Mobilization of Nitrogen in Fruiting Plants of a Cultivar of Cowpea

Patterns of flow of nitrogen were constructed for the post-anthesisdevelopment of symbiotically-dependent cowpea (Vigna unguiculataWalp. cv. Vita 3-Rhizobium CB756). Nitrogen fixed after floweringcontributed 40% of the fruits' total intake of N, mobilizationof N fixed before flowering the remaining 60%. Leaflets, nodulatedroot, stem plus petioles, and peduncles contributed mobilizedN in the approximate proportions 5: 2: 1: 1 respectively. Eachfruit drew on all available current sources of N, but N fromleaves was distributed preferentially to closest fruit(s), andlower fruits monopolized the N exported from nodulated rootsduring late fruiting. Rates of nitrogen fixation declined parallel with decreasingnet photosynthesis of shoots. Leaflets at upper reproductivenodes mobilized 70–77% of their N and declined steeplyin net photosynthesis rate per unit chlorophyll or per unitribulose-l, 5-bisphosphate carboxylase (RuBPCase)² before abscisingduring mid- to late fruiting, whereas leaflets at lower vegetativenodes (1–3) mostly failed to abscise, lost 44–57%of their N and maintained photosynthetic activity throughoutfruiting. Peptide hydrolase activity was examined in extracts of leaflets,roots and nodules, by autodigestion of extracts, or in assaysusing bovine haemoglobin and purified RuBPCase isolated fromcowpea as substrates. Hydrolase activities during fruiting werebroadly related to N loss from plant organs, but asynchronyin peaks of activity against different protein substrates indicateddistinct groups of hydrolases within single organs. Hydrolaseactivity of leaflet extracts against RuBPCase was highly andpositively correlated with in vivo rates of loss of RuBPCasefrom the same leaflets, and preferential degradation of thisprotein occurred during leaflet senescence. Key words: Nitrogen, Mobilization: Cowpea     

Role of Cell-wall-degrading Enzymes in the Development of Leaf Spots Caused by Ascochyta pisi and Mycosphaerella pinodes

Extracts of limited and spreading lesions caused by Mycosphaerellapinodes on detached pea leaflets contained proteolytic, cellulolytic,and pectolytic enzymes although only in spreading lesions wasthere much degradation of cell walls. The brown tissue fromlimited M. pinodes lesions was resistant to maceration by enzymesfrom spreading lesions. Limited lesions contained water-soluble,95 per cent ethanol insoluble, partially dialysable, inhibitorsof pectin transeliminase which is probably the macerating enzyme. Green, spreading M. pinodes lesions developed only on leafletsfloating on water. Growth of these lesions was accompanied bycontinous loss of phenolic substances to the water while thephenol content in infected tissue remained similar to that inuninoculated controls. In contrast, the phenol content in mature,limited M. pinodes lesions on leaflets suspended just abovethe water level was about four times that in healthy tissue.It is suggested that loss of phenolics from floating leafletsprevents tissue browning and the development of resistance ofthe cell walls to maceration. But this type of resistance doesnot appear to be a major factor in the limitation of lesionson suspended tissue. Extracts of limited Ascochyta pisi lesions on leaflets floatingon water contained pectolytic and hemicellulolytic enzymes.Some cellulase (Cx) activity was detected although there waslittle evidence of cellulose degradation in cell walls in infectedtissue. The nature of the macerating factor remains uncertainbut it was found that extracts from lesions contained inhibitorsof pectic enzymes and that tissue just beyond that colonizedby the fungus was resistant to maceration; this resistance isprobably important in restricting the growth of the pathogenin the leaf.     

Studies on chloroplast development in Chlamydomonas reinhardtii IV. Control of rapid chlorophyll formation in greening y-1 cells

