Cell Cycle Changes in the Shoot Apex of Silene coeli-rosa During the Second and Third Days of Floral Induction

The cell cycle in Silene coeli-rosa shoot apices was measuredto test whether or not early components of the floral stimulus,produced during the 2nd and 3rd long days (LD) of an inductiveLD treatment, resulted in an increase in the duration of G₂phase in constant 20–24 h cell cycles. Plants were grownat 20°C in short days (SD) of 8 h light and 16 h darknessfor 28 d (day 0). Starting on day 0, plants were given SD or3 LD each comprising an identical 8 h day and 16 h photo-extension,or 3 dark-interrupted (d.i.) non-inductive LD, interrupted at1700 h of each day with 1 h of darkness. The cell cycle (percentagelabelled mitoses method) and changes in cell number were determinedin the shoot apical meristem. During days 1–2 of the SDtreatment, the cell cycle and mean cell generation time (MCGT)was 18 and 32 h, respectively, giving a growth fraction of 56%.During days 2–3, the cell cycle and MCGT shortened to15 and 23 h, respectively (growth fraction = 65%). During days1–2 of the LD and d.i. LD treatments, cell cycles andMCGTs were 9–10 and 27–29 h, respectively, resultingin smaller growth fractions (about 33%). Thus, shortened cellcycles and altered growth fractions occurred regardless of whetheror not the treatment was inductive. The LD treatment resultedin a marked shortening of G₁ and, to a lesser extent, S-phase,whilst G₂ remained constant. These changes were consistent withincreases in the proportion of cells in G₂ during the photoextensionof each LD which were suppressed during the comparable periodsof the d.i. LD treatment. The latter treatment resulted in eachphase occupying virtually identical proportions of the cellcycle as in the SD treatment. Thus, the unique cell cycle responsesto the initial part of the inductive LD treatment were increasesin the proportion of cells in G₂ coupled with G₁ and G₂ beingof similar duration. Cell cycle, mean cell generation time, shoot apex, Silene coeli-rosa     

Inhibitory effect of fusicoccin on cell division

Rapid interactions in cell division and cytodifferentiationare induced by hormone treatments in dark-cultured explantsof Jerusalem artichoke. Fusicoccin, at concentrations between10^–6 and 10^–5 M, markedly inhibited the division-promotingactivity induced by plant hormones. Further, fusicoccin-treatedmeristematic root tips of Vicia faba and Allium cepa showeda rapid decrease in the mitotic index. Fusicoccin seems to inhibitsome hormone-sensitive processes required during the inductionand regulation of cell division. (Received March 28, 1979; )     

Studies of pesticides effects on Chlorella metabolism III. Effect of isopropyl 3-chlorocarbanilate (chlorpropham) on cell cycle and biosynthesis

Isopropyl 3-chlorocarbanilate (chlorpropham) inhibited Chlorellagrowth by 50% at 1.3 µM under non-photosynthetic conditions.Average DNA content per cell was increased 2.5-fold by chlorprophamtreatment at 4.7 µM. Oxygen uptake was not significantlyaffected at 47 µM. Protein synthesis was more sensitiveto chlorpropham than the other biosynthetic processes tested,but it was not inhibited at 1.3 µM. In cell cycle studiesperformed under photosynthetic conditions, 50% inhibition ofgrowth occurred at 4 µM. At 14 µM, growth in termsof cell number was completely suppressed while inhibition ofgrowth in terms of average cell volume was partial, resultingin comparatively larger cell volume. This was accompanied by3.0-fold increase in average DNA content per cell. (Received March 8, 1976; )     

Characterization of the vacuolar-type H+-ATPase from guard cell protoplasts of Commelina

