The Effect of Adenine and Mevalonic acid on the Endogenous Cytokinins of a Cytokinin-dependent Strain of Soya Bean Callus

The combined application of 10^–6 M adenine and 10^–6M mevalonic acid to soya bean callus accelerated its growth.Two biologically active compounds that co-chromatographed withzeatin and isopentenyl adenine were extracted from this callus.Studies with labelled adenine and mevalonic acid indicated thatthe cytokinin-dependent soya bean callus incorporated only avery small amount of the radioactive precursors into the biologically-activecompounds, making it extremely difficult to determine whetherthese compounds were synthesized de novo or whether they aroseas by-products of tRNA turnover. As cytokinins do not accumulatein rapidly-growing cytokinin-dependent soya bean callus culturedon kinetin as a source of cytokinin it seems as if biosynthesisde novo occurs when the callus is supplied with adenine andmevalonic acid. Glycine max (L.) Merrill, soya bean, callus culture, adenine, mevalonic acid, endogenous cytokinins     

Carbon Exchange Rate of Intact Individual Soya Bean Pods. 2. Ontogeny of Temperature Sensitivity

Diurnal temperature fluctuations induced change in soya bean-pod[Glycine max (L.) Merr.] carbon exchange rate (CER, where positiveCER represents CO₂ evolution). CER appeared to depend linearlyon temperature. Linear regressions of CER on temperature interceptedthe temperature axis at 5°C (i.e. zero CER at 5°C).Slopes of these regressions (i.e. temperature sensitivity) changedover the season. The CER-temperature sensitivity coefficient,K, (calculated from observed values of CER. pod temperatureand temperature intercept) rose from less than 0·02 mgCO₂ h^–1 pod^–1 °C^–1 during early pod-flll,peaked at over 0·04 mg CO₂ h^–1 pod^–1 °C^–1at mid pod-fill, and then declined during late pod-fill andmaturation. Glycine max (L.) Merr., Soya bean, carbon exchange rate, temperature     

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     

Effect of Ethylene and Culture Environment on Rice Callus Proliferation

Modifications to the gaseous envelope by callus during culturein Petri dishes were shown to reduce growth and promote necrosisof several rice (Oryza sativa L.) cultivars. Incubatingcallusunder a continuous flow of gas mixtures of known compositionsuggested that the inhibition of growth was caused by the accumulationof ethylene, the depletion of oxygen and, to a lesser extent,the accumulation of carbon dioxide. In order to evaluate theimportance of ethylene accumulation aminoethoxyvinylglycine(AVG), 1-aminocyclopropane-l-carboxylic acid (ACC and silvernitrate (AgNO₃), were added to the nutrient medium and ethylenemeasurements performed during callus culture. Ethylene restrictedcallus growth particularly under high (35 °C) as comparedto moderate (25 °C) temperatures and under illuminated ascompared to darkened incubation. Under illuminated incubationat 25 °C AVG (5 mmol m^–3) and AgNO°(50 mmol m^–3)significantly improvedcallus growth (100 and 60% respectively)while ACC (200 mmol m^–3) significantly decreased growth(40%). AVG and AgNO₃ were less effective under dark incubationat 25 °C where ethylene production was lower. Furthermore,callus growth was significantly better in large as comparedto small culture vessels since the ethylene concentration wasdiluted and more oxygen was available for respiration. Bettercontrol of ethylene and increased oxygen availability couldbe a way ofproducing healthy callus for the formation of embryogenictissues of otherwise recalcitrant cultivars of rice (e.g. IndicaIR42) and may be a way of improving manipulation of other cerealspecies. Key words: 1-Aminocyclopropane-1-carboxylic acid, aminoethoxyvinylglycine, callus, ethylene, Oryza sativa, silver nitrate     

Stimulatory and Inhibitory Effects of Sucrose Concentration on Xylogenesis in Lettuce Pith Explants; Possible Mediation by Ethylene Biosynthesis

