Regeneration of Tissues in Wounded Stems: A Quantitative Study

When a dicotyledonous stem is wounded by longitudinally splittinga young internode into halves, cells near the cut surface proliferateto form a callus within which vascular tissues differentiateand tend to restore a vascular cylinder in each half. Threephases of regeneration after wounding were identified and quantifiedin stems of three Solanaceous species. (1) In an initial ‘lag’phase, lasting about 2 d, neither cell division nor enlargementwere detected, but mitotic figures were observed within about300 µm of the cut surface. (2) Throughout a second, ‘division’phase, from about days 2–10, cell division and enlargementoccurred. Both were initiated mainly in the two cell layersnearest the surface. A mass of callus formed, with new cellwalls mostly parallel to the surface. Cell enlargement laggedbehind cell division for the first few days, so that mean radialcell diameter decreased until day 6, thereafter remaining almostconstant at 30–40 µm. Towards the end of this phase,mitoses ceased within the callus except in the positions ofthe future vascular and cork cambia, where radial cell diameterfell towards a constant 15–20 µm. (3) During a third,‘differentiation’ phase, cell division was restrictedto the cambial zones, and derivatives differentiated into cork,phloem or xylem according to position. The rate of increasein cell number per transect was 1.5–2.0 cells d^–1,of which more than half was xylem. Capsicum annuum L., sweet pepper, Lycopersicon esculentum Mill., tomato, cambium, cell division, differentiation, regeneration, wounding of stems, xylem     

Induction of a 'Periderm-like' Tissue in Excised Roots of the Fern Ophioglossum petiolatum Hook

Basal applications of plant growth regulators to excised rootsof Ophioglossum induce modifications in the outer cortical cells.Benzyladenine initiates periclinal divisions in these outercells producing a ‘periderm-like’ tissue while 2,4-Dcauses cell enlargement only. Walls of the outer cells of the‘periderm-like’ tissue are resistant to sulphuricacid and stain with Sudan IV and presumably are suberized.     

A Compartment Model of the Effect of Early-season Temperatures on Potential Size and Growth of 'Delicious' Apple Fruits

A compartmental growth model was developed to describe expansionof ‘Delicious’ apple fruit diameter and the effectof early-season temperatures on potential size at harvest. Themodel was based on the assumption that growth may be describedas a function of transfer between two conceptual compartments.Under this scheme, the first compartment represented all tissuecontributing to the setting of potential fruit size (determinedas the integral of its output) whereas the second compartmentrepresented all other fruit tissue whose growth actualized thatpotential. Expansion of both compartments was assumed to havea temperature response with an optimum, whereas an aging processwith an asymptotic temperature response controlled transferto the second compartment. Model parameters were estimated byfitting to data from controlled environment experiments in whichearly-season temperature conditions were varied. Predicted fruitgrowth curves showed close agreement with measured diameterdata. The results were consistent with a two-fold impact ofearly-season temperatures on apple fruit size: an immediate,direct effect on growth rate and an enduring effect, mediatedthrough fruit cell number or resource allocation to young fruit,reflecting the establishment of a potential that subsequentgrowth actualizes.Copyright 1999 Annals of Botany Company. Malus domesticaBorkh., apples, ‘Delicious’, fruit growth, models, temperature, potential size, cell division.     

Apple Fruit Cuticles and the Occurrence of Pores and Transcuticular Canals

Developmental studies were made on the dewaxed thin-sectionedapple fruit cuticles of 10 Malus sylvestris Mill, cultivarsfrom 4 weeks before anthesis through fruit maturation and harvest.Cuticular development appears to correlate well with the generalgrowth of the fruit. However, no correlation exists betweenfruit size and cuticle thickness. Cuticular pores were evidentas early as 1 week before anthesis and transcuticular canalsbecame evident by 1 week following anthesis. Dewaxed thin-sectioned,as well as isolated, mature fruit cuticles of 16 cultivars andfour crab-apples consistently revealed the distinct presenceof ubiquitous pores and canals. Evidence is provided by lightmicroscopy and scanning electron microscopy (SEM). Measurementswere taken of cuticle thicknesses during development and ofcuticular pore dimensions, and calculations were made of poreand canal numbers. Fruit size alone is not directly indicativeof total pore numbers per surface area. Canal lengths are directlyrelated to the developmental thickness of the cuticle. No correlationwas found between the thickness of the mature cuticle and eitherthe number of pores present or the pore diameters. Malus sylvestris Mill, apple fruit, morphology, cuticle, cuticular flanges, cuticular pores, transcuticular canals, ultrastructure     