Characteristics of the vacuolar-type (V-type) H⁺-ATPase fromguard cell protoplasts of Commelina communis L. were investigatedusing a linked enzyme assay and nitrate inhibition as a diagnosticindicator of the enzyme activity. ATPase activity was completelyinhibited by about 50 mol m^–3 nitrate and activity wasoptimal near pH 8.0. The temperature optimum for activity wasabout 37 C and an Arrhenius plot indicated changes in activationenergy for the ATPase at 15C and possibly at about 30 C. Theenzyme was stimulated by Cl^– while Ca²⁺ inhibited activity(l₅₀ = 1.5 mol m^–3). The apparent K_m (MgATP) was 0.62mol m^–3. Incubation of guard cell protoplasts for up to 5 h in 50 µMabscisic acid (ABA) or 25µM fusicoccin (FC) did not affectsubsequent ATPase activity. In vitro assays with FC or ABA alsodid not affect enzyme activity. Activity was not affected bylight or potassium ferricyanide, two factors which are knownto influence stomatal activity. Beticoline was a potent inhibitorof activity (l₅₀ = 50 µM) while DCCD was less effective(l₅₀ = 90µM). On chlorophyll, protein and protoplast bases, V-type ATPaseactivity was greater in guard cell protoplasts than mesophyllcell protoplasts by 66, 13.9 and 1.9, respectively. On atonoplast surface area basis the enzyme activity was 5.6 timeshigher in guard cell protoplasts than in mesophyll cell protoplasts Thus, although the characteristics of the V-type, H ⁺-ATPaseof GCP are very similar to those found in other cell types,rates of activity and probably tonoplast enzyme density aremuch greater in guard cell protoplasts than mesophyll cell protoplastsof C. communis which corresponds with the large and rapid ionfluxes across the tonoplast associated with stomatal movements Key words: Guard cell protoplasts, stomata, V-type H ⁺-ATPase     

Expression of the gus A gene in embryogenic cell lines of Sitka spruce following Agrobacterium-mediated transformation

Transformation of Picea sitchensis (Bong) Carr. was investigatedby incubating embryogenic cell lines, initiated from immatureand mature zygotic embryos, with a supervirulent strain of Agrobacteriumtumefaciens. The latter carried a gus A-intron gene. Transientgene expression was determined histochemically by recordingthe number of distinct areas of ß-glucuronidase (GUS)activity. Maximum expression of the gus gene was achieved witha bacterial suspension with an OD₆₀₀ of 0.8–1.1 dilutedwith an equal volume of MPM medium, Inoculation of cells withbacteria for 30 mm, 72 h co-cultivation period and exposureof Agrobacterium and plant cells to 50 µM acetosyrmngone.These results are discussed in relation to Agrobacterium-mediatedgene delivery for the stable transformation of Sitka spruceand other conifers. Key words: Sitka spruce, Agrobacterium, transformation, embryonal suspensor masses, GUS activity     

A Comparison between the Uptake of Potassium by Plants from Solutions of Constant Potassium Concentration and during Depletion

A comparison was made between two methods of measuring the relationshipbetween the external [K⁺] and the flux of K⁺ into whole plantsof Lolium perenne and Raphanus sativus. The values of flux obtainedfrom solutions of 1.2 µM K⁺ held constant around the rootswere three and six times greater for Lolium and Raphanus respectivelythan the values obtained at the same concentration in a depletionexperiment in which the solutions, initially 100 µM K⁺,were depleted to below 1.2 µM K⁺ by plant uptake. In thedepletion experiment with Lolium, the flux was higher into plantsgrown at low [K⁺] than into plants grown at 100 µM eventhough [K⁺] within the plant was about the same for all groupsof plants. It is suggested that Lolium grown at low [K⁺] hasan efficient mechanism for K⁺ uptake which continues to operatefor some time after the plants have been transferred to a higherconcentration. With both species, K_m was 15–20 µMin the depletion experiment and below 1 µM when concentrationswere held constant.     

