STIGMA OF NICOTIANA: ULTRASTRUCTURAL AND BIOCHEMICAL STUDIES

The aim of the present study has been to elucidate further the cytology of the glandular stigma of Nicotiana tabacum, the mechanism of the secretory process during stigma development and the biochemical composition of the exudate. The stigma consists of two distinct zones: a glandular zone formed by the papillae and 2–3 layers of cells (basal cells) immediately below them, and a non-glandular region formed by vacuolated cells which are in continuity with the transmitting tissue. The stigmatic exudate is a complex mixture of different chemical compounds such as proteins, saccharides, fatty acids and phenols. The role of stigma secretion is discussed in relation to pollen activation, recognition and pollen tube growth.     

The Structure and Cytochemistry of the Pistil of Sternbergia lutea (Amaryllidaceae)

Studies were carried out on structural and cytochemical aspectsof the pistil of Sternbergia lutea (L.) KerGawl. The stigmais of the wet papillate type; the papillae are unicellular andare arranged densely around the rim of a funnel-shaped stigma.The stigma exudate is limited and is confined to the bases ofthe papillae and the inner lining of the stigma. The papillaeare smooth in the distal part and are covered with intact cuticle-pelliclelining. The cuticle is disrupted at places towards the baseof the papillae releasing the exudate. The exudate is rich inpectins and other polysaccharides but poor in proteins and lipids.The papillae show dense cytoplasmic profiles with extensiveendoplasmic reticulum (ER), abundant mitochondria, polyribosomesand active dictyosomes. The style is hollow. The stylar cavityis surrounded by two to four layers of glandular cells. In theyoung pistil the canal is lined with a continuous cuticle, butin the mature pistil the cuticle becomes disrupted and the canalis filled with the secretion produced by the cells of the surroundingglandular tissue. Ultrastructurally, the cells of the glandulartissue are very similar to the stigmatic papillae. The innertangential wall of the cells bordering the canal is uniformlythicker than other walls. The secretion in the stylar canal,as well as the intercellular spaces of the glandular tissue,stain intensely for pectins and polysaccharides but poorly forproteins and lipids. Pollen tubes grow through the stylar canal.Structural and cytochemical details of the pistil of Sternbergiaare compared with other hollow-styled systems. Pistil, Sternbergia lutea (L.) Ker-Gawl., stigma and style, structure and cytochemistry     

Structural and Cytochemical Characteristics of the Stigma and Style in Vitis vinifera L. var. Sangiovese (Vitaceae)

Studies were carried out on structural and cytochemical aspectsof the stigma and style ofVitis vinifera . The stigma is ofthe wet papillate type with a continuous cuticle and pellicle.During the development of the papillae, the cell walls increasein thickness and produce a secretion product constituted oflipids that pass through the wall forming the exudate. The styleis solid with a central core of transmitting tissue which hasconspicuous intercellular spaces that increase remarkably fromthe periphery to the centre where the cuticle is present. Theintercellular spaces, where the pollen tubes grow, contain amatrix that includes polysaccharides, pectic substances andscattered areas of lipidic nature. Cytochemistry; stigma; style; ultrastructure; Vitis vinifera     

The dry and wet stigmas ofPrimula obconica: Ultrastructural and cytochemical dimorphisms

Summary The stigmatic papillae of the distylous speciesPrimula obconica are studied by means of cytochemical, light and electron microscopic techniques. The papillae on thrum stigmas are smaller than those on pin stigmas. At the bud stage, secretory vesicles are not a conspicious part of the cytoplasm, although certain signs of secretory activity are present. The young papillae bear the thin, superficial pellicle typical to dry stigmas. Small vesicles are numerous in mature papillae of both morphs, and seem to originate from the ER. A layer of closely packed, osmiophilic globuli is present in the outermost part of mature walls of pin papillae. At sites with cuticle disruption, the globuli seem to migrate outwards to be incorporated with the copious, blistery exudate. Due to this exudate the pin stigma is characterized as wet. Cytochemical tests suggest that the exudate contains mainly lipids, and different carbohydrates and protein are detected. It reacts positively in tests for peroxidase, acid phosphatase, and non-specific esterases. The thrum stigma remains dry at maturity, with a distinct pellicle also reacting positively to the enzyme tests. Only a few, scattered osmiophilic globuli, sized and situated as those in the pin papillae walls, are found in the thrum walls, and they do not form a proper layer.Thus the generally accepted correlation between dry stigmas and the sporophytic kind of self-incompatibility system is not substantiated withinP. obconica, and the possible influence of the dimorphisms to the pollen/stigma interaction is discussed.     