Numbers of tracheary elements differentiating in lettuce pithexplants rose with increase in concentration of sucrose in themedium up to an optimal concentration of 0·2%, and fellwith further increase in concentration to about one-tenth maximalat 3% sucrose. Although a few tracheary elements formed withoutexogenous sucrose, a very low concentration of sucrose (0·001%)was sufficient to stimulate additional xylogenesis. Pretreatmentof explants with 3% sucrose caused a persisting inhibition ofxylogenesis, especially in tissue that had been near the siteof sucrose application (sandwich technique). The requirementfor adequate, but not inhibitory, concentrations of sucrosefor xylogenesis may underlie the development of xylem alongsidethe sucrose-rich phloem in normal apical morphogenesis. For callus growth the response to sucrose was different: theoptimal concentration was 3%, with a broad plateau from 1 to4% sucrose. Sucrose concentrations of 2 to 3%, used in manytissue culture media, are thus roughly optimal for callus growth,but ten times the optimum for xylogenesis in lettuce pith explants. It is surprising that 0·001% (0·03 mM) sucrose,applied exogenously, can stimulate xylogenesis: endogenous sugarconcentrations are normally higher. Perhaps the stimulationis mediated by ethylene biosynthesis, which is known to be xylogenic.Rates of ethylene production per explant rose with increasingsucrose concentration from about 0·1 nl h^-1 at 0% sucroseto a slightly (significantly) higher level at 0·004%sucrose and to about 0·5 nl h^-1 at 3% sucrose. D -glucoseresembled sucrose in its effects on xylogenesis and ethyleneproduction, but L-glucose yielded no xylogenesis and littlestimulation of ethylene biosynthesis.Copyright 1994, 1999 AcademicPress Lactuca sativa, Coleus blumei, Nicotiana tabacum, lettuce pith explants, tracheary element differentiation, sucrose, glucose, ethylene     

Comparisons of the Respiratory Effluxes of Nodules and Roots in Six Temperate Legumes

The specific respiration rates of nodulated root systems, ofnodules and of roots were determined during active nitrogenfixation in soya bean, navy bean, pea, lucerne, red clover andwhite clover, by measurements on whole plants before and afterthe removal of nodule populations. Similar measurements weremade on comparable populations of the six legumes, lacking nodulesbut receiving abundant nitrate-nitrogen, to determine the specificrespiration of their roots. All plants were grown in a controlled-environmentclimate which fostered rapid growth. The specific respiration rates of nodulated root systems ofthe three grain and three forage legumes during a 7–14-dayperiod of vegetative growth varied between 10 and 17 mg CO₂g^–1 (dry weight) h^–1. This mean value consistedof two components: a specific root respiration rate of 6–9mg CO₂ g^–1 h^–1 and a specific nodule respirationrate of 22–46 mg CO₂ g^–1 h^–1. Nodule respirationaccounted for 42–70 per cent of nodulated root respiration;nodule weight accounted for 12–40 per cent of nodulatedroot weight. The specific respiration rates of roots lackingnodules and utilizing nitrate nitrogen were generally 20–30per cent greater than the equivalent rates of roots from nodulatedplants. The measured respiratory effluxes are discussed in thecontext of nitrogen nitrogen fixation, nitrate assimilation. Glycine max, Phaseolus vulgaris, Pisum sativum, Medicago sativa, Trifolium pratense, Trifolium repens, soya bean, navy bean, pea, lucerne, red clover, white clover, nodule respiration, root respiration, fixation, nitrate assimilation     

Hormonal Control of Xylogenesis in Pith Parenchyma Explants of Lactuca

The time course of xylem differentiation was determined in explantsof lettuce pith parenchyma (Lactuca sativa L. cv. Romana) culturedon Murashige and Skoog (1962) medium using different concentrationsof auxin (IAA) and one cytokinin (zeatin or kinetin). Increasinglevels of auxin from I mg 1^–1 to 15 mg 1^–1 in thepresence of a constant level of a cytokinin (zeatin or kinetin)yielded up to 10 mg 1^–1 IAA, an increase in the numberof tracheary element formations. Cytokinin concentrations aboveand below o.1 mg 1^–1 interacting with an optimal xylogenicamount of auxin inhibited xylogenesis. The IAA (10 mg 1^–1)-zeatin(0.1 mg ^1–1) treatment produced the greatest number oftracheids, while kinetin compared to zeatin did not producesuch an effect. The different effectiveness of zeatin and kinetinin inducing tracheary element formations was not due to a differentcapacity of the two cytokinins to stimulate cell division butit seems likely that zeatin, because of interaction with IAA,is more active than kinetin in the determination of the dividingcells in a specific type of cytodifferentiation. The IAA (10mg 1^–1)-zeatin (0.1 mg 1^–1) treatment produced about6.9 per cent tracheids with respect to cell division while IAA(10 mg 1^–1)-kinetin (0.1 mg 1^–1) produced 4.2 percent. These results are discussed with reference to the problemsof hormonal control of xylem differentiation.     

