Die Milch macht's – auch bei Pflanzen. Milchröhren und Milchsaft

Laticifers and latex Many different plants have laticifers and produce latex. Latex flows out when the plant is injured and coagulates quickly thus protecting the wound. It is a mixture of vacuoles and cytoplasm and contains various secondary plant products, especially polyterpenes (usually in the form of latex particles) and often toxins. The diversity of the latex corresponds with the diversity of laticifers. Nonarticulated laticifers are long, single cells, articulated ones are derived from cell fusions. Both kinds of laticifers may be branched. Reputed/ill‐reputed is the latex of Papaver somniferum, the source of opium. Technically very important has become the rubber of Hevea brasiliensis, after vulcanization of polyterpenes had been invented.     

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: 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.     

Laticifers and the classification of Euphorbia: the chemotaxonomy of Euphorbia esula L.

Laticifers and the classification of Euphorbia: the chemotaxonomy of Euphorbia esula L. Articulated and non-articulated laticifer cells represent distinctive cell types of relatively recent origin and occur in only a few families. Both types are of separate phylogenetic origin, reflecting independent evolutionary trends in the Euphorbiaceae. Supra-generic groupings of this family can be segregated into three taxonomic units using the laticifer character; with either articulated laticifers, non-articulated laticifers, or no laticifers. Such units may reflect more natural assemblages than now represented in the classification of this family. Laticifers possess chemical and morphological features of potential application as taxonomic characters to aid in delimiting species and interpreting evolutionary trends. The triterpenoid profile from latex of Euphorbia species has been shown to be diagnostic for a taxon. The qualitative and quantitative composition show a high level of stability under diverse environmental and physiological conditions indicating a genetic basis for triterpenoid synthesis. Triterpenoid profiles of known accessions of European E. esula L. and related presumptive taxa from North America readily separated them into distinctive chemotaxa that include one for E. esula L., whereas morphological features were found inadequate for separating accessions to presumptive taxa. Identification of adventive spurges in North America requires diagnostic analyses of Eurasian leafy spurges for comparison. Laticifer characters used in conjunction with relevant morphological features will provide a broadened insight into phylogenetic relationships with the Euphorbiaceae.     

Got milk? The secret life of laticifers

Laticifers are specialized cells that occur in over 20 plant families in several unrelated angiosperm orders. Although laticifers are likely to be of polyphyletic origin, their occurrence is considered a morphological indicator of relatedness among species. The classification of laticifers is based on developmental patterns and overall morphology. The cytoplasmic latex exuded in response to damage often includes specialized metabolites, such as cardenolides, alkaloids and natural rubber. Laticifers provide an effective location to store defense metabolites, although not all latex-bearing plants accumulate bioactive natural products. Ecophysiological studies have shown that latex and its associated metabolites are vital for the defense of plants against insects. The anatomy, development and physiology of laticifers are discussed with a focus on evolutionary and ecological perspectives.     

Lysosome and proteasome pathways are distributed in laticifers of Euphorbia helioscopia L.

At present, the lysosome pathway (LP) and proteasome pathway (PP) are known as major clearance systems in eukaryotic cells. The laticifer, a secretory tissue, degrades some cytoplasm during development. In this study, we investigated the distribution of LP and PP in non‐articulated laticifers of Euphorbia helioscopia L. Electron microscopy revealed that, plastids, mitochondria and some cyotsol were degraded in the late development laticifers, where there were numerous vesicles originated from dicytosomes. Accordingly, some key proteins in LP and PP were detected in E. helioscopia latex using isobaric tags for relative and absolute quantitation (iTRAQ) proteomics. Further immunohistochemistry analysis revealed that the clathrin heavy chain (CHC) belonging to LP and the ubiquitin‐mediated proteasome degradation increases gradually as the laticifer develops. Immuno‐electron microscopy revealed that the cysteine protease, CHC and AP‐2 complex subunit beta‐1 belonging to LP were mainly distributed in vesicles deriving from dicytosomes, which we called lysosome‐like vesicles. Ubiquitin was widely distributed in the cytosol, and proteasome activity was significantly reduced when various concentrations of the inhibitor MG132 were added to the latex total protein. We hypothesize that LP and PP are distributed in E. helioscopia laticifers; and it was speculated that LP and PP might be involved in the degradation of organelles and some cytoplasmic matrix in E. helioscopia laticifers.     

