PLASTIDS IN LATICIFERS OF PAPAVER. I. DEVELOPMENT AND CYTOCHEMISTRY OF LATICIFER PLASTIDS IN P. SOMNIFERUM L. (PAPAVERACEAE)

Plastids were observed in all stages of laticifer differentiation in Papaver somniferum L. Plastids in laticifer initials were present as proplastids that later developed electron-dense inclusions, but never possessed the thylakoids or starch grains that characterize chloroplasts in other cells. Electron-dense inclusions in laticifer plastids were membrane-bound and appeared to arise from the accumulation of material within an invagination of the inner plastid membrane. Cytochemical studies of these plastid inclusions indicated that their matrix was not composed of crystalline protein, α-amylose, amylopectin or polysaccharide. The results suggest that the electron-dense, membrane-bound inclusions in laticifer plastids may be composed of lipoprotein.     

Correlation of the appearance of morphinan alkaloids and laticifer cells in germinating Papaver bracteatum seedlings

The course of alkaloid accumulation and laticifer cell appearance was compared in germinating P. bracteatum seedlings. Seedlings of various ages (0–14 days old) were analyzed for their dopamine, thebaine, morphinan alkaloid immunoreactivity, and benzophenanthridine alkaloid levels. Simultaneous electron microscopic studies revealed that seedlings were devoid of laticifer initials until day 3, where-upon their numbers increased with time. The appearance of appreciable amounts of thebaine only occurred after day 4 of germination. Conversely, dopamine was rapidly formed at the onset of germination and reached millimolar concentrations well before laticifer cells were detected. Benzophenanthridine alkaloid levels remained fairly constant over the period analyzed. These results support the theory that the presence of laticifer cells is necessary for the accumulation of morphinan but neither benzophenanthridine alkaloids nor their mutual precursor, dopamine.Abbreviations RIA radioimmunoassay     

ONTOGENY AND CYTOCHEMISTRY OF ALKALOIDAL VESICLES IN LATICIFERS OF PAPAVER SOMNIFERUM L. (PAPAVERACEAE)

Development of alkaloidal vesicles in laticifers of opium poppy, Papaver somniferum L., was investigated at the ultrastructural level. Laticifer initials possessed abundant endoplasmic reticulum throughout their dense cytoplasm. During differentiation the endoplasmic reticulum organized into long, folded sheets that were parallel to the longitudinal walls along the periphery of the cell. Vesicles appeared to be derived from dilation of endoplasmic reticulum. This relationship was confirmed through cytochemical data obtained with zinc iodide-osmium tetroxide and osmium tetroxide impregnation. Alkaloidal vesicles had electron-dense regions or caps that occurred early in laticifer differentiation, but these caps became less conspicuous in mature cells. Caps appeared to be derived from small particles which condensed along the inner surface of the vesicle membrane and subsequently accumulated at one or two positions along the membrane of the vesicle.     

Histochemical and immunohistochemical identification of laticifer cells in callus cultures derived from anthers of Hevea brasiliensis

Laticifers are highly specialized cells present in over 20 plant families. They are well defined in planta. In vitro development of laticifers was also observed in some plants, but uncertain in the callus cultures of rubber tree, one of the most economically important latex producing plants. In the present study, we provide evidence that laticifer cells present in the callus cultures of rubber tree by histochemical and immunohistochemical studies. They present in the callus mainly as separate non-elongated form, a novel morphology different from the morphology of laticifer cells in planta, excluding their origin from explants. The occurring frequency of laticifer cells in the callus was genotype-dependent and negatively correlated with the somatic embryogenetic ability, suggesting that the presence of laticifer cells in the callus inhibit somatic embryogenesis in tissue culture of rubber tree. The genotypes PR107, RRIM600, Reyan8-79, and Reyan7-33-97 with lower embryogenetic ability compared to Haiken 2 had more laticifer cells, and laticifer clusters were only observed in these genotypes.     