Tracheary Element Differentiation Induced in Isolated Cylinders of Lettuce Pith: a Bipolar Gradient Technique

To study the influence of morphogenetic gradients on vasculardifferentiation patterns, a new technique was developed whichallows different substances to be applied at opposite ends ofa tissue block. It yielded information on the mobility of particularmorphogens and on the dependence of callus formation and trachearyelement differentiation on their presence. Application of indol-3ylacetic acid (1AA) (10 mg l^–1), zeatin (0.1 mg l^–1)and sucrose (3 per cent, w/v) in various combinations to theends of cylindrical explants of lettuce pith (Lactuca sativaL.) showed that (a) callus formation was stimulated by IAA,whereas induction of tracheary elements required both IAA andzeatin; (b) callus was confined to a few millimetres at theends of the explants, and tracheary elements occurred mainlywithin the callus; (c) sucrose or its metabolic products diffusedthe 10 mm length of the explants, while IAA and zeatin wereeffective only close to the application site; and (d) some callusand tracheary elements formed when no sucrose was applied, butboth increased with sucrose application, though inhibition oftracheary elements formation occurred with high sucrose concentrations. differentiation, pith explant, tissue culture, xylogenesis, indol-3yl acetic acid, sucrose, zeatin, lettuce, Lactuca sativa     

In Vitro Regeneration from Seedling Organs of Brassica oleracea var. italica Plenck cv. Green Comet I. Effect of Plant Growth Regulators

The calabrese cultivar Brassica oleracea var. italica cv. GreenComet was used in a study of the effects of exogenous hormoneson the growth and differentiation of seedling organs in vitro.Four types of explants were tested: hypocotyl segments, rootsegments, primary leaf discs and cotyledon discs. These explantswere incubated on media containing factorial combinations ofBAP x IBA, BAP x NAA, KN x IBA and KN x NAA (all at 0, 0.1,10 and 10.0mg l^–1). Hypocotyls were the most regenerativeexplants; shoot production was favoured by cytokinin: auxinratios greater than one and was decreased by IBA at 10 mg l^–1when callus was produced. Shoot formation from root explantsoccurred either in the absence of hormones or with low concentrations;no shoot was produced when any hormone was present at 10 mgl^–1. In contrast, shoot production from primary leaf diseswas favoured by high concentrations of both auxin and cytokininwith the combination of BAP and IBA the most effective. Shootproduction from cotyledon discs was sporadic with no consistentresponse on any auxin/cytokinin combination. After further experimentson the optimization of hormone concentration, the followingcombinations were chosen as allowing reliable regeneration:0.1 mg l^–1 BAP+0.1mg l^–1 IBA for hypocotyl segments,0.075 mg l^–1 KN +0.025 mg l^–1 IBA for root segments,and 5.0 mg l^–1 BAP+5.0 mg l^–1 IBA for leaf discs. Brassica oleracea var. italica, calabrese, tissue culture, seedling, auxin, cytokinin     

Callus, Organogenesis and Plantlet Formation in Tissue Cultures of Clitoria ternatea

Decoated seeds of Clitoria ternatea L. germinated on Murashigeand Skoog (Physiologia Plantarum 1962, 15, 473–97) basalmedium (BM) and differentiated callus and bipolar embryoids(two-step method) in low frequency. Calluses developed on lateralroots [BM+KN(0.1 mg 1^–1)], on roots and hypocotyls [BM+KN(0.5mg 1^–1)], and on roots [BM+KN+IAA (0.5 mg 1^–1 ofeach)]. On basal medium with KN (0.5 mg 1^–1) and withKN+IAA (0.5 mg1^–1 of each), multiple shoot buds and embryoids(one-step method) were differentiated directly on split hypocotylsand roots. In the former, shoot buds developed even on unsplithypocotyls. Rhizogenesis on isolated shoot buds occurred efficientlyin BM+indole butyric acid (IBA 0.1 mg 1^–1) and BM+IAA(0.1 mg 1^–1 and 0.5 mg 1^–1). Profuse direct embryoidsand shoot buds developing on root systems are interesting morphogeneticphenomena rarely reported. Clitoria ternatea L., callus, embryoids, multiple shoot buds, regeneration     

Regulation of Stem Abscission and Callus Growth in Shoot Explants of Sweet Orange [Citrus sinensis (L.) Osbeck]

