Ultrastructural changes in the cell walls of ripening apple and pear fruit

Ultrastructural changes in the cell walls of “Calville de San Sauveur” apples (Malus sylvestris Mill) and “Spadona” pear (Pyrus communis L.) fruit were followed during ripening. In apple, structural alterations in cell walls became apparent at advanced stages of softening and showed predominantly dissolution of the middle lamella. In pears softening was also associated with the dissolution of the middle lamella, and in addition a gradual disintegration of fibrillar material throughout the cell wall. In fully ripe fruit almost all of the fibrillar arrangement in the cell wall was lost. Application of enzyme solutions containing polygalacturonase and cellulase to tissue discs from firm pear fruit led to ultrastructural changes observed in naturally ripening pears. In apple polygalacturonase alone was sufficient to dissolve the middle lamella region of the cell walls, as was also found to occur in naturally ripening fruit. In both apple and pear the cell wall areas containing plasmodesmata maintained their structural integrity throughout the ripening process. At advanced stages of ripening vesicles appeared in the vicinity of plasmodesmata.     

Dehiscence of Fruit in Oilseed Rape (Brassica napus L.): I. ANATOMY OF POD DEHISCENCE

The development sequence of anatomical changes taking placewithin the pericarp tissues of Brassica napus siliquae havebeen studied at a fine- and ultra-structural level. Tissue differentiationoccurred during the initial 20 d after anthesis (DAA) and allowedthe identification of dehiscence zone cells. This descrete tissuewas subsequently further delineated by extensive lignificationof adjacent valve edge and replar vascular cells. Concomitantwith the onset of pericarp lignification, cytoplasmic contentsof the thin-walled dehiscence zone cells exhibited progressivesenescence and degradation. Wall breakdown, initially evidentin pods by 60 DAA, exclusively affected cells within the dehiscencezone, and eventually extended throughout this tissue from theepidermal suture to the locule, thus precipitating valve detachment.Ultrastructural examination confirmed that this loss of cellularcohesion was primarily attributable to middle lamella degradationand, furthermore, the dissolution of wall material was apparentlydependent on rupture of the dehiscence zone protoplast. Thesignificance of dehiscence zone cell modifications in relationto autolytic cell wall breakdown, together with possible implicationsfor the regulation of pod shatter, are discussed. Key words: Oilseed rape, Brassica napus, pod shatter, dehiscence zone, cell wall breakdown     

Measuring Cell Wall Dimensions using the Distance Transform

A method has been developed to measure various features of cellsand cell walls. It offers possibilities for quantitative analysisof microscopic images which were not feasible before. The methodis not based on a model and the only assumption required isthat the middle lamella is exactly in the middle of the cellwall of two neighbouring cells. Intercellular spaces are handledcorrectly with this method. Features can be measured both per individual cell and per frame.It is also possible to measure the variation of the cell wallwithin a single cell. Hence the variation of the cell wall thicknesswithin cells can be compared to variation between cells. The method is demonstrated for Gerbera but is generally applicableto anatomical studies of plant tissues.Copyright 1995, 1999Academic Press Gerbera jamesonii Bolus ex Adlam, cell wall, image analysis, cultivar, quantitative microscopy, distance transform     

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.     

Development, Ultrastructure and Secretion of Gum Ducts in Lannea coromandelica (Houtt.) Merrill (Anacardiaceae)

In Lannea coromandelica (Houtt.) Merrill, the gum ducts in theprimary phloem of the stem initiate and develop schizogenously.Formation of an intercellular space amongst a group of denselystained phloem procambial cells signals the initiation of aduct. Initiation of a duct starts by dissolution of the middlelamella of the walls. It widens by separation of cells alongthe radial walls by swelling and dissolution of the middle lamella.During separation of radial walls dictyosomes and paramuralbodies are observed in the peripheral cytoplasm at the siteof dissolution. Plasmodesmata occur in the radial and innertangential walls of epithelial cells of developing gum ducts.The epithelial cells have a dense cytoplasm and contain roughendoplasmic reticulum, ribosomes, polysomes, mitochondria withswollen cristae, plastids with poorly developed membranes, dictyosomesand vesicles. Dictyosomes and rough endoplasmic reticulum seemto be involved in the secretion of the gum. The polysaccharidegum constituents apparently originate from the outer wall layersof the epithelial cells. Following gum secretion, the epithelialcells degenerate. Lannea coromandelica (Houtt.) Merrill, Anacardiaceae, gum ducts, ultrastructure, gum secretion     