Pollination and Pollen-pistil Interaction in Oil Palm, Elaeis guineensis

Structural and cytochemical aspects of the pistil and detailsof pollination and pollen-pistil interaction were investigatedin the African oil palm (Elaeis guineensis Jacq.), an importantperennial oil crop. The stigma is trilobed, wet and papillate.The branched papillae are confined to a narrow linear zone oneach stigmatic lobe. Each stigmatic lobe harbours a deep stigmaticgroove, which runs adaxially along the surface. The stigmaticgroove is bordered by a well-defined layer of glandular cells,each of which has a pectinaceous cap on the inner tangentialwall. The style is hollow. The canal cells show thickeningson the inner tangential wall. The stigmatic groove and stylarcanal contain an extracellular matrix secreted by the canalcells which is rich in proteins, acidic polysaccharides andpectins. The canal cells at the base of the style are papillateand loosely fill the stylar canal. The stigma becomes receptivewhen the stigmatic lobes separate, and remains so for 24 h.Pollination is mediated by weevils as well as by the wind. Undernatural conditions the pollination efficiency was 100%. Pollinationinduces additional secretion in the stigmatic groove and stylarcanal. During post-pollination secretion, the pectinaceous capsof the cells lining the stigmatic groove are degraded. Pollengrains germinate on the stigmatic papillae and tubes grow onthe surface of the papillae, entering the stigmatic groove andadvancing along it into the stylar canal to eventually gainaccess to the locules. Pollen tubes are seen in the ovules 18–20h after pollination. Copyright 2001 Annals of Botany Company Arecaceae, Elaeis guineensis, African oil palm, pollination, stigmatic grove, stylar canal, Tenera hybrid, weevil     

Papillar integrity as an indicator of stigmatic receptivity in kiwifruit (Actinidia deliciosa)

Stigmatic receptivity is a major factor limiting fruit set inkiwifruit (Actinidia deliciosa). The aim of this work was toknow what determines the cessation of stigmatic receptivityin this species. Stigmatic receptivity has been analysed inkiwifruit through the capacity of the stigma to sustain pollengermination and has been related to stigmatic development anddegeneration.The stigma of kiwifruit flowers has a papillatesurface which from anthesis is covered with an abundant exudate.Papillae are unicellular and contain a number of phenolic deposits.During the lifetime of the flower these papillae gradually loseturgidity, while stigmatic secretion increases until 5 d afteranthesis when papillae start rupturing and the papillar contentis liberated into the germination medium. Stigmatic receptivityis high at anthesis and lasts for the next 4 d. However, itdrastically decreases 5 d after anthesis and it is nil 2 d later.The pattern of stigmatic receptivity closely fits that of papillarintegrity, indicating that stigmatic receptivity relies on thisintegrity. Since papillar integrity can easily be evaluatedin stigmatic arms, this criterion can be used as a quick methodto estimate stigmatic receptivity. Key words: Actinidia deliciosa, kiwifruit, stigma, receptivity, pollen     

STIGMA OF SOLANUM TUBEROSUM CV SHEPODY: MORPHOLOGY,ULTRASTRUCTURE,AND SECRETION

The stigma of Solarium tuberosum L. cv Shepody has a bilobed papillate surface covered with a viscous secretion at anthesis. The secretion originates as osmiophilic droplets in the cytoplasm, accumulates in the intercellular space, and fills the base of the papillae, after lifting and rupturing the cuticular layer covering the stigma surface. Cytochemical evidence shows that the stigmatic secretion is lipidic in nature; it did not stain with the periodic acid-Schiffs reaction for carbohydrate or Coomassie Brilliant Blue R-250 for proteins, but did stain with Sudan black or Nile red, a fluorescence probe for lipids. Sodium dodecylsulfate-polyacrylamide gel electrophoresis has revealed that the secretion contained three polypeptides that appeared to be 'indigenous' to the stigmatic secretion, and not contaminants. Comparative analyses of this and other proteins found in stigma secretions may provide clues to their possible roles in pollen grain adhesion and germination.     

