Studies on Mature Seeds of Cuscuta pedicellata and C. campestris by Electron Microscopy

The seeds of Cuscuta pedicellata have been investigated by transmissionand scanning electron microscopy. Additional observations havebeen made on seeds of C. campestris by SEM only. The seed coatconsists of an outer single epidermis, two different palisadelayers, and an inner multiparenchyma layer. The outer epidermalwall in C. pedicellata has a thick cuticle and zones rich inpectic substances. The thicker ‘U-shaped’ cell wallsin the outer palisade layer are strengthened by a wall layerof hemicellulose. The inner palisade layer has thick walledcells with a ‘light line’. The inner cell wall ofthe compressed multiparenchyma layer has a thin cuticle. A fairlythick cuticle is positioned directly on the endosperm surface.The aleurone cell walls are different from the remaining endospermwalls. The latter are thick and believed to be of galactomannans.There is a ‘clear’ zone between the plasmalemmaand the cell wall in the aleurone cells. The embryo cells arepacked with lipids and proteins. In Cuscuta campestris mostendosperm has been absorbed during the seed development. Theembryo apex has two minute leaf primordia. The features of theCuscuta seeds are discussed in relation to functional and environmentalconditions. Cuscuta pedicellata, Cuscuta campestris, seed, seed coat, cuticle, cell walls, endosperm, aleurone cells, galactomannan, embryo, TEM, SEM     

Amitosis and Endocytogenesis in the Fruit of Malus sylvestris

Cyto-histological investigations on the initiation and developmentof a non-pathogenic physiological ‘cork spot’ necrosisin the outer cortex of the fruit of Malus sylvestris Mill. ‘YorkImperial’ revealed two distinct aberrancies, namely, amitoticnuclear division and intracellular or endogenous proliferationswithout hyperplasia per se The incipient necrotic conditionbecomes evident internally about 3 weeks after fruit set asminute isolated, discoloured, amorphous spots of disorganizedruptured cells within otherwise healthy cortical tissue Thedisorganization continues slowly with adjoining cells becomingsimilarly necrotic After the lysis of the initial cork spotcells, about 1 month later, sporadic cellular changes occurin various healthy vacuolated cortical cells contiguous to andencompassing the necrotic tissues. The nuclei increase in sizeand volume as they assume distinctive positions in preparationfor amitotic nuclear divisions. The enlarged nucleus or macronucleus,containing one or several nucleoli, divides by a distinct cleavagedeveloping from a constriction perpendicular to its longitudinalaxis The division results in the amitotic formation of two daughtermicronuclei that usually become separated by the formation ofa cell wall. No evidence of cell plate formation was observedand the method of cell wall formation could not be determined.Repeated amitotic divisions of the micronuclei result in anintracellular or endocytogenetic proliferation of parenchymatouscells that are invariably confined within the original mothercell until its wall ruptures The endogenous proliferations arereleased into lacunae or intercellular spaces, eventually becomedisorganized, and disintegrate, with the accumulated residualincrements resulting in an overall ‘cork spot’ appearance. Malus sylvestris Mill., apple, amitosis, endocytogenesis, multinucleate cells, macronuclei, micronuclei     

The Ultrastructure of the Alkaloidal Vesicles of Papaver somniferum latex

Papaver somniferum latex contains abundant small vesicles. Theirultrastructure was studied in tissue sections from adult plantsand in sections of sequential fractions of centrifuged latex.The vesicles were found to exist in two forms, the first witha smooth but progressively granulated outer membrane and thesecond, probably derived from the first, with adherent ‘cap-like’structures which in the heavier centrifuged fractions possesseda zonally-ordered interior. These vesicle fractions were active in synthesizing morphineand the name ‘alkaloidal vesicle’ is proposed forthem. Papaver somniferum latex also contains an organelle whichwas found to resemble a complex organelle present in the latexof Hevea brasiliensis. Its function is not yet known.     