Anatomy of the Latex System in the Dormant Embryo of Euphorbia marginata Purch : Correlative Analysis of the Histological Organization and Topographic Distribution of Laticifers

The anatomy, morphology and topography of laticifers in themature embryo of Euphorbia marginata have been studied by conventionaland fluorescence microscopy. The mature embryo consists of awell-developed embryonic axis and two cotyledons and it containsdifferentiating vascular tissue. In the hypocotyl, six procambialstrands are present which branch and extend into the cotyledons,whereas in the root they are fused in a triarch structure. Inthe embryonic axis, the latex system consists of stelar andcortical components that are first anastomized in a cotyledonaryplexus and then are joined above in a cotyledonary system. Thestelar and cortical systems are linked by horizontal branchesto a nodal latex plexus placed under the epicotylary meristem.All the vascular laticifers, 24 in number, end unbranched inthe apex of the radicle. The cortical laticifers are profuselybranched toward the epidermis in tiny veins as far as the sub-epidermallevel. The cotyledonary laticifers follow the vascular tissueand form a network surrounding the mesophyllary areoles. Thetopography of the latex system leads one to hypothesize thatthe criteria which determine the constitution of the latex systemlareonly the essential peculiarities of idioblasts, but also correlativephenomena between growing laticifers and surrounding tissues.The stelar system appears to be under the control of vascularpolar growth, while the cortical and cotyledonary latex systemsseem to be subjected both to endogenous polar stimuli and toextra-embryonal influences. Euphorbia marginata, anatomy, embryo, laticifers     

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.     

Cellular localization of tyrosine decarboxylase expression in transgenic opium poppy and tobacco

Opium poppy, Papaver somniferum, is cultivated for its alkaloid-rich latex. Tyrosine decarboxylase (TyDC) is the first enzyme in poppy alkaloid biosynthesis and is encoded by a small gene family. A 2,060-bp promoter fragment of TyDC5 was translationally fused to the #-glucuronidase (GUS) reporter gene and introduced into poppy and tobacco (Nicotiana tabacum). Transgenic seedlings were stained for GUS activity which localized to the xylem parenchyma in the shoots of poppy and tobacco. Roots of both species had similar expression patterns with staining in the vascular cylinder surrounding the xylem. No staining was observed in poppy laticifers suggesting that other TyDC genes may be expressed in latex or that alkaloid precursors are supplied to laticifers by adjacent cells.     

Laticifer distribution in fig inflorescence and its potential role in the fig-fig wasp mutualism

Although in Moraceae the presence of laticifers is considered to be a synapomorphy, little is known about the distribution and morphology of this type of secretory structure in the reproductive organs of its species. Ficus, the largest genus of Moraceae, is characterized by an inflorescence known as syconium and by an obligate mutualistic interaction with pollinating wasps. The objectives of the present study were to evaluate the distribution and morphology of laticifers in syconia of 36 species belonging to different Ficus sections and to survey traits of taxonomic and adaptive value for the group. Syconia containing flowers in a receptive state were collected, fixed and processed for anatomical analysis. All species studied have branched laticifers distributed in the syconium receptacle, in the ostiolar bracts and in the pedicel of staminate flowers. Almost all species show laticifers in the pedicel of shorter-styled flowers. Laticifers also occur in the pedicel of longer-styled flowers in most Ficus sections, except F. curtipes (Conosycea section) and more than 75% of the studied species of the Americanae section. Laticifers are observed in the sepals of 25 of the 36 species studied and occasionally in the pistil. The presence of laticifers in the pedicel of shorter-style flowers and its absence in the pistil suggest that the distribution of this secretory structure in the fig flower was selected by pressures imposed by the fig-fig wasp mutualism. The laticifers in the pedicel of shorter-styled flowers may confer protection to the developing wasp larvae against natural enemies. However, the absence of laticifers in the pistil of most Ficus species studied was probably selected by the mutualistic relationship with the agaonid pollinating wasps since the latex could interfere with oviposition through the style, with the larval development of the pollinating fig wasps, and the emergence of pollinator offspring from their galls.     

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.     

Sucrose importation into laticifers of Hevea brasiliensis, in relation to ethylene stimulation of latex production

Background and Aims

The major economic product of Hevea brasiliensis is a rubber-containing cytoplasm (latex), which flows out of laticifers (latex cells) when the bark is tapped. The latex yield is stimulated by ethylene. Sucrose, the unique precursor of rubber synthesis, must cross the plasma membrane through specific sucrose transporters before being metabolized in the laticifers. The relative importance of sucrose transporters in determining latex yield is unknown. Here, the effects of ethylene (by application of Ethrel^®) on sucrose transporter gene expression in the inner bark tissues and latex cells of H. brasiliensis are described.

Methods

Experiments, including cloning sucrose transporters, real time RT-PCR and in situ hybridization, were carried out on virgin (untapped) trees, treated or untreated with the latex yield stimulant Ethrel.