Ultrastructure and histochemistry of Citrus limon (L.) stigma

Citrus limon has a wet stigma which can be divided in two zones: a glandular superficial one formed by papillae, and a non-glandular one formed by parenchymatic cells. The stigmatic exudate is produced by the papillae after the latter have reached their ultimate size. The papillae of the mature pistil are of varying size and composition. Both the unicellular and multicellular ones are present. The cells at the base of the papillae are rich in cytoplasm, whereas the tip cells are vacuolated. Histochemical analysis has shown that the exudate of Citrus is composed of lipids, polysaccharides, and proteins. Our results indicate that the lipidic component is produced and secreted first, followed by production and secretion of the polysaccharidic component. The lipidic component of the exudate is produced in the basal papillae cells and accumulates as droplets in dilated parts of the smooth endoplasmic reticulum (SER). Subsequently the lipid droplets are transported to the plasma membrane, and transferred by the latter into the cell walls. Then the exudate component is accumulated in the intercellular spaces and in the middle lamellar regions of the walls. Subsequently, the polysaccharidic component of the exudate is produced and secreted by the tip cells of the papillae.Abbreviations RER rough endoplasmic reticulum - SER smooth endoplasmic reticulum     

Developmental Aspects of Ultrastructure and Histochemistry of the Stylar Transmitting Tissue of Nicotiana sylvestris

The structure and histochemistry of the solid style of Nicotiana sylvestris Speg. and Comes have been studied by light and electron microscopy. The transmitting tissue develops large intercellular spaces filled with secretions rich in proteins and carbohydrates during maturation. The cells possess large nuclei, numerous plastids with starch grains, mitochondria, ribosomes and well developed endoplasmic reticulum and golgi apparatus. The plastids in the stylar region immediately below the stigma produce electron-dense osmiophilic substances which are probably transferred into the cytoplasm by a process resembling budding-off of vesicles. The Golgi apparatus may use the starch grains as a source of sugars for the synthesis and secretion of extracellular polysaccharides. The structural and cytochemical differences between the glandular cells of the stigma and the stylar transmitting tissue are discussed.     

Structural analysis of stigma development in relation with pollen–stigma interaction in sunflower

Structural analysis of stigma development in sunflower highlights the secretory role of papillae due to its semi-dry nature. Production of lipid-rich secretions is initiated at the staminate stage of the flowers in stigma development and increases at the receptive stage, coinciding with an extensive development of elaioplasts and endoplasmic reticulum network in the basal region of the papillae. Transfer cells, earlier identified only in the wet type of stigma, are also present in the transmitting tissue of the sunflower stigma. Attainment of physiological maturity by the stigmatic tissue, accompanying development from bud to pistillate stage, appears to affect the initial steps of pollen–stigma interaction. The nature of self-incompatibility in Helianthus has also been investigated in relation with pollen adhesion, hydration and germination. Pollen adhesion to the stigma is a rapid process in sunflower and stigma papillae exhibit greater affinity for pollen during cross pollination as compared to self-pollination. Components of the pollen coat and the pellicle on the surface of stigmatic papillae are critical for the initial phase of pollen–stigma interaction (adhesion and hydration). The lipidic components of pollen coat and the proteinaceous and lipidic components from the surface of the papillae coalesce during adhesion, leading to the movement of water from stigma to the pollen, thereby causing pollen hydration and its subsequent germination. Pollen germination (both in self-and cross-pollen) on the stigma surface and the growth of the pollen tube characterize the flexibility of self-incompatibility in sunflower. Compatible pollen grains germinate and the pollen tube penetrates the stigma surface to enter the nutrient-rich transmitting tissue. The pollen tube from incompatible pollen germination, however, fails to penetrate the stigmatic tissue and it grows parallel to the papillae. Present findings provide new insights into structural and functional relationships during stigma development and pollen–stigma interaction.     