Two-node explants from Sweet Orange cv. St Ives Valencia orangeshoots produced prolific callus and formed secondary abscissionzones within internodes when cultured in vitro with abscisicacid (ABA, 5 µM) or -naphthaleneacetic acid (NAA, 5 µM).Benzyladenine (BA, 1 µm) induced callus but had littleeffect on abscission. Secondary abscission zone formation wasassociated with ABA-induced and auxin-induced ethylene formation.Treatment of explants with inhibitors of ethylene synthesis[aminoethoxyvinyl glycine (AVG), Co²⁺, PO₄^3–] preventedformation of secondary abscission zones but had variable effectson callus formation. Newly made explants contained high concentrationsof endogenous ABA (up to 6000 ng g^–1 f.wt), as measuredby GC/MS/SIM. Long-term subculture of explants (two years) inmedia containing BA (1 µm) led to a reduction in endogenousABA level (40 ng g^–1 f. wt) and to loss of capacity toform extensive callus and secondary abscission zones. Citrus sinensis (L.) Osbeck cv. St Ives Valencia, sweet orange, secondary abscission zones, in vitro, ethylene, endogenous ABA, endogenous IAA     

The Effect of Inorganic Nutrients on the Hormonal Requirements of Normal, Habituated, and Crown-gall Tissue 4

The addition of Braun and Wood's inorganic supplements (845mg l^–1 KCl, 1800 mgl^–1 NaNO₃, 300 mg l^–1 NaH₂PO₄.2H₂O,790 mg l^–1 (NH₄)₂SO₄) to White's medium caused markedincreases in the growth of normal tissues of Helianthus annuus,Nicotiana rustica, Daucus carota, and Vinca rosea and crown-galltumour tissues of H. annuus. However, no evidence was obtainedwhich suggested that the presence of these extra salts markedlyinfluenced the essential requirements of normal callus for auxinsand kinetin. In contrast their presence significantly influencedthe hormonal requirements of certain habituated cultures ofH. annuus and V. rosea. These habituated cultures had specificauxin requirements on White's medium while either an auxin orkinetin was sufficient on high-salts medium. These results arediscussed in relation to previous reports which suggested thatthe biosyntheses of auxins and other growth factors in normaland crown-gall cultures are specifically activated by certaininorganic ions.     

Use of the Plastochron Index to Evaluate Effects of Light, Temperature and Nitrogen on Growth of Soya Bean (Glycine max L. Merr.)

The plastochron index (PI) has been compared with leaf growthand biomass accumulation in young soya bean plants of severalcultivars that were grown in controlled environments with differentirradiance levels and durations, temperatures, and nitrogen(N) regimes. Increasing the photoperiod from 10 to 16 h day^–1 increasedthe plastochron rate (PR) and the proportion of axillary growth.Doubling the photosynthetic photon flux density (PPFD) to 1000µmol m^–2S^–1, increased PR and the proportionof roots to total plant weight, but decreased the proportionof stems plus petioles to total. In a series of experiments,the plants were grown in an 8 h photoperiod at constant temperaturesof 17, 20, 26 or 32 °C. As temperature increased, PR increased,but the duration of leaf expansion decreased. Leaves were largestat 20 and progressively smaller at 26, 32 and 17 °C. Biomasswas greatest for a given PI at 20 °C and decreased in theorder of 26, 32, and 17 °C. The proportion of axillary growthalso was greatest at 20 °C. When plants were grown in a15 h photoperiod at temperatures from 17.1 to 26.6 °C, leafsize continued to increase up to the highest temperature. At17 °C, the PR in the 15 h photoperiod (PPFD 390 µmol;m^–2S^–1) was about threefold greater than in 8 h(500 µmol m^–2 S^–1); biomass accumulation perday was about fivefold greater. Increasing N from 3 to 36 mMincreased PR about 10 per cent, altered biomass partitioningamong plant parts, and increased the biomass of the plants.The NO₂ form of N markedly stimulated axillary growth as comparedwith the NH₄⁺ form. Environment or cultivar had little influenceon the duration of leaf expansion in terms of PI. Cultivarsdid not differ consistently in biomass production and allocationin the different environments. Glycine max (L.) Merrill, soybean, soya bean, plastochron index, leaf development, growth analysis, partitioning, light, nitrogen, temperature     

Carbon Exchange Rate of Intact Individual Soya Bean Pods. 1. Response to Step Changes in Light and Temperature