Induction of Cell Division Synchrony and Variation of Cytokinin Contents through the Cell Cycle in Tobacco Cultured Cells

Cell division synchrony was induced in tobacco {Nicotiana tabacum)cultured cells by several treatments. Very high synchrony throughouttwo cell cycles was induced by aphidicolin treatment (inhibitorof DNA polymerase , 10 µg/ml) and by treatment with lowtemperature (4°C) and hydroxyurea (50 µg/ml). Themitotic index reached its maximum (52% and 40% in aphidicolinand hydroxyurea treatments, respectively) at 11 h after removalof the added chemical. During the treatments, the cells werearrested in the G₁/S phase of the cell cycle. In the aphidicolin-inducedsystem, incorporation of ¹⁴C-thymidine confirmed that DNA synthesiswas started immediately after removal of the chemical. The aphidicolin-induced synchronous cells were used to studythe contents of butanol-soluble cytokinins during the cell cycle.Cytokinin contents increased conspicuously at the G₂/M boundary. ¹Present address: Department of Biology, Otsuma Women's University,Chiyodaku, Tokyo 102, Japan. (Received May 14, 1985; Accepted November 8, 1985)     

Flower Induction by Nitrogen Deficiency in Lemna paucicostata 6746

Lemna paucicostata 6746, a short-day plant, produced flowerbuds even under continuous light when cultured in nitrogen-deficientmodified Hoagland medium with 1% sucrose for 3 days or morefollowed by culture on nitrogen-rich medium (either nitrateor ammonium). Flowering was also induced by culture on mediumcontaining 20–100 µM nitrate as the sole nitrogensource for 10 days or more, but not on medium with a low ammoniumconcentration. However, if plants cultured on medium containing5–20 µM ammonium as the sole nitrogen source for10 days were grown in a nitrogen-rich medium for a further 4days, they produced flower buds. Thus, nitrogen deficiency caninduce day length-independent flowering in Lemna paucicoslata6746, but nitrogen is required for the manifestation of flowering. (Received January 31, 1986; Accepted April 24, 1986)     

On the causes of interspecific differences in the growth-irradiance relationship for phytoplankton. Part I. A comparative study of the growth-irradiance relationship of three marine phytoplankton species: Skeletonema costatum, Olisthodiscus luteus and Gonyaulax tamarensis

Three marine phytoplankton species (Skeletonema costatum, Olisthodiscusluteus andGonyaulax tamarensis) were grown in batch culturesat 15°C and a 14:10 L:D cycle at irradiance levels rangingfrom 5 to 450 µEinst m^–2 s^–1. At each irradiance,during exponential growth, concurrent measurements were madeof cell division, carbon-specific growth rate, photosyntheticperformance (both O₂ and POC production), dark respiration,and cellular composition in terms of C, N and chlorophyll a.The results indicate that the three species were similar withrespect to chemical composition, C:N (atomic) = 6.9 ±0.4, photo-synthetic quotient, 1.43 ± 0.09, and photosyntheticefficiency, 2.3 ±0.1 x 10^–3 µmol O₂ (µgChl a)^–1 h^–1 (µEinst m^–2 s^–1)^–1.Differences in maximum growth rate varied as the –0.24power of cell carbon. Differences in growth efficiency, werebest explained by a power function of Chl a:C at µ = 0.Compensation intensities, ranged from 1.1 µEinst m^–2s^–1 for S. costatum to 35 forG. tamarensis and were foundto be a linear function of the maintenance respiration rate.The results indicate that interspecific differences in the µ–Irelationship can be adequately explained in terms of just threeparameters: cell carbon at maximum growth rate, the C:Chl aratio (at the limit as growth approaches zero) and the respirationrate at zero growth rate. A light-limited algal growth modelbased on these results gave an excellent fit to the experimentalµ–I curves and explained 97% of the observed interspecificvariability. ¹Present address: Lamont-Doherty Geological Observatory Columbiaof University, Palisades, NY 10964, USA     

Effects of Zinc on Meristem Size and Proximity of Root Hairs and Xylem Elements to the Root Tip in a Zinc-tolerant and a Non-tolerant Cultivar of Festuca rubra L.