Effects of protein synthesis inhibitors, CAP and CHI, on diegreening of Chlamydomonas reinhardtii y-1 cells, particularlyon die P-factor formation (19) in the early phase, were studied.Chlorophyll synthesis in the normal greening process, whichis divided into three phases, was strongly inhibited by bothantibiotics, although the inhibition by CAP was weaker in themiddle and late phases. The development of potential for rapidchlorophyll formation (P-factor formation) that takes placein dark-grown cells during dark incubation following brief illuminationwas completely blocked by CHI, but not by CAP. A "CHI-sensitive"period for the P-factor formation was restricted to the initial30 min during the dark incubation following brief illumination(10 min). This initial 30-min period appeared to correspondto the time of protochlorophyll(ide) formation which was inhibitedby CHI. Light-dependent conversion of protochlorophyll(ide) to chlorophylland also the subsequent protochlorophyll(ide) synthesis, whichis "CHI-sensitive" seem to be prerequisite for the inductionof P-factor synthesis. A possible control mechanism involvedin the early phase of the greening process in y-1 cells is discussed. (Received February 12, 1976; )     

THE MORPHOLOGY OF THE OSPHRADIUM IN RELATION TO FEEDING HABITS IN MESO- AND NEOGASTROPODS

The morphology of the osphradium in nine species of meso- andneogastropods was examined using scanning electron microscopy.Herbivores and detritivores (Littorina, Aporrhais) have simpleosphradia consisting of long ridges, covered in ciliary tufts,and flanked on either side by bands of densely ciliated epithelium.Another herbivore Rhinoclavis has the central ridge dividedinto lobes. In carnivorous and predatory species (Polinices,Cypraea, Nucella, Nassarius, and Conus) the osphradium is shorter,larger and divided bi- or triserially, into leaflets. Conusstri-atus which feeds upon the most mobile and least predictableprey (fish) has the largest and most elaborate osphradium, withthe leaflets divided into digitiform processes. The sedentary,ciliary-feeding, Crepidula is anomalous in having an osphradiumdivided into leaflets, although these are vestigial along oneside.     

Lack of Influence of the Epidermis on Underlying Cell Development in Leaflets ofPisum sativumvar.argenteum(Fabaceae)

We explored whether epidermal pressure regulates cell and organgrowth in leaflets ofPisum sativumvar.argenteum,a mutant cultivarof the garden pea characterized by reduced adhesion betweenthe epidermis and subjacent mesophyll. Developing leaflets ofleaves arising at three positions on the seedling axis werepeeledin situand grown to maturity in humidity chambers. Themature anatomy and morphology could be accurately assessed becausewound responses normally associated with peeling were preventedby theArgmutation that permitted peeling without damage to themesophyll and by the humidity chambers that protected peeledareas from desiccation. The mesophyll cell size, state of differentiation,and layering pattern as well as the overall morphology of mature,peeled leaflets were indistinguishable from those of mature,intact leaflets grown under the same conditions. The epidermisexerted no detectable regulatory effect on the expansion ofthe leaflets as a whole or on the tissue layers and cells withinthe leaflets.Copyright 1999 Annals of Botany Company. Biomechanics, compression, epidermis, leaf development, mesophyll, pressure, wound response,Pisum sativumvar.argenteum.     

Gametogenesis in Chlamydomonas II. Effect of cycloheximide on the induction of sexuality

An inhibitor of protein synthesis, CHI, was used to study therelation of protein synthesis to the induction of sexualityin Chlamydomonas. (1) Freshly prepared CAP and DSm did not inhibitinduction, but CAP solution which had been illuminated causedstrong inhibition. (2) CHI inhibited the induction of sexualityremarkably, but not completely. The induction of sexuality maybe of two types, CHI-sensitive and CHI-insensitive. (3) Vigorousagitation of gametogenic cultures resulted in a marked inhibitionof induction in the presence of CHI, and less in its absence.(4) The amounts of light required for CHI-sensitive and CHI-insensitiveinduction differed; respectively, 5 min of illumination at ca.8,000 lux and 3 hr of illumination at the same intensity. (5)For complete blockage of CHI-sensitive induction, the additionof CHI 50 min before illumination was needed. (6) The activitiesof CHI-sensitive and CHIinsensitive induction changed with theage of the culture. (7) It was supposed that some material(s)inhibiting gametogenesis was present in the medium of the agedculture. Based on these results, a hypothetical scheme for gametogenesisis presented. (Received March 7, 1973; )     

Regulation of Leaf Shape in the Solanifolia Mutant of Tomato (Lycopersicon esculentum) by Plant Growth Substances