Histology of Callogenesis and Somatic Embryogenesis Induced in Stem Fragments of Cork Oak (Quercus suber) Cultured In Vitro

Calluses able to produce somatic embryos were formed duringin vitro culture of shoot fragments of cork oak (Quercus suberL.).Histological monitoring of these fragments during cultureshowed that it was the cortical parenchyma cells which underwentdedifferentiation before calluses were formed by repeated divisions.The calluses consisted of parenchyma cells surrounded by a fewlayers of meristematic cells. Proembryos formed in groups aroundthe edge of some calluses. Histological examination showed thatthey were produced by the evolution of two different categoriesof cell: one category had the appearance of ‘embryogenic’cells with very thick walls, a small vacuole rich in starchand a well-developed nucleus with a prominent nucleolus. Theother cells were very bulky with large vacuoles; their morphologywas similar to that of suspensor cells encountered in embryogenesisin gymnosperms. The ontogenic stages were similar to those describedin zygotic embryos of the genus Quercus. Nevertheless, mostof the embryonic structures deviated from normal developmentand at all stages produced secondary proembryos. Cork-oak, Quercus suber L, histology, callogenesis, somatic embryogenesis, embryogenic cells, starch, secondary embryogenesis     

Effect of Applied and Endogenous Auxin on Callus and Root Formation of In Vitro Shoots of the Apple: Rootstocks M26 and A2

The influence of exogenous IBA (indol-3yl-butyric acid) on rootand callus formation was studied in shoots of the apple rootstocksA2 and M26. The shoots grown in vitro were derived originallyfrom meristems of both juvenile and adult trees. Endogenousindol-3yl-acetic acid (IAA) concentrations in leaves and stemswere correlated with the responses to applied IAA. After 30 subcultures shoots from A2 and M26 rooted easily, butA2 did so more readily and even without IBA. Treatment withIBA improved percentage rooting and number of roots in bothrootstocks. Ex-adult and ex-juvenile shoots of A2 formed rootsto the same extent. However, ex-adult shoots of A2 showed ahigher IBA optimum for root number than ex-juvenile A2 and werealso less sensitive to supra-optimal IBA concentrations. Incontrast, in M26, there were no differences between ex-adultand ex-juvenile shoots. The results imply that rooting ability is associated more withdifferences between cultivars than with the origin of the explants.The best rooting occurred in ex-adult shoots of A2 which hadthe lowest endogenous IAA concentration, while callus formationwas correlated with high endogenous auxin concentration. Ex-adultA2 produced almost no callus even after exposure to high IBAconcentrations (25µM) whereas ex-adult M26 formed muchmore callus at 1/10 of the IBA concentration. Malus sylvestris (L.) Mill. var. domestica Borkh., Malus pumila Mill., apple rootstocks A2 and M26, in vitro culture, root and callus formation, HPLC analyses of IAA     

Abortive Meiosis in Plasmodial Pollen Mother Cells of Helianthemum

The anthers of two garden varieties of Helianthemum containnot only normal pollen mother cells but also a large proportionof plasmodia with varying numbers of nuclei of varying nuclearcomplements. These abnormal cells probably resulted from a splitspindle associated with a failure of wall formation during severalpremeiotic mitoses. Chromosome pairing at meiosis is higherthan might be expected and in abnormal tissues such as theseit provides evidence for ‘somatic pairing’ havingtaken place during the premeiotic mitoses. Meiosis stops atthe end of the first division in the plasmodia and althoughit is completed in the normal pollen mother cells viable pollenis never produced.     

Mechanism of Bruise Resistance in Pepino (Solanum muricatum) Fruit

Freshly harvested fruit of two cultivars and 16 selections ofthe pepino (Solanum muricatum Ait.) were given a compressiveloading. Bruise indices, calculated as the product of the resultantbruise length and width, varied significantly among clones.The cultivar ‘Suma’ was most susceptible to bruiseinjury, while selection S15/14 was most resistant. Exocarp cellsand interstitial air-spaces were larger in ‘Suma’than in S15/14. Exocarp cells were compressed, but did not ruptureunder the loading. Mesocarp cells separated from one another,ruptured and collapsed in compressed ‘Suma’ fruit.Mesocarp cells of S15/14 were unaffected by the treatment. Therole of the exocarp in the distribution of mechanical stressis discussed. Pepino, Solanum muricatum Ait, anatomy, bruising, fruit, mechanical stress, plant breeding     

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.     

