The Effects of Cytokinins and ABA on Stomatal Behaviour of Maize and Commelina

Epidermal strips and leaf fragments of Commelina and leaf fragmentsof maize were incubated on solutions containing naturally-occurringor synthetic cytokinins and/or ABA. The effects of these treatmentson stomatal behaviour were assessed. Cytokinins alone did notpromote stomatal opening in either species but concentrationsof both zeatin and kinetin from 10^–3 to 10^–1 molm^–3 caused some reversal of ABA-stimulated closure ofmaize stomata. The reversal of the ABA effect increased withincreasing cytokinin concentration. Cytokinins had no effecton ABA-stimulated closure of Commelina stomata. When appliedalone, at high concentration (10^–1 mol m^–3), toCommelina epidermis or leaf pieces both zeatin and kinetin restrictedstomatal opening. Key words: ABA, Cytokinins, Stomata, Maize, Commelina     

Studies on Foliar Penetration: VIII. EFFECTS OF CHLORINATION ON THE MOVEMENT OF PHENOXYACETIC AND BENZOIC ACIDS THROUGH CUTICLES ISOLATED FROM THE FRUITS OF LYCOPERSICON ESCULENTUM L.

The effects of chlorine substitution on the movement of phenoxyaceticand benzoic acids through enzymatically-isolated cuticles ofLycopersicon fruits were determined by following the transferof each acid containing ¹⁴C from a donor to a receiver solution.This cuticle is characterized by an isotropic cutin matrix,within which patches of birefringent cuticular waxes are foundnear the outer surface. The outer, morphological surface isrelatively smooth while at the junction with the outer wallsof the epidermal cells there is extensive cuticular developmentextending down between the anticlinal walls. The epicuticularwax appears as a soft sheet-like covering of which the surfaceis relatively featureless. Chlorination of phenoxyacetic acid results in an enhanced transferacross the isolated cuticle. The order was 2,4,5- and 2,4,6-trichlorophenoxyacetic> 2,4- and 3,5-dichlorophenoxyacetic > 2-chlorophenoxyacetic> phenoxyacetic acid. Removal of the epicuticular wax resultedin greater permeability for all compounds; transfer of the morepolar acids was favoured. In contrast, chlorination of benzoicacid decreases passage through the cuticle; the rate is highestfor benzoic acid followed in descending order by 2-chlorobenzoic,2,4- and 2,5-dichlorobenzoic and 2,3,6-trichlorobenzoic acid.Chlorination also depresses the passage of both phenoxyaceticand benzoic acid through a dialysis membrane. The effects ofchlorination on the lipid solubility of both series of compoundsare discussed in relation to differences in transfer acrossthe cuticle.     

Interacting Effects of Light and Day and Night Temperatures on the Growth of Four Species in the Vegetative Phase

A comparative examination has been made under controlled conditionsof the interacting effects on growth in the vegetative phasewhich takes place when the diurnal temperature is either maintainedconstant at 20 and 25 ?C or reduced at night by 5 and 10 ?Cand the plants subjected daily to either 2.16 or 4.32 ? 10⁴lx for 14 h. For each species (Gossypium hirsutum, Hclianthusannuus, Phaseolus vulgaris and Zea mays) the changes in netassimilation rate, the leaf area ratio and the relative growthrates in plant weight and leaf area were recorded. Contrary to some previous findings, none of these componentsof growth are favoured by cool nights. Rather such reductionsin temperature retard the growth processes to a varying degreeaccording to the species and the component. In general, significantreductions are more evident for the two relative growth rateswhen a drop from 20 to 10 ?C is combined with the lower levelof illumination. The implications of these results are considered in relationto a prior study where for similar light conditions the samespecies plants were subjected to constant temperatures rangingfrom 10 to 25 ?C.     