Fungicide sprays can injure the stigmatic surface during receptivity in almond flowers

Fungicides can be detrimental to flower development, pollen function and fruit set in a number of crops. Almond is a self-incompatible nut crop that has a fruit set of only approx. 30 % of the total number of flowers. Thus, interference of pollination and fertilization by fungicide sprays is of concern, and identification of chemicals having the least detrimental effects would be desirable. The objective of this study was to evaluate the effect of fungicide sprays on stigma morphology in almond using a laboratory spray apparatus that simulated field applications. Four fungicides (azoxystrobin, myclobutanil, iprodione and cyprodinil) were applied, and fresh, unfixed stigmatic surfaces were observed using a scanning electron microscope at 4 and 24 h after spraying. Increased exudate accumulation was induced by azoxystrobin at both time periods, and localized damage and collapse of stigmatic cells were observed after 24 h. Damaged stigmatic papillae exhibited wrinkling, surface distortion or collapse. Likewise, myclobutanil caused significant damage to and collapse of papillae; these were more extensive at later observations. Iprodione had no effect on exudate accumulation but caused marked and severe collapse of stigmatic papillae which was pronounced at 24 h. Cyprodinil promoted a copious increase in exudate secretion and caused the most severe collapse of stigmatic cells of all the fungicides evaluated. Damage was somewhat localized at 4 h but more global at 24 h. This study has verified that certain fungicide sprays have direct detrimental effects on stigma morphology and enhance exudate production in almond flowers.     

The Anatomy, Histochemistry and Ultrastructure of Stigmas and Styles in Commelinaceae

The anatomy and ultrastructure of stigmas in 37 species of 13genera of Commelinaceae are described. The stigmas are papillate,papillae forming a dense fringe of cells around the mouth ofthe stylar canal in most species. The papillar cell wall iscovered by an unstructured cuticle of variable thickness andis of variable thickness because of small wall ingrowths. Thecuticle and the external surface of the papillar cell wall arevariably disrupted, particularly in the mid and basal regionsof the cell. This was not found in species of the genus Aploleiaor Callisia. The cell cytoplasm possesses all major organellesexcept chloroplasts and each cell is vacuolate. In all species except Aploleia mulitiflora the style comprisesan epidermis, a cortex and a hollow, tripartite canal whichis continuous into the ovary cavity. The three vascular strandsare positioned at the apex of each canal lobe. The canal cellsare elongate and tabular and the wall abutting the canal hasingrowths. The style in Aploleia is solid and the transmittingtissue comprises cells whose walls are electron opaque. Thecytoplasms of both types of cell are similar in content althoughthere is a single, large vacuole in canal cells and many smallvacuoles in transmitting tissue. The morphology, position and histochemistry of stigmatic andstylar exudate was similar in all ‘wet’ stigmas.Most of the exudate originates from the stylar canal althoughsignificant contributions are made by the papillae in stigmasof Coleotrype, Dichorisandra and Thyrsanthemum. There is no apparent relationship between stigma structure andthe presence of self-incompatibility. Stigma papillae, stylar canal, transmitting tissue, Commelinaceae     

Structure of the style and wet non-papillate stigma of Colophospermum mopane, Caesalpinioideae: Detarieae

The capitate stigma of Colophospermum mopane (Kirk ex Benth.) Kirk ex J. Leonard is an intensely folded bilobed structure. The epidermal layer of the stigma consists of non-papillate cells. Before anthesis the epidermis is covered with a cuticle and thin proteinaceous layer. Elongated subepidermal cells constitute the secretory zone. Cell disintegration in the central region of each stigma lobe leads to cavities that become connected to the central cavity in the style. During early anthesis it appears as if the receptive surface of the stigma is confined to the depressions of the stigma surface and to the cleft between the two stigma lobes as the secretory product and pollen grains are mainly confined to these areas. The secretory products of the stigma and style are released during five different stages from prior to anthesis to late anthesis. The stigmatic exudate appears complex and consists of carbohydrates, proteins and lipids. The style has a hollow, lysigenous, fluid-filled canal that is not lined with an epidermal layer or cuticle. The stylar canal is continuous with the opening between the two stigma lobes and provides an open route for the passage of exudate. The stylar exudate is PAS-positive. The dorsal and ventral bundles that supply the style branch in such a way as to almost form a cylinder around the central transmitting tissue and stylar canal. New sieve elements proliferate before anthesis.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 295–304.     