Localization of Silicon in Specific Cell Wall Layers of the Stomatal Apparatus of Sugar Cane by Use of Energy Dispersive X-ray Analysis

By use of combined light, transmission electron, and scanningelectron microscopy with energy dispersive analysis of X-rays,silicon was localized in specific cell wall layers of the stomatalapparatus of sugar cane. The silicon occurs beneath the cuticlein layers of wall which correspond to the middle lamella andfirst-formed primary wall region of interior plant cells. Inother plants these regions are also reported high in pectinsand hemicellulose. It is suggested that the silicon may interactwith the pectins and hemicellulose and also with the ligninsand phenolics which may also be deposited in these wall layers. Saccharum officinarum L, sugar cane, stomata, silicon, X-ray analysis     

Endosperm Development in Solanum nigrum L. Formation of the Zone of Separation and Secretion

Endosperm development in Solanum nigrum was ab initio cellular.During early seed development, a Zone of Separation and Secretion(ZSS) differentiated within the endosperm. There were threephases in the formation of the ZSS. Phase I—stainabilityof the cell walls and middle lamella increased followed by numeroussmall plasmalemma invaginations (blebs) which became filledwith a fibrillar material. Phase II—the plasmalemma withdrewfrom the cell wall as a fibrillar lipo-carbohydrate matrix accumulatedoutside the plasmalemma. The middle lamella was gradually removedfrom between the cells forming the central axis of the cone.Phase III—the lipo-carbohydrate matrix continued to accumulateoutside the plasmalemma and also within the developing intercellularspaces. Some axial cells were completely separated from theremaining ZSS cells and became embedded in the matrix. The formationof the ZSS did not entail the destruction of the endosperm cellsand cell divisions were frequent. The ZSS was initially cone-shaped,capping the globular embryo. As the embryo sac enlarged, theZSS continued to differentiate. This resulted in a narrow curvedcorridor through the peripheral region of the endosperm whichterminated above the vascular trace. The embryo grew throughthe centre of the ZSS and pushed aside the separated axial cells.The ZSS facilitated the growth of the embryo through the restof the endosperm.Copyright 1993, 1999 Academic Press Lipo-carbohydrate matrix, extracellular matrix, endosperm development, Solanum nigrum, Zone of Separation and Secretion     

Pollen--stigma Interactions: Development and Cytochemistry of Stigma Papillae and their Secretions in Annona squamosa L. (Annonaceae)

The development and cytochemical features of the stigma andstyle have been investigated in Sugar apple, Annona squamosaL., using light and electron microscopy. The pistil is a syncarpwith an open stylar canal. Papillae of epidermal origin lineboth the surface of the stigma and the inner face of the stylarcanal. The papillae contain organelles characteristic of secretorycells with a highly thickened cellulosic wall. The wall is multi-layered,the zones differing in their microfibrillar stacking and orientation.The stigma is of the ‘wet’ type and the surfaceexudate is heterogeneous in microscopic appearance and reactscytochemically for proteins, carbohydrates and lipids. The surfacecuticle undergoes dissolution prior to anthesis. A secretionalso appears in the thickened middle lamella of the sub-epidermalcell layer which reacts cytochemically for pectinaceous acidicpolysaccharides. Esterase activity of papillae is indicative of the receptiveareas, and it is also related to the onset of receptivity. Acidphosphatase activity is intense in the sub-epidermal cell layerswhich probably reflects their secretory activity. Pollinationtriggers a copious flow of secretion onto the stigma surfacewhich engulfs the pollen grains. It appears that most of theacidic polysaccharides of this secretion come from the middlelamella of the sub-epidermal cell layer. Compatible pollen tubes have no apparent barriers to overcomeon their route to the embryo sac and the inherent protogynousdichogamy seems to control the acceptance or rejection of compatiblepollen. Annona squamosa L., sugar apple, stigma, style, secretions     

On Vessel Member Differentiation in the Bean (Phaseolus vulgaris L.)