A 4 d exposure to zinc (0.1 and 02 µg Zn cm^–3) reducedthe length of the root apical meristem in a Zn-sensitive cultivar(S59) of Festuca rubra L. to a much greater extent than in aZn-tolerant cultivar (Merlin). In S59, Zn treatment also inducedroot hair and xylem formation much closer to the root cap boundarythan in control roots, whereas Merlin was only marginally affectedby Zn treatment. The data are discussed in relation to previouslyestablished effects of Zn on the cell cycle and other cellularcharacters of the two cultivars. zinc, meristem size, root hair, xylem, Festuca rubra     

Microelectrophoretic and 9-Aminoacridine Fluorescence Study of the Surface Electrical Properties of Suspension Cultured Catharanthus roseus Cells and Isolated Protoplasts: Effects of Abscisic Acid Treatment

The effects of abscisic acid (ABA) treatments on the surfaceelectrical properties of cells and isolated protoplasts fromCatharanthus roseus cell suspension cultures were studied byelectrophoretic mobility and 9-aminoacridine (9AA) fluorescencemeasurements. The surface charge densities of the cells andprotoplasts estimated from electrokinetic data were –0.064Cm^–2and –0.048 C m^–2 respectively. These values wereclose to that estimated by 9AA fluorescence technique i.e.,–0.053 Cm^–2 for the cells and –0.041 Cm^–2for the isolated protoplasts accordingly. The net negative surfacecharge density decreased after application of 10 µM and50 µM ABA in both cells and protoplats, the more pronouncedeffect being observed at 10 µM ABA. When 100 µMABA was supplemented to the cell suspension culture the oppositeeffect was observed. The average charge density increased to–0.074 C m^–2 for the cells, and to –0.055C m^–2 for protoplasts, as revealed from the 9AA measurements.The results are discussed in terms of specific concentrationdependent ABA-induced alterations of the electrostatic propertiesof cell and protoplast membranes. (Received December 12, 1994; Accepted April 3, 1995)     

Modification of the root plasma membrane lipid composition of cadmium-treated Pisum sativum

The effects of in vivo Cd treatments on pea root plasma membrane(PM) lipid composition were studied. In the long-term experiment,plants were supplied with Cd: moderate stress (10 µM)or strong stress (50 µM) for 10 d. Growth of root andshoot was severely affected in 50 µM Cd-treated plants,as evidenced by the approximately 7-fold reduction in theirRelative Growth Increment (RGI). Treatment with Cd (10 µM)resulted in changes to the lipid composition of the pea rootPM, including increases in the degree of unsaturation of phospholipid-associatedfatty acids and in the relative amount of stigmasterol (30–42%).This change was accompanied by a reduction in sitosterol content(26.8 to 17.4 µg mg^–1 protein). However, the sterolcomposition was not altered in plants treated with 50 µMCd for 10 d. The content of phosphatidylethanolamine and phosphatidylcholine(major phospholipids present in pea root PM) decreased as Cdlevel increased, but the ratio between them remained unaffected.In the short-term experiment, plants exposed to Cd (50 µM)accumulated less sitosterol (from 27.7 to 14.0 µg g mg^–1protein) over 72 h, but no significant effect on other measuredlipids was observed. The physiological repercussions of changesin plasma membrane lipid composition, as a result of Cd exposureare discussed. Key words: Cadmium, lipids, pea, Pisum sativum, plasma membranes     

Phosphorus Deficiency Impairs Early Nodule Functioning and Enhances Proton Release in Roots of Medicago truncatula L.