The Ultrastructure and Analytical Microscopy of Silicon Deposits in the Roots of Saccharum officinarum (L.)

Silicon accumulation in the endodermis of the ‘set’and ‘shoot’ roots of Saccharum officinarum (L.)were investigated by scanning electron microscopy and electron-probemicroanalysis. Silicon microassay was also carried out by meansof the Corinth analytical microscope (CORA). Aggregates arelargely associated with the inner tangential wall (ITW) of theendodermis and their formation is basically similar to thoseseen in Sorghum bicolor (L.) Moench. In contrast to Sorghumthe earliest deposits in Saccharum appear in wall strata wellwithin the cell wall cytoplasm interface. An additional layerof silicon was also located along the endodermal pericycle boundaryextending some distance along the middle lamella of the radialwalls. The results are discussed in relation to those of previous studiesof silicon accumulation in endodermal cells and to possiblefactors affecting such accumulations.     

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.     

Leaf Anatomy, Emphasizing Unusual 'Concertina' Mesophyll Cells, of Two East African Legumes (Caesalpinieae, Caesalpinioideae, Leguminosae)

Cordeauxia edulis (Somalia and Ethiopia), andStuhlmannia moavii(Tanzania, Kenya and Madagascar) are evergreen shrubs or smalltrees of dry areas. They have similar leaf anatomy as revealedby resin sectioning and scanning electron microscopy. The cuticleis extremely thick and all vascular bundles lack bundle sheathextensions. The most unusual feature is the mesophyll, threeto seven layers consisting entirely of cylindrical palisadecells with lateral walls capable of changing vertical lengthby folding in a concertina-like manner. The matching outwardfolds of two adjacent cells always remain attached by meansof a row of wall thickenings (‘pegs’). The pegscan elongate, especially so between the widely separated mesophyllcells that occupy the substomatal chamber area. The unattachedflexible inward wall folds enable these ‘concertina’cells to shorten or lengthen vertically without disrupting cellinterconnections in the interior of each relatively long-livedleaf as it periodically loses and gains water. Concertina cellsmay be an anatomical adaptation allowing these leaves to remainevergreen and survive extended periods of drought and yet tostore water quickly when it becomes available. Leguminosae; Caesalpinioideae; Cordeauxia ; Stuhlmannia ; ‘concertina’ mesophyll cells; desert adaptation; hollow glandular trichomes; leaf anatomy; wall thickenings     

Spontaneous Chromosome Breakage in Lilium Endosperm

Abnormal mitoses, resulting from chromosome breakage and reunion,occurred in Lilium regale and in the hybrid L. ‘PhyllisCox’ 4 weeks after fertilization. The abnormalities appearedto commence with a single chromatid bridge and in ‘PhyllisCox’ abnormalities accumulated resulting in the formationof multiple bridges, ring chromosomes, fragments, dicentricchromosomes, and micronuclei. Failure of anaphase separationfollowed by restitution resulted in the formation of highlypolyploid nuclei in which mass spontaneous chromosome breakageoccurred. The accumulation of abnormalities in the hybrid endosperm reducedthe mitotic activity and culminated in the failure of the endosperm.This led to the death of the otherwise normal embryo. This sequenceof events is probably of wide importance in the failure of interspecifichybridization.     

Variable Cell Lineages form the Functional Pea Epidermis

Evidence was sought for cellular programs and cellular interactionsacting during the formation of stomatal spacing patterns. Dailyreplicas of the surfaces of Pisum sativum leaves were used toreconstruct the cellular development of specific regions ofthe epidermis. During the period studied small primordia becamemature leaves; this involved a 250-fold increase in area anda 20-fold increase in cell number. The earliest event correlatedwith the development of a stoma was an unequal division, andsuch divisions were common in neighbouring and even within thesame cells. A distinct cell lineage started with these unequaldivisions, forming both a stoma and most of the cells that separatedit from its neighbours. Both products of an unequal divisionbecame regular epidermal cells only where such development preventedthe formation of two stomata that would have been in directcontact with one another. Neighbouring stomata often developedand matured together, indicating that there was no mutual inhibitionbetween developing stomata that were more than one cell apart.It is concluded that stomata are products of an intracellularprogram which generates stomatal patterns during rather thanpreceding development. This program can be modified and evenstopped during its entire course, allowing for the correctionof local ‘mistakes’ of stomatal patterning. Cell lineages, cell determination, cellular interactions, epidermal development, garden peas, immature stomata, pattern formation, Pisum sativum, spacing patterns, stomata, unequal divisions     

