Stomatal Responses and the Senescence of Leaves

Guard cell responses were examined in green and senescing leavesof Victa faba using detached epidermal strips to eliminate influencesfrom the mesophyll. Stomatal opening was greater in epidermalstrips from mature leaves than from senescing leaves althoughthe latter still retained the ability to respond to CO₂ andto kinetin. It was concluded that the decline in stomatal activityduring senescence is an independent but parallel process tochanges occurring in the mesophyll. Vicia faba, leaf senescence, stomata, kinetin     

RNA Synthesis in Phaseolus Chloroplasts: II. RIBONUCLEIC ACID SYNTHESIS IN CHLOROPLASTS FROM DEVELOPING AND SENESCING LEAVES

The rate of RNA synthesis in chloroplasts from the primary leavesof Phaseolus vulgaris L. cv. Canadian Wonder was measured invitro as plant age increased. The rate per leaf began to fallbefore the leaf was 70% expanded. At full expansion, activityhad fallen by 70%. Chloroplast RNA synthesis per unit chlorophyllwas falling before the leaf was 25% expanded. When all parts of the plant above the mature primary leaveswere removed (detopping) chloroplast RNA synthesis in theseleaves rose within 36 h. The rate increased to a maximum 3–4d after detopping, when it was 5–10 times control values;thereafter it fell again. The chlorophyll content began to increaseabout 4 d after detopping, eventually rising by 100%. Detoppingcaused a 3-fold increase in the Triton X-100-soluble DNA contentof chloroplast preparations, measured after 3.5 d. At that timethe rate of RNA synthesis per unit Triton-soluble DNA was thesame in chloroplasts from the primary leaves of intact and detoppedplants. Detopping also resulted in an increase in the depthof the leaf palisade layer. The effects of detopping on chloroplasts were prevented by darknessand reduced by shading. Increased chloroplast RNA polymerase activity was also inducedin the primary leaves by placing a polythene bag over intactplants, enclosing everything above these leaves. Removal ofthe roots from detopped plants prevented the rise in the rateof chloroplast RNA synthesis.     

Characterisation of Non-Uniform Photosynthesis Induced by Abscisic Acid in Leaves Having Different Mesophyll Anatomies

The effects of abscisic acid (ABA) on photosynthesis in leavesof Helianthus annuus L. were compared with those in leaves ofVicia faba L. After the ABA treatment, the response of photosyntheticCO₂ assimilation rate, A, to calculated intercellular partialpressure of CO₂, P_i, (A(p_i) relationship) was markedly depressedin H. annuus. A less marked depression was also observed inV.faba. However, when the abaxial epidermes were removed fromthese leaves, neither the maximum rate nor the CO₂ responseof photosynthetic oxygen evolution was affected by the applicationof ABA. Starch-iodine tests revealed that photosynthesis was not uniformover the leaves of H. annuus treated with ABA. The starch contentwas diffferent in each bundle sheath extension compartment (thesmallest subdivision of mesophyll by veins with bundle sheathextensions, having an area of ca. 0.25 mm² and ca. 50 stomata).In some compartments, no starch was detected. The distributionof open stomata, examined using the silicone rubber impressiontechniques, was similar to the pattern of starch accumulation.In V.faba leaves, which lack bundle sheath extensions, distributionof starch was more homogeneous. These results indicate that the apparent non-stomatal inhibitionof photosynthesis by ABA deduced from the depression of A(pⁱ)relationship is an artifact which can be attributed to the non-uniformdistribution of transpiration and photosynthesis over the leaf.Intercellular gaseous environment in the ABA-treated leavesis discussed in relation to mesophyll anatomy. ¹ Present address: Department of Botany, Duke University, Durham,NC 27706, U.S.A. (Received September 30, 1987; Accepted January 13, 1988)     

Membrane-bound Plastid Inclusions and Chloroplast Thylakoid Formation in Sunflower (Helianthus annuus L.)