Factors Affecting the Abscission of Reproductive Organs in Yellow Lupins (Lupinus luteus L.): III. ENDOGENOUS GROWTH SUBSTANCES IN VIRUS-INFECTED AND HEALTHY PLANTS AND THEIR EFFECT ON ABSCISSION

Extracts of small and mature-size lupin pods yielded four substancesaffecting the growth of wheat-coleoptile sections: one acidpromotor (A), two acid inhibitors(B and X), and one neutralinhibitor(Y). Inhibitor B was extremely active, however, coleoptile sectionsshowed no signs of toxic effects; they resumed growth at a rapidrate after rinsing them and adding ß-indolylaceticand (IAA) to the medium. 1 µg of IAA was required to counteractthe effect of ‘B’ extracted from 230 mg. Of tissue.On an equal fresh weight basis the inhibiting action of ‘B’in lupin pods was 500–1,500 times more potent than thatof ‘inhibitor ß’ in etiolated pea seedlings. Small pods of plants infected with pea-mosaic virus yielded3 times the amount of ‘A’ of healthy plants (equivalentto 1 µg. IAA 0.3 µg. IAA per 25 g. of tissue respectively),and approximately the amount of ‘B’. Mature podsof virus-infected plants again yielded more‘A’,but also 2? times more ‘B’ than pods of healthyplants. Healthy pods yielded more ‘A’ than virus-infectedpods, and there was no difference in ‘X’. A lupin abscission test was developed and the effects of proximaland distal application of -naphthyl acetic acid (NAA) are presented,and discussed with respect to results of other abscission tests. ‘A’ accelerated abscission when applied proximally,and delayed or prevented it when applied distally. ‘B’strongly accelerated abscission when applied in either way.A possible mechanism explaining the abscission-inducing effectof developing pods on later flowers is discussed in terms ofthe substances ‘A’ and ‘B’. The partlyprevented abscission observed on virus-infected plants was foundto agree well with the proposed mechanism.     

Pressure and Solute Potentials in Stomatal Cells of Tradescantia virginiana

The postulated mechanical advantage of subsidiary cells overguard cells has been estimated using leaves of Tradescantiavirginiana. The turgor pressures of subsidiary cells were adjusted to bezero or maximal by plasmolytic treatments, and the resultingstomatal apertures were measured. The ‘mechanical advantage’was calculated from two mathematical models which define itas an ‘antagonism ratio’. The discussion deals withmethods of preparing the tissue, the validity of the plasmolytictreatments, and the function of the antagonism ratio in relationto the Spannungsphase     

Accumulation of Cytokinin-Induced Betacyanin in Specific Cells of Amaranthus tricolor Seedlings

The accumulation of betacyanin, in dark-grown Amaranthus tricolorseedlings, in response to cytokinins or red light, occurs mainlyin two specific tissues, the lower epidermal cells of the cotyledons(with the exception of guard cells), and the endodermis of thehypocotyl. The possible significance of this ‘spatialpattern of competence’ is discussed, together with theconcept of target cells in relation to plant hormones. The effect of removing exogenously supplied cytokinin at varioustimes during a 24 h induction period is reported. There is noevidence that cytokinins act by a ‘triggering’ effectwith a long half life, the response in the target cells beingthe same as that expected from the amount of cytokinin and cytokininmetabolite remaining in the tissue at the time of extraction.Either continuous presence of cytokinin is needed or any triggeraction is short lived, and continuous ‘re-triggering’is needed to achieve maximum response. Key words: Amaranthus tricolor, Betacyanin synthesis, Cytokinin action, Target cells     

Effects of Inorganic Phosphate on the Utilization of Sucrose by Suspension-cultured Catharanthus roseus Cells

The metabolic fate of [U-¹⁴C]sucrose in suspension culturesof Catharanthus roseus cells was monitored for 96 h after thecells were transferred to fresh complete (‘+Pi’)or to phosphate-deficient Murashige and Skoog (‘–Pi’)medium. Sucrose was hydrolysed extracellularly to glucose andfructose. The rate of uptake of sugars by the cells was 1.5–3times higher in ‘+Pi’ culture than in ‘–Pi’culture. Little difference in the rate of incorporation of radioactivityinto the ethanol-soluble fraction was found between the ‘+Pi’and ‘– Pi’ cultures during the initial 24h of culture, but after 48 h the rate in ‘ +Pi’cultures was higher than that in ‘–Pi’ cultures.Incorporation of radioactivity into ethanol-insoluble macromoleculeswas always significantly higher in the cells in ‘+Pi’cultures than in those in ‘–Pi’ cultures.The results suggest that Pi strongly affects the utilizationof sugars by cultured plant cells through the stimulation oftransport of sugars as well as through the activation of metabolism. Catharanthus roseus, Madagascar periwinkle, suspension culture, inorganic phosphate, sucrose, transport, metabolism     