Key Results

Seven putative full-length cDNAs of sucrose transporters were cloned from a latex-specific cDNA library. These transporters belong to all SUT (sucrose transporter) groups and differ by their basal gene expression in latex and inner soft bark, with a predominance of HbSUT1A and HbSUT1B. Of these sucrose transporters, only HbSUT1A and HbSUT2A were distinctly increased by ethylene. Moreover, this increase was shown to be specific to laticifers and to ethylene application.

Conclusion

The data and all previous information on sucrose transport show that HbSUT1A and HbSUT2A are related to the increase in sucrose import into laticifers, required for the stimulation of latex yield by ethylene in virgin trees.Key words: Hevea brasiliensis, laticifers, latex production, ethylene, sucrose transporters     

Localization and Behaviour of Actin Microfilaments in Laticiferous Cells of Hevea brasiliensis in Latex Exploitation

Actin microfilaments of laticiferous cells and bark wounds in Hevea brasiliensis were studied using TRITC-phalloidin fluorescent microscopy. Actin in latex from mature rubber trees was also investigated using SDS-PAGE and western-boltting. TRITC-fluorescent substance plugged the end of laticifers when latex flow stopped. Actin was detected only in C serum among the four latex fractions. Higher actin content was found in the latex collected at the beginning of tapping than in that collected just before latex flow stopped. Lower actin content was detected in the latex from rubber trees with more intensive exploitation. The present study indicated that actin microfilaments might play an important role in regulation of latex flow and plugging of the laticifers wounds.     

CYTOCHEMICAL LOCALIZATION OF CELLULASE ACTIVITY IN ARTICULATED,ANASTOMOSING LATICIFERS OF PAPAVER SOMNIFERUM L. (PAPAVERACEAE)

Cellulase activity was localized at the ultrastructural level in the articulated, anastomosing laticifers of Papaver somniferum. Electron-dense crystalline deposits indicating the presence of cellulase activity were restricted to discrete patches along the laticifer wall in regions of recently formed perforations. This report presents the first direct evidence for the involvement of cellulases in the cell wall perforation process in articulated laticifers.     

Effect ofCalotropis latex on laticifers differentiation in callus cultures ofCalotropis procera

Laticifers differentiation in callus cultures of Calotropis procera (Asclepiadaceae) as affected by own latex and its fractions incorporated in Murashige and Skoog (MS) medium is described. Callus cultures have been maintained on MS medium with 2.3 ΜM 6-furfurylaminopurine (FAP) and 3.0 ΜM 1-naphthylacetic acid (NAA). Marked increase in laticifers differentiation (from 10.1 to 28.4 %) was observed on this medium supplemented with 1 % (v/v) of latex. Latex fractions containing proteins + complex polysaccharides or inorganic salts also increased laticifers differentiation (by 21.8 % and 24.1 %, respectively). Other fractions (free amino acid + saccharides, phenols and terpenes + sterols) had no marked effect on laticifers differentiation while alkaloid fraction inhibited it. Effect of latex on laticifers differentiation was much more profound than the reported optimal concentration of plant growth regulators (4.6 ΜM FAP + 1 ΜM IAA). This research was supported by grant-in-aid for research from the University Grants Commission (No. F3-65/91SR II), New Delhi, to Dr. KG. Ramawat.     

Ontogeny of anastomosed laticifers in the stem apex of <Emphasis Type="Italic">Hancornia speciosa</Emphasis> (Apocynaceae): a topographic approach

Latex is a complex plant secretion with both ecological and economic importance. There is little information currently available on the cytological aspects of the ontogenesis of anastomosed laticifers, the ducts originating through the lysis of adjacent cell walls. Hancornia speciosa is a tree typical of the Cerrado (neotropical savanna) biome. Its latex has medicinal value and is also used to produce rubber. The ontogenesis of its laticifers and the process of latex synthesis are described here. Structural, cytochemical, and ultrastructural analyses of the stem apex and phytochemical analyses of the latex were performed. Laticifer ontogenesis begins early in promeristem cells and subsequently extends through the procambial region. The laticifer precursor cells demonstrate intense metabolic activity, evidenced by starch accumulation and the proliferation of mitochondria, dictyosomes, endoplasmic reticulum, and ribosomes—resulting in the thickening of the cell walls and accumulations of oil droplets in the cytoplasm and fibrous materials in the vacuoles. The ontogenetic process culminates with the partial dissolution of adjacent cell walls and the collapse of the cytoplasm, giving rise to anastomosed laticifers distributed throughout the phloem and adjacent regions of the cortex and medulla. The latex itself is composed of terpenes, mucilage, proteins, alkaloids, and organelle residues that form an emulsion. Laticifer development takes place in three phases: (1) the formation of the emulsion in the promeristem, (2) anastomosis and the collapse of the cytoplasm in the distal region of the procambium, and (3) the maturation of laticifers and latex storage in a central vacuole in the proximal region of the procambium.     