Morphology and Anatomy of Stems and Pedicels of Spring Flush Shoots Associated with Citrus Fruit Set

Flowering and vegetative shoots of ‘Shamouti’ orange[Citrus sinensis (L.) Osbeck] and ‘Marsh’ seedlessgrapefruit (Citrus paradisi Macf.) were examined for correlationof their morphology and anatomy with fruit set. Fruit set isfavoured on leafy inflorescences whereas abortion is nearlycomplete on leafless inflorescences. Leafless inflorescencesof ‘Shamouti’ with one flower were found to havea very thin stem which contained few vascular bundles, whereasthose with three flowers had better-developed vascular systems.The vascular system of leafy inflorescences is significantlydifferent from that of leafless ones and contains a distinctcentral xylem cylinder. The vascular area of leafless inflorescencesis only about one-quarter of that of the leafy ones. The vascularsystem of grapefruit resembles that of the ‘Shamouti’orange. This study emphasizes the importance of the dimensionof the vascular system for fruit set and provides a possibleexplanation for the better fruit set on both leafy and leaflessinflorescences with several flowers compared with single-floweredinflorescences. Anatomy; citrus; fruit set; leafless inflorescence; leafy inflorescence; pedicel; vascular system; vegetative shoot     

Fruit Position and Fruit Set Sequence in a Truss as Factors Determining Final Size of Tomato Fruits

The position of fruits in a truss and the induction sequenceof fruit set has been examined for the effects on the finalsize of parthenocarpic fruits in tomato. Fruits at two developmentalstages were examined to measure the change of d. wt and theimport of ¹⁴C leaf assimilates. The sequence of induction largely determined the import rateinto fruits of the same truss and first-induced fruits are thereforelarger within the same truss. The position of fruits in thetruss modifies the extent of this ‘primigen dominance’effect. Possible mechanisms for this dominance phenomenon arediscussed. Position, set sequence, size, fruit, Lycopersicon esculentum Mill     

Ethylene Production by Fe-deficient Roots and its Involvement in the Regulation of Fe-deficiency Stress Responses by Strategy I Plants

Species that showed marked morphological and physiological responsesby their roots to Fe-deficiency (Strategy I plants) were comparedwith others that do not exhibit these responses (Strategy IIplants). Roots from Fe-deficient cucumber (Cucumis sativusL.‘Ashley’), tomato (Lycopersicon esculentumMill.T3238FER) and pea (Pisum sativumL. ‘Sparkle’) plantsproduced more ethylene than those of Fe-sufficient plants. Thehigher production of ethylene in Fe-deficient cucumber and peaplants occurred before Fe-deficient plants showed chlorosissymptoms and was parallel to the occurrence of Fe-deficiencystress responses. The addition of either the ethylene precursorACC, 1-aminocyclopropane-1-carboxylic acid, or the ethylenereleasing substance, Ethephon, to several Fe-sufficient StrategyI plants [cucumber, tomato, pea, sugar beet (Beta vulgarisL.),Arabidopsis(Arabidopsis thaliana(L.) Heynh ‘Columbia’), plantago(Plantago lanceolataL.)] promoted some of their Fe-deficiencystress responses: enhanced root ferric-reducing capacity andswollen root tips. By contrast, Fe-deficient roots from severalStrategy II plants [maize (Zea maysL. ‘Funo’), wheat(Triticum aestivumL. ‘Yécora’), barley (HordeumvulgareL. ‘Barbarrosa’)] did not produce more ethylenethan the Fe-sufficient ones. Furthermore, ACC had no effecton the reducing capacity of these Strategy II plants and, exceptin barley, did not promote swelling of root tips. In conclusion,results suggest that ethylene is involved in the regulationof Fe-deficiency stress responses by Strategy I plants.Copyright1999 Annals of Botany Company. Arabidopsis (Arabidopsis thaliana(L.) Heynch), barley (Hordeum vulgareL.), cucumber (Cucumis sativusL.), ethylene, iron deficiency, maize (Zea maysL.), pea (Pisum sativumL.), plantago (Plantago lanceolataL.), ferric-reducing capacity, sugar beet (Beta vulgarisL.), tomato (Lycopersicon esculentumMill.), wheat (Triticum aestivumL.).     

