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.     

Fine structure and development of laticifers inGnetum gnemon L.

Summary Articulated laticifers are a regular component of stem cortex and pith ofGnetum gnemon L. Differentiation of laticifers starts very early and is completed within the very first centimeters adjacent to the apical meristem.Ultrastructurally, young laticifers ofGnetum can be distinguished from surrounding cells by the presence of characteristic cytoplasmic inclusions, called spherule complexes and composed of 40–70 nm light spherules and up to 0.5 by 1.0 m large particles. While there is no record on their first beginnings, a connection between rER and the large particles can be demonstrated during many steps of laticifer differentiation which includes an increase in spherule complex areas. On account of positive Sudan staining and responses to EM fixations and in comparison to other laticifers it is concluded that the spherule complexes are terpenoid-containing. A transition from spherules to larger particles (orvice versa) is discussed, but could not be documented.Relative to the formation of the spherule complexes the other changes during laticifer maturation are inferior. The vacuolar system, in close connection to an extensive ER system, does largely expand and finally takes over the almost entire cell lumina. Nuclei do persist for an extended period, even after breakdown of the end walls and during disorganization of the cytoplasm. Plastids in all stages of laticifer ontogeny are very rarely encountered.Supplemented part of an investigation presented in 1976 by S. H. to the Fakultät für Biologie der Universität Heidelberg in partial fulfilment of the requirements for the degree of a Diplom-Biologe.     

Immunolocalization of beta-D-glucans,pectins, and arabinogalactan-proteins during intrusive growth and elongation of nonarticulated laticifers in Asclepias speciosa Torr

Nonarticulated laticifers are latex-containing cells that elongate indefinitely and grow intrusively between the walls of meristematic cells. To identify biochemical mechanisms involved in the growth of nonarticulated laticifers, we have analyzed the distribution of various polysaccharides and proteoglycans in walls of meristematic cells in contact with laticifers, nonadjacent to laticifers, and in laticifer walls. In the shoot apex of Asclepias speciosa, the levels of callose and a (1-->4)-beta-galactan epitope are lower in meristematic walls in contact with laticifers than in nonadjacent walls. In contrast, we did not detect a decline in xyloglucan, homogalacturonan, and arabinogalactan-protein epitopes upon contact of meristematic cells with laticifers. Laticifer elongation is also associated with the development of a homogalacturonan-rich middle lamella between laticifers and their neighboring cells. Furthermore, laticifers lay down walls that differ from those of their surrounding cells. This is particularly evident for epitopes in rhamnogalacturonan I. A (1-->5)-alpha-arabinan epitope in this pectin is more abundant in laticifers than meristematic cells, while the opposite is observed for a (1-->4)-beta-galactan epitope. Also, different cell wall components exhibit distinct distribution patterns within laticifer walls. The (1-->5)-alpha-arabinan epitope is distributed throughout the laticifer walls while certain homogalacturonan and arabinogalactan-protein epitopes are preferentially located in particular regions of laticifer walls. Taken together, our results indicate that laticifer penetration causes changes in the walls of meristematic cells and that there are differences in wall composition within laticifer walls and between laticifers and their surrounding cells.     

Localization of cell wall polysaccharides in nonarticulated laticifers ofAsclepias speciosa Torr.