Differentiation of Non-articulated Laticifers in Poinsettia (Euphorbia pulcherrima Willd.)

Differentiation of non-articulated laticifers in poinsettia(Euphorbia pulcherrima Willd.) was studied ultra-structurally.Growing laticifers show: (1) a multinucleate apical region containingabundant ribosomes but few other differentiated organelles and(2) a sub-apical zone where the cytoplasm is dominated by vacuolesof diverse morphology with latex particles. These particlesappear first within narrow tubular vacuoles developed especiallyin the peripheral cytoplasm. During vacuolation of the laticifer,portions of cytoplasm, including some of the nuclei, becomeisolated by the enlarging and fusing vacuoles; eventually thesebecome lysed, except the latex particles which remain in thecentral vacuole. During differentiation of a laticifer branch,the cytoplasm contains the usual organelles, including a fewmicrobodies and coated vesicles. The plastids that lie withinthe peripheral cytoplasm differentiate into amyloplasts witha single elongated starch grain. Towards the end of differentiationthe cytoplasm becomes restricted to a thin parietal layer, withthe remaining organelles reduced or degenerate, surroundinga central vacuole filled with latex particles. Euphorbia pulcherrima Willd, poinsettia, ultrastructure, differentiation, laticifers     

Hormonal treatment of the bark of rubber trees (Hevea brasiliensis) increases latex yield through latex dilution in relation with the differential expression of two aquaporin genes

Natural rubber is synthesized in laticifers in the inner liber of the rubber tree (Hevea brasiliensis). Upon bark tapping, the latex is expelled due to liber turgor pressure. The mature laticifers are devoid of plasmodesmata; therefore a corresponding decrease in the total latex solid content is likely to occur due to water influx inside the laticifers. Auxins and ethylene used as efficient yield stimulants in mature untapped rubber trees, but, bark treatments with abscisic acid (ABA) and salicylic acid (SA) could also induce a transient increase latex yield. We recently reported that there are three aquaporin genes, HbPIP2;1, HbTIP1;1 and HbPIP1;1, that are regulated differentially after ethylene bark treatment. HbPIP2;1 was up-regulated in both the laticifers and the inner liber tissues, whereas HbTIP1;1 was up-regulated in the latex cells, but very markedly down-regulated in the inner liber tissues. Conversely, HbPIP1;1 was down-regulated in both tissues. In the present study, HbPIP2;1 and HbTIP1;1 showed a similar expression in response to auxin, ABA and SA, as seen in ethylene stimulation, while HbPIP1;1 was slightly regulated by auxin, but neither by ABA nor SA. The analysis of the HbPIP1;1 promoter region indicated the presence of only ethylene and auxin responsive elements. In addition, the poor efficiency of this HbPIP1;1 in increasing plasmalemma water conductance was confirmed in Xenopus oocytes. Thus, an increase in latex yield in response to all of these hormones was proposed to be the major function of aquaporins, HbPIP2;1 and HbTIP1;1. This study emphasized that the circulation of water between the laticifers and their surrounding tissues that result in latex dilution, as well as the probable maintenance of the liber tissues turgor pressure, favor the prolongation of latex flow.     

Sabotaging behaviour and minimal latex of Asclepias curassavica incur no cost for larvae of the southern monarch butterfly Danaus erippus

1. The southern monarch, Danaus erippus, uses mainly Asclepias curassavica as its host in the Neotropics, a plant species bearing articulated anastomosing laticifers. When artificially severed, A. curassavica has been shown to release significantly less latex than other Asclepias species. 2. The present study tested the hypothesis that sabotaging behaviour changes during the ontogeny of D. erippus and recorded latex outflow of A. curassavica during sabotaging and feeding. Larvae displayed vein‐cutting behaviour, which was initially observed in the second instar, became more pronounced in the third and fourth instars, and less frequent in the fifth instar. When present, latex outflow was never more than 1 µl at a time during either vein cutting or feeding, regardless of the instar. 3. Mandibular and midrib morphometrics revealed that larvae selected thicker midrib sites for severing as instars progressed; however, no correlation between mandibular size and midrib size severed was found within instars. 4. Costs of sabotaging behaviour and the effects of A. curassavica latex outflow on D. erippus larvae were also examined. Sabotaging behaviour did not incur growth costs for larvae, and only latex exudation volumes at least 10‐fold greater than those observed due to D. erippus sabotaging or feeding, caused significantly higher larval mortality than controls. 5. Since latex outflow is minimal or non‐existent in A. curassavica, sabotaging behaviour in D. erippus is mostly limited by morphological constraints and is probably driven by chemical stimulants rather than latex defence. In turn, latex does not constitute a major defence of A. curassavica against D. erippus.