The effect of phosphorus supply (1–15 µM) on protonrelease and the role of P in symbiotic nitrogen fixation inmedic (Medicago truncatula L. ‘Jemalong’) was investigated.As P concentration in the nutrient solution increased, shootand root growth increased by 19 and 15%, respectively by day35, with maximal growth at 4 µM P. A P concentration of15 µM appeared to be toxic to plants. Phosphorus supplyhad no influence on nodule formation by day 12 but increasednodule number by day 35. Nitrogenase activity was estimatedby in situ measurement of acetylene reduction activity (ARA)in an open-flow system. During the assay, a C₂H₂-induced declineof ARA was observed under all P concentrations except 4 µM.Specific ARA (per unit nodule weight) doubled when P supplywas increased from 1 to 8 µM. This effect of P was muchgreater than the effects of P on nodulation and host plant growth.Concentrations of excess cations in plants decreased with increasingP concentration in the nutrient solution. Phosphorus deficiencystimulated uptake of excess cations over anions by the plantsand hence enhanced proton release. The results suggest thatP plays a direct role in nodule functioning in medic and thatP deficiency increases acidification which may facilitate Pacquisition. Copyright 2001 Annals of Botany Company Medicago truncatula L. (medic), P deficiency, C₂H₂-ID, nitrogenase activity, proton release, cation-anion balance, open-flow system     

Cryopreservation of Shoot Tips of Tetraploid Potato (Solanum tuberosumL.) Clones by Vitrification

In vitro-grown shoot tips of five tetraploid potato (SolanumtuberosumL.) clones were cryopreserved by vitrification. Excisedshoot tips (0.5–0.7 mm) were pre-cultured on filter paperdiscs over half strength liquid Murashige and Skoog (MS) mediumsupplemented with 8.7 µMGA₃and different combinationsof sucrose (0.3, 0.5 and 0.7M) plus mannitol (0, 0.2 and 0.4M)for 2 d under a 16 h photoperiod at 24 °C. The pre-culturedshoot tips were either successively loaded with 20 and 60% PVS2 solutions or directly exposed to concentrated vitrificationsolution before physical vitrification during liquid nitrogentreatment. The vitrified shoot tips were warmed rapidly andtreated with dilution mixture (MS+1.2Msucrose) for 30 min beforeplating on regrowth medium. Addition of mannitol to the pre-culturemedium improved survival of vitrified shoot tips. Direct dehydrationof pre-cultured shoot tips with concentrated PVS 2 was detrimentalto survival of vitrified shoot tips. Shoot tips pre-culturedon medium containing 0.3Msucrose plus 0.2Mmannitol, and loadedwith 20% PVS 2 for 30 min followed by 15 min incubation in 60%PVS 2 and 5 min incubation in 100% PVS 2 at 0 °C resultedin up to 54% survival after vitrification. About 50% of vitrifiedand warmed shoot tips formed shoots directly. Post-thaw culturingof vitrified shoot tips on medium containing an elevated levelof sucrose (0.2M) under diffuse light for the first week enhancedthe survival rate. Continuous culturing of vitrified shoot tipson high-sucrose medium induced multiple shoot formation.Copyright1998 Annals of Botany Company Solanum tuberosumL., potato, cryopreservation, germplasm conservation,in vitroconservation, meristems, shoot tips, tissue culture, vitrification.     

Uptake of zinc from chelate-buffered nutrient solutions by wheat genotypes differing in zinc efficiency

Zinc-efficient Triticum aestivum (cv. Warigal) and Zn-inefficientTriticum turgidum conv. durum (cv. Durati) were grown in chelate-buffered,complete nutrient solutions providing either deficient or sufficientZn supply. When transferred to fresh chelatebuffered nutrientsolutions containing a wide range of Zn supplies (0–1.28µmol m^–3 Zn²⁺ activity) for 24–48 h, bothgenotypes increased net Zn uptake linearly with an increasein solution Zn²⁺ activities. Zincefficient Warigal accumulatedZn at a greater rate than Zn-inefficient Durati. The greaterrate of net Zn uptake was observed by plants of both genotypeswhen pretreated at deficient Zn supply. Net loss of Zn to thesolution was higher in plants pretreated with sufficient Znand was inversely related to Zn²⁺ activity in the external solution.When continuously supplied with 40 nmol m^–3 Zn²⁺, netZn uptake by Zn-efficient Warigal was significantly greaterthan that of Zn-inefficient Durati, but the difference diminishedwith plant age. Shoot concentrations of Fe, Mn and Cu were higherwhen plants were grown at deficient than at sufficient Zn supply.The Zn-efficient genotype transported less Zn and Fe to shootsand had higher Fe concentrations in roots than the Zn-inefficientgenotype, supporting the hypothesis that Zn efficiency may beconnected with inefficient transport of Fe from roots to shootsand thus initiation of the Fe-deficiency response resultingin increased release of Zn- and Fe-binding phytosiderophores.It is concluded that differential Zn efficiency of wheat genotypesis at least partly due to a greater ability of efficient genotypesto accumulate Zn. Key words: Chelate-buffering, genotypes, micronutrients, Triticum spp., uptake, zinc efficiency     