Certain ultrastructural features of vessel member differentiationwere examined in the primary xylem of petiole of bean (Phaseolusvulgaris L.). The cells used had helical secondary wall thickeningsand simple perforation plates. The primary cell wall increasesin thickness before the helices of secondary wall develop. Ina common wall between two vessel members of different ages,theprimary thickening occurs first in the older cell so thatfor a time the middle lamella is located closer to the youngercell rather than medianly. Apparently the helix is depositedafter the primary wall of a given cell reaches maximum thickness.The perforation of the end wall is preceded by primary thickeningof the part of the wall that is later removed. The marginalregion remains relatively thin and becomes covered with a rimof secondary wall. Vesicles with fibrous content appear nearthe surface within the end wall shortly before the perforationoccurs. A highly vacuolated protoplast with a much enlargednucleus and numerous organdIes is present during cell wall differentiation.After that process is completed, the protoplast disintegratesand the primary wall bearing the helix is hydrolysed where itis exposed to the cell lumen and, under certain conditions,also under the secondary wall.     

Development of the Sieve Plate in Saxifraga sarmentosa L.

The differentiating sieve plate in the phloem of the stolonof Saxifraga sarmentosa L. was studied with the electron microscope.Development of the pore site begins with differentiation ofa pair of collar-like areas around the plasmodesma which canbe seen in the youngest identifiable sieve plates. Further growthof the collars occurs by deposition of an amorphous substance,presumably caflose. Although the growth of the collars is simultaneouswith the growth of the surrounding cell wall it is rapid atfirst and the pore sites appear asdome-shaped protuberances.It also involves deposition of callose over an increasinglywider area of the cell wall and since the thickening of thenormal cell wall continues only where notcovered by callose,the collars assume a conical form. There seems to be no displacementor lysis of normal cell wall material during growth of the collars.Eventually the growth of the cell wall in thickness overtakesthe pore sites so that when the growth of the cell wall is completethe pore sites appear as depressions in the sieve plate. Theperforation of a pore site is accomplished by widening of theplasmodesmatal cylinder which begins at the middle lamella byremoval of callose. Endoplasmic reticulum is found in closeproximity to the plasmodesma andis believed to penetrate it.     

An Electron Microscope Investigation into the Structure of the Floridean Pit

Ultra-thin sections of the red algae Rhodymenia palmata andLaurencia caespitosa were examined in the electron microscopein order to determine the fine structure of the Floridean pits.The pits found in the two species examined are shown to be quitedifferent from each other, in Rhodymenia the pit is open sothat the cytoplasm is continuous through the cell wall, whilstin Laurencia the pit is completely closed by a lamella of thecell wall. These two types are shown to be equivalent to thosedescribed by Jungers (1933) on the basis of light microscopeobservations.     

Tensile growth stress and lignin distribution in the cell walls of yellow poplar, Liriodendron tulipifera Linn.

Five specimens that contained a continuous gradient of wood, from normal to tension wood regions, were collected from an inclined yellow poplar (Liriodendron tulipifera), and the released strain of tensile growth stress was quantified. Ultraviolet (UV) microspectrophotometry was used to examine the relationship between lignin distribution in the cell wall and the intensity of tensile growth stress. The UV absorption of the secondary wall and the cell corner middle lamella decreased with increasing tensile released strain (i.e., tensile growth stress). The UV absorption in the compound middle lamella region remained virtually constant, irrespective of the tensile released strain. The absorption maximum (5_max) remained virtually constant in the secondary wall, the cell corner middle lamella, and the compound middle lamella region at 273-274, 277-278, and 275-278 nm respectively, irrespective of the tensile released strain. The ratios of the UV absorbance at 280 to 260 nm and 280 to 273 nm of the secondary wall decreased with increasing tensile released strain. The ratios in the cell corner and compound middle lamella region remained constant, irrespective of the tensile released strain. The lignin content of the secondary wall decreased, while the syringyl/guaiacyl ratio increased with increasing tensile released strain. Gelatinous fibers were not observed in the tension wood regions, but the secondary wall became gelatinous-layer-like, i.e., the lignin content and microfibrillar angle decreased and the cellulose content increased. A definite gelatinous layer seems to be important for generating greater tensile growth stress. It is concluded that a decrease in lignin and an increase in cellulose microfibrils parallel to the fiber axis in the secondary wall are necessary to produce large tensile growth stress.     