Developmental control of xylem hydraulic resistances and vulnerability to embolism in Fraxinus excelsior L.: impacts on water relations

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

Freezing tolerance, protein composition, and abscisic acid localization and content of pea epicotyl, shoot, and root tissue 3

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

The Potassium Relations of Lemna minor L.: I. POTASSIUM UPTAKE AND PLANT GROWTH

A simple expression has been derived to predict the rate ofnet K uptake into exponentially growing plants of Lemna minor.Net uptake predictions are in good quantitative agreement withmeasurements of ‘steady-state’ K influx, indicatingthat, in the ‘steady state’, K movements in theplant are essentially undirectional and that efflux is small.This close matching of inward K movement to the demands of theexpanding tissue is temporarily disturbed if plants are transferredto media of different K status. Uptake rates in the ‘step-up’are initially enhanced and then fall gradually towards a new‘steady-state’ rate. In contrast, the ‘step-down’causes an initial depression of uptake and then rates increasegradually towards the new ‘steady-state’ rate. Itis argued that these changes in uptake rates are associatedwith alterations in the cytoplasmic K content.     

Seed Coat Structure and Histochemistry of Abelmoschus esculentus. Chalazal Region and Water Entry

The seed coat structure and histochemistry of Abelmoschus esculentuswere studied by bright-field, fluorescence and scanning electronmicroscopy. The seed coat was typical of species of the Malvaceae.The endotesta cells had inner tangential walls which were verythick and autofluorescent. The occurrence of phenolic substancesat this level has been related to seed coat imposed dormancy.The palisade cells were composed of three differently shapedparts: an upper ‘prismatic part’, a medium ‘transitionpart’ and a lower ‘twisted part’. The formerwas rich in hydrophilic substances, the latter was lignified.The swelling of the ‘prismatic parts’ was relatedto seed coat cracks. The region controlling onset of water entrywas thought to be the chalazal area. Thanks to the presenceof a large amount of highly acidic polysaccharide, water wasable to penetrate from the permeable maternal tissue, throughthe chalazal cap and plug as far as the boundary between thepalisade and underlying mesophyll. During imbibition of watera kidney-shaped ‘blister’ was seen to rise, formedby separation of the palisade cells from an underlying singlelayer of subpalisade cells. The palisade layer forming the blisterroof showed the same histochemical characteristic of other seedregions. The single layer of the blister floor showed an affinitywith Toluidine Blue O and Alcian blue. Both blister roof andfloor were strongly autofluorescent. Abelmoschus esculentus (L.) Moench, okra, seed coat, chalazal region, water entry, structure, histochemistry     

Influence of Flooding on Net CO2Assimilation, Growth and Stem Anatomy of Annona Species

A series of experiments was conducted to assess net CO₂assimilationand growth responses to waterlogging of grafted and seedlingtrees in the genus Annona. Seedlings of A. glabra, A. muricataandA. squamosa L., and scions of ‘Gefner’ atemoya(A. squamosaxA. cherimola Mill.), ‘49-11’ (‘Gefner’atemoyaxA. reticulata L.), ‘4-5’ (‘Priestley’atemoyaxA. reticulata), A. reticulata grafted onto either A.glabra, A. reticulata orA. squamosa rootstocks were floodedfor up to 60 d. Soil anaerobiosis occurred on the third dayof flooding. Seedlings ofA. glabra and A. muricata, and thescions ‘49-11’, ‘Gefner’ atemoya, andA. reticulata grafted onto A. glabra rootstock were consideredflood tolerant based on their ability to survive and grow inflooded conditions. Scions of the normally flood-sensitive A.reticulata, ‘Gefner’ atemoya, and ‘49-11’tolerated root waterlogging when grafted onto the flood-tolerantspecies, A. glabra. In contrast, flooding of A. squamosa seedlingsand rootstocks, and A. reticulata rootstocks greatly reducedgrowth and net CO₂assimilation rates, and resulted in 20–80%tree mortality. Stem anatomical responses to long-term flooding(12 continuous months) were assessed in seedlings of A. glabraand A. muricata, and trees of ‘49-11’ grafted ontoA. glabra. Flooded trees developed hypertrophied stem lenticels,particularly in A. glabra, and enlarged xylem cells resultingin thicker stems with reduced xylem density. Flooding did notincrease air spaces in pre-existing xylem near the pith or inxylem tissue that was formed during flooding. Thus, flood tolerancedid not involve aerenchyma formation in the stem. Copyright1999 Annals of Botany Company Flood tolerance, net CO₂assimilation, photosynthesis, stem anatomy, shoot growth, anaerobiosis, Annonaceae.     