The Nuclear Endoreduplication Cycle in Metaxylem Cells of Primary Roots of Zea mays L.

The nuclear DNA content of metaxylem cells in roots of Zea mayscv. Golden Bantam reaches 16C or 32C by successive rounds ofDNA endoreduplication. Each phase of endoreduplication (endo-S)is separated by a non-DNA synthetic phase (endo-G). These phasesseem to occur in zones at fixed distances from the root tip.The duration of the phases in two of the endoreduplication cycles(4C–8C, 8C–16C) has been estimated in two ways.The first makes use of the rate of movement of cells throughthe positions along the root where the different phases of thecycle are occurring, the second uses labelling with methyl-[³H]thymidineand autoradiography. Both methods indicate that the endo-S phaseswhich cause the nuclear DNA content to rise from 4C to 8C andfrom 8C to 16C last 8–10 h, and that the intervening endo-Gphase lasts 8–12 h. DNA endoreduplication keeps pace withthe increase of nuclear volume; cell volume increases at a morerapid rate, however. Comparison of the endoreduplication cyclein the metaxylem with the mitotic cycle in the adjoining filesof parenchyma cells shows that the mitotic cells complete theircycle more slowly. DNA synthesis, endoreduplication cycle, mitotic cycle, root apex, Zea mays     

Induction of 1-Aminocyclopropane-1-carboxylic Acid Synthase in Wounded Mesocarp Tissue of Winter Squash Fruit and the Effects of Ethylene

1-Aminocyclopropane-1-carboxylic acid (ACC) synthase activityincreased rapidly after wounding of mesocarp tissue of wintersquash fruit (Cucurbita maxima Duch.) and reached a peak at16 h after excision and then declined sharply. The rise in ACCsynthase activity was followed by increases in the endogenousACC content and the rate of ethylene production. The activityof ethylene forming enzyme (EFE) also increased rapidly in theexcised discs of mesocarp of winter squash fruit. ACC synthase activity was strongly inhibited by aminoethoxyvinylglycinewith a Ki value of 2.1 µM. Michaelis-Menten constant ofACC synthase for S-adenosylmethionine was 13.3 µM. Ethylene suppressed the induction of ACC synthase in the woundedmesocarp tissue. The suppression by ethylene increased withthe increasing concentrations of applied ethylene and the maximumeffect was obtained at about 100 µl 1^–1 ethylene,at which point the induction was suppressed by 54%. Ethylenedid not inhibit ACC synthase activity, nor did it suppress theinduction of EFE, but rather it slightly enhanced the latter. (Received August 24, 1984; Accepted October 29, 1984)     

Effects of monocerin on cell cycle progression in maize root meristems synchronized with aphidicolin

Summary Monocerin is a benzopyran fungal toxin with broad activity on plants, fungi and insects. Its effect upon cell cycle progression has been analyzed in maize roots. Meristematic cells were synchronized by treatment with aphidicolin. Flow cytometric DNA analysis and mitotic indices indicated durations of 1.5 h, 5 h, 2 h and 1 h for respectively G1, S, G2 and M phases of the normal cell cycle at 25°C. Treatment of these synchronized meristems with 0.5 mM monocerin during release after an aphidicolin block produced a short delay in S phase and then a more important delay (about 2.5 h) in entry into mitosis. Treatments for similar durations (3 h) during progression through the cycle revealed two periods of action of monocerin. The first appears to be mid to late S and the second one G2, before the transition point between G2 and M. Action on either one of these target periods could lead to a delay in the G2/M transition, but these two responses did not appear to be additive.Abbreviations APH Aphidicolin - CV Coefficient of variation - DAPI Diamidinophenylindole - DMSO Dimethyl sulfoxide - EDTA Ethylenediaminetetraacetic acid - HPLC High pressure liquid chromatography - MI Mitotic index - SD Standard deviation - UV ultraviolet light     