黄瓜雌花发育过程中柱头的腺特征

利用透射电镜技术研究了黄瓜(Cucumis sativus L.)雌花柱头发育过程中传递组织、分泌组织和乳突细胞的超微结构.在整个发育过程中,乳突细胞和分泌组织细胞的细胞质内密布很多管状及槽库膨大的内质网,产生很多分泌囊泡;在成熟柱头的传递组织和分泌组织细胞间观察到大量的胞间连丝;乳突细胞和分泌细胞高度液泡化,质膜内折;在柱头发育过程中分泌组织细胞的核周腔扩大形成裂瓣状核,到柱头成熟阶段裂瓣状核更加明显.进一步的研究显示,在成熟柱头的不同组织细胞中,ATPase的活性呈现在质膜和液泡膜上,随着柱头的发育,PM-H -ATPase的比活性明显增强.结果表明,黄瓜雌花柱头的腺特征随发育进程而趋于显著.     

Stigma-surface Esterase Activity and Stigma Receptivity in Some Taxa Characterized by Wet Stigmas

Stigma-surface esterase activity and stigma receptivity througha sequence of developmental stages of the pistil have been studiedin four taxa characterized by having wet stigmas — Petuniahybrida, Nicotiana tabacum, Crinum defixum and Amaryllis vittata.The style is solid in the first two and hollow in the lattertwo taxa. In all the taxa, stigma—surface esterase couldbe detected in a thin surface layer (pellicle) from a very earlystage of pistil development, irrespective of the presence orabsence of the exudate. However, the taxa showed variation instigma receptivity. In Petunia and Nicotiana, stigmas from pistilsof all the stages supported pollen germination and tube growth.In Amaryllis and Crinum, stigmas of only the mature pistils,when the exudate is present on the stigma, supported normalpollen germination and tube growth. It is inferred that in taxacharacterized by a wet stigma and solid style, the factors requiredfor pollen germination are present from an early stage of pistildevelopment and the exudate per se is not involved in pollengermination. In taxa characterized by a wet stigma and hollowstyle, however, the pellicle does not carry the factors requiredfor pollen germination and tube growth; they appear to be presentin the exudate. Petunia hybrida Vilm, Nicotiana tabacum L., Crinum defixum, Ker-Gawl, Amaryllis vittata Ait., tobacco, pollination, pollen germination, stigmatic exudate, stigma receptivity, stigma-surface esterase, esterase activity     

Structural features of <Emphasis Type="Italic">Rhododendron luteum</Emphasis> flower

The flower of Rhododendron luteum (L.) Sweet has a pentamerous structure with radial symmetry. The anthers filament surface is covered by dense non-glandular hairs to the half of the height. The tubular anther dehisces along creating two openings in the anther-sac walls and the viscous pollen is released through two splits along the anther lobes. The pistil is pentamerous and the axial channel is filled with a mucilaginous secretion product which is continuous with the exudate on the stigma surface. The stigmatic papillae are densely packed and their exudate is stained intensively red for carbohydrates, while pollen grains are stained positively for lipids. The five-locular ovary has isomerous carpels (syncarpous gynoecium) and the ovary surface is covered by numerous, densely-packed glandular and non-glandular hairs protecting the nectar against transpiration. Numerous ovules per locule occur with one integument and a thin-walled megasporiangium. In carpels, oil cells occur sporadically as solitary idioblasts, located around the vascular bundles. Transmitting tissue cells contain a large central, electron translucent vacuole, filling most of the cell containing dark osmiophilic bodies homogenous or granular in appearance.     