The ultrastructure, distribution and frequency of membrane-boundplastid inclusions present in the epidermal cells of leavesof intact sunflower plants (Helianthus annuus L.) and in theepidermal and mesophyll cells of sunflower leaf discs culturedin darkness have been studied. These inclusions appear to bedilated thylakoids containing a granular material which, undernormal conditions, is probably involved in chloroplast membraneformation. It is suggested that this material accumulates, andinclusions form, in the chloroplasts of sunflower leaves intwo specific situations. Firstly, in the completely differentiatedcells of the epidermis where the chloroplasts, although at arelatively immature stage, have nevertheless reached a terminalstage of development. Secondly, in the mesophyll cells of youngleaves when chloroplast development has been arrested at animmature stage by a 5-day dark period. In the latter situationthe material can be remobilized if plastid development is restimulated.The plastids of sunflower leaf discs cultured in darkness containboth membrane-bound inclusions and prolamellar bodies, indicatingthat they are separate and distinct structures possibly containingdifferent membrane components. Helianthus annuus L., sunflower, chloroplast, ultrastructure, plastid inclusions, thylakoid formation     

Experimental and Analytical Studies of Pteridophytes: XXXVII. A Note on the Inception of Microphylls and Macrophylls

In an attempt to shed new light on the nature of microphyllsand macrophylls, a study has been made of leaf inception inselected materials, including Psilotum, Tmesipteris and otherpteridophytes, and flowering plants such as Cuscuta with greatly‘reduced’ leaves. It is shown that the incipientprimordia of the small scale-like leaves of Psilotum and ofthe quite substantial, though microphyllous, laminate leavesof Tmesipteris are closely comparable and that there are noessential differences in the histological organization of incipientmicrophylls and incipient macrophylls. In parasitic speciessuch as Cuscuta, with small scale-like leaves, the organizationof the apical meristem and the inception of primordia are asin normal autotrophic species. The so-called ‘reduced’leaves and the microphyllous condition of some pteridophytesare attributable to a physiological-genetical limitation ofgrowth of primordia in the subapical regions of the shoot. Theconclusions which may be drawn from these observations are discussed.     

Observations on the Senescence of a Mutant Non-yellowing Genotype of Phaseolus vulgaris L.

A non-yellowing mutant of Phaseolus vulgaris L. was used toinvestigate factors involved in chlorophyll breakdown duringfoliar senescence. The mutant showed physiological changes similarto those of the normal yellowing type during senescence exceptthat leaf chlorophyll did not decline. Transmission electronmicroscope studies did not reveal appreciable differences inchloroplast ultrastructure between the two genotypes, suggestingthat chloroplast membrane integrity was not the factor preventingchlorophyll degradation in the mutant. However, the lack ofplastoglobuli in senescent mutant chloroplasts suggested thatthe lipid environment may be different from that of senescentnormal chloroplasts. Banding patterns of total soluble protein,resolved by sodium dodecyl sulphate-poly aery lamide gel electrophoresisshowed few, if any, differences between mature non-senescentnormal and mutant leaves; however, bands at 14 kD and 58 kDdiminished in senescent normal leaves, but remained in senescentmutant non-yellowing leaves. Key words: Non-yellowing mutant, Phaseolus vulgaris, senescence, chlorophyll degradation     

Intra-Leaf and Intracellular Gradients in Chloroplast Ultrastructure of Dorsiventral Leaves Illuminated from the Adaxial or Abaxial Side during their Development

The number of thylakoids per granum, which is smaller in sun-typechloroplasts than in shade-type chloroplasts, was counted forthe chloroplasts at various positions within the single leavesof Spinacia oleracea L. and Glycine max (L.) Merrill. The thylakoidnumber increased with depth from the adaxial surface, but thistrend was not evident within respective cells. Therefore, photosyntheticproperties of chloroplasts should be similar within a cell butdifferent among cell layers. The similarity within a cell maybe due to the nuclear control of the chloroplast developmentand/or to chloroplast movement along the cell walls. Illumination of the leaves of G. max from the abaxial side duringtheir expansion resulted in the complete inversion of the intra-leafgradient in the thylakoid number, indicating that the formationof the intra-leaf gradient in chloroplast properties is influencedby the intra-leaf light environment during the later phase ofleaf development. ¹presennt address: Department of Environmental Biology, ResearchSchool of Biological Sciences, Australian National University,P.O. Box 475, Canberra, A.C.T. 2601, Australia. (Received March 13, 1986; Accepted May 30, 1986)     

The Influence of Prior Water Stress and Abscisic Acid Foliar Spraying on Stomatal Responses to CO2, IAA, ABA, and Calcium in Leaves of Solanum melongena