Summary Asclepias speciosa Torr, has latex-containing cells known as nonarticulated laticifers. In stem sections of this species, we have analyzed the cell walls of nonarticulated laticifers and surrounding cells with various stains, lectins, and monoclonal antibodies. These analyses revealed that laticifer walls are rich in (1→4) β-D-glucans and pectin polymers. Immunolocalization of pectic epitopes with the antihomogalacturonan antibodies JIM5 and JIM7 produced distinct labeling patterns. JIM7 labeled all cells including laticifers, while JIM5 only labeled mature epidermal cells and xylem elements. Two antibodies, LM5 and LM6, which recognize rhamnogalacturonan I epitopes distinctly labeled laticifer walls. LM6, which binds to a (l→5) α-arabinan epitope, labeled laticifer walls more intensely than walls of other cells. LM5, which recognizes a (1→4) β-D-galac-tan epitope, did not label laticifer segments at the shoot apex but labeled more mature portions of laticifers. Also the LM5 antibody did not label cells at the shoot apical meristem, but as cells grew and matured the LM5 epitope was expressed in all cells. LM2, a monoclonal antibody that binds to β-D-glucuronic acid residues in arabinogalactan proteins, did not label laticifers but specifically labeled sieve tubes. Sieve tubes were also specifically labeled byRicinus communis agglutinin, a lectin that binds to terminal β-D-galactosyl residues. Taken together, the analyses conducted showed that laticifer walls have distinctive cytochemical properties and that these properties change along the length of laticifers. In addition, this study revealed differences in the expression of pectin and arabinogalactan protein epitopes during shoot development or among different cell types.     

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.     

Laticifers: An historical perspective

This review describes the development of the laticifer concept, with emphasis upon the nonarticulated type, from early observations of plant exudates and “juices” to the presentation of laticifers by Esau (1953). Classical writers and herbalists described practical applications of these substances. With the advent of the microscope early investigators believed that these substances occurred in structures present in most, if not all, plants and, wrongly, equated these structures to the circulatory system in animals. Introduction of the term, latex, into botany derived from its early use as a term for a blood component by physicians, and not for analogy to milk. However, the origin of the terms, laticifer and laticiferous, remains uncertain. Initial studies of laticifers were marked by the controversy of whether they represented intercellular spaces or elongated cells. Confirmation of their cellular character led to the designation of nonarticulated and articulated laticifers. Nonarticulated laticifers were shown to arise during early embryogeny in some plants. The ontogenetic origin of the articulated laticifer was unclear to early workers, but new laticifers were detected to be formed by cambium activity. Nonarticulated laticifers were described to develop by intrusive growth whereby tips of the cell penetrated between adjacent cells. The coenocytic condition of the nonarticulated laticifer resulted from nuclear divisions along the cell positioned in the growth region of the shoot and the subsequent distribution of the daughter nuclei along the length of the cell.     

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.     

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.     

Laticifers in Camptotheca acuminata Decne: distribution and structure

Summary. In this paper, a system of laticifers in Camptotheca acuminata Decne (Nyssaceae) is described. Laticifers were already present in the leaf primordia of the shoot apex. In the mature leaves, laticifers were found in the midrib and in the larger veins, both in the parenchymatic region delimited by vascular bundles and in the cortex just external to the phloem. In the stem, laticifers were present in both the primary and secondary body, running parallel to the longitudinal axis. They were located in the pith and in the cortex proximal to the phloem. No laticifers were found in the roots. The histochemical analyses indicated that the main compounds accumulated in laticifers were phenols. Neutral lipids and fatty acids were also present. Ultrastructural observations showed osmiophilic globules both in the vacuoles and in the peripheral regions of the cytoplasm of the laticifer cells. Plastids were present, although altered, with some parallel membranes and lacking starch grains. The discovery in C. acuminata of a laticifer system, which had never been described for the order Cornales, could be of taxonomic value, also considering that this order has traditionally represented one of the most problematic groups of flowering plants. Correspondence and reprints: Dipartimento di Biologia Vegetale, Università “La Sapienza”, Piazzale Aldo Moro 5, 00185 Rome, Italy.     