EMBRYOGENY AND HISTOGENESIS IN NERIUM OLEANDER II. ORIGIN AND DEVELOPMENT OF THE NON-ARTICULATED LATICIFER

Mahlberg , P. G. (U. Pittsburgh, Pittsburgh, Pa.) Embryogeny and histogenesis in Nerium oleander. II. Origin and development of the non-articulated Iaticifer. Amer. Jour. Bot. 48(1): 90–99. Illus. 1961.—Laticifer initials, collectively considered as a laticifer system, are differentiated in the globular embryo from meristematic cells which occupy a position within the potential procambial tissue. A total of usually 28 initials, in Nerium oleander, arise as an irregular ring of cells directly below the embryonic shoot apex, during initiation of the cotyledonary primordia. No anastomoses occur between laticifer initials. During subsequent development of the embryo, the laticifer initials grow in a bi-directional manner and penetrate into the root, cotyledons and toward the shoot apex. Upon enlargement the initials bifurcate repeatedly, many branches penetrate into the cotyledons, others grow into the cortex of the hypocotyl or penetrate between cells of the procambium. Repeated nuclear divisions within each initial result in the formation of a multinucleated protoplast in this cell type. The tips of laticifers occupy intercellular spaces during their growth; they do not penetrate into or through adjacent cells. A plexus of laticifer branches is formed within the cotyledonary node of the mature embryo. No new initials are formed during subsequent growth of the plant, rather certain branches from the cotyledonary nodal plexus penetrate into the enlarging shoot system. The nature of their growth habit and branching suggests that the tips of laticifer initials exhibit an intrusive form of growth.     

STRUCTURE AND ONTOGENY OF LATICIFERS IN CICHORIUM INTYBUS (COMPOSITAE)

The branched anastomosed laticifer system in the primary body of Cichorium intybus L. originates in embryos from files of laticiferous members at the boundary between phloic procambium and ground meristem. Upon seed germination, laticiferous members develop perforations in the end walls which become entirely resorbed. Perforations also develop in the longitudinal walls of contiguous laticiferous members and from lateral connections between developing laticifer branches. Additional laticiferous members originate as procambium differentiation proceeds, and their differentiation follows a continuous acropetal sequence in leaf primordia of the plumule. In roots, laticifers closely associated with sieve tubes in the secondary phloem originate from derivatives of fusiform initials in the vascular cambium. These laticifers develop wall perforations and in a mature condition resemble laticifers in the primary body. As the girth of the root increases, laticifers toward the periphery, unlike associated sieve tubes, resist crushing and obliteration. Laticifers vary in width from about 4 to 22 μm; the widest ones occur in involucral bracts and the narrowest ones in florets. There was no evidence that intrusive growth occurs during development of the laticifer system, although such growth may occur during development of occasional branches which extend through ground tissue independent of phloem and terminate in contact with the epidermis. Presence of amorphous callose deposits is related to aging of laticifers and mechanical injury.     

Observations on the Development and Fine Structure of the Articulated Laticifers of Papaver somniferum

The development and fine structure of articulated anastomosinglaticifers in Papaver somniferum were studied. Laticifers arenot present in the embryos but differentiate soon after germinationand are found in the phloem areas 18–30 h after the seedis sown. Laticifers and sieve elements are generally separatedby at least one cell layer in the roots, but in cotyledons,stems, and leaves they usually occur adjacent to each other. As the laticifers differentiate an abundance of vesicles formsin the cytoplasm. This process appears to involve the endoplasmicreticulum and it is suggested that the vesicles may be a specializedform of vacuole. Substances present in the vesicles react stronglywith iodine-potassium iodide. Laticifer-cytoplasm persists peripherallyand between the vesicles. It contains the usual cell organelles,the presence of which substantiates an active metabolic rolefor the laticifer contents.     

A comparative study of latex-producing tissues in genera of Liabeae (Asteraceae)

Laticifers are highly specialized living plant cells which produce and contain latex. Occurrence of latex was used to establish morphological affinities (i) between Liabeae and other Asteracean tribes, (ii) among the Liabean genera, and (iii) in order to obtain phylogenies within Liabeae. However, structures and types of latex-producing tissues in this tribe have not yet been studied anatomically. In the present paper latex-producing structures of aerial parts in species of Microliabum, Munnozia, and Paranephelius (Liabeae), from open areas in mid-elevation Andean forests and in Andean high-elevation habitats, were studied. In all the analyzed species, latex secretion was easily observed in stem and leaf blade hand sections. Laticifers accompanied vascular tissues in all the cases, throughout stems and leaves, and they were of the articulated anastomosed type, at least in fully developed stages. Laticifers were found facing both, the xylem and the phloem, except for Paranephelius stems, in which they occur merely next to the phloem. Leaf laticifers form a reticulum accompanying the vein system. The type of latex-producing tissue shared by Microliabum and Munnozia could be a character shared by common ancestry whereas the laticifer system of Paranephelius stems could represent an evolutionary novelty for this genus. The laticifer type described in this study in aerial parts of Liabeae may allow establishing morphological affinities with tribes Cichorieae and Arctoteae.     

