting the relevance of ABP1 for the formation of developmentally important, PIN-dependent auxin gradients.     

Ligand Specificity of Bean Leaf Soluble Auxin-binding Protein

The soluble bean leaf auxin-binding protein (ABP) has a high affinity for a range of auxins including indole-3-acetic acid (IAA), α-napthaleneacetic acid, phenylacetic acid, 2,4,5-trichlorophenoxyacetic acid, and structurally related auxins. A large number of nonauxin compounds that are nevertheless structurally related to auxins do not displace IAA from bean ABP. Bean ABP has a high affinity for auxin transport inhibitors and antiauxins. The specificity of pea ABP for representative auxins is similar to that found for bean ABP. The bean ABP auxin binding site is similar to the corn endoplasmic reticulum auxin-binding sites in specificity for auxins and sensitivity to thiol reagents and azide. Qualitative similarities between the ligand specificity of bean ABP and the specificity of auxin-induced bean leaf hyponasty provide further evidence, albeit circumstantial, that ABP (ribulose 1,5-bisphosphate carboxylase) can bind auxins in vivo. The high incidence of ABP in bean leaves and the high affinity of this protein for auxins and auxin transport inhibitors suggest possible functions for ABP in auxin transport and/or auxin sequestration.     

Mechanism of auxin interaction with Auxin Binding Protein (ABP1): a molecular dynamics simulation study

Auxin Binding Protein 1 (ABP1) is ubiquitous in green plants. It binds the phytohormone auxin with high specificity and affinity, but its role in auxin-induced processes is unknown. To understand the proposed receptor function of ABP1 we carried out a detailed molecular modeling study. Molecular dynamics simulations showed that ABP1 can adopt two conformations differing primarily in the position of the C-terminus and that one of them is stabilized by auxin binding. This is in agreement with experimental evidence that auxin induces changes at the ABP1 C-terminus. Simulations of ligand egress from ABP1 revealed three main routes by which an auxin molecule can enter or leave the ABP1 binding site. Assuming the previously proposed orientation of ABP1 to plant cell membranes, one of the routes leads to the membrane and the other two to ABP1's aqueous surroundings. A network of hydrogen-bonded water molecules leading from the bulk water to the zinc-coordinated ligands in the ABP1 binding site was formed in all simulations. Water entrance into the zinc coordination sphere occurred simultaneously with auxin egress. These results suggest that the hydrogen-bonded water molecules may assist in protonation and deprotonation of auxin molecules and their egress from the ABP1 binding site.     

Binding Affinities of Oxidized and Reduced Forms of Tetrahalogenated Benzoquinones to the QB Site in Oxygen-Evolving Photosystem II Particles from Synechococcus elongatus

Binding affinities of the Q_B site for four tetrahalogenatedbenzoquinones (THBQs) were investigated by measuring their abilityto serve as electron acceptors or act as inhibitors of oxygenevolution in Synechococcus photosystem II particles. Iodanil,bromanil and chloranil but not fluoranil induced a rapid oxidationof Q_A^– and doubled the area over the fluorescence inductioncurve, indicating dark oxidation of Q₄₀₀. Analyses of thesetwo THBQ-induced reactions and inhibition of the acceleratedQ_A^– oxidation by DCMU yielded binding constants of thequinones comparable to those determined from measurements ofoxygen evolution. Generally, THBQs bound tightly to the Q_B site.However, the binding affinity varied in a wide range with THBQs.The Q_B site bound iodanil with an extremely high affinity butfluoranil relatively weakly. The hydroquinone forms of the THBQsalso bound to the Q_B site and inhibited Q_A^– oxidationby Q_B. The concentrations of the hydroquinones required for50% inhibition of Q_A^– oxidation suggest that the Q_B sitebinds the hydroquinones more weakly than the corresponding quinonesexcept for fluoranil, which binds to the Q_B site more tightlyin its reduced form than in oxidized one. The abilities of THBQsto function as electron acceptors or inhibitors of oxygen evolution,and as oxidants of Q₄₀₀ in the dark, are discussed in relationto the binding affinities of the quinones to the Q_B site. ⁴Present address: Department of Biology, Faculty of Science,Toho University. Miyama 2-2-1, Funabashi, 274 Japan     

Relation between Ultra-Violet Spectral Change and Nucleotide Binding of the Chloroplast Coupling Factor 1