The solanifolia mutant (sf/sf) of tomato (Lycopersicon esculentum)produces leaves consisting of leaflets with entire margins,unlike the lobed leaflets of normal plants. Normal plants treatedwith gibberellic acid (GA₃) produced leaves with entire marginswhereas mutant plants exposed to 2-chloroethyl-trimethyl ammoniumchloride (CCC)—an inhibitor of gibberellin biosynthesis—producedlobing of leaflets. The leaf area of the mutant was significantlygreater than that of the normal, but was not significantly differentfrom GA₃-treated normal leaves. Similarly, in CCC-treated mutantleaves the leaf area was not different from that of normal untreatedleaves. These observations suggest that the sf/sf mutation affectsthe leaf shape through its effect on endogenous gibberellinsand/or inhibitory substances. Leaf shape, Lycopersicon esculentum, plant growth substances, tomato     

Distribution and utilization of chlorogenic acid in Coffea seedlings

Ripe coffee seeds (Coffea arabica L.) contain large quantitiesof the purine alkaloid caffeine and the depside chlorogenicacid (5-O-caffeoyl-quinic acid). Directly after germination,both compounds were recovered almost exclusively in the cotyledonsand surrounding endosperm and hardly at all in the hypocotyland root of coffee seedlings. During the first 10 weeks of development,the cotyledons invaded the endosperm and expanded. The caffeinecontent in the developing cotyledons changed little, but thechlorogenic acid content dropped to one-third of the originallevel. The loss of chlorogenic acid was not recovered in theother organs of the seedling. The drop in chlorogenic acid contentcoincided with an increase in the amount of cell wall-boundphenolic polymers in the developing cotyledons, which couldbe extracted after thioglycolic acid derivatization. In thedeveloping cotyledons, phenylalanine ammonia lyase (PAL) activitywas hardly detectable. These results suggest that the chlorogenicacid stored in coffee seeds is used for the deposition of phenolicpolymers, presumably lignin, in cotyledonary cell walls duringexpansion. The situation in cotyledons was found to be in markedcontrast to that in foliage leaflets, in which PAL activitywas 900-fold higher than in cotyledons. In the leaflets, bothchlorogenic acid and caffeine accumulated continuously duringleaf expansion. Key words: Coffea arabica, seedling, chlorogenic acid, caffeine     

Abscisic acid – an intraleaf water-stress signal

Physiological aspects of abscisic acid (ABA) as a drought signal directed specifically at guard cells are topical research foci. Most investigations concentrate on the importance of remote sources of ABA against the background knowledge that leaves are also a source of ABA. Foliar compartmentation of ABA and water-stress-induced release of cellular ABA into the apoplast imply additional levels of complexity. In the present study, we established that the pre-existing foliar ABA pool can be moved to guard cells. We detached leaflets of Vicia faba L. to eliminate an external source of ABA. The leaflets were then pretreated with cycloheximide (CHI), an inhibitor of protein biosynthesis that abolished ABA accumulation in the leaflets. Next, the leaflets were water stressed. After incubation, leaflets were snap-frozen and freeze-dried. Guard cells were individually dissected from the leaflets; such guard-cell samples contain both apoplastic and symplastic ABA. The ABA content of guard cells of CHI-treated, water-stressed leaflets was less than that of control water-stressed leaflets, but higher than that of control water-sufficient leaflets, indicating that guard cells are targets of intraleaf ABA redistribution under stress.     

Effects of Light and Growth Substances on the Diurnal Movements of the Leaflets of Mimosa pudica

The leaflets of Mimosa pudica show diurnal opening and closingmovements for some days when they are removed in pairs fromthe parent plant along with the pulvini and a small sectionof the rachis, and floated in water. The movements can be measuredfrom changes in the angle of the blades. When the leaflets arekept in darkness they show a diurnal movement of opening inthe morning at the same time as opening occurs in light. Light,however, is necessary to prevent the leaflets from closing duringthe day after an initial opening. Blue light was most effectivein maintaining the open condition throughout the day. Low concentrationsof indole-3-acetic acid (IAA) and gibberellic acid (GA) promotedopening of the leaflets while higher concentrations preventedtheir closure at night. IAA and GA acted synergistically inreducing dark-closure. Growth substances also prevented daylightclosure of the leaflets in the dark. The results arre discussedin relation to the changes in the osmotic relations of pulvinarcells regulating leaf movements.     