ORIGIN AND EARLY DEVELOPMENT OF NONARTICULATED LATICIFERS IN EMBRYOS OF EUPHORBIA MARGINATA

In E. marginata 12 nonarticulated laticifer initials arise in the cotyledonary node of the young embryo during the early heart stage. The initials arise progressively in the developing embryo, the first laticifers differentiating simultaneously with or shortly before the elements of the pro-cambium. The laticifers occupy a position lateral to the six procambial strands which are formed in the embryo. Upon subsequent growth each laticifer becomes vacuolated and nuclear division unaccompanied by cytokinesis results in the formation of a coenocytic protoplast. The enlarging laticifer produces several branches, one growing into the cotyledon, another growing down along the hypocotyl penetrating toward the root meristem, and one or several growing along intercellular spaces of adjacent cells. No fusion of these branches with one another or adjoining parenchyma cells was observed.     

ULTRASTRUCTURE OF NON-ARTICULATED LATICIFERS IN MATURE EMBRYOS AND SEEDLINGS OF ASCLEPIAS SYRIACA L. (ASCLEPIADACEAE)

The protoplast of the non-articulated branched laticifer in the embryo and seedling of Asclepias syriaca L. was studied at the ultrastructural level and was found to differ from that of adjacent cell types. Embryonal laticifers possess numerous vesicles with electron-dense contents, but lack a large organized central vacuole. Plastids have few lamellae, possess phytoferritin, and accumulate small amounts of starch. Other organelles and membrane systems are similar to those in other cells. After germination, laticifers develop numerous elongated vacuoles by dilation of endoplasmic reticulum. Nuclei in laticifers within the hypocotyl of seedlings are highly lobed and possess dilated perinuclear spaces. Plastids and other organelles are similar to those observed in the protoplast of laticifers in the embryo. The latex or rubber component of the laticifer is not apparent in mature embryos of 72-hr seedlings.     

Laticifer Ultrastructural and Immunocytochemical Studies of Papain in Carica papaya

Cotyledons of germinating papaya (Carica papaya L. ) seeds and exocarp of young fruits were used as materials for study. The ultrastructural changes occurring during differentiation of laticifer and the ultrastructural environment of papain synthesis were studied by means of TEM and immunocytochemistry. Electron microscopic observations showed that the differentiating laticiferous cells were rich in ribosomes and mitochondria. Endoplasmic reticulum (ER) was well developed and apparently active, forming secretory vesicles of various sizes. With further development, organelles were gradually degenerated and autophagy of cytoplasm within vacuole was evident. ER was dilated and split into fragments. Cell wall perforations occurred at several sites of adjacent laticifer elements. Towards maturity, laticifer was fully filled with vesicles containing electron-dense materials. Organelles disappeared thoroughly but plasmalemma remained. Sections were incubated with anti-chymopapain antibodies followed by goat-anti-rabbit IgG-gold. Labeled gold was found predominantly in ER and the associated vesicles of differentiating laticifer. Several controls were used to establish the specificity of the immunolaheling pattern. Investigations led to the conclusions that ER and polyribosomes were involved in papain synthesis. Papain was stored in the vesicles of ER origin temporally before reorganized into laticiferous vesicles with other components of latex.     

ULTRASTRUCTURE OF DEVELOPING AND MATURE NONARTICULATED LATICIFERS IN THE MILKWEED ASCLEPIAS SYRIACA L. (ASCLEPIADACEAE)

The ultrastructure of developing and mature nonarticulated laticifers in Asclepias syriaca L. (the common milkweed) was studied by conventional fixation and staining techniques and by osmium impregnation techniques. The mature laticifer protoplast in A. syriaca possesses a large central vacuole with an intact vacuolar membrane. Formation of this vacuole apparently results from dilation and subsequent enlargement of endoplasmic reticulum and possibly in part by fusion of smaller vacuoles and limited cellular-lytic autophagy. Widespread digestion or autophagy of cytoplasm within vacuoles is not evident. Nuclei, mitochondria, dictyosomes, and small vesicles are the most prominent components distributed in the peripheral cytoplasm. Plastids appear to degenerate as the laticifer matures. The specialized cellular component, latex, which is the vacuolar content of the laticifer, is interpreted to be produced in the cytoplasm and subsequently incorporated into the large central vacuole. Rubber globules, the most prominent latex component, are surrounded by a membrane that does not have a trilaminate structure. Globules are associated with an electron-dense fibrillar component in the vacuole.     

Ultrastructure and development of nonarticulated laticifers in seedlings ofEuphorbia maculata L.