Studies on Foliar Penetration: VII. FACTORS CONTROLLING THE PENETRATION OF CHLORIDE IONS INTO THE LEAVES OF PHASEOLUS VULGARIS

The pattern of penetration of chloride ions into the abaxialsurfaces of the primary leaves of Phaseolus vulgaris has severalfeatures in common with those previously recorded for 2,4-dichlorophenoxyaceticacid (2,4-D), 2,2-dichloropropionic acid (dalapon), and 4-amino-3,5,6-trichloropicolinicacid (picloram). In the dark the rate of entry of chloride ionsup to 24 h is constant, but in the light entry is at first slowand then more rapid. This acceleration does not occur at lowtemperature or when the tissue is treated with 3-(3,4-dichlorophenyl)-l,l-dimethylureaor phenylmercuric chloride. Neither does it take place at theadaxial surface or when the leaves are more mature. The most distinguishing feature of the pattern of penetrationof chloride ions is its dependence on external pH. In darknessentry is unaffected by changes in pH but in the light the rateof entry is increased as the pH falls, but this response isrestricted to young leaves. No binding of chloride seems tooccur within the tissue. These findings support the view that penetration of the abaxialsurface in young leaves of Phaseolus is largely determined bya membrane system. This system decreases in importance as theleaf matures and the overlying cuticle thickens. At the adaxialsurface the thicker cuticle is seemingly a major barrier topenetration even in very young leaves.     

The Uptake of Growth Substances: XI. VARIATIONS IN THE ACCUMULATION OF SUBSTITUTED PHENOXYACETIC ACIDS OF DIFFERING PHYSIOLOGICAL ACTIVITY BY SEGMENTS OF AVENA MESOCOTYL

An examination has been made of the phase of continuous accumulationof phenoxyacetic acid (POA) and the 2-, 4-, 2,4-, 2,6-, and2,4,5-chloro-derivatives, containing carbon-14 in the carboxylgroup, by segments of the Avena mesocotyl. On the basis of previousfindings to eliminate the initial transient components of uptake(Type I processes) the segments were pretreated for 13 to 18h in buffer or buffer containing the respective non-radioactivecompound. For five of the compounds the relationship between the rateof uptake and the external concentration takes the form of arectangular hyperbola, but for the sixth, 2,4,5-T, this relationshipdoes not hold. The data, except those for 2,4,5-T, have beenevaluated as linear regressions of rate of uptake against thequotient of rate over concentration. From each regression equationtwo constants have been derived: the point ‘B’ wherethe line intersects the rate axis (the theoretically maximumrate) and the slope of the regression ‘K’, whichcan alternatively be expressed as the concentration at whichthe observed rate equals half the value of ‘B’. The calculated values of B and K for POA are approximately twiceas great as the corresponding values for 2-CPA, and about 25times greater than for 4-CPA. The values for 2,4-D are closeto those for 4-CPA, and 2,6-D is intermediate between 2-CPAand 4-CPA. Although the constants for 2,4,5-T could not be calculatedprecisely, the rates of accumulation are about one-fourth ofthose measured for 2,4-D at equivalent concentrations. The uptake of radioactive 2,4-D is slightly depressed in thepresence of nonradioactive POA. Greater reductions are causedby 2-CPA or 2,6-D, and 2,4-D or 2,4,5-T are even more inhibitory.The pattern of inhibition caused by 2,4,5-T indicates competitionfor common sites of uptake, while POA appears not to be competitive.In corresponding experiments with POA, the presence of the otherregulators only caused small inhibitions and the order was different. Earlier work showed that in Avena accumulation is accompaniedby the conversion to a varying degree of the individual substitutedphenoxyacetic acids to conjugated derivatives. It is suggestedthat the variation between compounds in their rates of accumulationis in part due to differences in the stability of the conjugatedderivatives, and that the facility of conversion is a factorin determining physiological activity.     