Structural and Histochemical Changes in Palisade Cells of Prosopis juliflora Seed Coat in Relation to its Water Permeability

Histochemical investigations on the Prosopis juliflora seedcoat indicate the occurrence of a hydrophobic ‘strip’as the primary water barrier. Its position and the structureand histochemistry of the palisade cells of the seed coat differaccording to their location on the seed. These differences maybe responsible for differences in the water permeability ofvarious parts of the seed coat. In particular, parts of theseed coat in which the hydrophobic ‘strip’ is locatedmore superficially tend to be more water impermeable than partslike the chalaza, in which the ‘strip’ is more deeplylocated within the palisade cells. Prosopis juliflora, seed coat impermeability, palisade cells, hydrophobic ‘strip’     

The Growth and Carbon Economy of Selection Lines of Lolium perenne cv. S23 with Differing Rates of Dark Respiration: 1. Grown as Simulated Swards During a Regrowth Period

Simulated swards of each of two selection lines of Lolium perennecv. S23 with ‘fast’ and ‘slow’ ratesof ‘mature tissue’ respiration were establishedin growth rooms at 20/15 °C day/night temperatures and studiedover four successive regrowth periods of 46, 30, 26 and 53 daysduration. The ‘slow’ line outyielded the ‘fast’,both in harvestable shoot (above a 5 cm cut) and in root andstubble. Its advantage increased over successive regrowth periodsto 23 per cent (total biomass). Gas analysis measurements onthe entire communities (including roots), during the final regrowthperiod, showed that the ‘slow’ line had a 22–34per cent lower rate of dark respiration per unit dry weight.This enabled it to maintain its greater mass of tissue for thesame cost in terms of CO₂ efflux per unit ground area. Halfthe extra dry weight produced by the ‘slow’ line,relative to the ‘fast’, could be attributed to itsmore economic use of carbon. The rest could be traced to a 25per cent greater tiller number which enabled the ‘slow’line to expand leaf area faster (though not at a greater rateper tiller), intercept more light and fix more carbon, earlyin the regrowth period. Lolium perenne L., ryegrass, respiration, maintenance respiration, tiller production, simulated swards, canopy photosynthesis, carbon economy     

The Developmental Anatomy of the Root System in Yam Bean, Pachyrhizus erosus Urban

Investigations revealed that the anatomy of the primary radicularroot of yam bean (Pachyrhizus erosus L.) was typically dicotyledonousexcept that the xylem was not completely developed centripetally.Most of the roots had tetrarch xylem, although a few triarchand pentarch roots were also observed. In both tuberous andnon-tuberous roots, secondary thickening occurred by the formationof the meristematic vascular cambium which formed secondarytissues in a normal fashion. Subsequently, tuberization wasinitiated in the secondary xylem by the development of anomalous‘secondary’ cambia from parenchyma cells surroundingvessel elements. Anomalous ‘secondary’ cambia alsodeveloped from parenchyma cells not associated with vessels.Subsequently, anomalous ‘tertiary’ cambia differentiatedfrom tissues produced by the anomalous ‘secondary’cambia. Activities of these anomalous cambia resulted in theproduction of parenchyma storage cells and were chiefly responsiblefor the growth of the mature tuber. Pachyrhizus erosus L., yam bean, tuberous root, anatomy, anomalous ‘secondary’ cambia, anomalous ‘tertiary’ cambia, centripetal xylem development     

Pollen Wall Development of Xiphidium coeruleum (Haemodoraceae) and its Systematic Implications