The ultrastructure of nonarticulated laticifers in the seedlings ofEuphorbia maculata was studied at various developmental stages. The apical regions of the seedling laticifers growing intrusively contained large nuclei with mainly euchromatin and dense cytoplasm possessing various and many organelles such as rich ribosomes, several small vacuoles, giant mitochondria with dense matrices, rough endoplasmic reticulum, dictyosomes, and proplastids. This result suggested that the apical regions of laticifers were metabolically very active. Laticifers in seedlings at the first-leaf developmental stage did not contain latex particle. In seedlings at second-leaf growth stage, the laticifer cells contained numerous and elongated small vacuoles. These vacuoles appeared to arise by dilation of the endoplasmic reticulum and frequently possessed osmiophilic or electron-dense latex particles. The small vacuoles fused with the large vacuole occupying the central portion of the subapical region of laticifers, and then the latex particles were released into the large central vacuole. The latex particles varied in size and were lightly or darkly stained. Proplastids with a dense matrix and a few osmiophilic plastoglobuli were filled with an elongated starch grain and thus were transformed into amyloplasts. Latex particles were initially produced in the laticifers after seedlings had developed their second young leaves. In seedlings at forth-leaf stage, latex particles with an alveolated rim were found in the laticifers.     

Studies on Differentiation of Laticifers through Light and Electron Microscopy in Calotropis gigantea (Linn.) R.Br.

The distribution, cytological organization and differentiationof non-articulated laticifers in the primary and mature tissuesof Calotropis gigantea (Linn.) R.Br., were studied by the useof optical and electron microscopy. Laticifers occur in thecortex, vascular bundle and pith of the plant axis. At the earliestdetectable stage a laticifer is a cell which undergoes rapidelongation and nuclear division. This results in a multinucleateelongated cell which undergoes further increase in length withgradual degeneration of the cytoplasm. At the electron microscopiclevel the presumptive laticifer cell shows increasing vacuolationwhich forms a large central vacuole. Simultaneously the cytoplasmicorganelles undergo degeneration by autophagic processes. Laternumerous vesicles can be observed in the large central vacuole,the remaining cytoplasm being pushed to a thin layer. Maturelaticifers show three types of spherical structures of whichthe highly electron dense globules are the latex particles. Calotropis gigantea (Linn.), R.Br., laticifers, ultrastructure, differentiation     

Developmental regulation of benzylisoquinoline alkaloid biosynthesis in opium poppy plants and tissue cultures

Summary Opium poppy (Papaver somniferum L.) contains a number of pharmaceutically important alkaloids of the benzylisoquinoline type including morphine, codeine, papaverine, and sanguinarine. Although these alkaloids accumulate to high concentrations in various organs of the intact plant, only the phytoalexin sanguinarine has been found at significant levels in opium poppy cell cultures. Moreover, even sanguinarine biosynthesis is not constitutive in poppy cell suspension cultures, but is typically induced only after treatment with a funga-derived elicitor. The absence of appreciable quantities of alkaloids in dedifferentiated opium poppy cell cultures suggests that benzylisoquinoline alkaloid biosynthesis is developmentally regulated and requires the differentiation of specific tissues. In the 40 yr since opium poppy tissues were first culturedin vitro, a number of reports on the redifferentiation of roots and buds from callus have appeared. A requirement for the presence of specialized laticifer cells has been suggested before certain alkaloids, such as morphine and codeine, can accumulate. Laticifers represent a complex internal secretory system in about 15 plant families and appear to have multiple evolutionary origins. Opium poppy laticifers differentiate from procambial cells and undergo articulation and anastomosis to form a continuous network of elements associated with the phloem throughout much of the intact plant. Latex is the combined cytoplasm of fused laticifer vessels, and contains numerous large alkaloid vesicles in which latex-associated poppy alkaloids are sequestered. The formation of alkaloid vesicles, the subcellular compartmentation of alkaloid biosynthesis, and the tissue-specific localization and control of these processes are important unresolved problems in plant cell biology. Alkaloid biosynthesis in opium poppy is an excellent model system to investigate the developmental regulation and cell biology of complex metabolic pathways, and the relationship between metabolic regulation and cell-type specific differentiation. In this review, we summarize the literature on the roles of cellular differentiation and plant development in alkaloid biosynthesis in opium poppy plants and tissue cultures.     

THE STRUCTURE AND DEVELOPMENT OF AN UNUSUAL TYPE OF ARTICULATED LATICIFER IN MAMMILLARIA (CACTACEAE)

Although the laticifers of several species of Mammillaria can technically be classified as being of the articulated type, they differ significantly from all other reported articulated laticifers. They are derived from cells which differentiate only in older tissues, never in meristematic or young regions. The development involves the complete lysis of masses of cells, not just the perforation or resorption of the end walls in a single file of cells. At maturity, the laticifer lumen is lined with a one-to-several layered epithelium which may be quite thick. The laticifers increase in diameter with age, apparently by the lysis of the inner epithelial cells. Laticifers occur in the pith, cortex and tubercles of the vegetative body but were not observed in the roots, flower parts or in seedlings up to eight months old. Seven species were studied, all of which have “milky sap.” and the laticifers of each were virtually identical to the laticifers of the others.     