Properties of the nucleotide binding sites on chloroplast couplingfactor 1 (CF₁) were studied by equilibrium dialysis and UV spectroscopy.From our direct binding studies, we identified at least fourkinds of ADP binding sites on CF₁; a barely dissociable ADPbinding site (site A), a slowly exchangeable high affinity sitewith dissociation constant (Kd) 0.021 µM (site B), anotherslowly exchangeable high affinity site with Kd 1.6 µM(site C) and several low affinity (Kd {small tilde}30 µM)sites. The Kd values for sites B and C of the other nucleotidestested were 0.5 µM and 16 µM (GDP), 8 µM and34 µM (CDP), 17 µM and 20 µM (UDP) and 1.4µM and 1.4 µM (PP₁). From a comparison of the observed UV spectral change and theamount of nucleotide bound to these sites, as calculated fromthe above Kd values, we concluded that the nucleotide bindingto site B or G induces UV spectral changes that are almost thesame in shape and magnitude. The estimated difference molarabsorption coefficient () was 3.4?10³M^–1_ADP cm^–1for ADP at 278 nm. Our conclusions were strengthened by thegood agreement between the observed spectra and the calculatedspectra (derived from the Kd and values of ADP and GDP) whenADP and GDP were added together to CF₁. The cause of the unusual behavior of GDP in the UV differencespectrum which was unexplained in our previous report was shownto be competition between the GDP added and previously boundADP at sites B and C; this distorted the real spectrum inducedby GDP. (Received October 3, 1983; Accepted February 13, 1984)     

NaCl-and Gibberellic Acid-induced Changes in the Content of Auxin and the Activities of Cellulase and Pectin Lyase During Leaf Growth in Rice (Oryza sativa)

The interactive effect of NaCl salinity and gibberellic acidin the activities of cellulase and pectin lyase, and on thecontent of auxin and chlorophyll, has been determined duringleaf growth (fifth from base) in rice. The linear growth, chlorophyllcontent, activity of cellulase, and the auxin level of leaveswere markedly decreased when plants were exposed to salt stress(12 dS m^–1). However pectin lyase activity did not registerany significant alteration in the leaves of salt-stressed plantscompared with the control. Treatment of plants with gibberellicacid (GA₃) (10 ppm) increased the leaf growth and chlorophyllcontent with a concomitant rise in the activity of cellulaseunder stressed as well as non-stressed conditions. A markedincrease in the content of auxin was discernible in the leavesof salt-stressed plants treated with GA₃ compared with non-treatedsalinized ones. An appreciable increment in the activity ofpectin lyase in response to GA₃ administration was detectedonly in the leaves of non-stressed plants. These results indicatethat enhancement of cellulase activity and the augmentationof endogenous auxin content may be involved in the stimulationof rice leaf growth by GA₃ under saline conditions. Oryza sativa, rice, leaf growth, NaCl salinity, gibberellic acid, cellulase, pectin lyase, auxin     

The Uptake of Gaseous Ammonia by the Leaves of Italian Ryegrass

Lockyer, D. R. and Whitehead, D. C. 1986. The uptake of gaseousammonia by the leaves of Italian ryegrass.—J. exp. Bot.37: 919–927. Plants of Italian ryegrass (Lolium multiflorum Lam.) grown insoil with two rates of added ¹⁵N-labelled nitrate were exposed,in chambers, for 40 d to NH₃ in the air at concentrations of16, 118 and 520 µg m^–3. At the highest concentrationof NH₃, this source provided 47?3% of the total nitrogen inplants grown with the lower rate of nitrate addition (100mgN kg^–1 dry soil) and 35?2% with the higher rate (200mgN kg^–1 dry soil) At the intermediate concentration ofNH₃, the contributions to total plant N were 19?6% and 10?8%,respectively, at low and high nitrate while, at the lowest concentrationof NH₃, they were 5?1% and 32%. Most of the N derived from theNH₃ remained in the leaves, but some was transported to theroots. The amount of N derived from the NH³ that was presentin the leaves was not reduced by washing the leaves in waterat pH 5?0 before harvesting, indicating that the N was assimilatedby the plant and not adsorbed superficially. Rates of uptakeof NH₃ per unit leaf area ranged from 1?7 µg dm^–2h^–1 at a concentration of 16 µg m^–3 to 29?0µg dm^–2 h^–1 at a concentration of 520 µgm^–3 and with the lower rate of nitrate addition. Increasingthe supply of nitrate to the roots slightly reduced the rateof uptake of NH₃ per unit leaf area. Uptake of N from the higherrate of nitrate was reduced at the highest concentration ofNH₃ in the air. Key words: Ammonia, nitrogen, leaf sorption, Lolium multiflorum