A Comparison of Abscission of Rubber (Hevea brasiliensis) Leaves Infected with Microcyclus ulei with Leaf Abscission Induced by Ethylene Treatment, Deblading and Senescence

Studies on the histology and on effects of growth substancesand phenols as well as changes in activities of pectinmethylesterase indicated that the mechanism of abscission of Hevealeaflets infected with Microcyclus ulei differed from the mechanismof abscission of debladed, ethylene treated and senescent leaves.An abscission layer which was formed during abscission of debladed,ethylene-treated and senescent leaves was absent during abscissionof heavily diseased leaves. The ratio of pectinmethyl esteraseactivities in tissues distal to the abscission zone to activitiesin tissues proximal to the zone decreased in debladed and ethylenetreated leaves but such decreases were not detected during abscissionof Hevea leaves infected with M. ulei. Hevea brasiliensis Muell. Arg., rubber, leaf abscission, Microcyclus ulei, ethylene, indol-3-ylacetic acid, kinetin     

The initiation of growth in isolated lima bean axes. Physiological and fine structural effects of actinomycin D, cycloheximide and chloramphenicol.

Excised lima bean axes were germinated for 24 hr in water, variousconcentrations of CHI, CAP or actinomycin D. None of the inhibitorsaffected water uptake before onset of growth. Growth was completelyinhibited by CHI and CAP, but only partly by actinomycin D.Rate of ³H-leucine incorporation rose during the lag phase inwater uptake and increased further during growth. The incorporationwas inhibited by CAP and CHI at all stages, but by actinomycinD only during growth. Actinomycin D prevented developmentalfine structural changes. In CHI treated axes number of mitochondriaincreased, O₂2 uptake was reduced, and higher cytochrome oxidaseactivity induced. CAP caused changes in the appearance of proteinvacuoles, but apparently prevented loss of storage material.The relation between the physiological and structural effectsof the inhibitors is discussed. (Received June 13, 1970; )     

In vitro degradation of three-dimensional chitosan/apatite composite rods prepared via in situ precipitation

Three-dimensional (3D) compact rods with multilayer structure made from chitosan (CHI) and apatite (Ap) have been prepared. The cytocompatibility assay revealed that the CHI/Ap composite could promote cell proliferation. In vitro degradation behaviors of the rods have been systematically investigated for up to 6 weeks in phosphate buffer saline (PBS) solution at 37°C. The properties of the composite rods were measured by means of weight loss, swelling ratio, and the changes in mechanical properties, etc. The pH of the PBS solution during the first 2 weeks of degradation was also detected. Results showed that the medium of CHI/Ap composite rods exhibited more stable pH change compared with that of CHI rods. Weight loss as well as the changes in mechanical properties happened more often to CHI rods than CHI/Ap rods. The presence of Ap could effectively reduce the degradation rate of the composite rods. All the results suggested that the composite rods could keep the initial shapes and mechanical properties longer than the pure CHI rods.     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture V. The transition point for the effect of 6-methyl purine in the cell cycle

By following the nuclear division, chloroplast division, cytokinesisand zoospore liberation of Chlamydomonas reinhardi cells exposedto a high concentration of 6-methyl purine (6-MP), corroborativeevidence was obtained for our previous conclusion that 6-MP-exposedcells are brought to the starting point of a new round of thecell cycle with abortion of the cycle in process ("return-to-start"effect). This effect did not occur after cells had passed acritical stage (transition point) which seemed to be situatedshortly prior to the onset of nuclear division under the conditionsused. When 6-MP was applied to cells after the transition point,it caused an advancing effect on their zoospore liberation.A cycloheximide (CHI)-inhibition step existed shortly alterthe transition point for 6-MP. A model was proposed for theeffects of 6-MP and CHI. (Received August 8, 1977; )     

Effects of Jasmonic Acid on Motor Cell Physiology in Mimosa pudica L. and Cassia fasciculata Michx