Histogenesis of the transmitting tract in Strelitzia reginae

Organisation and development of the stigmatic, stylar and ovarian parts of the transmitting tract in Strelitzia reginae were evaluated. They were characterised by 1) cell shape, 2) appearance of distal cell wall, 3) type of plastid, 4) and vacuolar system. The long stigmatic trichomes have a secondary irregular wall layer separated from the primary wall. Cell structures include pleomorphic plastids with vesiculated thylakoids and frequently a crystal or a lipidic globule. In early stages of bud development the extensive endoplasmic reticulum (ER) is smooth, whereas it is mainly rough in older buds. Coated vesicles are frequent, as are dictyosomes. Prominent invaginations along the plasma membrane contain floccular deposits in the older flower buds. These deposits are similar in structural appearance to material in the large vacuoles. The basal part of the stigma has wedge-shaped cells with wall ingrowths. Three stages of stigmatic secretion during the development were characterised. The stylar canal is initially narrow but widens subsequently. The cuticle is detached and the apical cell walls show a fringed surface; from this wall inbuddings develop shortly before anthesis. At this stage sheets of rough ER are evident in the cell cortex. The plastids have a few vesiculated thylakoids, proteinaceous crystals and starch grains. The epithelial cells of young buds have numerous vacuoles, the volume of which decreases in more mature cells. The stylar canal is filled with a secretion at all stages of bud development. The face of the ovarian transmitting tract, lining the placenta, is smooth in young buds but lobed in older ones due to the division pattern of the epithelial cells. These cells are large, elongated and culumnar as young but narrow and wedge-shaped when more mature. Cell wall inbuddings are formed late during bud development.     

Structural and Cytochemical Analysis of the Stigma and Style in Tibouchina semidecandra Cogn. (Melastomataceae)

Structural and cytochemical aspects of the pistil of Tibouchinasemidecandra Cogn. were studied. The stigma is of the wet-papillatetype and is structurally divisible into a papillar zone anda stigmatic zone. The papillar zone consists of loosely arrangedpapillae which are matchstick-shaped, unicellular, and producelipid droplets that remain entrapped below the thick cuticle.The bulk of cell volume is made up of large vacuoles rich intannin. The stigmatic zone consists of layers of secretory cellswith dense cytoplasm, actively secreting dictyosomes and numerousrough endoplasmic reticulum (RER) profiles. Free-flowing lipidexudate, produced by these cells, is initially stored in theintercellular spaces, and subsequently extruded out to coverthe surface. The style is solid with a core of transmittingtissue traversing its whole length. The transmitting tissueconsists of loosely arranged cells with numerous organellesand conspicuous intercellular substance rich in polysaccharidesand pectins. Ultrastructural details indicate that the intercellularsecretion is accompanied with fraying of the wall component.Both the transverse and longitudinal walls contain plasmodesmata.Copyright1995, 1999 Academic Press Cytochemistry, stigma and style, ultrastructure, Tibouchina semidecandra     

Evidence for Stigmatic Self-incompatibility, Pollination Induced Ovule Enlargement and Transmitting Tissue Exudates in the Paleoherb, Saururus cernuus L. (Saururaceae)

Saururus cernuus, a species belonging to the primitive herbaceousangiosperm family Saururaceae, exhibits high rates of self-sterility.We investigated the structural and functional aspects of pollen-carpelinteractions following cross and self pollination to assessthe tissue specific site and timing of self-sterility and factorsimportant for successful cross pollen tube growth. Self-sterilitywas due to inhibition of self pollen germination at a dry stigma.Self pollination was associated with anomalous foot formation,reduced cell wall expansion and secretory activity of stigmaticpapillae, and callose production in stigmatic papillae. Followinggermination, cross compatible pollen tubes entered a solid coreof transmitting tissue and grew to the base of a short style.Entry of cross pollen tubes into the ovary was coincident withovule enlargement which placed the micropyle in the proximityof cross pollen tube tips. Ovule enlargement also occurred followingself pollination. Cross pollen tubes either entered an exudate-filledmicropyle directly from the style, or growth in the ovary waslocalized to the epidermis of the locule and outer integumentprior to entry into the micropyle. Prior to pollination, thetransmitting tract was void of secretions except for exudatein the micropyle. Growth of pollen tubes on the locule and integumentwas associated with exudate apparently arising from transmittingcells adjacent to growing pollen tubes. The present study providesthe first evidence in a primitive herbaceous species of stigmaticself-incompatibility (SI) in association with a dry stigma,pollination-induced signalling events affecting developmentof carpellary tissues, and micropylar exudates. Copyright 1999Annals of Botany Company SI evolution, dry stigma, exudates, pollen-carpel signalling, Saururaceae.