Carbon exchange rates (CER) of individual intact field-grownsoya bean [Glycine max (L.) Merr.] pods were measured continuouslywith a mobile gas analysis laboratory. Conditions in pod chamberssimulated those experienced normally by pods except for experimentalmodification of incident radiation or pod temperature. Undernormal conditions, CER (where positive CER represents CO₂ evolution)fluctuated diurnally with a morning rise followed by a slowafternoon and evening decline which was similar among pods whichwere measured simultaneously. The frequency of measurementspermitted detection of rapid CER responses to step changes inlight and pod temperature. CER rapidly decreased and increasedwhen the chamber was alternately exposed to full sunlight andcomplete darkness, respectively. CER responded similarly tosteps up [from ambient to elevated (+ 10°C) temperature]and steps down (from elevated to ambient temperature), respectively.Thus, a temperature sensitive process which regulated pod CERwas located within the pod. CER ranged from less than 0·1to more than 1·2 mg CO₂ h^–1 pod^–1 over theperiod of rapid dry-matter accumulation. Glycine max (L.) Merr., Soya bean, carbon exchange rate, light, temperature     

Effects of Sequential Exposure to Auxin and Cytokinin on Xylogenesis in Cultured Explants of Jerusalem Artichoke (Helianthus tuberosus L.)

During the course of a 4-d culture period, explants of Jerusalemartichoke tuber were exposed to auxin (0.2 mg 1^–1 2, 4-dichlorophenoxyaceticacid), and cytokinin (5.0 mg 1^–1 benzyl-amino purine),under a range of sequential regimes, to study the influenceof each hormone on tracheary element formation. The resultsindicate that auxin was necessary early in the culture periodand was primarily involved in cell proliferation. Cytokininstimulated xylogenesis when present late in the culture period,concomitant with the phase of cytodifferentiation, but not whenrestricted to the early period. The implications for a sustainedperiod of commitment to differentiation are discussed. Xylem differentiation, Jerusalem artichoke, auxin, cytokinin, tissue culture     

In Vitro Somatic Embryogenesis and Plant Regeneration in Clitoria ternatea

A sustainable plant regeneration system in vitro through somaticembryos from mature sexual embryos has been reported in Clitoriaternatea. Somatic embryos developed through callus from seedlingroots on hormone-free MS medium (MS₁). Addition of growth hormones,KN 0.5 mg dm^–3 (MS₂) or KN+1AA 0·5 mg dm^–3of each (MS₃) induced direct somatic embryos, in high frequency,on split root and hypocotyl systems. The embryogenic potentialvaried with the organ, roots or hypocotyls, and also with themedium. The morphogenetic capacity of the somatic embryos isretained for more than 2 years by subculturing at intervalsof 4 weeks on MS₃ in complete darkness. Somatic embryos, underthe appropriate subculture conditions (16 h light/8 h dark photoperiodat 24± 1 °C on media MS₃, MS₄ and MS₅), resultedin recurrent-somatic embryogenesis and was profuse at the shootand root apices of the somatic embryos. Mature somatic embryoswere transplanted to MS₁ to stimulate germination and plantletregeneration. Plantlets, developed from primary and secondaryembryos on MS₁ were successfully hardened and grown in naturaloutdoor conditions. The morphology and histology of the somaticembryo and plantlet and the culture conditions for continuousproduction of plantlets through direct somatic embryogeny arediscussed Key words: Clitoria ternatea, somatic embryos, plant regeneration     

Ethylene and Ethane Production in 2,4-D Treated and Salt Treated Tobacco Tissue Cultures

Gas chromatography was used to measure ethylene (ethene) andethane production by tobacco (Nicotiana tabacum cv. Wisconsinno. 38) callus tissues grown on media containing inorganic saltsaccording to Murashige and Skoog (1962), sucrose, myo-inositol,thiamine-HCl kinetic according to Linsmaier and Skoog (1965),and either 2,4-dichiorophenoxyacetic acid (2,4-D) in the range0–100 mgl^–1 or 2 mgl^–1 indoi-3-ylacetic acidplus NaCl in the range 0–200 Meq l^–1. Ethylene productionrates were high (> 500 nl h^–1 g^1– fresh weight)initially in all treatments. Subsequently, ethylene productiondeclined in rapidly growing cultures but remained high in moderatelyand severely 2,4-D (> 0·5 mgl^–1) stressed andin severely NaCl (150 Meql^–1) stressed cultures. Highinitial rates of ethane production (> 200 nl h^–1 g^–1fresh weight) were obtained under conditions of severe stresscaused by 2,4-D or NaCl but not in control or moderately inhibitedcultures. With further incubation ethane production declinedin the severely stressed cultures. It is concluded that ethyleneproduction can be used as an index of moderate 2,4-D stressand severe NaCl stress by virtue of the high persisting ratesof ethylene production in stressed cultures. Ethane productioncan be used as an early index of severe stress caused by either2,4-D or NaCl in vitro. Nicotiana tabacum L., tobacco, ethylene, ethenen, ethane, 2,4-dichlorophenoxyacetic acid, auxin, stress, callus tissue     