The influence of a water stress or foliar ABA spraying pretreatmenton stomatal responses to water loss, exogenous ABA, IAA, Ca²⁺,and CO₂ were studied using excised leaves of Solanum melongena.Both pretreatments increased stomatal sensitivity of water loss,in the presence and absence of CO₂, but decreased stomatal sensitivityto exogenous ABA. CO₂ greatly reduced the effect of exogenouslyapplied ABA. IAA decreased leaf diffusion resistance for controland ABA sprayed leaves, but did not influence the LDR of previouslywater-stressed leaves. CA²⁺ did not influence LDR of any leavesof any treatments. Key words: Water stress, stomatal response, pretreatments     

Phosphoenolpyruvate Carboxylase from Mesembryanthemum crystallinum: Its Isolation and Inactivation in vitro

Phosphoenolpyruvate carboxylase (PEPC) was isolated from leavesof Mesembryanthemum crystallinum, which performed Crassulaceanacid metabolism. PEPC was much more stable when extracted from expanded thanfrom expanding leaves. The inactivation of PEPC in desaltedextracts from expanding leaves was much faster at 25 than at0 °C, was stimulated by raising Mg²⁺ from 0.1 to 3.0 mM,and was reduced by bovine serum albumin. The same type of inactivationwas found after mixing extracts from the two types of leaves,and the decrease in PEPC activity then also included inactivationof the ’stable’ PEPC from the expanded leaves. Afterelution from DEAE-cellulose, PEPC from expanding leaves wasmuch more stable than in desalted, crude extracts. It is suggested that another enzyme is involved in this inactivationof PEPC, but this needs verification.     

Fatty Acids in Chloroplasts and Leaves

The fatty acid composition of green and white leaf tissue inAcer negundo, Zea mais, and Ilex aquifolium, of green and yellowtissue in Ligustrum ovatifolium and of etiolated and green tissuein Vicia faba has been determined. The mesophytic green leavesexamined show a general similarity in fatty acid composition,characterized by a high concentration of non-conjugated octadecatrienoicacid. Chloroplasts were isolated from Vicia and Acer and containan even higher concentration of this acid and only traces ofnon-conjugated octadecadienoic acid. Conjugated diene and trieneacids occur in traces in chioroplasts, but are also found innon-green leaf tissue. The fats of non-green leaves are in generalmore saturated than those from green tissue but vary considerablyin composition. The relationship between fat composition andplastid development is discussed.     

Effect of Osmotic Stress Upon Endogenous Hormone Levels in Euphorbia lathyrus L. and Vicis faba L.

Ethylene concentrations rise in stems, lacunae and roots ofEuphorbia Iathyrus subjected to osmotic stress. In contrastto other species, however, no such changes are observed in leaves.Similarly, ABA concentrations rise in the leaves of osmotically-stressedplants but little change was observed in roots. In no case wasthere any strong evidence for the existence of a threshold ofosmotic potential above which a response will not occur. Instead,marked increases in concentrations of both ABA and ethylenewere obtained in response to osmotic stresses which did notresult in significant changes in water saturation defecit (WSD)during the experiment. In Vicia faba, time course studies at constant osmotic potentialshowed that ABA concentrations increased with duration of stressand increased WSD but the effect was most marked in youngertissues for a given level of stress. Concentrations of boundABA showed a similar pattern with time but here the change wasleast marked in the younger tissues. Euphorbia Iathyrus, Vicia faba, endogenous growth regulators, osmotic stress     

Photosynthetic Electron Transport During Senescence of the Primary Leaves of Phaseolus vulgaris L.: III. KINETICS OF CHLOROPHYLL FLUORESCENCE EMISSION FROM INTACT LEAVES

The kinetics of 685 nm chlorophyll fluorescence emission weremeasured at 20 °C following illumination of primary leavesof P. vulgaris. During foliar senescence, a large reductionwas observed in the maximal level of fluorescence emission (P)of the induction curve, normalized with respect to the minimallevel (O), and in the time taken to reach P. This suggests thatfewer plastoquinone (PQ) molecules were able to accept electronsfrom each photosystem two (PS II) reaction centre in older leaves.Measurements of fluorescence emission at 77 °K indicatedthat the primary photochemical quantum yield of the PS II reactioncentres remained constant during senescence. The redox stateof the PQ pool was estimated throughout the induction curveat 20 °C. In both mature and senescent leaves PQ was highlyreduced at P. There followed a reoxidation of PQ in the matureleaves, but in the old leaves the PQ pool remained reduced.This indicates that the rate of electron flow from PQ to photosystemone (PS I) decreased considerably during senescence. Fluorescencewas quenched from P to a steady state level (T) in leaves ofall ages, and this was associated with a redistribution of excitationin favour of PS I. Since, in senescent leaves, changes in theredox state of PQ were absent, it is suggested that quenchingresulted from the generation of proton and ion gradients acrossthe thylakoid membranes, and the synthesis of ATP.     