Identification and characterization of latex-specific proteins in opium poppy

Latex from the opium poppy, Papaver somniferum L., was analyzed by polyacrylamide gel electrophoresis (PAGE). Two latex-specific bands were identified in protein samples of poppy latex using one-dimensional native PAGE. Second dimension analysis with SDS-PAGE indicates that these proteins have a relative molecular weight of approximately 20 kilodaltons. We have termed these polypeptides the major latex proteins (MLPs). Polyclonal antibodies prepared against the MLPs were used to probe protein gel blots of latex and poppy tissues known to lack laticifers. Laticifer-free tissues showed no reaction with anti-MLP immunoglobulin G indicating that MLPs are found only in poppy latex. MLP distribution was also examined in mature opium poppy tissues by immunocytochemistry. Laticifers were differentially labeled by fluorescein isothiocyanate secondary labeling of anti-MLP immunoglobulin G and could easily be identified in both transverse and longitudinal section. Fractionation studies of isolated latex showed that MLPs are concentrated in the latex cytosol and not in alkaloidal vesicles. Analysis of latex proteins by conventional two-dimensional electrophoresis indicates that the two MLP bands are composed of several distinct polypeptides with similar relative molecular weights. The pIs of these molecules range from 6.0 to 3.5. The role(s) of MLPs in laticifer metabolism has not been determined.     

Ultrastructure and Development of the Laticifers of Ficus carica L

Laticifers of Ficus caricaL. are of the branched, non-articulatedtype. They occur in the cortex and pith of the plant axis andpenetrate leaves and inflorescences. Observations were madeon laticifers located in shoot apices. Growing regions of laticifers undergo a sequence of ultrastructuralchanges. These are: (a) a pronounced increase in the vacuolarspace which divides the cytoplasm into separated masses; (b)a development of numerous vesicular structures in the cytoplasm.The vesicular structures are released into the vacuolar space.The whole process is accompanied by disintegration of cytoplasm.Apparently isolated masses of cytoplasm occur in the luminaof laticifer tips in sections taken from dormant apices. Itis assumed that these masses have a role in the initiation ofnew laticifer regions in the next growing season. Ficus caricaL., laticifers, ultrastructure, development differentiation     

Electron microscopy of infection of nematodes byDactylaria haptotyla

Infection of nematodes byDactylaria haptotyla, a nematode-trapping hyphomycete, was studied by electron microscopy. The cytoplasm of the adhesive knob in the fungus contained a number of electron-dense, membrane-bound vesicles, 0.2–0.5 µm in diam. The vesicles were rarely seen in the stalk cell or vegetative cell cytoplasm. When the adhesive knob came into contact with the nematode's cuticle, it secreted an adhesive which was seen in ultrathin sections between the knob and the cuticle as an amorphous mass. At the same time, electron-dense vesicles in the cytoplasm were reduced in number and many small vacuoles developed. Soon after capture of a nematode, the cell wall of the adhesive knob became obscure at the prospective site of penetration, where a vesicle, 0.7 µm in diam, was found in serial thin sections of the knob's cytoplasm. At the site facing the vesicle, the peripheral part of the nematode's cell exhibited a high electron density. The vesicle, which appeared to be derived from smaller electron-dense vesicles coalesced with each other, released its enzymic contents toward the captured nematodes before penetration by the fungus.     

The Terminal Protonephridial Complex of Haplopharynx rostratus (Platyhelminthes,Haplopharyngida)

The terminal protonephridial complex of Haplopharynx rostratus consists of three terminal cells. There are no weirs consisting of ribs connected by a filtration “membrane”, but some cytoplasmic outgrowths into the lumen of the terminal cells. Excretion is by exocytotic vesicles. The terminal cells also contain Golgi complexes and large membrane-bound vacuoles containing electron-dense material. The ciliary bundles (flames) of terminal cells 2 and 3 protrude into the lumen of the centrally located terminal cell I. The complex is surrounded by a sheath containing numerous filaments. The terminal complex of H. rostratus resembles that of the macrostomid Paromalostomum proceracauda, lending support to the view that the two taxa are closely related. © 1998 The Royal Swedish Academy of Sciences. Published by Elsevier Science Ltd. All rights reserved