When applied to pulvini of Mimosa pudica, jasmonic acid (JA)affected neither proton fluxes nor the membrane potential ofthe motor cells. When added to leaflets of Cassia fasciculata,JA increased the rate of dark-induced pulvinar movements ina concentration-dependent manner. This effect was observed withinas little as 15 min after a 1-h treatment that preceded theinducing signal. Treatments in buffered media at acidic pH resultedin the greatest physiological responses. Light-induced pulvinarmovements were considerably reduced under the same conditions.With continuous illumination, JA induced a closing movementof the leaflets in a concentrationdependent manner. These resultsare discussed in relation to the ionic changes in the pulvinarmotor cells and in relation to results obtained previously upontreatment of Cassia plants with ABA. Although ABA and JA havesimilar physiological effects on the dark-induced closure, theydiffer in the type of response elicited by brief treatment andwith respect to light-induced opening. (Received September 27, 1993; Accepted January 15, 1994)     

Site of photo-reception to opening response in Mimosa leaflets

A pair of leaflets excised from Mimosa pinna and floated onwater for a day, responded to light and mechanical stimuli.Only when a small spot of light was shone on the abaxial surfaceof pulvinule, did the opening response of the leaflets occur.No translocatable effect from suchirradiation was observed. ¹ Present address: Biological Institute, Faculty of Science,Kobe University, Kobe 657, Japan. (Received May 17, 1973; )     

Leaf Spots Induced by Ascochyta pisi and Mycosphaerella pinodes

The leaf-spot pathogens, Ascochyta pisi and Mycosphaerella pinodes,both cause limited, necrotic lesions in detached pea leafletssuspended above water. When leaflets were floated on water A.pisi lesions were still limited, but those caused by M. pinodesspread rapidly to occupy all the leaflet tissue. Increasing the numbers of spores in inocula decreased numbersof mature lesions caused by A. pisi, but increased numbers ofspreading lesions caused by M. pinodes. Older leaflets weremore susceptible to both pathogens. Studies of penetration and colonization of leaves with the aidof light and electron microscopy showed that cell-wall-degradingenzymes were involved in the formation of A. pisi lesions andin spreading lesions caused by M. pinodes. There was littleevidence of cell-wall degradation in limited M. pinodes lesionsin which penetration of walls by hyphae seemed to be mechanicalin nature. No physical barriers developed in tissues surrounding limitedlesions. Nevertheless, A. pisi was rarely found beyond the necroticarea. This suggested that tissues beyond the lesion had becomeresistant to the parasite. In contrast, M. pinodes often grewoutside the necrotic area, sometimes many days after this hadstopped growing, but when it did so it caused no more necrosisunless leaflets were placed in conditions in which the spreadingtype of lesion could develop.     

TheAfGene Regulates Timing and Direction of Major Developmental Events during Leaf Morphogenesis in Garden Pea (Pisum sativum)

The wildtype leaf of the garden pea possesses proximal pairsof leaflets and distal pairs of tendrils in the blade region.Theafila (af) mutation causes leaflets to be replaced by compound(branched) tendrils. We characterized the morphological variationin leaf form along the plant axis and leaf development in earlyand late postembryonic leaves onafilaplants to infer the roleof theAfgene. Leaf forms are more diverse early in shoot ontogenyonafilaplants.Afinfluences pinna length and pinna branchingin addition to pinna type. Pinna initiation in the proximalregion ofafilaleaf primordia is basipetal and delayed comparedto wildtype plants. In addition, pinna development in the proximalregion ofafilaleaves occurs for a longer period of time thanon wildtype leaf primordia. Therefore,Afregulates the timingand direction of leaf developmental processes in the proximalregion of the leaf, but has little effect on the distal region.These data support the heterochronic model of pea leaf morphogenesisproposed by Luet al. (International Journal of Plant Science157:311–355, 1996).Copyright 1999 Annals of Botany Company. afila,Fabaceae, garden pea, heterochrony, leaf morphogenesis,Pisum sativum.     

The Roles of Cell Division and Cell Expansion in the Growth of Alfalfa Leaf Mesophyll

Leaf mesophyll of Medicago sativa (L.) was investigated to determinethe roles of cell division and cell expansion in tissue growth.Samples of leaf tissue were macerated, stained, and squashed.The slides were studied under a phase microscope to determinethe percentage of recently divided cells and the average celldiameter for leaflets of varying lengths. Cell division wasgreatest in young leaflets and virtually ceased as a leaf lengthof 12 mm was attained. For leaflets less than 12 mm in length,the rate of increase in cell size appeared to be inversely associatedto the degree of cell division. For alfalfa leaflets greaterthan 12 mm in length, the mean cell size increased in proportionto leaf length since cell division had virtually ceased.