The Effect of Growth Regulators, CCC and B9, on Protein and Total Nitrogen of Bean Leaves (Phaseolus vulgaris) during Development

Total-N and protein-N were greatest in developing primary leavesof intact plants of Phaseolus vulgaris before the leaves werefully expanded. Treating plants with CCC or B9 delayed the timewhen total-N and protein-N diminished. Removing the growingpoint made nitrogen accumulate in the leaves and the total-Ndid not diminish until they were fully expanded. Treatment withCCC did not affect the amount of total-N or protein-N in decapitatedplants. Apparently nitrogen moves more slowly from primary leavesof CCC-treated plants because the shoot grows more slowly anddemands less nitrogen. Similarly, leaves of decapitated plantshave more nitrogen because there is no shoot. When primary leaveswere detached and rooted, protein continued to increase in themalthough they were fully expanded before they developed roots,again probably because the demand for nitrogen was small. Changesin protein were correlated with changes in chlorophyll duringdevelopment. Maximum photosynthesis per leaf probably occurs before leavesare fully expanded, and the protein changes in the leaf maybe closely related to the trend of photosynthesis during thelife of a leaf.     

Studies on nucleic acids in plastids of the pigment mutant C-2A{acute} of Scenedesmus obliquus

Pigment mutant C-2A{acute} of Scenedesmus obliquus whose chlorophyllformation and chloroplast development are light dependent, wasstudied for the nucleic acid content of its plastids. The ribosomalRNA of plastids of the achlorophyllous or greened mutant C-2A{acute},did not show any difference from that of the wild type. Incorporationof [5-³H] uridine into mutant cells was partially inhibitedby rifampicin, indicating this part as being plastidial incorporation.Since there were no significant differences in the ribosomalRNA of plastids between the mutant and the wild type of Scenedesmus,the ribosomal system in the plastids of mutant C-2A' seemednot to be affected by the mutation. C_sCl gradient patterns ofScenedesmus mutant and wild-type DNA were almost identical withthose of Chlorella DNA. A peak at a buoyant density of 1.69g/cm³, the same as that of Chlorella chloroplast DNA, couldbe identified in Scenedesmus also as plastid DNA because itdisappeared after prolonged treatment with myxin and hybridizedwith rifampicin-sensitive pulse-labelled RNA. This peak waspresent to nearly the same degree in the mutant and the wildtype, indicating that a larger deficit of plastid DNA did notoccur in the mutant. Whether or not the mutation might be localizedin the plastid genome is discussed. (Received March 19, 1976; )     

Leaf Heteroblasty in Pseudopanax crassifolius: Functional Significance of Leaf Morphology and Anatomy

Pseudopanax crassifolius (A. Cunn.) C. Koch. is a strongly heteroblastictree indigenous to New Zealand. Changes in the morphology andanatomy of leaves are described for seedling, juvenile, transitionaland adult phases of development. Seedlings produce five leaftypes; all are relatively small, thin, anthocyanic, have a lowspecific weight, and are anatomically comparable to the leavesof many shade plants. Juvenile leaves are long, linear, deflexed,coriaceous and sharply-toothed. They have a high specific weight,a thick, ornamented cuticle, a multiseriate hypodermis composedof collenchyma, well developed palisade and many spongy mesophylllayers. Juvenile leaves are strong relative to adult leavesand to the leaves of other species. Leaf strength is providedby fibres associated with the midrib. Adult leaves are shorter,broader and less massive than juvenile leaves, and are orientedhorizontally. Transitional leaves are morphologically intermediatebetween juvenile and adult leaves. The anatomy of juvenile,transitional and adult leaves is similar. The heteroblasticseries may be associated with changes in leaf construction costs,light interception and heat dissipation.Copyright 1993, 1999Academic Press Lancewood, Pseudopanax crassifolius (A. Cunn.) C. Koch., leaf heteroblasty, anatomy, morphology, structural adaptations     