Nuclear and microtubular cycles in heterophasic multinuclearTriticum root-tip cells induced by caffeine

Summary Nuclear and microtubular cycles were studied in large heterophasic multinuclear cells induced in root tips ofTriticum turgidum by caffeine treatment. Multinuclear cells and cells with polyploid nuclei exhibited various configurations of multiple and complex preprophase microtubule (Mt) bands (PPBs), including helical ones. The developmental stages of PPBs in some heterophasic cells did not comply with the cell cycle stages of the associated nuclei, a fact indicating that these events are not directly controlled by the associated nuclei. The heterophasic cells exhibited asynchronous nuclei at different stages of mitosis. In cells displaying prophase and interphase nuclei, the prophase spindle was either absent or developed around both of them or developed around the prophase nuclei earlier than around the interphase ones. During prometaphase-metaphase of the advanced nuclei the lagging interphase nuclei were induced to form prematurely condensed chromosomes (PCCs) along with spindle formation around them. These observations suggest that the mitotic transition in heterophasic cells is delayed but is ultimately achieved due to the effect of the advanced nuclei, which induces a premature mitotic entry of the lagging nuclei. Although kinetochore Mt bundles were found associated with PCCs, their metaphase and anaphase spindles were abnormal resulting in abnormal or abortive anaphases. In some heterophasic cells, metaphase-anaphase transition did not take place simultaneously in different chromosome groups, signifying that the cells do not exit from the mitotic state after anaphase initiation of the advanced nuclei. Asynchronous pace of mitosis of different chromosome groups was also observed during anaphase and telophase. Implications of these observations in understanding plant cell cycle regulation are discussed.Abbreviations cdk cyclin dependent kinase - Mt microtubule - PCC prematurely condensed chromosome - PPB preprophase band     

THE COMPARATIVE CELL CYCLE AND METABOLIC EFFECTS OF CHEMICAL TREATMENTS ON ROOT TIP MERISTEMS. I. IOXYNIL

In this study we investigated the cell cycle response of Vicia faba and Pisum sativum root tip meristems to ioxynil treatments at two concentrations, (10^−-4m and 10^−-6m ). After 24 h of treatment at 10^−-4m concentration, O₂ uptake and ATP concentrations were significantly reduced. The mitotic index was reduced and the cell cycle population position was shifted to indicate that previously inhibited cells reformed their nuclei and became tetraploid. Prolonged treatment at this concentration resulted in cell death. Treatment with ioxynil at 10^−-6m reduced the rate of entry into mitosis. Abnormal mitotic figures in all stages were observed, and the ploidy level of mitotically inhibited cells was doubled. These observations indicated that at 10^−-6m concentration ioxynil acts as a preprophase inhibitor, that is, it does not act directly on the mitotic apparatus but does affect processes on which mitosis depends.     

Accumulation of non-utilizable starch in laticifers of Euphorbia heterophylla and E. myrsinites

Starch biosynthesis and degradation was studied in seedlings and mature plants of Euphorbia heterophylla L. and E. myrsinites L. Mature embryos, which lack starch grains in the non-articulated laticifers, develop into seedlings that accumulate starch rapidly when grown either in the light or the dark. Starch accumulation in laticifers of dark-grown seedlings was ca. 47 and 43% of total starch in light-grown controls in E. heterophylla and E. myrsinites, respectively. In light-grown seedlings, starch was present in laticifers as well as parenchyma of stems and leaves, whereas in dark-grown seedlings starch synthesis was almost exclusively limited to laticifers. In 7-month-old plants placed into total darkness, the starch in chyma was depleted within 6 d, whereas starch in laticifers was not mobilized. The starch content of latex in plants during development of floral primordia, flowering, and subsequent fruit formation remained rather constant. The results indicate that laticifers in seedlings divert embryonal storage reserves to synthesize starch even under stress conditions (darkness) in contrast to other cells, and that starch accumulated in laticifers does not serve as a metabolic reserve. The laticifer in Euphorbia functions in the accumulation and storage of secondary metabolites yet retains the capacity to produce, but not utilize starch, a primary metabolite.     