The development of the pollen grain wall in Xiphidium coeruleum(Haemodoraceae) was studied using TEM and cytochemical stainingtechniques. Microsporocyte ontogeny initiates with the degradationof the cellulosic cell wall and subsequent deposition of a thickcallosic cell wall. Following callose deposition, successivemeiosis occurs, resulting in a tetragonal tetrad of microspores.during meiosis, the cell walls of the tapetum break down, releasingthe syncytial periplasmodium. Irregular non-sporopollenous globularbodies are deposited in this peripheral periplasmodium, whichis rich in ER, golgi bodies, vesicles, and characteristic starchplastids. Within the microspore cytoplasm, vesicles, golgi bodies,and plastids are plentiful during the early tetrad stage. Atthis time the plasma membrane of the microspore develops characteristicevaginations. An extracellular membrane, the ‘white line’,is secreted outside the microspore plasma membrane, followedby callose wall degradation. Bead-like deposits of exine orprimexine are deposited at points along the ‘white line’simultaneously on inner and outer surfaces and opposite theoriginal plasma membrane evaginations. The bead-like exine depositscontinue to grow during the release of the microspores and developinto laterally appressed, rod-shaped ektexinous elements havinga tangentially oriented commissure, the vestige of the original‘white line’. The mature intine is two-layered,the outer exintine containing radially oriented vesicular structures,which are apparently derived from plasma membrane extensions.Exine development in Xiphidium is similar to ‘nexine 1’development in Lilium and may have evolved from an ancestraltectate-columellate condition by the loss of the sexine. Walldevelopment in members of the Zingiberales is strikingly similarto that reported here for the Haemodoraceae—evidence ofa possible relationship between the two taxa. Xiphidium coeruleum, Haemodoraceae, pollen, tapetum, development, exine     

Determinate Floral Buds of Plantain (Musa AAB) as a Site of Adventitious Shoot Formation

Floral buds of the ‘False Horn’ plantain clonesMusa (AAB) ‘Harton Verde’, ‘Harton Negra’,and ‘Currare’ terminate in a large single floralstructure. The apices of these floral buds are here designatedas determinate since they have lost the ability to produce additionalfloral initials or buds. Terminal peduncle segments can be culturedin a modified Murashige and Skoog (1962) medium supplementedwith N⁶-benzyl-aminopurine (5 mg I^–1). Under these conditions,this apparent inability to yield buds can be overcome as vegetativeshoot clusters form in the axils of the bracts. Rooted plantletsare obtainable by treating shoots with naphthaleneacetic acid(1 mg I^–1) and activated charcoal (0.025%). The adventitiousorigin of the shoots has been established. Musa cultivars, plantains, floral bud, adventitious buds, tissue culture     

CORRECTIONS, VOLUME 29

Part 1, under the frontispiece portrait of Dr. N. B. Eales,the words ‘President 1948–1951’ should havebeen added. Page 103, line 49, for ‘Newton Collection’ read‘Norman Collection (Canon Norman)’. 185, line 37, for ‘capillaris’ read ‘capillacca’. 188, Table 1, for ‘bemoralis’. read ‘nemoralis’. 188, Table 2, for ‘Cochlicella acuta (Müll)? ventrosa(Fér.)’ read ‘Cochlicella ventrosa (Fér.)’. 191, line 24, for ‘araheo-’ read ‘archeo-’.     

Pyrimidine Nucleotide Biosynthesis in Vinca rosea Cells: Changes in the Activity of the de novo and Salvage Pathways during Growth in a Suspension Culture

Marked changes in the activity of the ‘de novo’and ‘salvage’ pathways of pyrimidine biosynthesisduring growth of Vinca rosea cells in a batch suspension culturewere observed. The activity of these pathways was investigated by determiningthe contribution of ¹⁴C of [2-¹⁴Cluracil, 12-¹⁴Cluridine. and[6-¹⁴Clorotate to the cell constituents and by measuring theactivity of the several enzymes of these pathways. During the lag phase of the culture, ‘uracil-’ and‘uridine-salvage’ pathways made the predominantcontribution to nucleotide biosynthesis, but, following theinitiation of cell division, the ‘de novo’ pathwayfor nucleotide biosynthesis operated appreciably. These results suggest that nucleotide synthesis during cellgrowth in a suspension culture can be divided into two stages:a ‘turnover stage’, during the lag phase of cellgrowth, and a ‘true biosynthetic stage’, which isinitiated in the cell division phase.