The initiation of Phaeocystis colonies

This study was designed to elucidate the sequence of eventsthat leads to the formation of new colonies of Phaeocystis sp.(strain PCC 540) starting from single cells released from maturecolonies. Colonies were first isolated by filtration onto a10 µm mesh. Colonial cells were then liberated by shakingand inoculated into individual culture wells containing mediumwith a PO₄^2– concentration of {small tilde}1 µM.Cell size and shape were determined daily by image analysis,while chlorophyll and DNA distributions were estimated by flowcytometry. Released cells were non-flagellated and mostly locatedin the G₁ phase of the cell cycle. They developed flagella andup to 90% became motile within 24 h. Swarmers lost motilityrapidly, became elongated, began to cycle again, excreted amucilaginous compound and divided leading to new colonies withina few days. During this reproducible process, no change of ploidycould be observed. Colonies initially adhered to the bottomof culture wells. Frequent mixing drastically reduced the fractionof colonies produced and their volume. High initial PO₄^2–concentrations (5 µM) delayed colony appearance, whereaslow concentrations (0.3 µM) prevented colony formation.The two main conclusions of this study are: (i) under favorableconditions ({small tilde}1 µM PO₄^2– no mixing),a large percentage of released colonial cells give back coloniesafter going through a flagellated stage; (ii) sexuality doesnot appear to be involved in this process. ¹Present address: CREMA BP 5, F-17137 L'Houmeau, France     

Ethylene Biosynthesis and Xylogenesis in Lactuca Pith Explants Cultured In Vitro in the Presence of Auxin and Cytokinin: The Effect of Ethylene Precursors and Inhibitors

The possible involvement of ethylene in the induction of xylemdifferentiation was studied in lettuce (Lactuca saliva L. cv.Romaine) pith parenchyma explants. The addition of the ethyleneprecursors L-methionine (0.25 µM), S-adenosylmethionine(25 µM) and 1-aminocyclopropane-l-carboxylic acid (0.01µM), or the ethylene-releasing agent 2-chloroethylphosphonicacid (1.0 µM), to a standard IAA-kinetin-containing mediumenhanced xylogenesis compared to control explants cultured inthe absence of these compounds. In the presence of the ethyleneinhibitors aminoethoxyvinylglycine, Co(NO₃)₂ and AgNO₃, xylogenesiswas inhibited. Inhibition of xylogenesis by aminoethoxyvinylglycine(75 µM), Co(NO₃)₂ (50 µM) and AgNO₃ (6.0 µM)was reversed by exogenous 1-aminocyclopropane-l-carboxylic acid(0.01 µM), 2-chloroethylphosphonic acid (5.0 µM)and L-methionine (0.25 µM), respectively. Ethylene productionby explants cultured on media containing L-methionine or 1-aminocyclopropane-l-carboxylicacid was greater than the biosynthesis of ethylene by explantscultured in the absence of these compounds. The incorporationof 2-chloroethylphosphonic acid into the culture medium resultedin higher rates of ethylene production compared to explantscultured on the IAA-kinetin medium. The presence of either aminoethoxyvinylglycineor Co(NO₃)₂ inhibited ethylene production by explants culturedon the IAA-kinetin medium. The data support the hypothesis thatethylene plays a positive role in the initiation of xylem differentiation. Key words: Xylogenesis, Differentiation, Ethylene, IAA, Kinetin, Lactuca sativa