Ultrastructure of the Cell Wall of Vessel Contact Cells in the Xylem of Tomato Stems

Mature contact cells in the metaxylem of tomato stems are characterizedby a distinctive wall deposit occurring mainly over the areaof the pit membrane that separates the cell from an adjoiningvessel member but also extending as a layer on the secondarywall round the interior of the cell. Cytochemical tests showthe layer contains a low concentration of pectin, no lignin,and a high concentration of polysaccharides. It can be distinguishedfrom the primary and secondary walls by quantitative differencesin the cytochemical staining reactions and by its loose-texturedappearance under the electron microscope. The layer is similarin composition and structure to the ‘protective layer’described in woody plants. The layer has also been found inpea and cotton and may be a characteristic feature of the contactcells in herbaceous plants. Cell wall ultrastructure, contact cells, Lycopersicon esculentum, Mill., protective layer, tomato, xylem     

Ontogenesis, Differentiation and Precocious Germination in Anther-derived Somatic Embryos of Grapevine (Vitis vinifera L.): Proembryogenesis

Histological steps of callogenesis and proembryogenesis in anthercultures ofVitis vinifera L. ‘Grenache noir’ aredescribed. Embryogenic calli were obtained on Nitsch and Nitschmedium supplemented with 1mgl^-12,4-dichlorophenoxyacetic acid(2,4-D) and 0.25mgl^-1benzylaminopurine (BAP). Calli were initiatedfrom anther connective cells only and no division of microsporesoccurred. The embryos were hence of somatic origin. Proembryosdeveloped either directly (i.e. without intervening callus)from the endothecium, or indirectly from the connective-derivedcallus. In both cases, proembryos originated from single cells.They developed from starchy differentiated cells of a predeterminedtype. The polarity of the somatic proembryo was establishedfrom the first divisions and it was marked by precocious developmentof an easily recognizable suspensor. Other analogies with thedevelopment of the zygote are also emphasised. Vitis vinifera L.; grapevine; somatic embryogenesis; proembryogenesis; histology     

Alteration of the Cell Surface during the Vegetative Cell Cycle of the Unicellular Green Alga Chlamydomonas reinhardtii

Alterations of the cell surface during the vegetative cell cycleof the unicellular green alga Chlamydomonas reinhardtii wereinvestigated using polyclonal antibodies against the purifiedand subsequently deglycosylated insoluble cell wall componentand against a 100 kDa polypeptide of the deglycosylated, chaotrope-solublewall fraction, respectively. Both antibodies recognized epitopeswithin the non-glycosylated domains of a ‘150 kDa’chaotrope-soluble glycoprotein (=GP3B) localized in the outerlayers of the C. reinhardtii cell wall. Immunofluorescence studiesindicated that both antibodies reacted with the surface of ‘late’sporangia (harvested 1 h before liberation of the zoospores),but not with the cell surfaces of released zoospores, growingcells and young sporangia, respectively. After pretreatmentwith aqueous LiCl, however, the cell surfaces of zoospores,growing cells and young sporangia became accessible to theseparticular antibodies. Highly purified preparations of the insolublewall fraction revealed strong immunofluorescence with both antibodiesbut not with the corresponding preimmune sera. Based on thesedata, we concluded that the antigenic sites of the insolubleglycoprotein framework of the C. reinhardtii wall are maskedby LiCl-soluble glycoproteins in single cell stages and youngsporangia, but not or to a lesser extent in the case of themother walls of ‘late’ sporangia. The conclusionwas supported by findings that (I) the multilayered structureof the mother-cell wall was disturbed in ‘late’,but not in young sporangia and that (II) the amounts of chaotropesolublecell wall glycoproteins present in the LiCl-extracts from intactsporangia decreased during ripening of the sporangia. (Received January 10, 1996; Accepted May 27, 1996)