Studies in the Principles of Phytotoxicity: V. INTERRELATIONSHIPS BETWEEN SPECIFIC DIFFERENCES IN SPRAY RETENTION AND SELECTIVE TOXICITY

In a detailed study of the factors which control spray retentionby the shoot, five species with markedly different morphologicalcharacteristics have been examined (Helianthus annuus, Brassicaatha, Linum usitatisrnnwn, Hordeum vulgare, and Pisum sativum).The amount of spray retained, expressed as volume per unit weightof shoot or unit leaf area, is dependent on the stage of developmentof each species, the volume of spray applied, and the surfacetension of the spray solution. The interrelationships betweenthese factors are complex for all the second-order interactionsproved to be highly significant. Under one set of condi tionsthere may be a tenfold difference in retention between two specieswhile under another set the corresponding difference may beas little as twofold. Between a spray output of 14 and 143 ml./m.³ the level of retentionby P. sativum and H. vulgare is linearly related to output butfor H. annuus and B. alba the relationship is curvilinear. Whenthe surface tension is reduced below 40 dynes/cm. retentionby P. sativum and H. vulgare is increased, but for B. alba andH. annuus retention is decreased if the output exceeds 50 ml./m.For a given output, a reduction in the mean droplet size augmentsretention by H. vulgare but not by H. annuus. Differences in retention either between species or between stagesof develop ment are linked with the ratio of leaf area to shootweight, the nature of the leaf surfaces, the angle of incidenceat which the droplets strike the leaf, and localized accumulation.Retention is maximal when the leaf-area ratio is high, the surfacescompletely wetted, and the laminae set in a horizontal plane:once this maximum value is reached any additional spray receivedruns off the shoot. The relationship between retention and the concentration ofthe spray solution required to bring about some standard biologicalresponse has been examined for two groups of compounds—thosewhich are freely translocated (triethanolarnine salts of 2:4-dichloro-,2:4:5-trichloro-, and z-methyl-4-chloro-phenoxyacetic acids)and those which are not (arnrnonium 2:4-dinitro-6-sec-butylphenate).For the first group the equi-effective concentration is inverselyand linearly related to retention since the total amount receivedis of primary importance in determining the inhibition of growth,though the position of the droplets must be taken into account.Placing micro-drops containing the growth regulator on the cotyledonsof L. usitatimmwn inhibits the growth of the shoot less thana spray application of the same amount over the whole shoot:for H. annuus placing the growth regulator on either the firstor second pair of leaves or on the first internode induces moreinhibition of growth than application to the cotyledons. With ammonium dinitro-sec-butylphenate the relationship betweenretention and concentration is also inverse but approximatelylogarithmic for L. usitatisrimum. For some other species thechanges in retention consequent upon a reduction in the surfacetension are not matched by comparable changes in the equi-effectiveconcentration. For B. alba and H. annuus though lowering thesurface tension may decrease retention it may also Himiniahthe concentration because the fall in retention is more thanoffset by a greater penetration of the compound into the shoot. It is concluded that differences in spray retention betweenspecies can play a major role in determining selective actionand the conditions under which this factor may operate mosteffectively are discussed.     

Studies in the Principles of Phytotoxicity: III. THE pH FACTOR AND THE TOXICITY OF 3:5-DINITRO-o-CRESOL, A WEAK ACID

It has previously been shown that the concentration of 3:5-dinitro-ocresolrequired in the external medium to halve (a) the growth rateof the mould Trichoderma viride or (b) the respiration rateof yeast increased same 75–100 times as the pH changedfrom 4 to 7. In the present investigation it has been foundthat when solutions or suspensions of 3:5-dinitro-o-cresol areapplied as sprays to seedlings of Brassica alba the concentrationneeded to kill half the plants changes by a factor of less than2 over the same pH range. On the other hand, when Lemma minor is grown in nutrient solutioncontaining the toxicant, the concentration required to halvethe rate of frond multiplication increases sharply as the pHis raised from 5.4 to 6.8. Moreover, when leaf disks of B. albaare vacuum infiltrated with buffered solutions of 3:5-dinitro-o-cresolthe concentration needed to halve the respiration rate rises50-fold between pH 4 and 8. It is considered that the essential condition which determinesthe similarity of the result for L. minor, the leaf disks, andmicro-organisms is that a relatively large volume of solutioncontaining the toxicant is in direct contact with the tissues.In contrast, under the conditions of spraying the volume ofspray droplets relative to that of plant tissue is small. Thusthe pH effect ie masked since the buffering capacity of thecells exceeds that of the droplets, with the result that 3:5-dinitro-o-cresollargely acts upon the cells at a pH determined by the tissues,a pH which may bear little relation to that of the originalsolution. Supporting evidence is produced that the pH factor operatesin the same way for other weak acids, such as the chlorophenoxyaceticacids.     