Cellulase, Fruit Softening and Abscission in Red RaspberryRubus idaeusL. cv Glen Clova

The ripening of raspberry fruit (Rubus ideausL. cv Glen Clova)is associated with a climacteric rise in ethylene production.As the fruit pigments change from green to red there is a progressivesoftening, loss of skin strength and a breakdown of cell wallsin the mesocarp. An increase in cellulase (endo-1,4-ß-D-glucanase)in both drupelets and receptacles accompanies these changes.The localization of cellulase in the regions of the fruit associatedwith abscission zones suggest the enzyme may be involved infruit separation as well as softening. Rubus idaeusL; raspberry; fruit ripening; ethylene; abscission; cell wall breakdown; cellulase; endo-1,4-ß-D-glucanase     

The Role of Cell Expansion in the Abscission of Impatiens sultani leaves

The histological events occurring during the latter stages ofabscission were followed continuously in longitudinal slicesthrough the petiole base of Impatiens sultani Hook. It appearsthat the middle lamella of the cortical parenchyma cells isdegraded first. This is followed by an expansion of these cellsand a concomitant stretching and separation of the collenchymaand vascular trace. The parenchyma cells continue to enlargeuntil they become virtually spherical, a process which finallyruptures the last restraining xylem vessels. The increased volumeof the parenchyma cells appears to be principally due to a conformationalchange in cell shape from a near regular hexagonal prism toa sphere of similar surface area. The dimensions of the prismaticcells are such that most of them change into spheres whose diametersare the same as the transverse distance between the oppositesides of the six axially orientated faces of the prisms. Cellularexpansion is thus entirely directed along the axis of the petiole.The significance of these observations to the general anatomyand mechanism of fracture of abscission zones is discussed. Impatiens sultani Hook., abscission, cell expansion, cell wall degradation, cell shape     

An ultrastructural study of acid phosphatase localization in Phaseolus vulgaris xylem by the use of an azo-dye method.

The localization of acid phosphatase during xylem development has been examined in the bean, Phaseolus vulgaris. The azo dye, the final reaction product, is initially prominent in the dictyosomes, vesicles apparently participating in secondary wall formation, and in the middle lamella of the young vessel element. Final reaction particles are also present in mitochondria, chloroplasts, and certain vacuoles and are sparsely scattered in the cytoplasm. At a later stage of vessel differentiation, the azo dye is concentrated in the disintegrating cytoplasm and along the fibrils of the partially hydrolysed primary wall and middle lamella. In the mature vessel element, the azo dye is still present along the disintegrated primary wall at the side of the vessel and covers the secondary wall. In the parenchyma cell adjacent to the vessel element, acid phosphatase localization is found in the dictyosomes, endoplasmic reticulum, mitochondria, small vacuoles, and the middle lamella. The controls from all stages of vessel element development were free of azo dye particles. The concentration of acid phosphatase along the secondary walls of the mature vessels and in the middle lamella between other cells indicates that this enzyme has other functions besides autolysis of the cytoplasm and primary cell wall. Acid phosphatase may participate in the formation of the secondary wall and may also have a role in the secretion and transport of sugars.     

Ultrastructure of tomato fruit ripening and the role of polygalacturonase isoenzymes in cell wall degradation

Ultrastructural changes in the pericarp of tomato (Lycopersicon esculentum Mill) fruit were followed during ripening. Ethylene production was monitored by gas chromatography and samples analyzed at successive stages of the ripening process.

Changes in the cytoplasmic ultrastructure were not consistent with the suggestion that ripening is a `senescence' phenomenon. A large degree of ultrastructural organization, especially of the mitochondria, chromoplasts, and rough endoplasmic reticulum, was retained by ripe fruit.

Striking changes in the structure of the cell wall were noted, beginning with dissolution of the middle lamella and eventual disruption of the primary cell wall. These changes were correlated with appearance of polygalacturonase (EC 3.2.1.15) isoenzymes. Application of purified tomato polygalacturonase isoenzymes to mature green fruit tissue duplicated the changes in the cell wall noted during normal ripening. Possible roles of the polygalacturonase isoenzymes in cell wall disorganization are discussed.

     

Alfalfa Stem Tissues: Cell-wall Development and Lignification

Alfalfa stems contain a variety of tissues with different patternsof cell-wall development. Development of alfalfa cell wallswas investigated after histochemical staining and with polarizedlight using light microscopy and scanning electron microscopy.Samples of the seventh internode, from the base of stems grownon cut stems, were harvested at five defined stages of developmentfrom early internode elongation through to late maturity. Internodeseven was elongating up to the third sample harvest and internodediameter increased throughout the entire sampling period. Chlorenchyma,cambium, secondary phloem, primary xylem parenchyma and pithparenchyma stem tissues all had thin primary cell walls. Pithparenchyma underwent a small amount of cell-wall thickeningand lignification during maturation. Collenchyma and primaryphloem tissues developed partially thickened primary walls.In contrast to a recent report, the formation of a ring shaped,lignified portion of the primary wall in a number of cells inthe exterior part of the primary phloem was found to precedethe deposition of a thick, non-lignified secondary wall whichwas degradable by rumen microbes. In numerous xylem fibres fromthe fourth harvest date onwards, an additional highly degradablesecondary wall layer was deposited against a previously depositedlignified and undegradable secondary wall. The pattern of lignificationobserved in alfalfa stem tissues suggests that polymerizationof monolignols by peroxidases at the luminal border of the primarycell wall creates an impermeable zone which restricts lignificationof the middle lamella region of tissues with thick primary walls.Copyright1998 Annals of Botany Company Alfalfa,Medicago sativaL., stem tissue, cell wall, development, lignification, degradation.     