The distribution of plasmodesmata in the phloem ofHevea brasiliensis in relation to laticifer loading

Summary Cell to cell connections, including plasmodesmata and perforations, were examined in the non-conducting secondary phloem ofHevea brasiliensis. Samples were taken from trunks of numerous trees, from several clones, and prepared for thin sectioning and transmission or scanning electron microscopy and as optical sections for fluorescence microscopy. Numerous plasmodesmata were found clustered in primary pit-fields between the ray and axial parenchyma cells. Between the laticifers and adjacent parenchyma sheath cells, structures corresponding to functional plasmodesmata were not observed. But some unusual structural features were occasionally seen in these walls. These observations are discussed in relation to the possible function of the cell types, and to the loss of latex on the tapping ofHevea. It is suggested that the loading of the laticifer might first require a symplastic pathway for the transport of metabolites, at the end of which the assimilates must enter the apoplast. A transmembrane active transport system then transfers the metabolites in the laticifer. The presumable role of parenchyma cells in the loading of laticifers is emphasized.     

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.     

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.     

FOSSIL LATICIFERS FROM EOCENE BROWN COAL DEPOSITS OF THE GEISELTAL

Thick mats of cellular remains from Eocene brown coal deposits of the Geiseltal near Halle, DDR, were determined to be fossil nonarticulated laticifers. Nuclear magnetic resonance analyses of intact strands showed they consisted of eis-1,4-configuration rubber representing the polymerized isoprenoid contents of individual laticifers. Only remains of laticifers are present; other cells are absent as a result of biodegradation. The long laticifers, often with a surrounding cell wall, retained a tubular shape during their preservation. The isoprenoid content, which filled the entire lumen, possessed a cribriform structural character. The interstices within the rubber represent areas of former protoplasm of the cell. Various configurations in the protoplasm molded by the rubber during the initial phase of fossilization appear as negative images of former nuclei, organelles, and possibly membrane surfaces. The laticifer axes possess branches of several configurations comparable in morphology to those in branched, nonarticulated laticifers in extant plants. Acetone extracts of the rubber contents analyzed by gas-liquid chromatography identified the presence of several hydrocarbons which form a characteristic profile for the laticifer. It is suggested that the distinctive cellular micromorphology, rubber configuration, and hydrocarbon profile of these laticifers can be employed as markers in comparative studies with extant plants to identify the generic or species origin of these laticifers.     

夹竹桃科2种引种植物分泌结构的解剖学研究

为了解夹竹桃科(Apocynaceae)植物乳汁管的发生发育,对爱之蔓(Ceropegia woodii)和百万心(Dischidia ruscifolia)营养器官中的分泌结构进行了显微观察。结果表明,爱之蔓和百万心营养器官中均有无节分枝乳汁管的分布,茎皮层中的乳汁管大部分具有明显的分枝,叶中乳汁管具明显分枝,分布与走向多与叶脉维管组织平行。另外,爱之蔓营养器官中的分泌结构除乳汁管外,还有分泌腔。这为夹竹桃科植物的系统分类研究提供了解剖学依据。     

Histochemical study of detailed laticifer structure and rubber biosynthesis-related protein localization in <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using spectral confocal laser scanning microscopy

In Hevea brasiliensis, laticifers produce and accumulate rubber particles. Despite observation using histochemical methods, development stage structure and structures with ceasing functions have rarely been described. Spectral confocal laser scanning microscopy with Nile red staining simplifies laticifer structure observation in tangential sections while enhancing the resolution. Laticifer and ray images were extracted from unmixed images and used to monitor changes during growth. A laticifer network structure developed from increased anastomoses between adjoining laticifers outside of the conducting phloem, but because of increased radial division and growth of rays, the network structure ruptured and disintegrated. We also investigated immunohistochemical localization of two rubber particle-associated proteins in the laticifers: small rubber particle protein (SRPP) and rubber elongation factor (REF). Mature bark test results show that SRPP is localized only in the laticifer layers in the conducting phloem; REF is localized in all laticifer layers. Because SRPP plays a positive role in rubber biosynthesis, results show that the rubber biosynthesis capability of laticifers is concentrated where rays and the sieve tube actively transport metabolites.