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; )     

Exogenous Auxin and N-1 -Naphthylphthalamic Acid Effects on Populus deltoides Xylogenesis

A method was developed for evaluating the empirical alterationof xylem vessel differentiation in the central leaf trace ofPopulus deltoides, a species that exhibits helical phyllotaxis.Effects of experimental treatments for a period of six plastochronswere evaluated by vessel parameter ratios = 2.P_T/ (P_T+1 + P_T–1),where P was either vessel number or mean transverse vessel areameasured at mid-intern ode at Leaf Plastochron Indices of T– 1, T, and T + 1. Excising leaf laminae reduced vesselnumber and mean vessel area in the associated central leaf traceby 50% and 70%, respectively, compared to unexcised laminaecontrols. Replacing excised laminae with a concentration seriesof exogenous indoleacetic acid (IAA) resulted in a 5% increaseper log mol m^–3 of IAA in the number of vessels differentiatingin the associated central leaf traces compared to excised controls.Mean vessel areas within these leaf traces were 50% of thoseof intact leaf traces. No significant effects of different concentrationsof exogenously applied IAA on mean vessel area could be demonstrated.A lanolin paste ring of N-1 -naphthylphthalamic acid (NPA),an auxin transport inhibitor, around the petioles of intactleaves reduced the number of differentiating vessels by 7% andmean vessel area by 29% per log mol m^–3 of NPA comparedto central leaf traces of leaves ringed with plain lanolin paste.The results suggest that NPA treatments may be used to distinguishexperimentally, at least in part, the cell division from thecell enlargement phases of primary xylogenesis within centralleaf traces of P. deltoides stems. Key words: Auxin transport, Vessel area, Vessel number     

Effects of Na2SO4, K2SO4 and KCl on Growth and Ion Uptake of Callus Cultures of Vaccinium corymbosum L. cv. Blue Crop

Non-selected and Na₂SO-, K₂SO₄- or KCl-selected callus culturesof Vaccinium corymbosum L. cv. Blue Crop were grown on mediasupplemented with 0, 25 and 50 mM Na₂SO₄ (non-selected and Na₂SO(-selectedonly), 0, 25 and 50mMK₂SO₄ (non-selected and K₂SO₄-selectedonly) or 0, 50 and 100 mM KCl (non-selected and KCl-selectedonly). On all media, growth of selected callus (on a fresh-weightor dry-weight basis) was greater than that of non-selected callus,and selected callus grew optimally on the level and type ofsalt on which it was selected. Selected callus was friable andmaintained a higher f. wt:d. wt ratio. Tissue water potentialin selected callus was more negative than in non-selected callus. Flame photometry and chloridometry showed Na⁺, K⁺ and Cl^–accumulated in callus to concentrations equal to or greaterthan the initial concentration in the medium. Turbidometry showedthat tissue SO₄^2- concentration was lower than the concentrationin the medium. In most cases selected callus accumulated moreNa⁺, K^sup, SO₄^2– or Cl^– than non-selected callus.Vacuolar ion concentration was measured by electronprobe X-raymicroanalysis, and on most media selected callus had highervacuolar ion concentrations than non-selected callus. SO₄^2–and Cl^– were accumulated in the vacuoles at concentrationshigher than the external medium, but vacuolar Na⁺ concentrationdid not reach external concentration on Na₂SO₄ and on potassiumsalts was maintained between 12 and 17 mM. Vacuolar K⁺ concentration(approx. 142–191 mM on no salt) decreased on Na₂SO₄ andincreased on K₂SO₄ and KCl. There was no precise correlation between total or specific ionaccumulation (Na⁺, K⁺, SO₄^2– and Cl^– and fresh-weightyield. Results suggest that selection results in adaptationin response to decreased water potential of the medium. Vaccinium corymbosum, blueberry, electronprobe X-ray microanalysis, callus, in vitro selection, salt tolerance, KCl, K₂SO₄, Na₂SO₄