An Analysis of the Influence of Plant Density on the Growth of Vicia faba: I. THE INFLUENCE OF DENSITY ON THE PATTERN OF DEVELOPMENT

The competitive effects of varying the density on the developmentof Vicia faba have been studied in a series of multifactorialexperiments where the spacing both between and within rows wassimultaneously altered. Over densities ranging from 11 to 67plants per metre² seed production on an area basis tended tobe maximal at 35–45 plants per metre² in the winter typeand at the highest density for the spring type. As the densityincreased the number of pods per plant and the extent of branchingfell progressively but there was no appreciable change in eitherseed size or the number of seeds per pod: thus seed productionwas solely governed by the number of mature pods formed. Thenumber of flowers per plant was more dependent on the numberof inflorescences than on the number of flowers per inflorescence.Increasing the population diminished the number of nodes bearinginflorescences particularly in the upper part of the shoot,while the size of the inflorescence was decreased to a lesserextent. The number of flowers forming mature pods was very small(9–14 per cent.). At the top of the shoot the flowerswere infertile, while above the middle node most of the podsformed were shed while still immature. Thus the primary effectof increasing density was to depress the number of nodes onthe lower half of the stems which produced mature pods. By thetime the flowering stage was reached plant height was alreadycorrelated with density due more to a change in internode lengththan an alteration in the number of differentiated nodes; atmaturity the differences in height were smaller due to the greaternumber of nodes in the widely spaced plants. For a given density,alterations in the distance between rows had little influenceon development. The possible physiological factors responsiblefor the changing pattern of development brought about by varyingdensity are discussed.     

Interactions in the Physiological Effects of Growth Substances on Plant Development

The interrelated effects of 3-indolylacetic acid and sodium2 : 4-dichlorophenoxyacetate on the growth and development ofLemna minor and Helianthus annuus have been investigated bymeans of multifactorial experiments, involving three to fivelevels of each compound. A clonal population of L. minor wasmaintained under constant conditions of light (550 foot-candles)and temperature (25 C.) and the growth regulators were addedto a phosphate-buffered culture solution (pH 51), which waschanged every 2 days. In the H. annuus experiments the compounds,in aqueous solution, were placed by means of a micro-pipetteon the leaves of young plants grown in a greenhouse or in theopen. Indolylacetic acid, at concentrations up to 25 p.p.m., increasesthe relative growth rate of L. minor on either a dry-weightor frond-area basis. Sodium dichlorophenoxyacetate at 0025p.p.m. has little effect on growth, but at and above concentrationsof 02 p.p.m. growth is depressed. The ratio of frond area tototal dry weight is increased by indolylacetic acid but is depressedby the phenoxyacetate. In the presence of indolylacetic acid,more particularly at the higher levels, the relative inhibitionof growth caused by sodium dichlorophenoxyacetate is reduced. When sunflowers are treated daily with indolylacetic acid (0,6, 30, 150 µg./ plant) or initially with sodium dichlorophenoxyacetate(0, 20, 100 µ.g./plant) in all possible combinations,then the relative growth rates of the treated leaves, the leavesabove the treated leaves, the first internode, and the rootsare depressed when each compound is applied alone. On the otherhand, the growth rate of the hypocotyl is increased by bothgrowth regulators, while for the second internode indolylaceticacid decreases the growth rate but the phenoxyacetate at thelower level accelerates it. The combined action of the two compoundson the different parts tends to be cumulative, save for thehypocotyl, where the effect of indolylacetic acid is dependenton the level of the phenoxyacetate. It is concluded that these results cannot be explained on theconcept of wholly additive effects. There is evidence that thecompounds can mutually interfere in their actions and it wouldseem that more than one mechanism is involved. Many of the resultscan be satisfactorily interpreted on the basis that the twocompounds act on a common system and that they compete withone another, but the results are not compatible with the postulatethat the dichlorophenoxyacetic acid directly inactivates indolylaceticacid.