Chloroplast Protein Synthesis During Barley Leaf Growth and Senescence: Effect of Leaf Excision

Chloroplast protein synthesis was measured during the expansion,maturity and senescence of the oldest leaf of barley, Hordeumvulgare L., var. Hassan. A maximum rate of protein synthesisoccurred near the end of the expansion stage 9 d after sowing.Protein synthesis increased again at the beginning of senescenceand reached a new maximum at day 14 after sowing. Detachmentand incubation of leaves in the dark stimulated chioroplastprotein synthesis by fully expanded or by senescent leaves butnot by expanding leaves. If the detached leaves were kept inthe light, chloroplast protein synthesis was stimulated in fullyexpanded but not in senescent leaves. Short treatments (18 h)of leaf segments with growth substances in either light or indarkness, significantly changed the rate of protein synthesisshown by chloroplasts. The relationship between chloroplastprotein synthesis and leaf senescence is discussed. Key words: Hormones, light, maturity     

Acclimation of Respiratory Properties of Leaves of Spinacia oleracea L., a Sun Species, and of Alocasia macrorrhiza (L.) G. Don., a Shade Species, to Changes in Growth Irradiance

To clarify the way in which the light available for growth affectsrespiration in leaves of sun and shade plants, we examined therespiratory properties of mature leaves of Spinacia oleraceaL., a sun species, and of Alocasia macrorrhiza (L.) G. Don.,a shade species, that had been grown at various irradiances.In leaves of S. oleracea, the respiratory rates, on a dry massbasis, decreased with time during the night, and the higherwas the growth irradiance during the day, the higher was therespiratory rate. The marked decreases in the respiratory rateduring the night were accompanied by decreases in the concentrationof carbohydrates in the leaves. By contrast, the respiratoryrates of leaves of A. macrorrhiza were virtually constant throughoutthe night and the absolute rates were lower than those of S.oleracea even though the absolute value of the concentrationof carbohydrates and its decrease at night resembled to thosein S. oleracea. The maximum activities of respiratory enzymeswere also similar to those in S. oleracea. However, the leavesof A. macrorrhiza contained less soluble protein than thoseof S. oleracea. These results suggest that, in S. oleracea,the concentration of carbohydrates might determine the respiratoryrate while such is not the case in A. macrorrhiza. The lowerrespiratory rates in A. macrorrhiza might be due to a lowerdemand for ATP. (Received November 29, 1995; Accepted February 15, 1996)     

Studies on the Role of Photosynthesis in the Photoperiodic Induction of Flowering in the Short-Day Plants Kalanchoe blossfeldiana Poellniz and Xanthium pensylvanicum Wallr: II. THE EFFECT OF CHEMICAL INHIBITORS OF PHOTOSYNTHESIS

The role of photosynthesis in flower induction in the short-dayplants Kalanchoe blossfeldiana and Xanthium pensylvanicum wasinvestigated by chemical suppression of photosynthesis and preventionof chlorophyll formation in the induced leaf. ‘Bleaching’leaves with streptomycin completely prevented flowering in X.pensylvanicum at concentrations shown to reduce the chlorophylland carotenoid content of the leaf significantly. Such leaveswere unable to induce flowering even when supplied with sugarsand other photosynthetic products. Photosystem II inhibitors,DCMU and cadmium ion, inhibited induction in both species aswell as suppressing photosynthesis (as tested by O₂ evolutionand starch production) whereas the photosystem I inhibitor,metronidazole, had no effect. Antimycin A inhibited floweringin K. blossfeldiana and may have a similar site of action toDCMU. Neither ammonium ion nor DBMIB, which acts upon plastoquinone(i.e. between PS I and PS II in the ‘Z scheme’),had any effect on floral induction and it is argued that theinductive process is independent of photosynthetic phosphorylationbut a step in the electron transport pathway between the sitesof action of DCMU and DBMIB may be crucial. DSPD and its hydrolysisproduct, salicylaldehyde, suppressed flowering in K. blossfeldianabut the uncertainty regarding their chemistry precludes anyfirm conclusions regarding the nature of their action.