杜仲次生木质部分化过程中木质素与半纤维素组分在细胞壁中分布的动态变化

利用紫外光显微镜、透射电子显微镜结合免疫胶体金标记,研究了杜仲（Eucommia ulmoides Oliv.)次生木质部分化过程中木质素与半纤维素组分（木葡聚糖和木聚糖）在细胞壁分布的动态变化。在形成层及细胞伸展区域,细胞壁具有木葡聚糖的分布,而没有木聚糖和木质素沉积,随着次生壁S1层的形成,木质素出现在细胞角隅和胞间层,木聚糖开始出现在S1层中,此时木葡聚糖则分布在初生壁和胞间层;随着次生,壁S2层及S3层的形成和加厚,木质逐逐步由细胞角隅和胞间层扩展到S1、S2和S3层,其沉积呈现出不均匀的块状或片状沉积模式,在次生壁各层形成与其木质化的同时,木聚糖逐渐分布于整个次生壁中,而木糖聚糖仍局限分布于初生壁和胞间层。结果表明,随着细胞次生壁的形成与木质化,细胞壁结构发生较大变化。细胞壁的不同区域,如细胞角隅、胞间层、初生壁和次生壁各层,具有不同的半纤维素组成,其与木质等细胞壁组分结构构成不同的细胞壁分子结构。     

Characteristics of the Deposition of Microfibrils during Formation of the Polylamellate Walls in the Coenocytic Green Alga Chamaedoris orientalis

Characteristics of the deposition of cellulose microfibrilsduring formation of polylamellate walls and the arrangementof cortical microtubules in the tip-growing bipolar cells ofChamaedoris orientalis were examined by replica preparationmethods and indirect immunofluorescence microscopy. The polylamellatewall is made up of two or three kinds of wall lamella whichdiffer in terms of the orientation of microfibrils. Individuallamellae were periodically initiated one after another fromthe pole that was situated exactly at each growing apex of thecell and they were deposited basipetally. The orientation ofmicrofibrils in each lamella was constant during deposition.Microfibrils in different lamellae were deposited at the sametime through the cell wall but the timing of the depositionwas staggered between neighboring lamellae so that the microfibrilswould not be interwoven. By contrast, cortical microtubuleswere persistently arranged longitudinally all over the celland no focal points to which they converged helically were visible,even around the cell apices. The mechanisms that regulate theformation of the polylamellate wall are discussed and a modelfor interpreting the involvement of the cortical microtubulesin such mechanisms is proposed. (Received July 31, 1989; Accepted January 27, 1990)     

Dynamic Changes in Distribution of Lignin and Hemicelluloses in Cell Walls During Differentiation of Secondary Xylem in Eucommia ulmoides (in English)

The dynamic changes in the distribution of lignin and hemicelluloses (xylans and xyloglucans) in cell walls during the differentiation of secondary xylem in Eucommia ulmoides Oliv. were studied by means of ultraviolet light microscopy and transmission electron microscopy combined with immunogold labelling. In the cambial zone and cell expansion zone, xyloglucans were localized both in the tangential and radial walls, but no xylans or lignin were found in these regions. With the formation of secondary wall S1 layer, lignin occurred in the cell corners and middle lamella, while xylans appeared in S1 layer, and xyloglucans were localized in the primary walls and middle lamella. In pace with the formation of secondary wall S2 and S3 layer, lignification extended to S1, S2 and S3 layer in sequence, showing a patchy style of lignin deposition. Concurrently, xylans distributed in the whole secondary walls and xyloglucans, on the other hand, still localized in the primary walls and middle lamella. The results indicated that along with the formation and lignification of the secondary wall, great changes had taken place in the cell walls. Different parts of cell walls, such as cell corners, middle lamella, primary walls and various layers of secondary walls, had different kinds of hemicelluloses, which formed various cell wall architecture combined with lignin and other cell wall components.